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Sample records for fluorescence intensity multiple

  1. Intense fluorescence of Au20

    NASA Astrophysics Data System (ADS)

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

    2017-08-01

    Ligand-protected Au clusters are non-bleaching fluorescence markers in bio- and medical applications. Here we show that their fluorescence can be 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 Highest Occupied Molecular Orbital-Lowest Unoccupied Molecular Orbital (HOMO-LUMO) diabatic bandgap of the cluster. Au20 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 absorption and predict both main absorption peaks and intrinsic fluorescence in fair agreement with experiment.

  2. High intensity portable fluorescent light

    NASA Technical Reports Server (NTRS)

    Kendall, F. B.

    1972-01-01

    Eight high intensity portable fluorescent lights were produced. Three prototype lights were also produced, two of which were subsequently updated to the physical and operational configuration of the qualification and flight units. Positioning of lamp apertures and reflectors in these lights is such that the light is concentrated and intensified in a specific pattern rather than widely diffused. Indium amalgam control of mercury vapor pressure in the lamp gives high output at lamp ambient temperatures up to 105 C. A small amount of amalgam applied to each electrode stem helps to obtain fast warm-up. Shrinking a Teflon sleeve on the tube and potting metal caps on each end of the lamp minimizes dispersion of mercury vapor and glass particles in the event of accidental lamp breakage. Operation at 20 kHz allows the lamps to consume more power than at low frequency, thus increasing their light output and raising their efficiency. When used to expose color photographic film, light from the lamps produces results approximately equal to sunlight.

  3. Multiple frequency fluorescence lifetime imaging microscopy.

    PubMed

    Squire, A; Verveer, P J; Bastiaens, P I

    2000-02-01

    fitting algorithms were tested on a simulated data set and were successful in disentangling two populations having 1 ns and 4 ns fluorescence lifetimes. Spatial invariance of the lifetimes was exploited to improve the accuracy significantly. Multiple frequency fluorescence lifetime imaging microscopy was then successfully applied to resolve the fluorescence lifetimes and fluorescence intensity contributions in a rhodamine dye mixture in solution, and green fluorescent protein variants co-expressed in live cells.

  4. A model for multiexponential tryptophan fluorescence intensity decay in proteins.

    PubMed Central

    Bajzer, Z; Prendergast, F G

    1993-01-01

    Tryptophan fluorescence intensity decay in proteins is modeled by multiexponential functions characterized by lifetimes and preexponential factors. Commonly, multiple conformations of the protein are invoked to explain the recovery of two or more lifetimes from the experimental data. However, in many proteins the structure seems to preclude the possibility of multiple conformers sufficiently different from one another to justify such an inference. We present here another plausible multiexponential model based on the assumption that an energetically excited donor surrounded by N acceptor molecules decays by specific radiative and radiationless relaxation processes, and by transferring its energy to acceptors present in or close to the protein matrix. If interactions between the acceptors themselves and back energy transfer are neglected, we show that the intensity decay function contain 2N exponential components characterized by the unperturbed donor lifetime, by energy transfer rates and a probability of occurrence for the corresponding process. We applied this model to the fluorescence decay of holo- and apoazurin, ribonuclease T1, and the reduced single tryptophan mutant (W28F) of thioredoxin. Use of a multiexponential model for the analysis of the fluorescence intensity decay can therefore be justified, without invoking multiple protein conformations. Images FIGURE 1 PMID:8312471

  5. Fluorescence wavelength and intensity variations of cave waters

    NASA Astrophysics Data System (ADS)

    Baker, A.; Genty, D.

    1999-04-01

    The fluorescence properties of groundwaters percolating into four cave systems have been monitored over the period 1997-1998. Fluorescence was excited between 220 and 400 nm and the emission measured from 300 to 500 nm using a fluorescence spectrophotometer. Three fluorescence centres were observed; one at the excitation-emission pair of 290-340:395-430 nm, (humic-like, probably fulvic acid), one at 265-280:300-370 nm (protein like) and a less defined region of high fluorescence at 230-280:310-420 nm (humic and/or protein like). The most consistent fluorescence intensity was observed in the excitation-emission pair of 290-340:395-430 nm, attributed to a fulvic acid source. Subtle differences (±5%) in the fluorescence excitation and emission wavelength of this fluorescence peak in the groundwater were observed between the four sites, and the fluorescence intensity varied considerably (×60) between the four sites. Both the wavelength and the intensity variations in fluorescence are caused by the differences in the vegetation cover, soil type and humification. Data from the most intensely monitored site (Brown's Folly Mine, England; 9 sample stations, 10-20 days frequency sampling) revealed no spatial variability in the 290-340:395-430 nm (fulvic acid) fluorescence; in contrast time-series analysis suggests that the seasonal variations do occur, with a decrease in the emission wavelength correlating with the first (autumn) peak in fluorescence intensity, and a decrease in the excitation wavelength correlating with a second (winter) fluorescence intensity peak. Results demonstrate the potential of utilising fluorescence wavelength variations in sourcing karst groundwaters, and as a possible palaeoenvironmental proxy of the overlying soil conditions if trapped within the cave speleothems.

  6. Influence of vessel stenosis on indocyanine green fluorescence intensity assessed by near-infrared fluorescence angiography.

    PubMed

    Yamamoto, Masaki; Nishimori, Hideaki; Fukutomi, Takashi; Handa, Takemi; Kihara, Kazuki; Tashiro, Miwa; Sato, Takayuki; Orihashi, Kazumasa

    2017-07-01

    Although useful for visualizing blood flow during revascularization surgery, the permeability of near-infrared fluorescence (NIR) angiography using indocyanine green (ICG) does not allow for vessel stenosis visualization. We hypothesized that changes in ICG fluorescence intensity reflect vessel stenosis, and evaluated the influence of stenosis on blood flow by ex vivo experimentation. The vessel stenosis model comprised a silicon tube, a graft occluder, and artificial blood. During near-infrared angiography, the fluorescense intensity was calculated during pre- and post-stenosis of an artificial circuit, using a NIR angiography. We measured the maximum fluorescence intensity and the time to maximum fluorescence intensity. Severe stenosis (≥75%) attenuated the increase in ICG fluorescence intensity in the tube significantly, pre- and post-stenosis. The time to maximum fluorescence intensity did not differ between sites pre- and post-stenosis, irrespective of stenosis severity. Stenosis affected the ICG fluorescence intensity through the vessel. Thus, quantitative analysis using NIR angiography may detect severe vessel stenosis (≥75%), and the extinction curve of indocyanine fluorescence intensity may support the evaluation of blood flow. The absence of differences in the time to maximum fluorescence intensity for degrees of stenosis might suggest a limitation of previous conventional qualitative assessments.

  7. Intensive immunosuppression in multiple sclerosis.

    PubMed

    Zaffaroni, M; Ghezzi, A; Comi, G

    2006-03-01

    Immunosuppressive drugs have been used out of label in multiple sclerosis (MS) for over 30 years and around 10% of patients are actually under immunosuppressive treatment. The rationale for immunosuppression in MS lies in the hypothesis that MS is an inflammatory immune-mediated disease that can take advantage of strong anti-inflammatory activity. Azathioprine, methotrexate, cyclophosphamide and mitoxantrone are the most utilised agents, but only the latter has been approved for clinically active MS. Many of them are safe in combination with interferon-beta and are under investigation in controlled trials. Plasma exchange is limited to catastrophic attacks in refractory MS whilst bone marrow transplantation is considered in patients with an extremely severe, active disease as the final option in escalation therapy. Although immunosuppressants are best effective in induction therapy, their use is limited by toxicity and potential long-term risk.

  8. Complete suppression of the fluorophore fluorescence by combined effect of multiple fluorescence quenching groups: A fluorescent sensor for Cu²⁺ with zero background signals.

    PubMed

    Long, Lingliang; Wu, Yanjun; Wang, Lin; Gong, Aihua; Hu, Rongfeng; Zhang, Chi

    2016-02-18

    The reaction-based fluorescent sensors have attracted increasing attention in the past decades. However, the application of these sensors for accurate sensing was significantly retarded by the background fluorescence from the sensors themselves. In this work, we demonstrated a novel strategy that the background fluorescence of the sensor could be completely eliminated by the combined effect of multiple fluorescence quenching groups. Based on this new strategy, as proof-of-principle study, a fluorescent sensor (CuFS) for Cu(2+) was judiciously developed. In CuFS, three types of fluorescence quenching groups were directly tethered to a commonly used coumarin fluorophore. The fluorescence of coumarin fluorophore in CuFS was completely suppressed by the combined effect of these fluorescence quenching groups. Upon treatment with 22 μM Cu(2+), sensor CuFS achieved a dramatic fluorescence enhancement (fluorescence intensity enhanced up to 811-fold) centered at 469 nm. The detection limits was determined to be 12.3 nM. The fluorescence intensity enhancement also showed a good linearity with the Cu(2+) concentration in the range of 12.3 nM to 2 μM. By fabricating test strips, sensor CuFS can be utilized as a simple tool to detect Cu(2+) in water samples. Furthermore, the fluorescent sensor was successfully applied in detecting different concentration of Cu(2+) in living cells.

  9. Fluorescence intensity, lifetime, and anisotropy screening of living cells based on total internal reflection techniques

    NASA Astrophysics Data System (ADS)

    Bruns, Thomas; Angres, Brigitte; Steuer, Heiko; Strauss, Wolfgang S. L.; Schneckenburger, Herbert

    2009-02-01

    A setup for fluorescence measurements of surfaces of biological samples, in particular the plasma membrane of living cells, is described. The method is based on splitting of a laser beam and multiple total internal reflections (TIR) within the bottom of a microtiter plate, such that up to 96 individual samples are illuminated simultaneously by an evanescent electromagnetic field. Two different screening procedures for the detection of fluorescence arising from the plasma membrane of living cells by High Throughput Screening (HTS) and High Content Screening (HCS), are distinguished. In the first case a rapid measurement of large sample numbers based on fluorescence intensity, and in the second case a high content of information from a single sample based on the parameters fluorescence lifetime (Fluorescence Lifetime Screening, FLiS) and fluorescence anisotropy (Fluorescence Lifetime Polarization Screening, FLiPS) is achieved. Both screening systems were validated using cultivated cells incubated with different fluorescent markers (e. g. NBD-cholesterol) as well as stably transfected cells expressing a fluorescent membrane-associating protein. In addition, particularly with regard of potential pharmaceutical applications, the kinetics of the intracellular translocation of a fluorescent protein kinase c fusion protein upon stimulation of the cells was determined. Further, a caspase sensor based on Förster Resonance Energy Transfer (FRET) between fluorescent proteins was tested. Enhanced cyan fluorescent protein (ECFP) anchored to the inner leaflet of the plasma membrane of living cells transfers its excitation energy via a spacer (DEVD) to an enhanced yellow fluorescent protein (EYFP). Upon apoptosis DEVD is cleaved, and energy transfer is disrupted, as proven by changes in fluorescence intensity and decay times.

  10. Zeeman degeneracy effects in collisional intense-field resonance fluorescence

    NASA Technical Reports Server (NTRS)

    Cooper, J.; Ballagh, R. J.; Burnett, K.

    1980-01-01

    Resonance fluorescence due to intense laser fields from a Zeeman degenerate atom being perturbed by collisional interactions is calculated in the impact regime by using the quantum-fluctuation-regression theorem. Various interesting effects are found. For example, the scattered intensity spectrum for a J = 0 to J = 1 transition for polarization parallel to the laser polarization is essentially an asymmetric triplet, whereas for a perpendicular polarization due to collisions the spectrum is essentially a doublet (whose frequencies do not correspond with any of those of the triplet). Further, the width of the fluorescent component (whose frequency is close to the unperturbed frequency) actually decreases with increasing laser power.

  11. Two-dimensional fluorescence intensity distribution analysis: theory and applications.

    PubMed

    Kask, P; Palo, K; Fay, N; Brand, L; Mets, U; Ullmann, D; Jungmann, J; Pschorr, J; Gall, K

    2000-04-01

    A method of sample analysis is presented which is based on fitting a joint distribution of photon count numbers. In experiments, fluorescence from a microscopic volume containing a fluctuating number of molecules is monitored by two detectors, using a confocal microscope. The two detectors may have different polarizational or spectral responses. Concentrations of fluorescent species together with two specific brightness values per species are determined. The two-dimensional fluorescence intensity distribution analysis (2D-FIDA), if used with a polarization cube, is a tool that is able to distinguish fluorescent species with different specific polarization ratios. As an example of polarization studies by 2D-FIDA, binding of 5'-(6-carboxytetramethylrhodamine) (TAMRA)-labeled theophylline to an anti-theophylline antibody has been studied. Alternatively, if two-color equipment is used, 2D-FIDA can determine concentrations and specific brightness values of fluorescent species corresponding to individual labels alone and their complex. As an example of two-color 2D-FIDA, binding of TAMRA-labeled somatostatin-14 to the human type-2 high-affinity somatostatin receptors present in stained vesicles has been studied. The presented method is unusually accurate among fluorescence fluctuation methods. It is well suited for monitoring a variety of molecular interactions, including receptors and ligands or antibodies and antigens.

  12. Multispectral excitation based multiple fluorescent targets resolving in fluorescence molecular tomography

    NASA Astrophysics Data System (ADS)

    Zhou, Yuan; Guang, Huizhi; Pu, Huangsheng; Zhang, Jiulou; Bai, Jing; Luo, Jianwen

    2016-04-01

    Fluorescence molecular tomography (FMT) can visualize biological activities at cellular and molecular levels in vivo, and has been extensively used in drug delivery and tumor detection research of small animals. The ill-posedness of the FMT inverse problem makes it difficult to reconstruct and resolve multiple adjacent fluorescent targets that have different functional features but are labeled with the same fluorochrome. An algorithm based on independent component analysis (ICA) for multispectral excited FMT is proposed to resolve multiple fluorescent targets in this study. Fluorescent targets are excited by multispectral excitation, and the three-dimensional distribution of fluorescent yields under the excitation spectrum is reconstructed by an iterative Tikhonov regularization algorithm. Subsequently, multiple fluorescent targets are resolved from mixed fluorescence signals by employing ICA. Simulations were performed and the results demonstrate that multiple adjacent fluorescent targets can be resolved if the number of excitation wavelengths is not smaller than that of fluorescent targets with different concentrations. The algorithm obtains both independent components that provide spatial information of different fluorescent targets and spectral courses that reflect variation trends of fluorescent yields along with the excitation spectrum. By using this method, it is possible to visualize the metabolism status of drugs in different structure organs, and quantitatively depict the variation trends of fluorescent yields of each functional organ under the excitation spectrum. This method may provide a pattern for tumor detection, drug delivery and treatment monitoring in vivo.

  13. Intensity calibration and flat-field correction for fluorescence microscopes.

    PubMed

    Model, Michael

    2014-04-01

    Standardization in fluorescence microscopy involves calibration of intensity in reproducible units and correction for spatial nonuniformity of illumination (flat-field or shading correction). Both goals can be achieved using concentrated solutions of fluorescent dyes. When a drop of a highly concentrated fluorescent dye is placed between a slide and a coverslip it produces a spatially uniform field, resistant to photobleaching and with reproducible quantum yield; it can be used as a brightness standard for wide-field and confocal microscopes. For wide-field microscopes, calibration can be further extended to absolute molecular units. This can be done by imaging a solution of known concentration and known depth; the latter can be prepared by placing a small spherical lens in a diluted solution of the same fluorophore that is used in the biological specimen.

  14. Fluorescence intensity analysis through simplex optimization in flow cytometry.

    PubMed

    Takase, K; Iwaki, K; Gunji, T; Yata, J

    1989-03-10

    Fluorescence intensity analysis in flow cytometric surface immunophenotyping has recently been appreciated in clinical applications. A curve fitting method to estimate the mean and SD values of fluorescence intensity is described in this report. A Gaussian distribution is aimed to be adapted for a specified distribution in logarithmically scaled histogram data through the simplex optimization, one of the non-linear least squares methods. In comparison with the conventional methods which include the detection of peak point and the direct calculation, this fitting method has demonstrated exceeding precisions in the estimation of both parameters with limited involved cell counts in typical lymphocytic phenotyping. The actual estimation for a precise SD value will develop the quality control approaches based on the fluorescence intensity analysis. While this method is not suitable for distributions that involve extremely small cell counts or that deviate markedly from a symmetric Gaussian, it has additional advantages of loose requirements, namely, narrow fitting regions, ordinarily small cell counts, practical computational periods and a simple programming.

  15. Lidar Tracking of Multiple Fluorescent Tracers: Method and Field Test

    NASA Technical Reports Server (NTRS)

    Eberhard, Wynn L.; Willis, Ron J.

    1992-01-01

    Past research and applications have demonstrated the advantages and usefulness of lidar detection of a single fluorescent tracer to track air motions. Earlier researchers performed an analytical study that showed good potential for lidar discrimination and tracking of two or three different fluorescent tracers at the same time. The present paper summarizes the multiple fluorescent tracer method, discusses its expected advantages and problems, and describes our field test of this new technique.

  16. Strongly intensive measures for multiplicity fluctuations

    NASA Astrophysics Data System (ADS)

    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.

  17. Estimation of fluorescence-tagged RNA numbers from spot intensities.

    PubMed

    Häkkinen, Antti; Kandhavelu, Meenakshisundaram; Garasto, Stefania; Ribeiro, Andre S

    2014-04-15

    Present research on gene expression using live cell imaging and fluorescent proteins or tagged RNA requires accurate automated methods of quantification of these molecules from the images. Here, we propose a novel automated method for classifying pixel intensities of fluorescent spots to RNA numbers. The method relies on a new model of intensity distributions of tagged RNAs, for which we estimated parameter values in maximum likelihood sense from measurement data, and constructed a maximum a posteriori classifier to estimate RNA numbers in fluorescent RNA spots. We applied the method to estimate the number of tagged RNAs in individual live Escherichia coli cells containing a gene coding for an RNA with MS2-GFP binding sites. We tested the method using two constructs, coding for either 96 or 48 binding sites, and obtained similar distributions of RNA numbers, showing that the method is adaptive. We further show that the results agree with a method that uses time series data and with quantitative polymerase chain reaction measurements. Lastly, using simulated data, we show that the method is accurate in realistic parameter ranges. This method should, in general, be applicable to live single-cell measurements of low-copy number fluorescence-tagged molecules. MATLAB extensions written in C for parameter estimation and finding decision boundaries are available under Mozilla public license at http://www.cs.tut.fi/%7ehakkin22/estrna/ CONTACT: andre.ribeiro@tut.fi. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  18. Toward instrument-independent quantitative measurement of fluorescence intensity in fiber-optic spectrometer systems

    NASA Astrophysics Data System (ADS)

    Zhao, Jianhua; Lui, Harvey; McLean, David I.; Zeng, Haishan

    2007-10-01

    Fluorescence has been widely used in biological research and clinical diagnosis. One challenge facing the rapid growth of fluorescence application is the inability to make comparable fluorescence intensity measurements during a long period of clinical study and across laboratories. We propose a method to implement system-independent fluorescence intensity calibration in fiber-optic fluorescence spectrometer systems. This method is based on a National Institute for Standards and Technology traceable standard light source for system spectral response calibration, and a fluorescence reference standard for fluorescence intensity calibration. Human skin in vivo and a fluorescence phantom made of fluorescent microspheres were measured with two different system configurations and at different probe-sample distances. Experimental results showed very good agreement with theory after system-independent fluorescence intensity calibration.

  19. Multiple stimulated emission fluorescence photoacoustic sensing and spectroscopy

    SciTech Connect

    Li, Gaoming; Gao, Fei; Feng, Xiaohua; Zheng, Yuanjin; Qiu, Yishen

    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.

  20. Multiple stimulated emission fluorescence photoacoustic sensing and spectroscopy

    NASA Astrophysics Data System (ADS)

    Li, Gaoming; Gao, Fei; Qiu, Yishen; Feng, Xiaohua; Zheng, Yuanjin

    2016-07-01

    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.

  1. Intense quenching of fluorescence intensity of poly(vinyl pyrrolidone) molecules in presence of gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Behera, M.; Ram, S.

    2013-12-01

    We study the quenching of fluorescence intensity of 40 g/L poly(vinyl pyrrolidone) PVP molecules by varying the content of gold nanoparticles (GNPs) from 1 to 5 μM in 1-butanol. A profound exponential decay of the emission band intensity in the π ← nπ* band of the PVP molecules at ~392 nm upon gradual addition of the GNPs demonstrates an existence of an excited state interaction of NPs with the PVP molecules in a gold colloid in 1-butanol. Such quenching is caused by the non-bonding electron transfer from the O-atom of carbonyl group of the PVP molecules to the surface of the GNP. X-ray photoemission spectroscopy (XPS) study corroborates the spectroscopic results. A linear Stern-Volmer plot with a quenching constant of 2.23 × 106 M-1 reveals dynamic quenching in a non-aqueous NF. A mechanism of fluorescence quenching was proposed in support of XPS and images taken from hybrid nanostructure using transmission electron microscope. Study on quenching of fluorescence intensity of PVP fluorophore in the presence of GNPs is useful for optoelectronic devices and biosensors.

  2. Competition of multiple filaments during the propagation of intense femtosecond laser pulses

    SciTech Connect

    Hosseini, S.A.; Luo, Q.; Ferland, B.; Liu, W.; Chin, S.L.; Kosareva, O.G.; Panov, N.A.; Kandidov, V.P.; Akoezbek, N.

    2004-09-01

    We observed a universal phenomenon of the competition among multiple filaments generated during the propagation of intense femtosecond laser pulses in air. We show that the fluorescence signal from the excitation of nitrogen molecules inside the plasma channel contains important information pertaining to the formation and interaction of multiple filaments. The detected backscattered nitrogen fluorescence from inside the filaments yielded irregular changes from shot to shot which cannot be explained by fluctuation arising from the initial laser pulse itself. Numerical simulations reveal a complex dynamics of multiple filament propagation and interaction dynamics that depends strongly on the initial perturbations of the laser beam. The irregular changes of the fluorescence signal are attributed to the interference between adjacent hot spots that evolve into filaments which give rise to new hot spots (filaments) in between, and thus give the appearance of the fusion or branching of filaments.

  3. Fluorescence of indocyanine green in blood: intensity dependence on concentration and stabilization with sodium polyaspartate.

    PubMed

    Maarek, J M; Holschneider, D P; Harimoto, J

    2001-12-31

    Indocyanine green (ICG) has been widely used in cardiovascular, hepatic, and ophthalmologic studies. Application of this fluorescent dye has been handicapped by its poor stability in solution and by the complex dependence of its fluorescence intensity on concentration. Noncovalent interactions between ICG and sodium polyaspartate (PASP) stabilize ICG fluorescence in aqueous solution, but the effect of PASP on ICG fluorescence in blood has not been described. The current study had two main goals: first, to characterize in vitro in blood the relationship between fluorescence intensity and concentration of ICG-PASP (ICG) and the stability of this relationship over time; second, to test a new phenomenological model describing the dependence of ICG fluorescence on concentration. Freshly-prepared ICG and ICG-PASP solutions produced the same fluorescence intensity over a wide range of concentrations (0.0005-0.1271 mg/ml). The peak fluorescence of ICG was reduced by 11% after 10 h and by 72% at 7 days. In contrast, the peak fluorescence intensity of ICG-PASP solutions was nearly unchanged for up to 14 days. The dependence of the fluorescence intensity on concentration was accurately represented by our model that accounted for the generation of fluorescence following light absorption, and for the reabsorption of the emitted fluorescence by ICG.

  4. Precise microinjection into living cells by summation of fluorescence intensity

    NASA Astrophysics Data System (ADS)

    Taninaka, Kiyoshi; Yabuki, A.; Ito, A.; Harada, T.

    2007-02-01

    It is difficult to introduce a specific amount of a substance into cells by existing injection methods because there is no appropriate method of directly measuring the quantity of the injected substance. Although radioisotopes can be used, there is currently no apparatus that can practically handle such radioisotopes. The measurement of the diameter of a liquid droplet in air or oil is affected by surface tension if the liquid droplet is very small; but this issue does not occur with microinjection, in which a water solution is discharged under pressure through a capillary and into a cell. It is also difficult to measure the density or mass of the injected substance because of the low discharge rate, unlike the case of inkjet printers. To solve these problems, we propose a method of precise microinjection by summation of fluorescence intensity. In addition, we developed a new pressure pulse injection device that generates pressure with a rectangular waveform and a precise amplitude and pulse width to improve controllability of the discharge amount. Lastly, when the above device and method are combined, the coefficient of correlation between the specified number of pressure pulses per unit of time and the actual discharge amount exceeded 0.999. This research paper describes in detail the measurement system, standalone performance, and quantities of substances introduced into living cells.

  5. A portable fluorescent sensing system using multiple LEDs

    NASA Astrophysics Data System (ADS)

    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.

  6. Protein-like fluorescence intensity as a possible tool for determining river water quality

    NASA Astrophysics Data System (ADS)

    Baker, Andy; Inverarity, Roger

    2004-10-01

    The results of a comparison between chemical water quality determinants and river water fluorescence on the River Tyne, NE England, demonstrate that tryptophan-like fluorescence intensity shows statistically significant relationships between nitrate, phosphate, ammonia, biochemical oxygen demand (BOD) and dissolved oxygen. Tryptophan-like fluorescence intensity at the 280 nm excitation/350 nm emission wavelength fluorescence centre correlates with both phosphate (r = 0.80) and nitrate (r = 0.87), whereas tryptophan-like fluorescence intensity at the 220 nm excitation/350 nm emission wavelength centre correlates with BOD (r = 0.85), ammonia (r = 0.70) and dissolved oxygen (r = -0.65). The strongest correlations are between tryptophan-like fluorescence intensity and nitrate and phosphate, which in the Tyne catchment derive predominantly from point and diffuse source sewage inputs. The correlation between BOD and the tryptophan-like fluorescence intensity suggests that this fluorescence centre is related to the bioavailable or labile dissolved organic matter pool. The weakest correlations are observed between tryptophan-like fluorescence intensity and ammonia concentration and dissolved oxygen. The weaker correlation with ammonia is due to removal of the ammonia signal by wastewater treatment, and that with dissolved oxygen due to the natural aeration of the river such that this is not a good indicator of water quality. The observed correlations only hold true when treated sewage, sewerage overflows or cross connections, or agricultural organic pollutants dominate the water quality - this is not true for two sites where airport deicer (propylene glycol, which is non-fluorescent) or landfill leachate (which contains high concentrations of humic and fulvic-like fluorescent DOM) dominate the dissolved organic matter in the river. Mean annual tryptophan-like fluorescence intensity agrees well with the General Water Quality Assessment as determined by the England and Wales

  7. Quantitative measurement of binary liquid distributions using multiple-tracer x-ray fluorescence and radiography

    SciTech Connect

    Halls, Benjamin R.; Meyer, Terrence R.; Kastengren, Alan L.

    2015-01-01

    The complex geometry and large index-of-refraction gradients that occur near the point of impingement of binary liquid jets present a challenging environment for optical interrogation. A simultaneous quadruple-tracer x-ray fluorescence and line-of-sight radiography technique is proposed as a means of distinguishing and quantifying individual liquid component distributions prior to, during, and after jet impact. Two different pairs of fluorescence tracers are seeded into each liquid stream to maximize their attenuation ratio for reabsorption correction and differentiation of the two fluids during mixing. This approach for instantaneous correction of x-ray fluorescence reabsorption is compared with a more time-intensive approach of using stereographic reconstruction of x-ray attenuation along multiple lines of sight. The proposed methodology addresses the need for a quantitative measurement technique capable of interrogating optically complex, near-field liquid distributions in many mixing systems of practical interest involving two or more liquid streams.

  8. Quantitative measurement of binary liquid distributions using multiple-tracer x-ray fluorescence and radiography

    SciTech Connect

    Halls, Benjamin R.; Meyer, Terrence R.; Kastengren, Alan L.

    2015-01-23

    The complex geometry and large index-of-refraction gradients that occur near the point of impingement of binary liquid jets present a challenging environment for optical interrogation. A simultaneous quadruple-tracer x-ray fluorescence and line-of-sight radiography technique is proposed as a means of distinguishing and quantifying individual liquid component distributions prior to, during, and after jet impact. Two different pairs of fluorescence tracers are seeded into each liquid stream to maximize their attenuation ratio for reabsorption correction and differentiation of the two fluids during mixing. This approach for instantaneous correction of x-ray fluorescence reabsorption is compared with a more time-intensive approach of using stereographic reconstruction of x-ray attenuation along multiple lines of sight. The proposed methodology addresses the need for a quantitative measurement technique capable of interrogating optically complex, near-field liquid distributions in many mixing systems of practical interest involving two or more liquid streams.

  9. Nonlinear and nonmonotonic nature of the intensity of fluorescence emission on a surface of turbid fluorescing biotissues

    NASA Astrophysics Data System (ADS)

    Rogatkin, Dmitry; Lapaeva, Ludmila; Guseva, Irina

    2014-05-01

    Many researchers during past 20 years have used the laser fluorescence spectroscopy (LFS) for in vivo tissue diagnosis. But in the up-to-date medical in vivo LFS there is a problem of quantification of the fluorophores concentrations in optically-turbid biotissues basing on measurements of the laser induced fluorescence on a surface of the tissues. The purpose of our work is both experimental and theoretical study of the character of dependences of measured fluorescence intensities on tissues' optical properties and on fluorophores concentrations in tissues. In the experimental part of our study the measurements of the superficial fluorescence on phantoms at different known concentration of fluorophores in them were carried out. As a result experimental dependences of the registered intensities of the laser induced fluorescence emission on fluorophores concentration were plotted. In the theoretical part of our study the analytical solution for a fluorescence emission by Kokhanovsky's method based on the well-known two-flux Kubelka-Munk approach (KMA) was used. Besides, in the study the Kokhanovsky's method was modified by its association with our improved KMA, allowing us to receive exact solutions for boundary intensities collected by an optical probe. As a result a set of theoretical curves describing the influence of fluorophore concentration in tissues on the registered intensities was obtained as well. Both experimental and theoretical results show a good qualitative agreement between each other. Also these results show that the dependence of the fluorescence intensity from tissues' optical properties and from the concentration of fluorophores can be both nonlinear and non-monotonic.

  10. Correlation of conformational heterogeneity of the tryptophyl side chain and time-resolved fluorescence intensity decay kinetics

    NASA Astrophysics Data System (ADS)

    Laws, William R.; Ross, J. B. Alexander

    1992-04-01

    The time-resolved fluorescence properties of a tryptophan residue should be useful for probing protein structure, function, and dynamics. To date, however, the non-single exponential fluorescence intensity decay kinetics for numerous peptides and proteins having a single tryptophan residue have not been adequately explained. Many possibilities have been considered and include: (1) contributions from the 1La and 1Lb states of indole; (2) excited-state hydrogen exchange; and (3) environmental heterogeneity from (chi) 1 and (chi) 2 rotamers. In addition, it has been suggested that generally many factors contribute to the decay and a distribution of probabilities may be more appropriate. Two recent results support multiple species due to conformational heterogeneity as the major contributor to complex kinetics. First, a rotationally constrained tryptophan analogue has fluorescence intensity decay kinetics that can be described by the sum of two exponentials with amplitudes comparable to the relative populations of the two rotational isomers. Second, the multiple exponentials observed for tyrosine-containing model compounds and peptides correlate with the (chi) 1 rotamer populations independently determined by 1H NMR. We now report similar correlations between rotamer populations and fluorescence intensity decay kinetics for a tryptophan analogue of oxytocin. It appears for this compound that either (chi) 2 rotations do not appreciably alter the indole environment, (chi) 2 rotations are rapid enough to average the observed dependence, or only one of two possible (chi) 2 populations is associated with each (chi) 1 rotamer.

  11. Differences in the intensity of light-induced fluorescence emitted by resin composites.

    PubMed

    Kim, Bo-Ra; Kang, Si-Mook; Kim, Gyung-Min; Kim, Baek-Il

    2016-03-01

    The aims of this study were to compare the intensities of fluorescence emitted by different resin composites as detected using quantitative light-induced fluorescence (QLF) technology, and to compare the fluorescence intensity contrast with the color contrast between a restored composite and the adjacent region of the tooth. Six brands of light-cured resin composites (shade A2) were investigated. The composites were used to prepare composite discs, and fill holes that had been prepared in extracted human teeth. White-light and fluorescence images of all specimens were obtained using a fluorescence camera based on QLF technology (QLF-D) and converted into 8-bit grayscale images. The fluorescence intensity of the discs as well as the fluorescence intensity contrast and the color contrast between the composite restoration and adjacent tooth region were calculated as grayscale levels. The grayscale levels for the composite discs differed significantly with the brand (p<0.001): DenFil (10.84±0.35, mean±SD), Filtek Z350 (58.28±1.37), Premisa (156.94±1.58), Grandio (177.20±0.81), Charisma (207.05±0.77), and Gradia direct posterior (211.52±1.66). The difference in grayscale levels between a resin restoration and the adjacent tooth was significantly greater in fluorescence images for each brand than in white-light images, except for the Filtek Z350 (p<0.05). However, the Filtek Z350 restoration was distinguishable from the adjacent tooth in a fluorescence image. The intensities of fluorescence detected from the resin composites varied. The differences between the composite and adjacent tooth were greater for the fluorescence intensity contrast than for the colors observed in the white-light images. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Multi-Photon Fluorescence Microscopy: Behavior of Biological Specimens Under High Intensity Illumination

    DTIC Science & Technology

    2000-07-01

    used in these types of microscopy. We have used maize leaf protoplast as a model system to evaluate the photo-induced response of living sample under...to fluorescence emission, second harmonic generation was observed in the maize protoplasts. Keywords: Multi-photon fluorescence microscopy, photon...damage, cell damage, high intensity illumination, maize 1. INTRODUCTION Multi-photon fluorescence microscopy has been cited for its advantage in the

  13. The Effects of Absorption and Self-Absorption Quenching on Fluorescent Intensities

    ERIC Educational Resources Information Center

    Henderson, Giles

    1977-01-01

    Describes an experiment that shows that fluorescence emission intensities are significantly modified by absorption processes and that these effects can be eliminated by positioning the sample cell such that the interrogation region corresponds to the corner of the cell. (MLH)

  14. Coordinate-targeted fluorescence nanoscopy with multiple off states

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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.

  16. Fluorescence detection of adenosine triphosphate through an aptamer-molecular beacon multiple probe.

    PubMed

    Zeng, Xiaodan; Zhang, Xiaoling; Yang, Wen; Jia, Hongying; Li, Yamin

    2012-05-01

    An aptamer-molecular beacon (MB) multiple fluorescent probe for adenosine triphosphate (ATP) assay is proposed in this article. The ATP aptamer was used as a molecular recognition part, and an oligonucleotide (short strand, SS) partially complementary with the aptamer and an MB was used as the other part. In the presence of ATP, the aptamer bound with it, accompanied by the hybridization of MB and SS and the fluorescence recovering. Wherever there is only very weak fluorescence can be measured in the absence of ATP. Based on the relationship of recovering fluorescence and the concentration of ATP, a method for quantifying ATP has been developed. The fluorescence intensity was proportional to the concentration of ATP in the range of 10 to 500 nM with a detection limit of 0.1 nM. Moreover, this method was able to detect ATP with high selectivity in the presence of guanosine triphosphate (GTP), cytidine triphosphate (CTP), and uridine triphosphate (UTP). This method is proved to be simple with high sensitivity, selectivity, and specificity.

  17. Multiple Excitations Multi-Spectral Fluorescence (MEMSF) Bio-Detector

    DTIC Science & Technology

    2005-10-01

    automatically preserved for further confirmatory and forensic analyses. The MEMSF detection process is based upon the intensities of multiple...wavelength excitation/emission pairs (EEP) selected to characterize and identify bioaerosols . Figure 1. The MEMSF bio-detector has two principle...stages – collect then detect . 1 Report Documentation Page Form ApprovedOMB No. 0704-0188 Public reporting burden for the collection of information is

  18. Fluorescent and high intensity discharge lamp use in chambers and greenhouses

    NASA Technical Reports Server (NTRS)

    Langhans, Robert W.

    1994-01-01

    Fluorescent and High Intensity Discharge lamps have opened up great opportunities for researchers to study plant growth under controlled environment conditions and for commercial growers to increase plant production during low/light periods. Specific technical qualities of fluorescent and HID lamps have been critically reviewed. I will direct my remarks to fluorescent and high intensity discharge (HID) lamps in growth chambers, growth rooms, and greenhouses. I will discuss the advantages and disadvantages of using each lamp in growth chambers, growth rooms and greenhouses.

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

    NASA Astrophysics Data System (ADS)

    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.

  20. [Determining the relative content of cisplatin in cells by acridine orange fluorescence intensity].

    PubMed

    Goroshinskaia, I A; Tarnopol'skaia, O V; Kachesova, P S

    2014-01-01

    The fluorescence intensity of acridine orange (AO) dye bound to living human leukocytes, rat leukocytes, and Pliss lymphosarcoma cells of rat has been studied by fluorescence microscopy. The cell suspension of each test was divided into two parts, to one of which the drug cisplatin was added to a final concentration of 0.1 μg/mL and the mixture was incubated for 20 - 60 min. Acridine orange was then added and the average intensity of red and green fluorescence of cells was determined. The brightness of AO in the green region of the spectrum was reduced approximately 3-fold (p = 0.001) in cells with cisplatin. In the red region of the fluorescence, differences in the brightness of cells with and without cisplatin were not observed. These data indicate that a decrease in the brightness of green fluorescence of AO can be a test for the presence of cisplatin in cells.

  1. Simultaneous confocal recording of multiple fluorescent labels with improved channel separation.

    PubMed

    Carlsson, K; Aslund, N; Mossberg, K; Philip, J

    1994-12-01

    Confocal microscopes are often used to study specimens labelled with fluorophores. A commonly used method for simultaneous recording of the distribution of multiple fluorophores is to divide the fluorescent light emitted by the specimen into different wavelength regions using dichroic and bandpass filters. These different wavelength regions are then distributed to multiple detectors. However, the broad and overlapping spectra of commonly used fluorophores often result in considerable crosstalk between channels. A new technique, intensity-modulated multiple-beam scanning (IMS) microfluorometry, can be used to reduce this cross-talk substantially. The IMS technique is implemented with two laser beams of different wavelengths, intensity-modulated at different frequencies, which illuminate the specimen simultaneously. The two laser wavelengths predominantly excite one fluorophore each. Fluorescent light from the specimen is divided into two wavelength regions (red and green) which are detected by two photomultiplier tubes. The output signals from the photomultiplier tubes are connected to lock-in amplifiers. The effect of using modulated laser beams, in combination with lock-in amplifiers, is strongly to reduce cross-talk between the channels. The performance of the IMS technique using various types of specimen is compared with the results obtained using the conventional multi-detector method.

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

    PubMed

    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.

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

    PubMed

    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.

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

    PubMed

    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.

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

    PubMed Central

    Agarwal, Krishna; Macháň, Radek

    2016-01-01

    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. PMID:27934858

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

    NASA Astrophysics Data System (ADS)

    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.

  7. Mathematical Modeling of Tear Film Break up Modes and Fluorescent Intensity

    NASA Astrophysics Data System (ADS)

    Siddique, Javed; Braun, Richard; Begley, Carolyn; Winkeler, Adam; King-Smith, Peter E.

    2013-11-01

    The purpose of this study is to develop mathematical model for variables of interest in tear film break up (TBU) to compare with experimental images of TBU to better predict local values of tear film osmolarity and fluorescence during and following the TBU. Models are developed for local changes tear film thickness, insoluble surfactant concentration as well as osmolarity and fluorescein concentration inside the tear film. Fluorescence concentration was converted to fluorescent intensity using the expression involving film thickness and the full range of fluorescence as described by Nichols et al. (IOVS 2012). The fluorescent intensity response is a primary tool for visualizing the tear film thickness, and it is qualitatively different in the dilute vs concentrated regimes. Computed results over a wide range of fluorescein concentrations show that elevated surfactant concentration or evaporation rate led to thinner regions where TBU first occurs. The model predicts locally elevated concentration of osmolarity within areas of TBU and fluorescence intensity patterns very similar to computed thickness and the observed experimental results. The osmolarity may increase from 50 % to 1300 % of the isosmolar value, depending sensitively on the corneal permeability. Supported by Simons Foundation Grant 281839 (JIS), NSF Grant 1022706, NEI Grant 1RO1EY021794 (Begley), and NEI Grant RO1EY17951 (King-Smith).

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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 < 0.05). Opallis was the only 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).

  10. Pre hospital intensive care in multiple trauma children.

    PubMed

    Hervé, C; Gaillard, M; Petit, J L; Geni, S; Huguenard, P

    1986-01-01

    From January 1979 to December 1984, 1,272 calls, concerning injured children, aged 11 days to 15 years, justified the intervention of a Mobile Medical Emergency and Intensive Care Service, in the department of "Val-de-Marne" near Paris. Three hundred and twenty-two were very serious trauma children (25%); 45 were in cardiac arrest, and 41 died on the scene of the accident despite the intensive cares delivered by the anesthetists or pediatricians. Two hundred and eighty-one children were hospitalized in an intensive multiple trauma pediatric unit (97 cases) or in a neurosurgical pediatric unit (184 cases). The mode of accident was traffic accident (252), fall (48), fire arms (4), knife wounds (7), hanging or strangulation (9), others (2). They concerned 119 females and 203 males. 126 were multiple trauma children (40%). 37% of these accidents happened between May and July, and 40% occurred between 3 to 6 p.m. The 322 children immediately received medical care but 26% died during their hospitalization (17% in the first 24 hours). Thus mortality rate is 35% (114 cases).

  11. Effect of dielectric spacer thickness on signal intensity of surface plasmon field-enhanced fluorescence spectroscopy.

    PubMed

    Murakami, Takashi; Arima, Yusuke; Toda, Mitsuaki; Takiguchi, Hiromi; Iwata, Hiroo

    2012-02-15

    Surface plasmon field-enhanced fluorescence spectroscopy (SPFS) combines enhanced field platform and fluorescence detection. Its advantages are the strong intensity of the electromagnetic field and the high signal/noise (S/N) ratio due to the localized evanescent field at the water/metal interface. However, the energy transfer from the fluorophore to the metal surface diminishes the fluorescence intensity, and this reduces the sensitivity. In this study, we tested whether polystyrene (PSt) could act as a dielectric layer to suppress the energy transfer from the fluorophore to the metal surface. We hypothesized that this would improve the sensitivity of SPFS-based immunoassays. We used α-fetoprotein (AFP) as a model tumor biomarker in the sandwich-type immunoassay. We determined the relationship between fluorescent signal intensity and PSt layer thickness and compared this to theoretical predictions. We found that the fluorescence signal increased by optimally controlling the thickness of the PSt layer. Our results indicated that the SPFS-based immunoassay is a promising clinical diagnostic tool for quantitatively determining the concentrations of low-level biomarkers in blood samples.

  12. Mathematical Modeling of Tear Film Break up Modes and Fluorescent Intensity

    NASA Astrophysics Data System (ADS)

    Siddique, Javed; Braun, Richard; Begley, C. G.; King-Smith, P. E.

    2014-11-01

    We develop a mathematical model for variables of interest in tear film break up (TBU) to compare with experimental images of TBU to better predict local values of tear film (TF) osmolarity and fluorescence during and following the TBU. Models are developed for local changes in TF thickness, osmolarity and fluorescein concentration. Fluorescence concentration was converted to fluorescent intensity using the expression involving film thickness and the full range of fluorescence (Nichols et al. (IOVS 2012). The fluorescent intensity response is a primary tool for visualizing the TF thickness, and it is qualitatively different in the dilute vs concentrated regimes. Computed results over a wide range of fluorescein concentrations show that evaporation rate led to thinner regions where TBU first occurs. The computed results will be closely compared with experimential fluorescence and other imaging techniques to help determine relevant parameters. The model predicts locally elevated concentration of osmolarity within areas of TBU and predicts osmolarity in these regions which can't be measured experimental results to date. The osmolarity may increase from 50% to 1300% of the isosmolar value, depending sensitively on the corneal permeability and diffusivity of solutes in tear film. J. I. Siddique would like to acknowledge the support from Simons Foundation.

  13. Quantitative measurement of binary liquid distributions using multiple-tracer x-ray fluorescence and radiography

    DOE PAGES

    Halls, Benjamin R.; Meyer, Terrence R.; Kastengren, Alan L.

    2015-01-23

    The complex geometry and large index-of-refraction gradients that occur near the point of impingement of binary liquid jets present a challenging environment for optical interrogation. A simultaneous quadruple-tracer x-ray fluorescence and line-of-sight radiography technique is proposed as a means of distinguishing and quantifying individual liquid component distributions prior to, during, and after jet impact. Two different pairs of fluorescence tracers are seeded into each liquid stream to maximize their attenuation ratio for reabsorption correction and differentiation of the two fluids during mixing. This approach for instantaneous correction of x-ray fluorescence reabsorption is compared with a more time-intensive approach of usingmore » stereographic reconstruction of x-ray attenuation along multiple lines of sight. The proposed methodology addresses the need for a quantitative measurement technique capable of interrogating optically complex, near-field liquid distributions in many mixing systems of practical interest involving two or more liquid streams.« less

  14. Quantitative measurement of binary liquid distributions using multiple-tracer x-ray fluorescence and radiography

    DOE PAGES

    Halls, Benjamin R.; Meyer, Terrence R.; Kastengren, Alan L.

    2015-01-23

    The complex geometry and large index-of-refraction gradients that occur near the point of impingement of binary liquid jets present a challenging environment for optical interrogation. A simultaneous quadruple-tracer x-ray fluorescence and line-of-sight radiography technique is proposed as a means of distinguishing and quantifying individual liquid component distributions prior to, during, and after jet impact. Two different pairs of fluorescence tracers are seeded into each liquid stream to maximize their attenuation ratio for reabsorption correction and differentiation of the two fluids during mixing. This approach for instantaneous correction of x-ray fluorescence reabsorption is compared with a more time-intensive approach of usingmore » stereographic reconstruction of x-ray attenuation along multiple lines of sight. The proposed methodology addresses the need for a quantitative measurement technique capable of interrogating optically complex, near-field liquid distributions in many mixing systems of practical interest involving two or more liquid streams.« less

  15. Time-resolved and temperature tuneable measurements of fluorescent intensity using a smartphone fluorimeter.

    PubMed

    Hossain, Md Arafat; Canning, John; Yu, Zhikang; Ast, Sandra; Rutledge, Peter J; Wong, Joseph K-H; Jamalipour, Abbas; Crossley, Maxwell J

    2017-05-30

    A smartphone fluorimeter capable of time-based fluorescence intensity measurements at various temperatures is reported. Excitation is provided by an integrated UV LED (λex = 370 nm) and detection obtained using the in-built CMOS camera. A Peltier is integrated to allow measurements of the intensity over T = 10 to 40 °C. All components are controlled using a smartphone battery powered Arduino microcontroller and a customised Android application that allows sequential fluorescence imaging and quantification every δt = 4 seconds. The temperature dependence of fluorescence intensity for four emitters (rhodamine B, rhodamine 6G, 5,10,15,20-tetraphenylporphyrin and 6-(1,4,8,11-tetraazacyclotetradecane)2-ethyl-naphthalimide) are characterised. The normalised fluorescence intensity over time of the latter chemosensor dye complex in the presence of Zn(2+) is observed to accelerate with an increasing rate constant, k = 1.94 min(-1) at T = 15 °C and k = 3.64 min(-1) at T = 30 °C, approaching a factor of ∼2 with only a change in temperature of ΔT = 15 °C. Thermally tuning these twist and bend associated rates to optimise sensor approaches and device applications is proposed.

  16. Fluorescence intensity positivity classification of Hep-2 cells images using fuzzy logic

    NASA Astrophysics Data System (ADS)

    Sazali, Dayang Farzana Abang; Janier, Josefina Barnachea; May, Zazilah Bt.

    2014-10-01

    Indirect Immunofluorescence (IIF) is a good standard used for antinuclear autoantibody (ANA) test using Hep-2 cells to determine specific diseases. Different classifier algorithm methods have been proposed in previous works however, there still no valid set as a standard to classify the fluorescence intensity. This paper presents the use of fuzzy logic to classify the fluorescence intensity and to determine the positivity of the Hep-2 cell serum samples. The fuzzy algorithm involves the image pre-processing by filtering the noises and smoothen the image, converting the red, green and blue (RGB) color space of images to luminosity layer, chromaticity layer "a" and "b" (LAB) color space where the mean value of the lightness and chromaticity layer "a" was extracted and classified by using fuzzy logic algorithm based on the standard score ranges of antinuclear autoantibody (ANA) fluorescence intensity. Using 100 data sets of positive and intermediate fluorescence intensity for testing the performance measurements, the fuzzy logic obtained an accuracy of intermediate and positive class as 85% and 87% respectively.

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

    NASA Astrophysics Data System (ADS)

    Nikitin, Andrey G.

    2016-02-01

    The Editor and publisher have retracted "Manipulating fluorescence color and intensity with regular nanoparticle-based composite materials" Appl. Phys. Lett. 108, 051112 (2016). Because of a legal issue related to the publication of work covered under a confidentiality agreement, the electronic version of the paper has been removed from the website.

  18. Lifetime of fluorescence from light-harvesting chlorophyll a/b proteins: excitation intensity dependence

    SciTech Connect

    Nordlund, T.M.; Knox, W.H.

    1981-10-01

    The fluorescence from a purified, aggregate form of the light-harvesting chlorophyll a/b protein has a lifetime of 1.2 +/- 0.5 ns at low excitation intensity, but the lifetime decreases significantly when the intensity of the 20-ps, 5300nm excitation pulse is increased above about 10/sup 16/ photons/cm/sup 2/. A solubilized, monomeric form of the protein, on the other hand, has a fluorescence lifetime of 3.1 +/- 0.3 ns independent of excitation intensity from 10/sup 14/-10/sup 18/ photons/cm/sup 2//pulse. We interpret the lifetime shortening in the aggregates and the lack of shortening in monomers in terms of exciton annihilation, facilitated in the aggregate by the larger population of interacting chlorophylls.

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

    PubMed

    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

  20. Preparation of Novel Europium Complex Doped Ag@SiO2 Nanoparticles with Intense Fluorescence.

    PubMed

    Liu, Bing; Yin, Dongguang; Song, Kailin; Yang, Juan Ou; Wang, Chengcheng; Wu, Minghong

    2015-01-01

    In this study, a new europium complex of 4,4'-bis (1",1",1",2",2",3",3"-heptafluoro-4",6"- hexanedion-6"-yl)-o-terpheny-Eu(3+)-4,7-diphenyl-1,10-phenanthroline-2,9-dicarboxylic acid-(3-aminopropyl) trimethoxysilane (BHHT-Eu(3+)-DPPDA-APTMS) was prepared first. Then novel core-shell Ag@SiO2 nanoparticles with BHHT-Eu(3+)-DPPDA-APTMS doped in shell were synthesized by a facile water-in-oil microemulsion method. The properties of the prepared complex and nanoparticles, and the effect of metal enhanced fluorescence for the nanoparticles were investigated. The prepared nanopartilces exhibited intense fluorescence, uniform morphology and good water-solubility. The fluorescent intensities of silver core-present nanopartciles were significant higher than that of silver core-absent nanoparticles owing to the metal enhanced fluorescence of silver core. It is expectable that the as-prepared nanoparticles can serve as a potential fluorescent nanoprobe, applying in high sensitive biological and medical detections.

  1. Mapping fluorophore distributions in three dimensions by quantitative multiple angle-total internal reflection fluorescence microscopy.

    PubMed Central

    Olveczky, B P; Periasamy, N; Verkman, A S

    1997-01-01

    The decay of evanescent field intensity beyond a dielectric interface depends upon beam incident angle, enabling the 3-d distribution of fluorophores to be deduced from total internal reflection fluorescence microscopy (TIRFM) images obtained at multiple incident angles. Instrumentation was constructed for computer-automated multiple angle-TIRFM (MA-TIRFM) using a right angle F2 glass prism (n(r) 1.632) to create the dielectric interface. A laser beam (488 nm) was attenuated by an acoustooptic modulator and directed onto a specified spot on the prism surface. Beam incident angle was set using three microstepper motors controlling two rotatable mirrors and a rotatable optical flat. TIRFM images were acquired by a cooled CCD camera in approximately 0.5 degree steps for >15 incident angles starting from the critical angle. For cell studies, cells were grown directly on the glass prisms (without refractive index-matching fluid) and positioned in the optical path. Images of the samples were acquired at multiple angles, and corrected for angle-dependent evanescent field intensity using "reference" images acquired with a fluorophore solution replacing the sample. A theory was developed to compute fluorophore z-distribution by inverse Laplace transform of angle-resolved intensity functions. The theory included analysis of multiple layers of different refractive index for cell studies, and the anisotropic emission from fluorophores near a dielectric interface. Instrument performance was validated by mapping the thickness of a film of dihexyloxacarbocyanine in DMSO/water (n(r) 1.463) between the F2 glass prism and a plano-convex silica lens (458 mm radius, n(r) 1.463); the MA-TIRFM map accurately reproduced the lens spherical surface. MA-TIRFM was used to compare with nanometer z-resolution the geometry of cell-substrate contact for BCECF-labeled 3T3 fibroblasts versus MDCK epithelial cells. These studies establish MA-TIRFM for measurement of submicroscopic distances between

  2. Multiple fluorescence microscopy and optoelectronic imaging: possibilities and limits

    NASA Astrophysics Data System (ADS)

    Gundlach, Heinz

    1997-12-01

    The last 20 years have seen an unexpected great renaissance and a partial revolution in light microscopy. This recent progress is due to new design in optics and instrumentation as well as improvement of optical contrast enhancement techniques. Recent progress in fluorescence microscopy is achieved by multiparameter fluorescence techniques, by improvement of conventional photomicrography as well as by optoelectronic imaging, confocal laser scanning microscopy, image processing and analysis. Due to the increase in number of fluorescence dyes, double and triple bandpass filter sets permit a rapid changeover between different fluorochromes simultaneously.

  3. Effects of multiple scattering on fluorescence correlation spectroscopy measurements of particles moving within optically dense media

    NASA Astrophysics Data System (ADS)

    Zustiak, Silviya; Riley, Jason; Boukari, Hacène; Gandjbakhche, Amir; Nossal, Ralph

    2012-12-01

    Fluorescence correlation spectroscopy (FCS) is increasingly being used to assess the movement of particles diffusing in complex, optically dense surroundings, in which case measurement conditions may complicate data interpretation. It is considered how a single-photon FCS measurement can be affected if the sample properties result in scattering of the incident light. FCS autocorrelation functions of Atto 488 dye molecules diffusing in solutions of polystyrene beads are measured, which acted as scatterers. Data indicated that a scattering-linked increase in the illuminated volume, as much as two fold, resulted in minimal increase in diffusivity. To analyze the illuminated beam profile, Monte-Carlo simulations were employed, which indicated a larger broadening of the beam along the axial than the radial directions, and a reduction of the incident intensity at the focal point. The broadening of the volume in the axial direction has only negligible effect on the measured diffusion time, since intensity fluctuations due to diffusion events in the radial direction are dominant in FCS measurements. Collectively, results indicate that multiple scattering does not result in FCS measurement artifacts and thus, when sufficient signal intensity is attainable, single-photon FCS can be a useful technique for measuring probe diffusivity in optically dense media.

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

    PubMed

    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

  5. Multiple Stimuli-Responsive Fluorescence Behavior of Novel Polyamic Acid Bearing Oligoaniline, Triphenylamine, and Fluorene Groups.

    PubMed

    Yan, Ying; Sun, Ningwei; Li, Fangfei; Jia, Xiaoteng; Wang, Ce; Chao, Danming

    2017-02-22

    Multiple stimuli-responsive fluorescent materials have gained increasing attention for their fundamental investigation and intelligent applications. In this work, we report design and synthesis of a novel polyamic acid bearing oligoaniline, triphenylamine, and fluorene groups, which served as sensitive units and fluorescence emission unit, respectively. The resulting polymer exhibits multiple stimuli-responsive fluorescence switching behavior triggered by redox species, pH, electrochemical, and pressure stimuli. Every fluorescence switching mechanism upon each stimulus was studied in detail. The interactions and energy transfer between sensitive units and emission unit are largely responsible for this fascinating fluorescent switching behavior. This work provides a deep understanding of the optical switching essence upon these stimuli, opening the way for the development of new fluorescent sensing applications.

  6. Two-photon microscopy of cortical NADH fluorescence intensity changes: correcting contamination from the hemodynamic response

    PubMed Central

    Baraghis, Edward; Devor, Anna; Fang, Qianqian; Srinivasan, Vivek J.; Wu, Weicheng; Lesage, Frédéric; Ayata, Cenk; Kasischke, Karl A.; Boas, David A.; Sakadžić, Sava

    2011-01-01

    Quantification of nicotinamide adenine dinucleotide (NADH) changes during functional brain activation and pathological conditions provides critical insight into brain metabolism. Of the different imaging modalities, two-photon laser scanning microscopy (TPLSM) is becoming an important tool for cellular-resolution measurements of NADH changes associated with cellular metabolic changes. However, NADH fluorescence emission is strongly absorbed by hemoglobin. As a result, in vivo measurements are significantly affected by the hemodynamics associated with physiological and pathophysiological manipulations. We model NADH fluorescence excitation and emission in TPLSM imaging based on precise maps of cerebral microvasculature. The effects of hemoglobin optical absorption and optical scattering from red blood cells, changes in blood volume and hemoglobin oxygen saturation, vessel size, and location with respect to imaging location are explored. A simple technique for correcting the measured NADH fluorescence intensity changes is provided, with the utilization of a parallel measurement of a physiologically inert fluorophore. The model is applied to TPLSM measurements of NADH fluorescence intensity changes in rat somatosensory cortex during mild hypoxia and hyperoxia. The general approach of the correction algorithm can be extended to other TPLSM measurements, where changes in the optical properties of the tissue confound physiological measurements, such as the detection of calcium dynamics. PMID:22029350

  7. Two-photon microscopy of cortical NADH fluorescence intensity changes: correcting contamination from the hemodynamic response.

    PubMed

    Baraghis, Edward; Devor, Anna; Fang, Qianqian; Srinivasan, Vivek J; Wu, Weicheng; Lesage, Frédéric; Ayata, Cenk; Kasischke, Karl A; Boas, David A; Sakadzić, Sava

    2011-10-01

    Quantification of nicotinamide adenine dinucleotide (NADH) changes during functional brain activation and pathological conditions provides critical insight into brain metabolism. Of the different imaging modalities, two-photon laser scanning microscopy (TPLSM) is becoming an important tool for cellular-resolution measurements of NADH changes associated with cellular metabolic changes. However, NADH fluorescence emission is strongly absorbed by hemoglobin. As a result, in vivo measurements are significantly affected by the hemodynamics associated with physiological and pathophysiological manipulations. We model NADH fluorescence excitation and emission in TPLSM imaging based on precise maps of cerebral microvasculature. The effects of hemoglobin optical absorption and optical scattering from red blood cells, changes in blood volume and hemoglobin oxygen saturation, vessel size, and location with respect to imaging location are explored. A simple technique for correcting the measured NADH fluorescence intensity changes is provided, with the utilization of a parallel measurement of a physiologically inert fluorophore. The model is applied to TPLSM measurements of NADH fluorescence intensity changes in rat somatosensory cortex during mild hypoxia and hyperoxia. The general approach of the correction algorithm can be extended to other TPLSM measurements, where changes in the optical properties of the tissue confound physiological measurements, such as the detection of calcium dynamics.

  8. Two-photon microscopy of cortical NADH fluorescence intensity changes: correcting contamination from the hemodynamic response

    NASA Astrophysics Data System (ADS)

    Baraghis, Edward; Devor, Anna; Fang, Qianqian; Srinivasan, Vivek J.; Wu, Weicheng; Lesage, Frédéric; Ayata, Cenk; Kasischke, Karl A.; Boas, David A.; Sakadžić, Sava

    2011-10-01

    Quantification of nicotinamide adenine dinucleotide (NADH) changes during functional brain activation and pathological conditions provides critical insight into brain metabolism. Of the different imaging modalities, two-photon laser scanning microscopy (TPLSM) is becoming an important tool for cellular-resolution measurements of NADH changes associated with cellular metabolic changes. However, NADH fluorescence emission is strongly absorbed by hemoglobin. As a result, in vivo measurements are significantly affected by the hemodynamics associated with physiological and pathophysiological manipulations. We model NADH fluorescence excitation and emission in TPLSM imaging based on precise maps of cerebral microvasculature. The effects of hemoglobin optical absorption and optical scattering from red blood cells, changes in blood volume and hemoglobin oxygen saturation, vessel size, and location with respect to imaging location are explored. A simple technique for correcting the measured NADH fluorescence intensity changes is provided, with the utilization of a parallel measurement of a physiologically inert fluorophore. The model is applied to TPLSM measurements of NADH fluorescence intensity changes in rat somatosensory cortex during mild hypoxia and hyperoxia. The general approach of the correction algorithm can be extended to other TPLSM measurements, where changes in the optical properties of the tissue confound physiological measurements, such as the detection of calcium dynamics.

  9. Assessment of Cellular Redox State Using NAD(P)H Fluorescence Intensity and Lifetime

    PubMed Central

    Blacker, Thomas S.; Berecz, Tunde; Duchen, Michael R.; Szabadkai, Gyorgy

    2017-01-01

    NADH and NADPH are redox cofactors, primarily involved in catabolic and anabolic metabolic processes respectively. In addition, NADPH plays an important role in cellular antioxidant defence. In live cells and tissues, the intensity of their spectrally-identical autofluorescence, termed NAD(P)H, can be used to probe the mitochondrial redox state, while their distinct enzyme-binding characteristics can be used to separate their relative contributions to the total NAD(P)H intensity using fluorescence lifetime imaging microscopy (FLIM). These protocols allow differences in metabolism to be detected between cell types and altered physiological and pathological states. PMID:28286806

  10. Strain-independent temperature measurement by use of a fluorescence intensity ratio technique in optical fiber.

    PubMed

    Wade, S A; Collins, S F; Grattan, K T; Baxter, G W

    2000-06-20

    The strain sensitivity of the fluorescence intensity ratio temperature-sensing technique has been measured to be (2 +/- 3) x 10(-4)%/muepsilon in Yb3+-doped fiber, implying a temperature-to-strain cross sensitivity of (2 +/- 3) x 10(-4) degrees C/muepsilon. The near-zero strain sensitivity means that this optical-fiber sensor technique is well suited for temperature measurement in strain-affected environments.

  11. Use of Multiple Fluorescent Labels in Biological Sensing

    DTIC Science & Technology

    2006-05-01

    stanford.edu Abstract: We describe novel fluorescent N-deoxyribosides (1 and 2) having 2-pyrido-2-benzimidazole and 2- quino -2-benzimidazole as aglycones...with conjugated systems (Chart 1). We chose these pyrido- (and related quino -) benzimidazole heterocycles (3 and 4), both of which were previously

  12. Intra- and Inter-annual Fluorescence Intensity Variations in Drip Water, Heshang Cave, Central China: Implications for Speleothem Palaeoclimatology

    NASA Astrophysics Data System (ADS)

    Jin, L.; Hu, C.; Li, X.; Ruan, J.; Hartland, A.

    2015-12-01

    Cave drip water acts as a signal carrier for the soil-rock-air system leading to the capture of climatic and environmental information in stalagmites. This paper seeks to develop an understanding of the environmental and climatic factors which control fluorescence variations in dripwater from in Heshang Cave, Central China. This information is essential to unravelling the significance of organic fluorescence in stalagmites and its utility in quantitative paleoclimate reconstructions. On the seasonal time scale, drip water fluorescence is largely controlled by the decomposition and translocation of dissolved organic matter in the soil, related to climate factors like temperature and precipitation. On the inter-annual time scale, longer duration monitoring data in scarce, yet this is needed to fully comprehend the influence of climate in stalagmite fluorescence time series. This study presents nine consecutive years of monthly drip water fluorescence intensity and drip rate data from two perennial drip sites in Heshang Cave. Drip water fluorescence was generally characterized by intensities in spring/summer and low intensities in autumn/winter. In dry hydrologic years, little seasonality in fluorescence signals was observed, but the opposite was observed in wet years. On the inter-annual time scale, the annual mean intensities of drip water fluorescence positively correlated with local annual rainfall with a 1-year lag (R2HS4=0.94; R2HS6=0.74). This indicates that rainfall is the main control on total drip water fluorescence (integrating across a hydrologic year), despite significant degrees of intra-annual fluorescence variation being observed between wet and dry years. These findings are of direct relevance for paleoclimate reconstruction using fluorescence intensities in stalagmites from the Asian monsoon region. Key words: fluorescence; dissolved organic matter; drip water rates; seasonality; precipitation

  13. Life History Changes in Coral Fluorescence and the Effects of Light Intensity on Larval Physiology and Settlement in Seriatopora hystrix

    PubMed Central

    Roth, Melissa S.; Fan, Tung-Yung; Deheyn, Dimitri D.

    2013-01-01

    Fluorescence is common in both coral adult and larval stages, and is produced by fluorescent proteins that absorb higher energy light and emit lower energy light. This study investigated the changes of coral fluorescence in different life history stages and the effects of parental light environment on larval fluorescence, larval endosymbiotic dinoflagellate abundance, larval size and settlement in the brooding coral Seriatopora hystrix. Data showed that coral fluorescence changed during development from green in larvae to cyan in adult colonies. In larvae, two green fluorescent proteins (GFPs) co-occur where the peak emission of one GFP overlaps with the peak excitation of the second GFP allowing the potential for energy transfer. Coral larvae showed great variation in GFP fluorescence, dinoflagellate abundance, and size. There was no obvious relationship between green fluorescence intensity and dinoflagellate abundance, green fluorescence intensity and larval size, or dinoflagellate abundance and larval size. Larvae of parents from high and low light treatments showed similar green fluorescence intensity, yet small but significant differences in size, dinoflagellate abundance, and settlement. The large variation in larval physiology combined with subtle effects of parental environment on larval characteristics seem to indicate that even though adult corals produce larvae with a wide range of physiological capacities, these larvae can still show small preferences for settling in similar habitats as their parents. These data highlight the importance of environmental conditions at the onset of life history and parent colony effects on coral larvae. PMID:23544072

  14. Multi-color femtosecond source for simultaneous excitation of multiple fluorescent proteins in two-photon fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Wang, Ke; Liu, Tzu-Ming; Wu, Juwell; Horton, Nicholas G.; Lin, Charles P.; Xu, Chris

    2013-02-01

    Simultaneous imaging of cells expressing multiple fluorescent proteins (FPs) is of particular interest in applications such as mapping neural circuits, tracking multiple immune cell populations, etc. To visualize both in vivo and ex vivo tissue morphology and physiology at a cellular level deep within scattering tissues, two-photon fluorescence microscopy (2PM) is a powerful tool that has found wide applications. However, simultaneous imaging of multiple FPs with 2PM is greatly hampered by the lack of proper ultrafast lasers offering multi-color femtosecond pulses, each targeting the two-photon absorption peak of a different FP. Here we demonstrate simultaneous two-photon fluorescence excitation of RFP, YFP, and CFP in human melanoma cells engineered to express a "rainbow" pallet of colors, using a novel fiber-based source with energetic, three-color femtosecond pulses. The three-color pulses, centered at 775 nm, 864 nm and 950 nm, are obtained through second harmonic generation of the 1550 nm pump laser and SHG of the solitons at 1728 nm and 1900 nm generated through soliton self-frequency shift (SSFS) of the pump laser in a large-mode-area (LMA) fiber. The resulting wavelengths are well matched to the two-photon absorption peaks of the three FPs for efficient excitation. Our results demonstrate that multi-color femtosecond pulse generation using SSFS and a turn-key, fiber-based femtosecond laser can fulfill the requirements for simultaneous imaging of multiple FPs in 2PM, opening new opportunities for a wide range of biological applications where non-invasive, high-resolution imaging of multiple fluorescent indicators is required.

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

    NASA Astrophysics Data System (ADS)

    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.

  16. Enhancing fluorescence intensity of Ellagic acid in Borax-HCl-CTAB micelles.

    PubMed

    Wang, Feng; Huang, Wei; Zhang, Shuai; Liu, Guokui; Li, Kexiang; Tang, Bo

    2011-03-01

    Ellagic acid (C(14)H(6)O(8)), a naturally occurring phytochemical, found mainly in berries and some nuts, has anticarcinogenic and antioxidant properties. It is found that fluorescence of Ellagic acid (EA) is greatly enhanced by micelle of cetyltrimethylammonium bromide (CTAB) surfactant. Based on this effect, a sensitive proposed fluorimetric method was applied for the determination of Ellagic acid in aqueous solution. In the Borax-HCl buffer, the fluorescence intensity of Ellagic acid in the presence of CTAB is proportional to the concentration of Ellagic acid in range from 8.0×10(-10) to 4.0×10(-5) mol L(-1); and the detection limits are 3.2×10(-10) mol L(-1) and 5.9×10(-10) mol L(-1) excited at 266 nm and 388 nm, respectively. The actual samples of pomegranate rinds are simply manipulated and satisfactorily determined. The interaction mechanism studies argue that the negative EA-Borax complex is formed and solubilized in the cationic surfactant CTAB micelle in this system. The fluorescence intensity of EA enhances because the CTAB micelle provides a hydrophobic microenvironment for EA-Borax complex, which can prevent collision with water molecules and decrease the energy loss of EA-Borax complex. Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.

  17. Enhancing fluorescence intensity of Ellagic acid in Borax-HCl-CTAB micelles

    NASA Astrophysics Data System (ADS)

    Wang, Feng; Huang, Wei; Zhang, Shuai; Liu, Guokui; Li, Kexiang; Tang, Bo

    2011-03-01

    Ellagic acid (C 14H 6O 8), a naturally occurring phytochemical, found mainly in berries and some nuts, has anticarcinogenic and antioxidant properties. It is found that fluorescence of Ellagic acid (EA) is greatly enhanced by micelle of cetyltrimethylammonium bromide (CTAB) surfactant. Based on this effect, a sensitive proposed fluorimetric method was applied for the determination of Ellagic acid in aqueous solution. In the Borax-HCl buffer, the fluorescence intensity of Ellagic acid in the presence of CTAB is proportional to the concentration of Ellagic acid in range from 8.0 × 10 -10 to 4.0 × 10 -5 mol L -1; and the detection limits are 3.2 × 10 -10 mol L -1 and 5.9 × 10 -10 mol L -1 excited at 266 nm and 388 nm, respectively. The actual samples of pomegranate rinds are simply manipulated and satisfactorily determined. The interaction mechanism studies argue that the negative EA-Borax complex is formed and solubilized in the cationic surfactant CTAB micelle in this system. The fluorescence intensity of EA enhances because the CTAB micelle provides a hydrophobic microenvironment for EA-Borax complex, which can prevent collision with water molecules and decrease the energy loss of EA-Borax complex.

  18. Accounting for Photophysical Processes and Specific Signal Intensity Changes in Fluorescence-Detected Sedimentation Velocity

    PubMed Central

    2015-01-01

    Fluorescence detected sedimentation velocity (FDS-SV) has emerged as a powerful technique for the study of high-affinity protein interactions, with hydrodynamic resolution exceeding that of diffusion-based techniques, and with sufficient sensitivity for binding studies at low picomolar concentrations. For the detailed quantitative analysis of the observed sedimentation boundaries, it is necessary to adjust the conventional sedimentation models to the FDS data structure. A key consideration is the change in the macromolecular fluorescence intensity during the course of the experiment, caused by slow drifts of the excitation laser power, and/or by photophysical processes. In the present work, we demonstrate that FDS-SV data have inherently a reference for the time-dependent macromolecular signal intensity, resting on a geometric link between radial boundary migration and plateau signal. We show how this new time-domain can be exploited to study molecules exhibiting photobleaching and photoactivation. This expands the application of FDS-SV to proteins tagged with photoswitchable fluorescent proteins, organic dyes, or nanoparticles, such as those recently introduced for subdiffraction microscopy and enables FDS-SV studies of their interactions and size distributions. At the same time, we find that conventional fluorophores undergo minimal photobleaching under standard illumination in the FDS. These findings support the application of a high laser power density for the detection, which we demonstrate can further increase the signal quality. PMID:25136929

  19. Multidimensional fluorescence microscopy of multiple organelles in Arabidopsis seedlings

    PubMed Central

    Kato, Naohiro; Reynolds, Dexter; Brown, Matthew L; Boisdore, Marietta; Fujikawa, Yukichi; Morales, Andrea; Meisel, Lee A

    2008-01-01

    Background The isolation of green fluorescent protein (GFP) and the development of spectral variants over the past decade have begun to reveal the dynamic nature of protein trafficking and organelle motility. In planta analyses of this dynamic process have typically been limited to only two organelles or proteins at a time in only a few cell types. Results We generated a transgenic Arabidopsis plant that contains four spectrally different fluorescent proteins. Nuclei, plastids, mitochondria and plasma membranes were genetically tagged with cyan, red, yellow and green fluorescent proteins, respectively. In addition, methods to track nuclei, mitochondria and chloroplasts and quantify the interaction between these organelles at a submicron resolution were developed. These analyzes revealed that N-ethylmaleimide disrupts nuclear-mitochondrial but not nuclear-plastids interactions in root epidermal cells of live Arabidopsis seedlings. Conclusion We developed a tool and associated methods for analyzing the complex dynamic of organelle-organelle interactions in real time in planta. Homozygous transgenic Arabidopsis (Kaleidocell) is available through Arabidopsis Biological Resource Center. PMID:18489765

  20. Real-time micro-scale temperature imaging at low cost based on fluorescent intensity ratio

    PubMed Central

    Xiong, Jianghao; Zhao, Mingshu; Han, Xiaotian; Cao, Zhongmin; Wei, Xiantao; Chen, Yonghu; Duan, Changkui; Yin, Min

    2017-01-01

    Real-time temperature imaging with high spatial resolution has been a challenging task but also one with wide potential applications. To achieve this task, temperature sensor is critical. Fluorescent materials stand out to be promising candidates due to their quick response and strong temperature dependence. However, former reported temperature imaging techniques with fluorescent materials are mainly based on point by point scanning, which cannot fulfill the requirement of real-time monitoring. Based on fluorescent intensity ratio (FIR) of two emission bands of SrB4O7:Sm2+, whose spatial distributions were simultaneously recorded by two cameras with special filters separately, real-time temperature imaging with high spatial resolution has been realized with low cost. The temperature resolution can reach about 2 °C in the temperature range from 120 to 280 °C; the spatial resolution is about 2.4 μm and the imaging time is as fast as one second. Adopting this system, we observed the dynamic change of a micro-scale thermal distribution on a printed circuit board (PCB). Different applications and better performance could also be achieved on this system with appropriate fluorescent materials and high sensitive CCD detectors according to the experimental environment. PMID:28145482

  1. Real-time micro-scale temperature imaging at low cost based on fluorescent intensity ratio

    NASA Astrophysics Data System (ADS)

    Xiong, Jianghao; Zhao, Mingshu; Han, Xiaotian; Cao, Zhongmin; Wei, Xiantao; Chen, Yonghu; Duan, Changkui; Yin, Min

    2017-02-01

    Real-time temperature imaging with high spatial resolution has been a challenging task but also one with wide potential applications. To achieve this task, temperature sensor is critical. Fluorescent materials stand out to be promising candidates due to their quick response and strong temperature dependence. However, former reported temperature imaging techniques with fluorescent materials are mainly based on point by point scanning, which cannot fulfill the requirement of real-time monitoring. Based on fluorescent intensity ratio (FIR) of two emission bands of SrB4O7:Sm2+, whose spatial distributions were simultaneously recorded by two cameras with special filters separately, real-time temperature imaging with high spatial resolution has been realized with low cost. The temperature resolution can reach about 2 °C in the temperature range from 120 to 280 °C the spatial resolution is about 2.4 μm and the imaging time is as fast as one second. Adopting this system, we observed the dynamic change of a micro-scale thermal distribution on a printed circuit board (PCB). Different applications and better performance could also be achieved on this system with appropriate fluorescent materials and high sensitive CCD detectors according to the experimental environment.

  2. Enhancement of the quantum dot fluorescence intensity by Au nanoparticle decoration of a porous silicon photonic crystal

    NASA Astrophysics Data System (ADS)

    He, Lei; Jia, Zhenhong; Zhou, Jun; Zhang, Hongyan; Lv, Xiaoyi; Sun, Difei

    2017-05-01

    A new way to enhance the photoluminescence of CdSe/ZnS quantum dots embedded in porous silicon (PSi) is reported, which results in the double enhancement of the quantum dot (QD) fluorescence due to Au nanoparticle decoration and the porous silicon having a distributed Bragg reflector (DBR) structure. Through the dropwise addition of a solution of CdSe/ZnS QDs on Au nanoparticle-decorated porous silicon samples, the QDs successfully infiltrated the porous silicon substrate. Among the fluorescence intensities of QDs/single-layer PSi, QDs/multilayer PSi, QDs/Au/single-layer PSi and QDs/Au/multilayer PSi, we find that the fluorescence of the QD-infiltrated multilayer PSi is stronger than that of the single-layer sample, and the multilayer sample decorated with Au nanoparticles shows further improvement of the fluorescence intensity through plasmonic effects. PSi, as a substrate with a spongy structure, offers favorable conditions for enhancing fluorescence intensity.

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

    NASA Astrophysics Data System (ADS)

    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.

  4. Enhancing magnetic resonance imaging tumor detection with fluorescence intensity and lifetime imaging

    NASA Astrophysics Data System (ADS)

    Erten, Ahmet; Hall, David; Hoh, Carl; Tran Cao, Hop S.; Kaushal, Sharmeela; Esener, Sadik; Hoffman, Robert M.; Bouvet, Michael; Chen, James; Kesari, Santosh; Makale, Milan

    2010-11-01

    Early detection is important for many solid cancers but the images provided by ultrasound, magnetic resonance imaging (MRI), and computed tomography applied alone or together, are often not sufficient for decisive early screening/diagnosis. We demonstrate that MRI augmented with fluorescence intensity (FI) substantially improves detection. Early stage murine pancreatic tumors that could not be identified by blinded, skilled observers using MRI alone, were easily identified with MRI along with FI images acquired with photomultiplier tube detection and offset laser scanning. Moreover, we show that fluorescence lifetime (FLT) imaging enables positive identification of the labeling fluorophore and discriminates it from surrounding tissue autofluorescence. Our data suggest combined-modality imaging with MRI, FI, and FLT can be used to screen and diagnose early tumors.

  5. Improvement of fluorescence intensity of nitrogen vacancy centers in self-formed diamond microstructures

    SciTech Connect

    Furuyama, S.; Yaita, J.; Kondo, M.; Tahara, K.; Iwasaki, T.; Shimizu, M.; Kodera, T.; Hatano, M.

    2015-10-19

    We present umbrella-shaped diamond microstructures with metal mirrors at the bottom in order to improve the amount of collected photons from nitrogen vacancy centers. The metal mirrors at the bottom are self-aligned to the umbrella-shaped diamond microstructures which are selectively grown through holes created on a metal mask. By the finite-difference time-domain simulations, we found that the umbrella-shaped microstructures, which have an effect similar to solid immersion lens, could collect photons more efficiently than bulk or pillar-shaped microstructures. Improvement of the fluorescence intensity by factors of from 3 to 5 is shown experimentally.

  6. Theory of time-dependent intense-field collisional resonance fluorescence

    NASA Technical Reports Server (NTRS)

    Kleiber, P. D.; Cooper, J.; Burnett, K.; Kunasz, C. V.; Raymer, M. G.

    1983-01-01

    The time-dependent theory of Courtens and Szoke (1977) is generalized using the approach of Burnett et al. (1982) to derive time-dependent spectral intensities of resonance fluorescence from atoms driven by a pulsed laser in the presence of collisions. These results are valid both for laser detunings inside and outside the usual impact region of the spectrum, including Zeeman degeneracy effects. This theory is applied to a simple but important example (J = 0 to J = 1) to obtain quantitative predictions for the observable scattered-light spectrum which can be directly compared with recent experiments.

  7. Theory of time-dependent intense-field collisional resonance fluorescence

    NASA Technical Reports Server (NTRS)

    Kleiber, P. D.; Cooper, J.; Burnett, K.; Kunasz, C. V.; Raymer, M. G.

    1983-01-01

    The time-dependent theory of Courtens and Szoke (1977) is generalized using the approach of Burnett et al. (1982) to derive time-dependent spectral intensities of resonance fluorescence from atoms driven by a pulsed laser in the presence of collisions. These results are valid both for laser detunings inside and outside the usual impact region of the spectrum, including Zeeman degeneracy effects. This theory is applied to a simple but important example (J = 0 to J = 1) to obtain quantitative predictions for the observable scattered-light spectrum which can be directly compared with recent experiments.

  8. Revealing single emitter spectral dynamics from intensity correlations in an ensemble fluorescence spectrum.

    PubMed

    Brokmann, Xavier; Marshall, Lisa; Bawendi, Moungi

    2009-03-16

    We show that the single emitter linewidth underlying a broadened ensemble emission spectrum can be extracted from correlations among the stochastic intensity fluctuations in the ensemble spectrum. Spectral correlations can be observed at high temporal and spectral resolutions with a cross-correlated pair of avalanche photodiodes placed at the outputs of a scanning Michelson interferometer. As illustrated with simulations in conjunction with Fluorescence Correlation Spectroscopy, our approach overcomes ensemble and temporal inhomogeneous broadening to provide single emitter linewidths, even for emitters under weak, continuous, broadband excitation. (c) 2009 Optical Society of America

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

    PubMed

    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. Copyright © 2011 Elsevier B.V. All rights reserved.

  10. Beau's lines and multiple periungueal pyogenic granulomas after long stay in an intensive care unit.

    PubMed

    Guhl, Guillermo; Torrelo, Antonio; Hernández, Angela; Zambrano, Antonio

    2008-01-01

    A child developed multiple Beau's lines and periungueal pyogenic granulomas after admission to the intensive care unit. Immobilization, hypoxia, and drugs might have acted as potential causative factors.

  11. Apparatus and method for measuring fluorescence intensities at a plurality of wavelengths and lifetimes

    DOEpatents

    Buican, T.N.

    1993-05-04

    Apparatus and method is described for measuring intensities at a plurality of wavelengths and lifetimes. A source of multiple-wavelength electromagnetic radiation is passed through a first interferometer modulated at a first frequency, the output thereof being directed into a sample to be investigated. The light emitted from the sample as a result of the interaction thereof with the excitation radiation is directed into a second interferometer modulated at a second frequency, and the output detected and analyzed. In this manner excitation, emission, and lifetime information may be obtained for a multiplicity of fluorochromes in the sample.

  12. Apparatus and method for measuring fluorescence intensities at a plurality of wavelengths and lifetimes

    DOEpatents

    Buican, Tudor N.

    1993-01-01

    Apparatus and method for measuring intensities at a plurality of wavelengths and lifetimes. A source of multiple-wavelength electromagnetic radiation is passed through a first interferometer modulated at a first frequency, the output thereof being directed into a sample to be investigated. The light emitted from the sample as a result of the interaction thereof with the excitation radiation is directed into a second interferometer modulated at a second frequency, and the output detected and analyzed. In this manner excitation, emission, and lifetime information may be obtained for a multiplicity of fluorochomes in the sample.

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

    PubMed

    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.

  14. 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.

    PubMed

    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.

  15. Structural and permeability sensitivity of cells to low intensity ultrasound: Infrared and fluorescence evidence in vitro.

    PubMed

    Domenici, Fabio; Giliberti, Claudia; Bedini, Angelico; Palomba, Raffaele; Udroiu, Ion; Di Giambattista, Lucia; Pozzi, Deleana; Morrone, Stefania; Bordi, Federico; Congiu Castellano, Agostina

    2014-04-01

    This work is focused on the in vitro study of the effects induced by medical ultrasound (US) in murine fibroblast cells (NIH-3T3) at a low-intensity of exposure (spatial peak temporal average intensity Ita<0.1Wcm(-2)). Conventional 1MHz and 3MHz US devices of therapeutic relevance were employed with varying intensity and exposure time parameters. In this framework, upon cells exposure to US, structural changes at the molecular level were evaluated by infrared spectroscopy; alterations in plasma membrane permeability were monitored in terms of uptake efficiency of small cell-impermeable model drug molecules, as measured by fluorescence microscopy and flow cytometry. The results were related to the cell viability and combined with the statistical PCA analysis, confirming that NIH-3T3 cells are sensitive to therapeutic US, mainly at 1MHz, with time-dependent increases in both efficiency of uptake, recovery of wild-type membrane permeability, and the size of molecules entering 3T3. On the contrary, the exposures from US equipment at 3MHz show uptakes comparable with untreated samples.

  16. Improvement of the fluorescence intensity during a flow cytometric analysis for rice protoplasts by localization of a green fluorescent protein into chloroplasts.

    PubMed

    You, Min Kyoung; Lim, Sun-Hyung; Kim, Min-Jin; Jeong, Ye Sol; Lee, Mi-Gi; Ha, Sun-Hwa

    2014-12-31

    Protoplasts have been a useful unicellular system for various molecular biological analyses based on transient expression and single cell analysis using fluorescence-activated cell sorting (FACS), widely used as a powerful method in functional genomics. Despite the versatility of these methods, some limits based on low fluorescence intensity of a flow cytometric analysis (FCA) using protoplasts have been reported. In this study, the chloroplast targeting of fluorescent proteins (FPs) led to an eight-fold increase in fluorescence intensity and a 4.5-fold increase of transfection ratio from 14.7% to 65.7% as compared with their targeting into the cytoplasm. Moreover, the plot data of FCA shows that 83.3% of the K-sGFP population is under the threshold level, regarded as a non-transgenic population with background signals, while 65.7% of the K-sGFP population is spread on overall intervals. To investigate the reason underlying this finding, mRNA/protein levels and transfection efficiency were analyzed, and results suggest that mRNA/protein levels and transfection ratio are not much different between K-sGFP and KR-sGFP. From those results, we hypothesized that the difference of fluorescence intensity is not only derived from cellular events such as molecular level or transfection efficiency. Taken together, we suggest that the translocation of FPs into chloroplasts contributes to the improvement of fluorescence intensity in FCA and, apparently, plays an important role in minimizing the loss of the transfected population. Our study could be usefully applicable for highly sensitive FACS and FCA-investigations of green tissue.

  17. Mapping tillage intensity by integrating multiple remote sensing data

    USDA-ARS?s Scientific Manuscript database

    Tillage practices play an important role in the sustainable agriculture system. Conservative tillage practice can help to reduce soil erosion, increase soil fertility and improve water quality. Tillage practices could be applied at different times with different intensity depending on the local weat...

  18. X-ray Methods in High-Intensity Discharges and Metal-Halide Lamps: X-ray Induced Fluorescence

    SciTech Connect

    Curry, John J.; Lapatovich, Walter P.; Henins, Albert

    2011-12-09

    We describe the use of x-ray induced fluorescence to study metal-halide high-intensity discharge lamps and to measure equilibrium vapor pressures of metal-halide salts. The physical principles of metal-halide lamps, relevant aspects of x-ray-atom interactions, the experimental method using synchrotron radiation, and x-ray induced fluorescence measurements relevant to metal-halide lamps are covered.

  19. Using water raman intensity to determine the effective excitation and emission path lengths of fluorophotometers for correcting fluorescence inner filter effect

    USDA-ARS?s Scientific Manuscript database

    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 ...

  20. Weighted PARAFAC and Non-linear Regression for Handling Intensity Changes in Fluorescence Spectroscopy caused by pH Fluctuations.

    PubMed

    Svendsen, Carina; Skov, Thomas; van den Berg, Frans W J

    2016-07-22

    Fluorescence spectroscopy is a sensitive and selective technique, which can be of great value in bioprocesses to provide online, real-time measures of chemical compounds. Although fluorescence spectroscopy is a widely studied method, not much attention has been given to issues concerning intensity variations in the fluorescence landscapes due to pH fluctuations. This study elucidates how pH fluctuations cause intensity changes in fluorescence measurements and thereby decreases the quality of the subsequent quantification. A photo-degradation process of riboflavin was investigated by fluorescence spectroscopy and used as a model system. A two-step modeling approach, combining weighted PARAllel FACtor analysis (PARAFAC) with weighted non-linear regression of the known reaction kinetics, is suggested as a way of handling the fluorescence intensity shifts caused by the pH changes. The suggested strategy makes it possible to compensate for uncertainties in the shifted data and thereby obtain more reliable concentration profiles for the chemical compounds and kinetic parameters of the reaction.

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

    NASA Astrophysics Data System (ADS)

    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).

  2. Sympathetic and sensory innervation of small intensely fluorescent (SIF) cells in rat superior cervical ganglion.

    PubMed

    Takaki, Fumiya; Nakamuta, Nobuaki; Kusakabe, Tatsumi; Yamamoto, Yoshio

    2015-02-01

    The sympathetic ganglion contains small intensely fluorescent (SIF) cells derived from the neural crest. We morphologically characterize SIF cells and focus on their relationship with ganglionic cells, preganglionic nerve fibers and sensory nerve endings. SIF cells stained intensely for tyrosine hydroxylase (TH), with a few cells also being immunoreactive for dopamine β-hydroxylase (DBH). Vesicular acetylcholine transporter (VAChT)-immunoreactive puncta were distributed around some clusters of SIF cells, whereas some SIF cells closely abutted DBH-immunoreactive ganglionic cells. SIF cells contained bassoon-immunoreactive products beneath the cell membrane at the attachments and on opposite sites to the ganglionic cells. Ganglion neurons and SIF cells were immunoreactive to dopamine D2 receptors. Immunohistochemistry for P2X3 revealed ramified nerve endings with P2X3 immunoreactivity around SIF cells. Triple-labeling for P2X3, TH and VAChT allowed the classification of SIF cells into three types based on their innervation: (1) with only VAChT-immunoreactive puncta, (2) with only P2X3-immunoreactive nerve endings, (3) with both P2X3-immunoreactive nerve endings and VAChT-immunoreactive puncta. The results of retrograde tracing with fast blue dye indicated that most of these nerve endings originated from the petrosal ganglion. Thus, SIF cells in the superior cervical ganglion are innervated by preganglionic fibers and glossopharyngeal sensory nerve endings and can be classified into three types. SIF cells might modulate sympathetic activity in the superior cervical ganglion.

  3. Sub-annual paleoenvironmental information evaluated from intensity variations of fluorescent annual layers in a stalagmite from Ryuo-do Cave, Nagasaki Prefecture, western Japan

    NASA Astrophysics Data System (ADS)

    Sasaki, Hana; Onishi, Yuri; Ishihara, Yoshiro; Yoshimura, Kazuhisa

    2017-04-01

    vary stratigraphically, and multiple types of fluorescence intensity pattern are observed in the stalagmite. When the co-precipitation of FA is governed by the hiatus model, it is suggested that a gradual increase in the annual layers will result from a large accumulation of calcite after the annual peak in the FA concentration, whereas there will be a gradual decrease if the main growth occurs before the annual peak in FA concentration. However, in the case of the PC model, a gradually increasing type of pattern is formed if the main growth occurs before the annual peak in FA concentration, and a gradually decreasing type is formed if the main growth occurs afterwards. If the annual peak of FA concentration occurs several months after high summer, it is suggested that intervals showing a gradually increasing type were formed in winter, and intervals showing a gradually decreasing type were formed in the early summer, in the case of the hiatus model. In the case of PC model, the seasons are reversed. In the climatic environment around the Ryuo-do Cave, the growth rates of stalagmites are affected by cave air circulation in winter and by rainfall (rainy season) in early summer.

  4. Intense Field-Matter Interactions: Multiple Ionization of Clusters

    SciTech Connect

    Snyder, E.M.; Buzza, S.A.; Castleman, A.W. Jr.

    1996-10-01

    We report the results for the production of highly charged atomic species (e.g., Xe{sup 20+}, Kr{sup 18+}, O{sup 5+}, and C{sup 4+}) resulting from the interaction of intense laser fields (up to {approximately}10{sup 15} W/cm{sup 2}) with atomic and multicenter molecular clusters. The processes are also investigated using ultrafast pump-probe techniques, showing distinct beating patterns for the ionization structure in the molecular system. A comparison of our results with predictions of several different theoretical models provides strong support for the ionization ignition mechanism. {copyright} {ital 1996 The American Physical Society.}

  5. State transitions and fluorescence quenching in the cyanobacterium Synechocystis PCC 6803 in response to changes in light quality and intensity.

    PubMed

    Zhao, Wenfeng; Xie, Jie; Xu, Xiuling; Zhao, Jingquan

    2015-01-01

    State transition and non-photochemical fluorescence quenching in cyanobacteria are short-term adaptations of photosynthetic apparatus to changes in light quality and intensity, however, the kinetic details and relationship are still not clear. In this work, time-dependent 77K fluorescence spectra were monitored for cyanobacterium Synechocystis PCC 6803 cells under blue, orange and blue-green light in a series of intensities. The characteristic fluorescence signals indicated state transition taking place exclusively under 430-450 or 580-600nm light or 480-550nm light at the intensities ⩽150μEm(-2)s(-1) to achieve a conserved level with variable rate constant. Under 480-500nm or 530-550nm light at the intensities ⩾160μEm(-2)s(-1), state transition took place at first but stopped as soon as the fluorescence quenching appeared. The dependence of appearance, induction period, level and rate constant for the quenching on light intensity suggests that a critical concentration of photo-activated OCPs is necessary and may be achieved by a dynamic equilibrium between the activation and deactivation under light.

  6. Fluorescent and high intensity discharge lamp use in chambers and greenhouses

    SciTech Connect

    Langhans, R.W.

    1994-12-31

    Fluorescent and High Intensity Discharge lamps have opened up great opportunities for researchers to study plant growth under controlled environment conditions and for commercial growers to increase plant production during low/light periods. This report describes the advantages and disadvantages of using each lamp in growth chambers, growth rooms and greenhouses. Growth Chambers are small (3m x 4/m and smaller) walk-in or reach-in enclosures with programmable, accurate temperature, relative humidity (RH) and irradiance control over wide ranges. The intent of growth chambers was to replicate sunlight conditions and transfer research results directly to the greenhouse or outside. It was realized that sunlight and outside conditions could not be mimicked. Growth chambers are also used to study irradiance and spectral fluxes. Growth Rooms are usually large rooms (larger than 3m x 4m) with only lamp irradiance, but providing relatively limited ranges of environmental control (i.e., 10 to 30 C temperature, 50 to 90% RH and ambient to 1000 ppm CO{sub 2}), and commonly independent of outside conditions. Irradiance requirements for growth rooms are similar to those of growth chambers. Growth rooms are also used for growing a large number of plants in a uniform standard environment condition and in commercial horticulture for tissue culture, seed germination (plugs) and seedling growth. Greenhouses are designed to allow maximum sunlight penetration through the structure. Initially greenhouses were used to extend the growing season. Then as heating systems, and cooling systems improved, they were used year round. Low light during the winter months reduced plant growth, but with the advent of efficient lamps (HID and fluorescent) it became possible to increase growth to rates close to that in summer months. Supplementary lighting is used during low light periods of the year and anytime to ensure consistent total daily irradiance for research plants.

  7. Application of a fluorescence intensity ratio technique for the intrinsic determination of pH using an optical fiber sensor

    NASA Astrophysics Data System (ADS)

    Thotath, Bhadra; Nguyen, T. Hien; Zhang, Weiwei; Wren, Stephen P.; Baxter, Gregory W.; Sun, Tong; Collins, Stephen F.; Grattan, Kenneth T. V.

    2015-09-01

    An intensity ratio technique has been used for characterizing fluorescence spectra from novel coumarin dyes for pH sensing, in the range of 0.5 - 6, providing results that are independent of possible fluctuations in the intensity of the excitation source, deterioration of the indicator and changes in optical coupling. The arrangement was determined to have a sensitivity of 25% per unit pH change (at a pH of 4).

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

    NASA Astrophysics Data System (ADS)

    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.

  9. mMaple: A Photoconvertible Fluorescent Protein for Use in Multiple Imaging Modalities

    PubMed Central

    McEvoy, Ann L.; Hoi, Hiofan; Bates, Mark; Platonova, Evgenia; Cranfill, Paula J.; Baird, Michelle A.; Davidson, Michael W.; Ewers, Helge; Liphardt, Jan; Campbell, Robert E.

    2012-01-01

    Recent advances in fluorescence microscopy have extended the spatial resolution to the nanometer scale. Here, we report an engineered photoconvertible fluorescent protein (pcFP) variant, designated as mMaple, that is suited for use in multiple conventional and super-resolution imaging modalities, specifically, widefield and confocal microscopy, structured illumination microscopy (SIM), and single-molecule localization microscopy. We demonstrate the versatility of mMaple by obtaining super-resolution images of protein organization in Escherichia coli and conventional fluorescence images of mammalian cells. Beneficial features of mMaple include high photostability of the green state when expressed in mammalian cells and high steady state intracellular protein concentration of functional protein when expressed in E. coli. mMaple thus enables both fast live-cell ensemble imaging and high precision single molecule localization for a single pcFP-containing construct. PMID:23240015

  10. Visible-light optical coherence tomography-based multimodal retinal imaging for improvement of fluorescent intensity quantification

    PubMed Central

    Nafar, Zahra; Jiang, Minshan; Wen, Rong; Jiao, Shuliang

    2016-01-01

    We developed a spectral-domain visible-light optical coherence tomography (VIS-OCT) based multimodal imaging technique which can accomplish simultaneous OCT and fluorescence imaging with a single broadband light source. Phantom experiments showed that by using the simultaneously acquired OCT images as a reference, the effect of light attenuation on the intensity of the fluorescent images by materials in front of the fluorescent target can be compensated. This capability of the multimodal imaging technique is of high importance for achieving quantification of the true intensities of autofluorescence (AF) imaging of the retina. We applied the technique in retinal imaging including AF imaging of the retinal pigment epithelium and fluorescein angiography (FA). We successfully demonstrated the effect of compensation on AF and FA images with the simultaneously acquired VIS-OCT images. PMID:27699094

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

    SciTech Connect

    Yan, Yiying Lü, Zhiguo Zheng, Hang

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed Central

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

    2016-01-01

    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. PMID:27527813

  14. Correlation analysis of intracellular and secreted cytokines via the generalized integrated mean fluorescence intensity.

    PubMed

    Shooshtari, Parisa; Fortuno, Edgardo S; Blimkie, Darren; Yu, Miao; Gupta, Arvind; Kollmann, Tobias R; Brinkman, Ryan R

    2010-09-01

    The immune response in humans is usually assessed using immunogenicity assays to provide biomarkers as correlates of protection (CoP). Flow cytometry is the assay of choice to measure intracellular cytokine staining (ICS) of cell-mediated immune (CMI) biomarkers. For CMI analysis, the integrated mean fluorescence intensity (iMFI) was introduced as a metric to represent the total functional CMI response as a CoP. iMFI is computed by multiplying the relative frequency (percent positive) of cells expressing a particular cytokine with the MFI of that population, and correlates better with protection in challenge models than either the percentage or the MFI of the cytokine-positive population. While determination of the iMFI as a CoP can readily be accomplished in animal models that allow challenge/protection experiments, this is not feasible in humans for ethical reasons. As a first step toward extending the iMFI concept to humans, we investigated the correlation of the iMFI derived from a human innate immune response ICS assay with functional cytokine release into the culture supernatant, as innate cytokines need to be released to have a functional impact. Next, we developed a quantitatively more correlative mathematical approach for calculating the functional response of cytokine-producing cells by incorporating the assignment of different weights to the magnitude (frequency of cytokine-positive cells) and the quality (the MFI) of the observed innate immune response. We refer to this model as generalized iMFI.

  15. Highly fluorescent rigid supramolecular polymeric nanowires constructed through multiple hydrogen bonds.

    PubMed

    Luo, Jia; Lei, Ting; Wang, Lei; Ma, Yuguo; Cao, Yong; Wang, Jian; Pei, Jian

    2009-02-18

    Supramolecular polymeric nanowires 1(n) constructed by a 3D shape-persistent hexaacid 1 through multiple hydrogen bonding interactions was developed. Single molecular nanoires were also obtained from its highly dilute solution.Hexaacid 1 containing pi-conjugated chromophores successfully self-assembled to afford these nanofibers with high solid quantum efficiency (22%), which provides us a pathway to fabricate optoelectronic devices using these highly fluorescent nanofibers.

  16. Open source software for quantification of cell migration, protrusions, and fluorescence intensities.

    PubMed

    Barry, David J; Durkin, Charlotte H; Abella, Jasmine V; Way, Michael

    2015-04-13

    Cell migration is frequently accompanied by changes in cell morphology (morphodynamics) on a range of spatial and temporal scales. Despite recent advances in imaging techniques, the application of unbiased computational image analysis methods for morphodynamic quantification is rare. For example, manual analysis using kymographs is still commonplace, often caused by lack of access to user-friendly, automated tools. We now describe software designed for the automated quantification of cell migration and morphodynamics. Implemented as a plug-in for the open-source platform, ImageJ, ADAPT is capable of rapid, automated analysis of migration and membrane protrusions, together with associated fluorescently labeled proteins, across multiple cells. We demonstrate the ability of the software by quantifying variations in cell population migration rates on different extracellular matrices. We also show that ADAPT can detect and morphologically profile filopodia. Finally, we have used ADAPT to compile an unbiased description of a "typical" bleb formed at the plasma membrane and quantify the effect of Arp2/3 complex inhibition on bleb retraction. © 2015 Barry et al.

  17. Noninvasive imaging of multiple myeloma using near infrared fluorescent molecular probe

    NASA Astrophysics Data System (ADS)

    Hathi, Deep; Zhou, Haiying; Bollerman-Nowlis, Alex; Shokeen, Monica; Akers, Walter J.

    2016-03-01

    Multiple myeloma is a plasma cell malignancy characterized by monoclonal gammopathy and osteolytic bone lesions. Multiple myeloma is most commonly diagnosed in late disease stages, presenting with pathologic fracture. Early diagnosis and monitoring of disease status may improve quality of life and long-term survival for multiple myeloma patients from what is now a devastating and fatal disease. We have developed a near-infrared targeted fluorescent molecular probe with high affinity to the α4β1 integrin receptor (VLA-4)overexpressed by a majority of multiple myeloma cells as a non-radioactive analog to PET/CT tracer currently being developed for human diagnostics. A near-infrared dye that emits about 700 nm was conjugated to a high affinity peptidomimmetic. Binding affinity and specificity for multiple myeloma cells was investigated in vitro by tissue staining and flow cytometry. After demonstration of sensitivity and specificity, preclinical optical imaging studies were performed to evaluate tumor specificity in murine subcutaneous and metastatic multiple myeloma models. The VLA-4-targeted molecular probe showed high affinity for subcutaneous MM tumor xenografts. Importantly, tumor cells specific accumulation in the bone marrow of metastatic multiple myeloma correlated with GFP signal from transfected cells. Ex vivo flow cytometry of tumor tissue and bone marrow further corroborated in vivo imaging data, demonstrating the specificity of the novel agent and potential for quantitative imaging of multiple myeloma burden in these models.

  18. [Theoretical study of fluorescence of photosynthetic pigments at complex dependence of intensity of exciting light on time].

    PubMed

    Alekseev, A A; Belov, A A; Kirzhanov, D V; Kukushkin, A K

    2012-01-01

    Now the methods using the radiance with complex dependence of light intensity on time are applied to research of photosynthesis by means of fluorescence, exciting photosynthetic pigments. One of these methods is applied in PAM-fluorometers--the commercial devices currently widely used to investigate a state of photosynthesizing systems. However, if excitation is performed in this way, the question, what components of fluorescence are registered at an output of such devices, remains to be open-ended. In this work an attempt to analyse this task has been made.

  19. Mapping grape berry photosynthesis by chlorophyll fluorescence imaging: the effect of saturating pulse intensity in different tissues.

    PubMed

    Breia, Richard; Vieira, Sónia; da Silva, Jorge Marques; Gerós, Hernâni; Cunha, Ana

    2013-01-01

    Grape berry development and ripening depends mainly on imported photosynthates from leaves, however, fruit photosynthesis may also contribute to the carbon economy of the fruit. In this study pulse amplitude modulated chlorophyll fluorescence imaging (imaging-PAM) was used to assess photosynthetic properties of tissues of green grape berries. In particular, the effect of the saturation pulse (SP) intensity was investigated. A clear tissue-specific distribution pattern of photosynthetic competence was observed. The exocarp revealed the highest photosynthetic capacity and the lowest susceptibility to photoinhibition, and the mesocarp exhibited very low fluorescence signals and photochemical competence. Remarkably, the seed outer integument revealed a photosynthetic ability similar to that of the exocarp. At a SP intensity of 5000 μmol m(-2) s(-1) several photochemical parameters were decreased, including maximum fluorescence in dark-adapted (F(m)) and light-adapted (F'(m)) samples and effective quantum yield of PSII (Φ(II)), but the inner tissues were susceptible to a SP intensity as low as 3200 μmol m(-2) s(-1) under light-adapted conditions, indicating a photoinhibitory interaction between SP and actinic light intensities and repetitive exposure to SP. These results open the way to further studies concerning the involvement of tissue-specific photosynthesis in the highly compartmentalized production and accumulation of organic compounds during grape berry development. © 2013 Wiley Periodicals, Inc. Photochemistry and Photobiology © 2013 The American Society of Photobiology.

  20. Fluorescence of the gamma, epsilon, and delta systems of nitric oxide - Polarization and use of calculated intensities for spectrometer calibration.

    NASA Technical Reports Server (NTRS)

    Poland, H. M.; Broida, H. P.

    1971-01-01

    Results of a study in which fluorescence of the gamma system of nitric oxide was obtained by excitation from both the 2144 A line of ionized cadmium and a continuum source. Individual rotational lines of the 2144 A excited fluorescence spectrum were found to be partially polarized and to have polarizations of differ ing sign. Measured relative vibrational band intensities from line and continuum excitation were compared to calculated Franck-Condon factors. Those Franck-Condon factors based on a single potential for the two spin states of the X super pi state agreed better with measured values than those based on separate potentials for the two spin states. Calculated intensities of the v prime = 3 progression were used to calibrate the instrument response in the wavelength region from 2000 to 2500 A and were checked with measured intensities of the v prime = 0.1, and 2 progressions. Fluorescence of the epsilon and delta bands obtained with continuum lamp excitation also were compared to calculated intensities.

  1. Development of pathological diagnostics of human kidney cancer by multiple staining using new fluorescent Fluolid dyes.

    PubMed

    Wuxiuer, Dilibaier; Zhu, Yun; Ogaeri, Takunori; Mizuki, Keiji; Kashiwa, Yuki; Nishi, Kentaro; Isobe, Shin-ichiro; Aoyagi, Tei-ichiro; Kiyama, Ryoiti

    2014-01-01

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

  3. Prediction of Complement-Binding Capacity of HLA Antibodies Based on Mean Fluorescence Intensity.

    PubMed

    Zhao, J; Fu, Y X; Yang, T; Shen, Z Y; Wu, C L

    2016-01-01

    Human leukocyte antigen (HLA) antibodies estimated by Luminex single-antigen beads, especially those that fix complement, are associated with antibody-mediated rejection and graft failure. However, the relationship between HLA antibody strength and complement-binding ability is controversial. Serum samples of 31 sensitized renal patients waiting for renal transplantation were retrospectively analyzed by IgG-Luminex to identify HLA antibodies and in parallel by C1q-Luminex to determine the complement binding of HLA antibodies. The percentage of HLA class I antibodies binding with C1q was lower than that of HLA class II antibodies (43.2% vs. 51.3%, P = .006). The mean fluorescence intensities (MFI) of IgG-Luminex correlated with the MFI of C1q-Luminex for the same antibodies (Spearman correlation; class I, r = 0.665, P < .01; class II, r = 0.761, P < .01). Receiver operating characteristic (ROC) curve analysis showed that the MFIs of HLA antibodies by IgG-Luminex predicted their C1q-binding abilities (area under the curve [AUC] class I = 0.917; AUC class II = 0.927). Using MFI cutoff values of 8238 and 6754 in IgG-Luminex for HLA class I and class II antibodies, respectively, the sensitivity and specificity for C1q binding were 82.4% and 87.4% for class I antibodies and 90.9% and 82% for class II antibodies. The MFI of HLA antibodies by IgG-Luminex predicts the complement-binding capability to a certain extent before transplantation. Copyright © 2016 Elsevier Inc. All rights reserved.

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

    NASA Astrophysics Data System (ADS)

    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.

  5. Multiple-pulse pumping for enhanced fluorescence detection and molecular imaging in tissue.

    PubMed

    Rich, Ryan M; Gryczynski, Ignacy; Fudala, Rafal; Borejdo, Julian; Stankowska, Dorota L; Krishnamoorthy, Raghu R; Raut, Sangram; Maliwal, Badri P; Shumilov, Dmytro; Doan, Hung; Gryczynski, Zygmunt

    2014-03-15

    Applications of fluorescence based imaging techniques for detection in cellular and tissue environments are severely limited by autofluorescence of endogenous components of cells, tissue, and the fixatives used in sample processing. To achieve sufficient signal-to-background ratio, a high concentration of the probe needs to be used which is not always feasible. Since typically autofluorescence is in the nanosecond range, long-lived fluorescence probes in combination with time-gated detection can be used for suppression of unwanted autofluorescence. Unfortunately, this requires the sacrifice of the large portion the probe signal in order to sufficiently filter the background. We report a simple and practical approach to achieve a many-fold increase in the intensity of a long-lived probe without increasing the background fluorescence. Using controllable, well separated bursts of closely spaced laser excitation pulses, we are able to highly increase the fluorescence signal of a long-lived marker over the endogenous fluorescent background and scattering, thereby greatly increasing detection sensitivity. Using a commercially available confocal microscopy system equipped with a laser diode and time correlated single photon counting (TCSPC) detection, we are able to enhance the signal of a long-lived Ruthenium (Ru)-based probe by nearly an order of magnitude. We used 80 MHz bursts of pulses (12.5 ns pulse separation) repeated with a 320 kHz repetition rate as needed to adequately image a dye with a 380 ns lifetime. Just using 10 pulses in the burst increases the Ru signal almost 10-fold without any increase in the background signal.

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

    NASA Astrophysics Data System (ADS)

    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.

  7. The correlation of the maximum intensity of fluorescence with pigment characteristics of leaves of Betula pendula

    NASA Astrophysics Data System (ADS)

    Zavoruev, V. V.; Zavorueva, E. N.

    2015-11-01

    Using fluorimeter Junior PAM (Heinz Walz GmbH, Germany) the fluorescence parameters of leaves of Betula pendula are investigated. A linear dependence of the maximum fluorescence (Fm) of leaves from the ratio of total chlorophylls concentration to concentration of carotenoids is obtained. Such dependence is found for samples collected during the period of vegetation and for simultaneous selection of colored leaves.

  8. Dynamic labelling of neural connections in multiple colours by trans-synaptic fluorescence complementation

    PubMed Central

    Macpherson, Lindsey J.; Zaharieva, Emanuela E.; Kearney, Patrick J.; Alpert, Michael H.; Lin, Tzu-Yang; Turan, Zeynep; Lee, Chi-Hon; Gallio, Marco

    2015-01-01

    Determining the pattern of activity of individual connections within a neural circuit could provide insights into the computational processes that underlie brain function. Here, we develop new strategies to label active synapses by trans-synaptic fluorescence complementation in Drosophila. First, we demonstrate that a synaptobrevin-GRASP chimera functions as a powerful activity-dependent marker for synapses in vivo. Next, we create cyan and yellow variants, achieving activity-dependent, multi-colour fluorescence reconstitution across synapses (X-RASP). Our system allows for the first time retrospective labelling of synapses (rather than whole neurons) based on their activity, in multiple colours, in the same animal. As individual synapses often act as computational units in the brain, our method will promote the design of experiments that are not possible using existing techniques. Moreover, our strategies are easily adaptable to circuit mapping in any genetic system. PMID:26635273

  9. Paper-based upconversion fluorescence resonance energy transfer biosensor for sensitive detection of multiple cancer biomarkers.

    PubMed

    Xu, Sai; Dong, Biao; Zhou, Donglei; Yin, Ze; Cui, Shaobo; Xu, Wen; Chen, Baojiu; Song, Hongwei

    2016-03-22

    A paper-based upconversion fluorescence resonance energy transfer assay device is proposed for sensitive detection of CEA. The device is fabricated on a normal filter paper with simple nano-printing method. Upconversion nanoparticles tagged with specific antibodies are printed to the test zones on the test paper, followed by the introduction of assay antigen. Upconversion fluorescence measurements are directly conducted on the test zones after the antigen-to-antibody reactions. Furthermore, a multi-channel test paper for simultaneous detection of multiple cancer biomarkers was established by the same method and obtained positive results. The device showed high anti-interfere, stability, reproducible and low detection limit (0.89 ng/mL), moreover it is very easy to fabricate and operate, which is a promising prospect for a clinical point-of-care test.

  10. Paper-based upconversion fluorescence resonance energy transfer biosensor for sensitive detection of multiple cancer biomarkers

    PubMed Central

    Xu, Sai; Dong, Biao; Zhou, Donglei; Yin, Ze; Cui, Shaobo; Xu, Wen; Chen, Baojiu; Song, Hongwei

    2016-01-01

    A paper-based upconversion fluorescence resonance energy transfer assay device is proposed for sensitive detection of CEA. The device is fabricated on a normal filter paper with simple nano-printing method. Upconversion nanoparticles tagged with specific antibodies are printed to the test zones on the test paper, followed by the introduction of assay antigen. Upconversion fluorescence measurements are directly conducted on the test zones after the antigen-to-antibody reactions. Furthermore, a multi-channel test paper for simultaneous detection of multiple cancer biomarkers was established by the same method and obtained positive results. The device showed high anti-interfere, stability, reproducible and low detection limit (0.89 ng/mL), moreover it is very easy to fabricate and operate, which is a promising prospect for a clinical point-of-care test. PMID:27001460

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

    ERIC Educational Resources Information Center

    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…

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

    ERIC Educational Resources Information Center

    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…

  13. [Retrospective, descriptive, observational study of treatment of multiple actinic keratoses with topical methyl aminolevulinate and red light: results in clinical practice and correlation with fluorescence imaging].

    PubMed

    Fernández-Guarino, M; Harto, A; Sánchez-Ronco, M; Pérez-García, B; Marquet, A; Jaén, P

    2008-12-01

    Actinic keratosis (AK) is one of the most common skin diseases seen in clinical practice. In the last 5 years, several studies assessing the efficacy of photodynamic therapy in the treatment of multiple AKs have been published. We aimed to assess the clinical outcomes of photodynamic therapy in patients with multiple AKs and the correlation of those outcomes with fluorescence imaging. In this retrospective, descriptive, observational study of 57 patients treated in our hospital with photodynamic therapy for multiple AKs, we recorded age, sex, and lesion site (face, scalp, and dorsum of the hands). All patients were treated in the same way: methyl aminolevulinic acid (Metvix) was applied for 3 hours and the skin then irradiated with red light at 630 nm, 37 J/cm(2), for 7.5 minutes (Aktilite). The response, remission duration, tolerance, number of sessions, and fluorescence images were recorded by site. The chi(2) test was used to assess between-site differences and the correlation between fluorescence imaging and clinical response. The greatest improvements were obtained for facial lesions; these required fewer sessions and remission lasted longer than lesions at other sites. The treatment was best tolerated on the dorsum of the hands. The fluorescence area and the reduction in intensity on applying treatment were found to be strongly and significantly correlated with the extent of clinical response. Overall, the outcomes of treatment of multiple AKs with photodynamic therapy are better for the face than for the scalp and dorsum of the hands. Fluorescence imaging may be an effective tool for predicting response to treatment.

  14. Up-converted fluorescence from photosynthetic light-harvesting complexes linearly dependent on excitation intensity.

    PubMed

    Leiger, Kristjan; Freiberg, Arvi

    2016-01-01

    Weak up-converted fluorescence related to bacteriochlorophyll a was recorded from various detergent-isolated and membrane-embedded light-harvesting pigment-protein complexes as well as from the functional membranes of photosynthetic purple bacteria under continuous-wave infrared laser excitation at 1064 nm, far outside the optically allowed singlet absorption bands of the chromophore. The fluorescence increases linearly with the excitation power, distinguishing it from the previously observed two-photon excited fluorescence upon femtosecond pulse excitation. Possible mechanisms of this excitation are discussed.

  15. The effect of a gas environment on the fluorescence intensity of quantum-dot composite systems

    NASA Astrophysics Data System (ADS)

    Danilov, V. V.; Khrebtovb, A. I.; Shtrom, I. V.; Tsyrlin, G. E.; Samsonenko, Yu. B.

    2016-09-01

    The fluorescence kinetics of a composite structure based on colloidal CdSe/ZnS core-shell quantum dots deposited on an array of GaAs nanowires in atmospheres of different gases is studied upon excitation by cw laser radiation. It is suggested that the fluorescence enhancement mechanism consists in the transfer of part of the vibrational energy of quantum dots to surrounding gas molecules due to inelastic collisions.

  16. Multiple Velocity Profile Measurements in Hypersonic Flows using Sequentially-Imaged Fluorescence Tagging

    NASA Technical Reports Server (NTRS)

    Bathel, Brett F.; Danehy, Paul M.; Inmian, Jennifer A.; Jones, Stephen B.; Ivey, Christopher B.; Goyne, Christopher P.

    2010-01-01

    Nitric-oxide planar laser-induced fluorescence (NO PLIF) was used to perform velocity measurements in hypersonic flows by generating multiple tagged lines which fluoresce as they convect downstream. For each laser pulse, a single interline, progressive scan intensified CCD camera was used to obtain separate images of the initial undelayed and delayed NO molecules that had been tagged by the laser. The CCD configuration allowed for sub-microsecond acquisition of both images, resulting in sub-microsecond temporal resolution as well as sub-mm spatial resolution (0.5-mm x 0.7-mm). Determination of axial velocity was made by application of a cross-correlation analysis of the horizontal shift of individual tagged lines. Quantification of systematic errors, the contribution of gating/exposure duration errors, and influence of collision rate on fluorescence to temporal uncertainty were made. Quantification of the spatial uncertainty depended upon the analysis technique and signal-to-noise of the acquired profiles. This investigation focused on two hypersonic flow experiments: (1) a reaction control system (RCS) jet on an Orion Crew Exploration Vehicle (CEV) wind tunnel model and (2) a 10-degree half-angle wedge containing a 2-mm tall, 4-mm wide cylindrical boundary layer trip. The experiments were performed at the NASA Langley Research Center's 31-inch Mach 10 wind tunnel.

  17. In vivo imaging of zebrafish digestive organ function using multiple quenched fluorescent reporters.

    PubMed

    Hama, Kotaro; Provost, Elayne; Baranowski, Timothy C; Rubinstein, Amy L; Anderson, Jennifer L; Leach, Steven D; Farber, Steven A

    2009-02-01

    Optical clarity of larvae makes the zebrafish ideal for real-time analyses of vertebrate organ function through the use of fluorescent reporters of enzymatic activities. A key function of digestive organs is to couple the generation of enzymes with mechanical processes that enable nutrient availability and absorption. However, it has been extremely difficult, and in many cases not possible, to directly observe digestive processes in a live vertebrate. Here we describe a new method to visualize intestinal protein and lipid processing simultaneously in live zebrafish larvae using a quenched fluorescent protein (EnzChek) and phospholipid (PED6). By employing these reagents, we found that wild-type larvae exhibit significant variation in intestinal phospholipase and protease activities within a group but display a strong correlation between the activities within individuals. Furthermore, we found that pancreas function is essential for larval digestive protease activity but not for larval intestinal phospholipase activity. Although fat-free (ffr) mutant larvae were previously described to exhibit impaired lipid processes, we found they also had significantly reduced protease activity. Finally, we selected and evaluated compounds that were previously suggested to have altered phospholipase activity and are known or suspected to have inflammatory effects in the intestinal tract including nonsteroidal anti-inflammatory drugs, and identified a compound that significantly increases intestinal phospholipid processing. Thus the multiple fluorescent reporter-based methodology facilitates the rapid analysis of digestive organ function in live zebrafish larvae.

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

    PubMed

    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.

  19. Fluorescence in situ hybridisation of multiple probes on a single microscope slide.

    PubMed Central

    Larin, Z; Fricker, M D; Maher, E; Ishikawa-Brush, Y; Southern, E M

    1994-01-01

    We report a method to analyse multiple samples by fluorescence in situ hybridisation on a single glass microscope slide. Wells were formed in which independent hybridisation reactions could proceed by sealing a silicon rubber gasket to the slide. In the largest format tested, different probes were hybridised simultaneously by applying them directly from a 96-well microtitre dish which was inverted on a glass plate. This technique will increase the rate of analysis of multiple probes against a standard set of chromosomes and could also be used to analyse different karyotypes using a panel of probes such as single chromosome paints during a single operation. It should be useful for both chromosomal mapping projects and screening for chromosome abnormalities in clinical diagnostic laboratories. Images PMID:7937078

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

    PubMed

    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.

  1. Intensity normalization of additive and multiplicative spatially multiplexed patterns with n encoded phases

    NASA Astrophysics Data System (ADS)

    Juarez-Salazar, Rigoberto; Robledo-Sanchez, Carlos; Guerrero-Sanchez, Fermin; Barcelata-Pinzon, Antonio; Gonzalez-Garcia, Jorge; Santiago-Alvarado, Agustin

    2016-02-01

    An efficient, robust and user-free intensity normalization method for patterns with n frequency-multiplexed phases of both additive and multiplicative types is proposed. For this, the parameter estimation approach by using the least-squares method is applied. The theoretical principles are given and the good performance of the proposal is verified by computer simulation and experimental results. Because the proposed method has good features such as robustness, simplicity, fast and user-free execution, it could be implemented in a wide variety of applications. This contribution motivates future generalization in both phase demodulation algorithms and experimental setups to exploit the benefits of additive/multiplicative patterns with multiple phases.

  2. [Detection of multiple human papillomavirus infection in cervical specimens by flow fluorescent hybridization].

    PubMed

    Chen, Qing-yun; Bian, Mei-lu; Zhang, Xiao-yan; Ou, Hua; Ma, Li

    2009-04-07

    To explore the clinical significance of detection of multiple human papillomavirus infection in cervical lesions detected by flow fluorescent hybridization technology with Luminex multi-analytic profiling (xMAP). Cervical exfoliated cell specimens were collected from 301 randomly selected women accepting opportunistic screening for cervical lesions with the cytological results and hybrid capture 2 (HC-2) assay>or=atypical squamous cells of uncertain significance (ASC-US), 48 with the pathological diagnosis>or=cervical intraepithelial neoplasia (CIN)2 (case group) and 253 with normal histological result or only inflammation (control group), aged (34+/-9) (21-59). The samples were tested with xMAP technology with blind method. The coincidence of the xMAP and HC-2 was assessed. The HPV genotype, multiple HPV infection rate, and their relationships to the patients' clinical-pathological features were analyzed. The rates of sensitivity, specificity, and accuracy of xMAP technology to detect>or=CIN2 cervical lesions were 80.49%, 80.00%, and 80.07% respectively. The positive and negative predictive values were 47.06% and 96.30% respectively. The Kappa Index for agreement between xMAP technology and HC-2 was 0.56. The prevalence rate of high-risk HPV infection was 28.24% (85/301). The prevalence rate of multiple HPV infection was 11.30% (34/301), significantly lower than that of single type high-risk HPV infection (16.94%, P<0.05). The proportion of multiple HPV infection in total positive HPV results was 35.05% (34/97). The proportion of duplex and treble HPV infection were 29.90% (29/97) and 5.15% (5/97) respectively. The multiple HPV infection rate of the case group was 37.50% (18/48), significantly higher than that of the control group (6.32%, 16/253, P<0.01). The common duplex HPV infection modes were 16+51/58 (n=4), 51/58+52/59 (n=4), 11+16 (n=3), and 11+52/59 (n=3), 18+52/59 (n=3). The common treble HPV infection modes were 11+16+52/59, 16+18+31, 16+18+52/59, 31

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

    PubMed

    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.

  4. Multiplicative Word Problems and Intensive Quantities: An Integrated Software Response. Technical Report 85-19.

    ERIC Educational Resources Information Center

    Kaput, James J.

    This document reports on a project that is examining some of the difficulties encountered in teaching word problems involving multiplication, division, and intensive quantities. Some of the various uses of these operations and their structures are considered. Described are discoveries and assumptions regarding students' cognitive models of these…

  5. Intensive (Daily) Behavior Therapy for School Refusal: A Multiple Baseline Case Series

    ERIC Educational Resources Information Center

    Tolin, David F.; Whiting, Sara; Maltby, Nicholas; Diefenbach, Gretchen J.; Lothstein, Mary Anne; Hardcastle, Surrey; Catalano, Amy; Gray, Krista

    2009-01-01

    The following multiple baseline case series examines school refusal behavior in 4 male adolescents. School refusal symptom presentation was ascertained utilizing a functional analysis from the School Refusal Assessment Scale (Kearney, 2002). For the majority of cases, treatment was conducted within a 15-session intensive format over a 3-week…

  6. Fluorescence colocalization microscopy analysis can be improved by combining object-recognition with pixel-intensity-correlation.

    PubMed

    Moser, Bernhard; Hochreiter, Bernhard; Herbst, Ruth; Schmid, Johannes A

    2017-01-01

    The question whether two proteins interact with each other or whether a protein localizes to a certain region of the cell is often addressed with fluorescence microscopy and analysis of a potential colocalization of fluorescence markers. Since a mere visual estimation does not allow quantification of the degree of colocalization, different statistical methods of pixel-intensity correlation are commonly used to score it. We observed that these correlation coefficients are prone to false positive results and tend to show high values even for molecules that reside in different organelles. Our aim was to improve this type of analysis and we developed a novel method combining object-recognition based colocalization analysis with pixel-intensity correlation to calculate an object-corrected Pearson coefficient. We designed a macro for the Fiji-version of the software ImageJ and tested the performance systematically with various organelle markers revealing an improved robustness of our approach over classical methods. In order to prove that colocalization does not necessarily mean a physical interaction, we performed FRET (fluorescence resonance energy transfer) microscopy. This confirmed that non-interacting molecules can exhibit a nearly complete colocalization, but that they do not show any significant FRET signal in contrast to proteins that are bound to each other. © 2017 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Expression of varied GFPs in Saccharomyces cerevisiae: codon optimization yields stronger than expected expression and fluorescence intensity

    PubMed Central

    Kaishima, Misato; Ishii, Jun; Matsuno, Toshihide; Fukuda, Nobuo; Kondo, Akihiko

    2016-01-01

    Green fluorescent protein (GFP), which was originally isolated from jellyfish, is a widely used tool in biological research, and homologs from other organisms are available. However, researchers must determine which GFP is the most suitable for a specific host. Here, we expressed GFPs from several sources in codon-optimized and non-codon-optimized forms in the yeast Saccharomyces cerevisiae, which represents an ideal eukaryotic model. Surprisingly, codon-optimized mWasabi and mNeonGreen, which are typically the brightest GFPs, emitted less green fluorescence than did the other five codon-optimized GFPs tested in S. cerevisiae. Further, commercially available GFPs that have been optimized for mammalian codon usage (e.g., EGFP, AcGFP1 and TagGFP2) unexpectedly exhibited extremely low expression levels in S. cerevisiae. In contrast, codon-optimization of the GFPs for S. cerevisiae markedly increased their expression levels, and the fluorescence intensity of the cells increased by a maximum of 101-fold. Among the tested GFPs, the codon-optimized monomeric mUkG1 from soft coral showed the highest levels of both expression and fluorescence. Finally, the expression of this protein as a fusion-tagged protein successfully improved the reporting system’s ability to sense signal transduction and protein–protein interactions in S. cerevisiae and increased the detection rates of target cells using flow cytometry. PMID:27782154

  8. Multiple ionization of rare gas atoms irradiated with intense VUV radiation.

    PubMed

    Wabnitz, H; de Castro, A R B; Gürtler, P; Laarmann, T; Laasch, W; Schulz, J; Möller, T

    2005-01-21

    The interaction of intense vacuum-ultraviolet radiation from a free-electron laser with rare gas atoms is investigated. The ionization products of xenon and argon atomic beams are analyzed with time-of-flight mass spectroscopy. At 98 nm wavelength and approximately 10(13) W/cm(2) multiple charged ions up to Xe6+ (Ar4+) are detected. From the intensity dependence of multiple charged ion yields the mechanisms of multiphoton processes were derived. In the range of approximately 10(12)-10(13) W/cm(2) the ionization is attributed to sequential multiphoton processes. The production of multiple charged ions saturates at 5-30 times lower power densities than at 193 and 564 nm wavelength, respectively.

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

    NASA Astrophysics Data System (ADS)

    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

  10. Assessment of natural fluorescence as a tracer of diffuse agricultural pollution from slurry spreading on intensely-farmed grasslands.

    PubMed

    Naden, Pamela S; Old, Gareth H; Eliot-Laize, Caroline; Granger, Steve J; Hawkins, Jane M B; Bol, Roland; Haygarth, Phil

    2010-03-01

    The value of natural fluorescence in tracing diffuse pollution, in liquid phase, following slurry application to land was assessed by field experiment using twelve one hectare lysimeters on a heavy clay soil in Devon, UK, during autumn 2007. A strong linear relationship was found between natural fluorescence intensity and slurry concentration. The ratio of indices of tryptophan-like and fulvic/humic-like fluorescence (TI:FI) varied between 2 and 5 for a range of slurries sampled from Devon farms and allowed slurry to be distinguished from uncontaminated drainage waters (TI:FI<1). Incidental losses of slurry, indicated by significantly enhanced TI:FI ratios, high TI and high ammonium levels, occurred via the drain flow pathway of the drained lysimeters during the first small event following slurry-spreading. The maximum estimated loss from a single lysimeter was 2-8kg or 0.004-0.016% of the applied slurry. In the second larger storm event, some five weeks later, significantly enhanced TI:FI ratios in the drain flows were not associated with high TI but with high nitrate levels and, compared to the earlier storm, an increase in the humification index. This implies the loss of slurry decomposition products during this event but further work is needed to validate this. There was no significant enhancement of TI:FI in the surface/throughflow pathways of the drained or undrained lysimeters in either of the events. The observed change over a period of weeks in the strength and nature of the fluorescence signal from spread slurry restricts quantification of slurry losses to those immediately after slurry spreading. Nonetheless, this study demonstrates the utility of fluorescence as an indicator of slurry in drainage waters and the importance of field drains in diffuse agricultural pollution. Copyright 2009 Elsevier Ltd. All rights reserved.

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

    NASA Technical Reports Server (NTRS)

    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.

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

    NASA Technical Reports Server (NTRS)

    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.

  13. Multiple Velocity Profile Measurements in Hypersonic Flows Using Sequentially-Imaged Fluorescence Tagging

    NASA Technical Reports Server (NTRS)

    Bathel, Brett F.; Danehy, Paul M.; Inman, Jennifer A.; Jones, Stephen B.; Ivey,Christopher b.; Goyne, Christopher P.

    2010-01-01

    Nitric-oxide planar laser-induced fluorescence (NO PLIF) was used to perform velocity measurements in hypersonic flows by generating multiple tagged lines which fluoresce as they convect downstream. For each laser pulse, a single interline, progressive scan intensified CCD (charge-coupled device) camera was used to obtain two sequential images of the NO molecules that had been tagged by the laser. The CCD configuration allowed for sub-microsecond acquisition of both images, resulting in sub-microsecond temporal resolution as well as sub-mm spatial resolution (0.5-mm horizontal, 0.7-mm vertical). Determination of axial velocity was made by application of a cross-correlation analysis of the horizontal shift of individual tagged lines. A numerical study of measured velocity error due to a uniform and linearly-varying collisional rate distribution was performed. Quantification of systematic errors, the contribution of gating/exposure duration errors, and the influence of collision rate on temporal uncertainty were made. Quantification of the spatial uncertainty depended upon the signal-to-noise ratio of the acquired profiles. This velocity measurement technique has been demonstrated for two hypersonic flow experiments: (1) a reaction control system (RCS) jet on an Orion Crew Exploration Vehicle (CEV) wind tunnel model and (2) a 10-degree half-angle wedge containing a 2-mm tall, 4-mm wide cylindrical boundary layer trip. The experiments were performed at the NASA Langley Research Center's 31-Inch Mach 10 Air Tunnel.

  14. Gateway Vectors for Simultaneous Detection of Multiple Protein−Protein Interactions in Plant Cells Using Bimolecular Fluorescence Complementation

    PubMed Central

    Hikino, Kazumi; Goto-Yamada, Shino; Nishimura, Mikio; Nakagawa, Tsuyoshi; Mano, Shoji

    2016-01-01

    Bimolecular fluorescence complementation (BiFC) is widely used to detect protein—protein interactions, because it is technically simple, convenient, and can be adapted for use with conventional fluorescence microscopy. We previously constructed enhanced yellow fluorescent protein (EYFP)-based Gateway cloning technology-compatible vectors. In the current study, we generated new Gateway cloning technology-compatible vectors to detect BiFC-based multiple protein—protein interactions using N- and C-terminal fragments of enhanced cyan fluorescent protein (ECFP), enhanced green fluorescent protein (EGFP), and monomeric red fluorescent protein (mRFP1). Using a combination of N- and C-terminal fragments from ECFP, EGFP and EYFP, we observed a shift in the emission wavelength, enabling the simultaneous detection of multiple protein—protein interactions. Moreover, we developed these vectors as binary vectors for use in Agrobacterium infiltration and for the generate transgenic plants. We verified that the binary vectors functioned well in tobacco cells. The results demonstrate that the BiFC vectors facilitate the design of various constructions and are convenient for the detection of multiple protein—protein interactions simultaneously in plant cells. PMID:27490375

  15. Simultaneous multiple-point velocity measurements using laser-induced iodine fluorescence

    NASA Technical Reports Server (NTRS)

    Mcdaniel, J. C.; Hiller, B.; Hanson, R. K.

    1983-01-01

    A technique is demonstrated for measuring velocity at multiple locations in a plane of a gaseous flowfield using Doppler-shifted absorption with fluorescence detection from iodine molecules, excited by a sheet of tunable single-axial-mode argon-ion laser radiation at 514.5 nm. Measurements were made simultaneously at 10,000 points in an iodine-seeded supersonic flow field with a 100 x 100 element photodiode array camera and were found to agree well with a numerical solution for the velocity field. The accuracy with which a component of velocity can be measured is limited, in the current approach, by the iodine linewidth to about 5 m/sec.

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

    PubMed

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

  18. Improved high-intensity microwave discharge lamp for atomic resonance absorption and fluorescence spectrometry.

    PubMed

    Lifshitz, A; Skinner, G B; Wood, D R

    1978-09-01

    An unusually good combination of high intensity and narrow line has been achieved in a microwave discharge lamp by placing the optical window in the center of the microwave cavity. Construction details and performance characteristics are described.

  19. A Model of Population and Subject (MOPS) Intensities with Application to Multiple Sclerosis Lesion Segmentation

    PubMed Central

    Tomas-Fernandez, Xavier; Warfield, Simon K.

    2015-01-01

    White matter (WM) lesions are thought to play an important role in multiple sclerosis (MS) disease burden. Recent work in the automated segmentation of white matter lesions from MRI has utilized a model in which lesions are outliers in the distribution of tissue signal intensities across the entire brain of each patient. However, the sensitivity and specificity of lesion detection and segmentation with these approaches have been inadequate. In our analysis, we determined this is due to the substantial overlap between the whole brain signal intensity distribution of lesions and normal tissue. Inspired by the ability of experts to detect lesions based on their local signal intensity characteristics, we propose a new algorithm that achieves lesion and brain tissue segmentation through simultaneous estimation of a spatially global within-the-subject intensity distribution and a spatially local intensity distribution derived from a healthy reference population. We demonstrate that MS lesions can be segmented as outliers from this intensity model of population and subject (MOPS). We carried out extensive experiments with both synthetic and clinical data, and compared the performance of our new algorithm to those of state-of-the art techniques. We found this new approach leads to a substantial improvement in the sensitivity and specificity of lesion detection and segmentation. PMID:25616008

  20. Laser-induced fluorescence thermometry of heating in water from short bursts of high intensity focused ultrasound.

    PubMed

    Al-Qraini, Moath M; Canney, Michael S; Oweis, Ghanem F

    2013-04-01

    Free field experimental measurements of the temperature rise of water in the focal region of a 2 MHz high intensity focused ultrasound (HIFU) transducer were performed. The transducer was operated in pulse-mode with millisecond bursts, at acoustic intensities of 5 to 18.5 kW/cm(2) at the focus, resulting in non-linear wave propagation and shock wave formation. Pulsed, planar, laser-induced fluorescence (LIF) was used as a fast rise-time, non-intrusive, temperature measurement method of the water present in the focal region. LIF thermometry is based on calibrating the temperature-dependent fluorescence intensity signal emitted by a passive dye dissolved in water when excited by a pulse of laser light. The laser beam was formed into a thin light sheet to illuminate a planar area in the HIFU focal region. The laser light sheet was oriented transverse to the acoustic axis. Cross-sectional, instantaneous temperature field measurements within the HIFU focal volume showed that the water temperature increased steadily with increasing HIFU drive voltage. Heating rates of 4000-7000°C/s were measured within the first millisecond of the HIFU burst. Increasing the length of the burst initially resulted in an increase in the water temperature within the HIFU focal spot (up to ∼3 ms), after which it steadied or slightly dropped. Acoustic streaming was measured and shown to be consistent with the reduction in heating with increased burst length due to convective cooling. LIF thermometry may thus be a viable non-invasive method for the characterization of HIFU transducers at high power intensities.

  1. Structural intensity in plates with multiple discrete and distributed spring-dashpot systems

    NASA Astrophysics Data System (ADS)

    Khun, M. S.; Lee, H. P.; Lim, S. P.

    2004-09-01

    The finite element method has been used to predict the structural intensity of a plate with multiple discrete and distributed dampers. Different combinations of dampers in different positions were employed to study the effects of multiple dampers in an individual plate model to examine the energy flow phenomenon in the presence of many dissipative elements. The structural intensity of plate structures connected by loosened bolts subjected to forced excitation is also presented in this paper. The loosened bolts are modelled by discrete as well as distributed spring-dashpot systems. The distributed springs and dampers are introduced to characterize the joint behavior of the plates. Numerical results are also presented for the plates connected by two bolts.

  2. Fluorescence spectroscopy to discriminate neoplastic human brain lesions: a study using the spectral intensity ratio and multivariate linear discriminant analysis

    NASA Astrophysics Data System (ADS)

    Nazeer, Shaiju S.; Saraswathy, Ariya; Gupta, Arun Kumar; Jayasree, Ramapurath S.

    2014-02-01

    Fluorescence spectroscopy is an emerging tool used to differentiate normal and malignant tissue based on the emission spectral profile from endogenous fluorophores. The goal of this study is to estimate the concentration of fluorophores using autofluorescence spectroscopy and try to utilize its diagnostic potential on samples of clinical importance. Brain tumor tissues from patients who received craniotomy for the removal of astrocytoma, glioma, meningioma and schwannoma were utilized in this study. Fluorescence emissions of the formalin fixed samples were recorded at excitation wavelengths of 320 and 410 nm. The emission characteristics of fluorophores such as collagen, nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD), phospholipids and porphyrins of tumor tissue and adjacent normal tissue were elicited. Exact tissue classification was carried out using the spectral intensity ratio (SIR) and multivariate principal component analysis-linear discriminant analysis (PCA-LDA). The diagnostic algorithm based on PCA-LDA provided better classification efficiency than SIR. Moreover, the spectral data based on an excitation wavelength of 410 nm are found to be more efficient in the classification than 320 nm excitation, using PCA-LDA. Better efficacy of PCA-LDA in tissue classification was further confirmed by the receiver operator characteristic (ROC) curve method. The results of this study establish the feasibility of using fluorescence spectroscopy based real time tools for the discrimination of brain tumors from the adjacent normal tissue during craniotomies, which at present faces a huge challenge.

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

    PubMed Central

    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

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

    SciTech Connect

    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

  5. Multiple fluorescent scattering of N2 ultraviolet emissions in the atmospheres of the earth and Titan

    NASA Astrophysics Data System (ADS)

    Conway, R. R.

    1983-06-01

    Multiple scattering in a plane-parallel radiative transfer model, and a line-by-line synthesis of individual bands, are used to calculate the absorption and reemission by N2 of the Lyman-Birge-Hopfield (LBH) and Birge-Hopfield (BH) band systems in both the earth aurora and the dayglow of Titan. Iteration through successive scatterings indicates that fewer than ten scatterings are important, even for the thickest bands. The excitation rate caused by multiple scattering is sensitive to the vibrational population distribution of the ambient N2, especially for the BH system in the earth aurora. The LBH system has an enhanced sensitivity to vibrational temperature on Titan, due to the rapidly changing photoabsorption cross section of CH4 around 1400 A. It is suggested that certain Rydberg bands should have an intensity profile similar to that of the BH bands.

  6. Solutions of Stress Intensity Factors of Multiple Internal Axial Cracks in Hollow Cylinders

    NASA Astrophysics Data System (ADS)

    Ismail, Al Emran

    2017-01-01

    This paper solved numerically the stress intensity factors of multiple internal axial cracks in hollow cylinders under internal pressure. Semi-elliptical shaped cracks are modelled using ANSYS finite element program and the stress intensity factors are calculated using interaction domain integral which is based on the J-integral. There are two important parameters are considered such as crack aspect ratio, a/t is 0.2, 0.4 and 0.75 and crack spacing distance, d/c is 0.4, 0.7 and 1.1. For multiple internal axial cracks under internal pressure, stress intensity factors along the crack front decreased when crack aspect ratio, a/t is increased. It is also revealed that the role of crack spacing on the stress intensity factor is insignificant especially for the shallow cracks (a/t < 0.4). However, for the crack having a/t = 0.75, increasing the distance of d/c capable to increase the SIFs due to interaction effect.

  7. Analysis on intensity demodulated strain sensing based on multiple phase-shifted FBG

    NASA Astrophysics Data System (ADS)

    Zhang, Xiaolei; Peng, Gangding; Zhang, Faxiang; Qi, Haifeng; Song, Zhiqiang; Li, Shujuan; Ni, Jiasheng; Wang, Chang

    2014-11-01

    An intensity demodulated strain sensing scheme based on multiple phase-shifted fiber Bragg grating (FBG) is investigated. Wavelength shifts related to the external strain signal will result in various power change received by the photodetector. The reflected spectrum of the sensing multiple phase-shifted FBG are designed using the transfer matrix method to and fabricated by dithering phase mask method. The strain sensing scheme is analyzed numerically by relate the power change to reflectivity spectrum, thus help us optimize the design and fabrication of multiple phase-shifted FBG to achieve maximum sensitivity. The experimental results show that the strain sensitivity can be significantly enhanced by over 10dB by using phase-shifted FBG with many phase shifts compared to a normal FBG.

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

    PubMed

    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 (P<0.05), but the level of serum albumin was significantly increased (P<0.05) at the day 3, 5, 7 and 14. In the meantime, the PaO2 was significantly elevated at the day 3, 5 and 7 (P<0.01) after intensive insulin therapy. The incidence of gastrointestinal dysfunction, the incidence of MODS, the length of hospital stay and the mortality were markedly decreased (P<0.01). The results suggest early treatment with intensive insulin therapy is effective for traumatic shock combined with MODS and can decrease the length of hospital stay and the mortality.

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

    PubMed

    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.

  10. Response of fish communities to multiple pressures: Development of a total anthropogenic pressure intensity index.

    PubMed

    Poikane, Sandra; Ritterbusch, David; Argillier, Christine; Białokoz, Witold; Blabolil, Petr; Breine, Jan; Jaarsma, Nicolaas G; Krause, Teet; Kubečka, Jan; Lauridsen, Torben L; Nõges, Peeter; Peirson, Graeme; Virbickas, Tomas

    2017-05-15

    Lakes in Europe are subject to multiple anthropogenic pressures, such as eutrophication, habitat degradation and introduction of alien species, which are frequently inter-related. Therefore, effective assessment methods addressing multiple pressures are needed. In addition, these systems have to be harmonised (i.e. intercalibrated) to achieve common management objectives across Europe. Assessments of fish communities inform environmental policies on ecological conditions integrating the impacts of multiple pressures. However, the challenge is to ensure consistency in ecological assessments through time, across ecosystem types and across jurisdictional boundaries. To overcome the serious comparability issues between national assessment systems in Europe, a total anthropogenic pressure intensity (TAPI) index was developed as a weighted combination of the most common pressures in European lakes that is validated against 10 national fish-based water quality assessment systems using data from 556 lakes. Multi-pressure indices showed significantly higher correlations with fish indices than single-pressure indices. The best-performing index combines eutrophication, hydromorphological alterations and human use intensity of lakes. For specific lake types also biological pressures may constitute an important additional pressure. The best-performing index showed a strong correlation with eight national fish-based assessment systems. This index can be used in lake management for assessing total anthropogenic pressure on lake ecosystems and creates a benchmark for comparison of fish assessments independent of fish community composition, size structure and fishing-gear. We argue that fish-based multiple-pressure assessment tools should be seen as complementary to single-pressure tools offering the major advantage of integrating direct and indirect effects of multiple pressures over large scales of space and time.

  11. Inverse identification of intensity distributions from multiple flux maps in concentrating solar applications

    NASA Astrophysics Data System (ADS)

    Erickson, Ben; Petrasch, Jörg

    2012-06-01

    Radiative flux measurements at the focal plane of solar concentrators are typically performed using digital cameras in conjunction with Lambertian targets. To accurately predict flux distributions on arbitrary receiver geometries directional information about the radiation is required. Currently, the directional characteristics of solar concentrating systems are predicted via ray tracing simulations. No direct experimental technique to determine intensities of concentrating solar systems is available. In the current paper, multiple parallel flux measurements at varying distances from the focal plane together with a linear inverse method and Tikhonov regularization are used to identify the directional and spatial intensity distribution at the solution plane. The directional binning feature of an in-house Monte Carlo ray tracing program is used to provide a reference solution. The method has been successfully applied to two-dimensional concentrators, namely parabolic troughs and elliptical troughs using forward Monte Carlo ray tracing simulations that provide the flux maps as well as consistent, associated intensity distribution for validation. In the two-dimensional case, intensity distributions obtained from the inverse method approach the Monte Carlo forward solution. In contrast, the method has not been successful for three dimensional and circular symmetric concentrator geometries.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    SciTech Connect

    Nikitin, Andrey G.

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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.

  16. Small intensely fluorescent (SIF) cells and sympathetic nerves in the adult rabbit portal vein and during perinatal development.

    PubMed

    Crowe, R; Burnstock, G

    1982-01-01

    The ontogenesis of small intensely fluorescent (SIF) cells and the adrenergic nerve plexus is described in stretch preparations of the rabbit portal vein. On the 25 to 26th days of gestation there was a predominance of SIF cells (8 to 30 microns in diameter), but a few nerve fibres in bundles were also present. Each portal vein preparation contained 6 to 9 groups of cells. The distribution and number of SIF cells and nerve bundles remained constant until the 31st day of gestation at which stage the number of SIF cells had decreased, while the density of the nerve plexus had increased approximately 4-fold. The adult portal vein exhibited a dense adrenergic plexus, but SIF cells were absent from nine out of ten preparations.

  17. A Novel Multiple Hypothesis Based Particle Tracking Method for Clathrin Mediated Endocytosis Analysis Using Fluorescence Microscopy

    PubMed Central

    Liang, Liang; Shen, Hongying; De Camilli, Pietro; Duncan, James S.

    2015-01-01

    In order to quantitatively analyze biological images and study underlying mechanisms of the cellular and subcellular processes, it is often required to track a large number of particles involved in these processes. Manual tracking can be performed by the biologists, but the workload is very heavy. In this paper, we present an automatic particle tracking method for analyzing an essential subcellular process, namely clathrin mediated endocytosis. The framework of the tracking method is an extension of the classical multiple hypothesis tracking (MHT), and it is designed to manage trajectories, solve data association problems, and handle pseudo-splitting/merging events. In the extended MHT framework, particle tracking becomes evaluating two types of hypotheses. The first one is the trajectory-related hypothesis, to test whether a recovered trajectory is correct, and the second one is the observation-related hypothesis, to test whether an observation from an image belongs to a real particle. Here, an observation refers to a detected particle and its feature vector. To detect the particles in 2D fluorescence images taken using total internal reflection microscopy, the images are segmented into regions, and the features of the particles are obtained by fitting Gaussian mixture models into each of the image regions. Specific models are developed according to the properties of the particles. The proposed tracking method is demonstrated on synthetic data under different scenarios and applied to real data. PMID:24808351

  18. Wave field reconstruction from multiple plane intensity-only data: augmented lagrangian algorithm.

    PubMed

    Migukin, Artem; Katkovnik, Vladimir; Astola, Jaakko

    2011-06-01

    A complex-valued wave field is reconstructed from intensity-only measurements given at multiple observation planes parallel to the object plane. The phase-retrieval algorithm is obtained from the constrained maximum likelihood approach provided that the additive noise is gaussian. The forward propagation from the object plane to the measurement plane is treated as a constraint in the proposed variational setting of reconstruction. The developed iterative algorithm is based on an augmented lagrangian technique. An advanced performance of the algorithm is demonstrated by numerical simulations.

  19. Intensity-resolved IR multiple photon ionization and fragmentation of C60.

    PubMed

    Bakker, Joost M; Lapoutre, Vivike J F; Redlich, Britta; Oomens, Jos; Sartakov, Boris G; Fielicke, André; von Helden, Gert; Meijer, Gerard; van der Meer, Alexander F G

    2010-02-21

    The sequential absorption of multiple infrared (IR) photons by isolated gas-phase species can lead to their dissociation and/or ionization. Using the newly constructed "Free-Electron Laser for IntraCavity Experiments" (FELICE) beam line at the FELIX facility, neutral C(60) molecules have been exposed to an extremely high number (approximately 10(23)) of photons/cm(2) for a total time duration of up to 5 micros. At wavelengths around 20 microm, resonant with allowed IR transitions of C(60), ionization and extensive fragmentation of the fullerenes are observed. The resulting photofragment distributions are attributed to absorption in fragmentation products formed once C(60) is excited to internal energies at which fragmentation or ionization takes place within the duration of the laser pulse. The high IR intensities available combined with the large interaction volume permit spatially resolved detection of the ions inside the laser beam, thereby disentangling the contributions from different IR intensities. The use of spatial imaging reveals intensity dependent mass distributions that are substantially narrower than what has been observed previously, indicating rather narrow energy distributions. A simple rate-equation modeling of the excitation process supports the experimental observations.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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.

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

    PubMed

    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

  3. Relationship between Fluorescence Intensity of GFP and the Expression Level of Prestin in a Prestin-Expressing Chinese Hamster Ovary Cell Line

    NASA Astrophysics Data System (ADS)

    Iida, Koji; Nagaoka, Tomoyuki; Tsumoto, Kouhei; Ikeda, Katsuhisa; Kumagai, Izumi; Kobayashi, Toshimitsu; Wada, Hiroshi

    Outer hair cells (OHCs) in mammals can elongate and contract at frequencies up to 100kHz in response to changes in their membrane potential. The origin of this unique motility is the motor protein prestin, which is densely packed in the lateral membrane of the OHCs. In a previous work, we constructed a prestin-expressing cell line using Chinese hamster ovary (CHO) cells to obtain a stable supply of prestin. When we research prestin using constructed cells, it is necessary to estimate the expression level of prestin in the cells easily and non-invasively. As the prestin gene and a green fluorescent protein (GFP) gene were introduced into constructed cells using the same vector, the expression level of prestin and fluorescence intensity of GFP are possibly correlated. Since this correlation is not clear, however, in this study, we therefore investigated whether the expression level of prestin evaluated by patch-clamp recording and the fluorescence intensity of GFP obtained from fluorescence images are correlated or not. As a result, it was demonstrated that they were correlated. The expression level of prestin can therefore be evaluated by measuring the fluorescence intensity of GFP.

  4. Diurnal Variability in Chlorophyll-a, Carotenoids, CDOM and SO42− Intensity of Offshore Seawater Detected by an Underwater Fluorescence-Raman Spectral System

    PubMed Central

    Chen, Jing; Ye, Wangquan; Guo, Jinjia; Luo, Zhao; Li, Ying

    2016-01-01

    A newly developed integrated fluorescence-Raman spectral system (λex = 532 nm) for detecting Chlorophyll-a (chl-a), Chromophoric Dissolved Organic Matter (CDOM), carotenoids and SO42− in situ was used to successfully investigate the diurnal variability of all above. Simultaneously using the integration of fluorescence spectroscopy and Raman spectroscopy techniques provided comprehensive marine information due to the complementarity between the different excitation mechanisms and different selection rules. The investigation took place in offshore seawater of the Yellow Sea (36°05′40′′ N, 120°31′32′′ E) in October 2014. To detect chl-a, CDOM, carotenoids and SO42−, the fluorescence-Raman spectral system was deployed. It was found that troughs of chl-a and CDOM fluorescence signal intensity were observed during high tides, while the signal intensity showed high values with larger fluctuations during ebb-tide. Chl-a and carotenoids were influenced by solar radiation within a day cycle by different detection techniques, as well as displaying similar and synchronous tendency. CDOM fluorescence cause interference to the measurement of SO42−. To avoid such interference, the backup Raman spectroscopy system with λex = 785 nm was employed to detect SO42− concentration on the following day. The results demonstrated that the fluorescence-Raman spectral system has great potential in detection of chl-a, carotenoids, CDOM and SO42− in the ocean. PMID:27420071

  5. Bead-based multiplex immuno-assays for cytokines, chemokines, growth factors and other analytes: median fluorescence intensities versus their derived absolute concentration values for statistical analysis.

    PubMed

    Breen, Edmond J; Polaskova, Veronika; Khan, Alamgir

    2015-02-01

    Within the scientific literature, analyses of data from bead based multiplex immunoassays are based on either median fluorescence intensities (MFI) or derived absolute concentration values (ACV) but no consideration of which set of data is the most appropriate for analysis has been published. Here we look at the variance of MFI versus their ACV from the expression of 14 analytes in plasma, using 6 commercially available kits, across 177 patients, recorded at two time points and the associated analyte standards. In total 60 micro titre plates were used resulting in 4965 MFI. In doing so we develop a new background subtraction procedure that reduced by 50% the number of out-of-range values observed in our data set. Using a linear mixed-effect model, which normalizes for assay-to-assay variation, MFI produced similar significant differences than that observed using absolute concentration values. We show that subtracting analyte blanks produces 15% negative MFI resulting in uncertainty of the data being analysed. We argue for analysis of protein expression values MFI are generally a better choice than absolute concentration values. It is argued that analyte standards are not required on each plate, or not at all, in multi-plate experiments, but knowledge of the concentration curve and the range of MFI values that fall within the limits of this curve for each analyte is required. The significance of using MFI over concentration values for the life scientist means higher statistical power and lower costs. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Platform for efficient switching between multiple devices in the intensive care unit.

    PubMed

    De Backere, F; Vanhove, T; Dejonghe, E; Feys, M; Herinckx, T; Vankelecom, J; Decruyenaere, J; De Turck, F

    2015-01-01

    This article is part of the Focus Theme of METHODS of Information in Medicine on "Managing Interoperability and Complexity in Health Systems". Handheld computers, such as tablets and smartphones, are becoming more and more accessible in the clinical care setting and in Intensive Care Units (ICUs). By making the most useful and appropriate data available on multiple devices and facilitate the switching between those devices, staff members can efficiently integrate them in their workflow, allowing for faster and more accurate decisions. This paper addresses the design of a platform for the efficient switching between multiple devices in the ICU. The key functionalities of the platform are the integration of the platform into the workflow of the medical staff and providing tailored and dynamic information at the point of care. The platform is designed based on a 3-tier architecture with a focus on extensibility, scalability and an optimal user experience. After identification to a device using Near Field Communication (NFC), the appropriate medical information will be shown on the selected device. The visualization of the data is adapted to the type of the device. A web-centric approach was used to enable extensibility and portability. A prototype of the platform was thoroughly evaluated. The scalability, performance and user experience were evaluated. Performance tests show that the response time of the system scales linearly with the amount of data. Measurements with up to 20 devices have shown no performance loss due to the concurrent use of multiple devices. The platform provides a scalable and responsive solution to enable the efficient switching between multiple devices. Due to the web-centric approach new devices can easily be integrated. The performance and scalability of the platform have been evaluated and it was shown that the response time and scalability of the platform was within an acceptable range.

  7. Tracking graphene by fluorescence imaging: a tool for detecting multiple populations of graphene in solution

    NASA Astrophysics Data System (ADS)

    Guidetti, G.; Cantelli, A.; Mazzaro, R.; Ortolani, L.; Morandi, V.; Montalti, M.

    2016-04-01

    Most methods used for the characterization of graphene produced by liquid phase exfoliation require the deposition of the liquid sample on a substrate and subsequent drying. Because of this or other post-synthetic treatments, the reliability of the data in describing the actual features of the graphene particles in the pristine solution becomes questionable. Hence there is a need for new methods that permit the study of graphene directly in solution. Fluorescence imaging is at present the most convenient and sensitive method to visualize nanosized objects in solution. Here we report the development of a new method for visualizing and tracking exfoliated graphene directly in solution using a conventional set-up for fluorescence microscopy. We functionalized a fluorescent surfactant typically used for exfoliating graphite in aqueous phase (Pluronic P123) with two different fluorophores, in order to make graphene detectable by fluorescence microscopy. The photophysical interactions between the fluorescent surfactant and graphene were investigated at the bulk level. Finally, fluorescence microscopy allowed us to track the carbon particles produced and to identify two different populations of particles with sizes of 265 +/- 25 and 1100 +/- 200 nm respectively. The correlation of these results with TEM and DLS data is discussed.Most methods used for the characterization of graphene produced by liquid phase exfoliation require the deposition of the liquid sample on a substrate and subsequent drying. Because of this or other post-synthetic treatments, the reliability of the data in describing the actual features of the graphene particles in the pristine solution becomes questionable. Hence there is a need for new methods that permit the study of graphene directly in solution. Fluorescence imaging is at present the most convenient and sensitive method to visualize nanosized objects in solution. Here we report the development of a new method for visualizing and tracking

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

    NASA Astrophysics Data System (ADS)

    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.

  9. Order dependence of the profile of the intensities of multiple-quantum coherences

    SciTech Connect

    Lundin, A. A.; Zobov, V. E.

    2015-05-15

    A modification of the widespread phenomenological model theory of multiple-quantum (MQ) nuclear magnetic resonance spectra of a single cluster of correlated spins has been developed. In contrast to the mentioned theory, the size distribution of such clusters has been consistently taken into account. To obtain the distribution, solutions for the amplitudes of the expansion in the complete set of orthogonal operators are used. Expressions specifying the dependence of the profile of the intensities of MQ coherences on their number n (order) have been obtained. The total form of the dependence has been evaluated by means of the numerical implementation of the resulting expressions. The asymptotic expressions for large n values (wings of the spectrum) have been obtained analytically by the saddle-point method. It has been shown that the dependence under study has a Gaussian central part and exponential wings. The results obtained are in agreement with the previous calculations for some model systems and existing experimental data.

  10. Hydrogen Balmer alpha intensity distributions and line profiles from multiple scattering theory using realistic geocoronal models

    NASA Technical Reports Server (NTRS)

    Anderson, D. E., Jr.; Meier, R. R.; Hodges, R. R., Jr.; Tinsley, B. A.

    1987-01-01

    The H Balmer alpha nightglow is investigated by using Monte Carlo models of asymmetric geocoronal atomic hydrogen distributions as input to a radiative transfer model of solar Lyman-beta radiation in the thermosphere and atmosphere. It is shown that it is essential to include multiple scattering of Lyman-beta radiation in the interpretation of Balmer alpha airglow data. Observations of diurnal variation in the Balmer alpha airglow showing slightly greater intensities in the morning relative to evening are consistent with theory. No evidence is found for anything other than a single sinusoidal diurnal variation of exobase density. Dramatic changes in effective temperature derived from the observed Balmer alpha line profiles are expected on the basis of changing illumination conditions in the thermosphere and exosphere as different regions of the sky are scanned.

  11. Exploration of the electron multiple recollision dynamics in intense laser fields with Bohmian trajectories

    NASA Astrophysics Data System (ADS)

    Jooya, Hossein Z.; Telnov, Dmitry A.; Chu, Shih-I.

    2016-06-01

    Electron multiple recollision dynamics under intense midinfrared laser fields is studied by means of the de Broglie-Bohm framework of Bohmian mechanics. Bohmian trajectories contain all the information embedded in the time-dependent wave function. This makes the method suitable to investigate the coherent dynamic processes for which the phase information is crucial. In this study, the appearance of the subpeaks in the high-harmonic-generation time-frequency profiles and the asymmetric fine structures in the above-threshold ionization spectrum are analyzed by the comprehensive and intuitive picture provided by Bohmian mechanics. The time evolution of the individual electron trajectories is closely studied to address some of the major structural features of the photoelectron angular distributions.

  12. Hydrogen Balmer alpha intensity distributions and line profiles from multiple scattering theory using realistic geocoronal models

    NASA Technical Reports Server (NTRS)

    Anderson, D. E., Jr.; Meier, R. R.; Hodges, R. R., Jr.; Tinsley, B. A.

    1987-01-01

    The H Balmer alpha nightglow is investigated by using Monte Carlo models of asymmetric geocoronal atomic hydrogen distributions as input to a radiative transfer model of solar Lyman-beta radiation in the thermosphere and atmosphere. It is shown that it is essential to include multiple scattering of Lyman-beta radiation in the interpretation of Balmer alpha airglow data. Observations of diurnal variation in the Balmer alpha airglow showing slightly greater intensities in the morning relative to evening are consistent with theory. No evidence is found for anything other than a single sinusoidal diurnal variation of exobase density. Dramatic changes in effective temperature derived from the observed Balmer alpha line profiles are expected on the basis of changing illumination conditions in the thermosphere and exosphere as different regions of the sky are scanned.

  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

    NASA Astrophysics Data System (ADS)

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

    NASA Astrophysics Data System (ADS)

    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)

  15. Supramolecular Construction of Multifluorescent Gels: Interfacial Assembly of Discrete Fluorescent Gels through Multiple Hydrogen Bonding.

    PubMed

    Ji, Xiaofan; Shi, Bingbing; Wang, Hu; Xia, Danyu; Jie, Kecheng; Wu, Zi Liang; Huang, Feihe

    2015-12-22

    Multifluorescent supramolecular gels with complex structures are constructed from discrete fluorescent gels, which serve as the building blocks, through hydrogen bonding interactions at interfaces. The multifluorescent gel can realize rapid healing within only ≈100 s.

  16. Step-rate thresholds for physical activity intensity in persons with multiple sclerosis.

    PubMed

    Agiovlasitis, Stamatis; Motl, Robert W

    2014-01-01

    This study examined whether the relationship between metabolic equivalent units (METs) and step-rate is altered in persons with multiple sclerosis (MS) and developed step-rate thresholds for activity intensity for these persons. Participants were 24 persons with MS (20 women; age = 44 ± 12) and 24 healthy persons without MS (20 women; age = 41 ± 11). The MS group was divided using the 12-item MS Walking Scale (MSWS-12) into two walking impairment subgroups: (a) minimal (n = 13, MSWS-12 ≤ 12.5) and (b) mild-moderate (n = 11, MSWS-12 > 12.5). METs were measured with spirometry and step-rate with hand-tally. Step-rate, height, group, the step-rate by group interaction, and the square of step-rate significantly predicted METs. At a given height, the step-rate thresholds at 3 and 6 METs were lower for persons with minimal impairment than persons without MS and even lower for persons with mild-moderate impairment. The relationship between METs and step-rate is altered in persons with MS, lowering their step-rate thresholds for activity intensity, especially for persons with MS who have higher levels of walking impairment.

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

    PubMed

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

  19. Temporal trends of postinjury multiple-organ failure: still resource intensive, morbid, and lethal.

    PubMed

    Sauaia, Angela; Moore, Ernest E; Johnson, Jeffrey L; Chin, Theresa L; Banerjee, Anirban; Sperry, Jason L; Maier, Ronald V; Burlew, C Cothren

    2014-03-01

    While the incidence of postinjury multiple-organ failure (MOF) has declined during the past decade, temporal trends of its morbidity, mortality, presentation patterns, and health care resources use have been inconsistent. The purpose of this study was to describe the evolving epidemiology of postinjury MOF from 2003 to 2010 in multiple trauma centers sharing standard treatment protocols. "Inflammation and Host Response to Injury Collaborative Program" institutions that enrolled more than 20 eligible patients per biennial during the 2003 to 2010 study period were included. The patients were aged 16 years to 90 years, sustained blunt torso trauma with hemorrhagic shock (systolic blood pressure < 90 mm Hg, base deficit ≥ 6 mEq/L, blood transfusion within the first 12 hours), but without severe head injury (motor Glasgow Coma Scale [GCS] score < 4). MOF temporal trends (Denver MOF score > 3) were adjusted for admission risk factors (age, sex, body max index, Injury Severity Score [ISS], systolic blood pressure, and base deficit) using survival analysis. A total of 1,643 patients from four institutions were evaluated. MOF incidence decreased over time (from 17% in 2003-2004 to 9.8% in 2009-2010). MOF-related death rate (33% in 2003-2004 to 36% in 2009-2010), intensive care unit stay, and mechanical ventilation duration did not change over the study period. Adjustment for admission risk factors confirmed the crude trends. MOF patients required much longer ventilation and intensive care unit stay, compared with non-MOF patients. Most of the MOF-related deaths occurred within 2 days of the MOF diagnosis. Lung and cardiac dysfunctions became less frequent (57.6% to 50.8%, 20.9% to 12.5%, respectively), but kidney and liver failure rates did not change (10.1% to 12.5%, 15.2% to 14.1%). Postinjury MOF remains a resource-intensive, morbid, and lethal condition. Lung injury is an enduring challenge and should be a research priority. The lack of outcome improvements suggests

  20. Temporal trends of postinjury multiple-organ failure: Still resource intensive, morbid, and lethal

    PubMed Central

    Sauaia, Angela; Moore, Ernest E.; Johnson, Jeffrey L.; Chin, Theresa L.; Banerjee, Anirban; Sperry, Jason L.; Maier, Ronald V.; Burlew, C. Cothren

    2014-01-01

    BACKGROUND While the incidence of postinjury multiple-organ failure (MOF) has declined during the past decade, temporal trends of its morbidity, mortality, presentation patterns, and health care resources use have been inconsistent. The purpose of this study was to describe the evolving epidemiology of postinjury MOF from 2003 to 2010 in multiple trauma centers sharing standard treatment protocols. METHODS “Inflammation and Host Response to Injury Collaborative Program” institutions that enrolled more than 20 eligible patients per biennial during the 2003 to 2010 study period were included. The patients were aged 16 years to 90 years, sustained blunt torso trauma with hemorrhagic shock (systolic blood pressure < 90 mm Hg, base deficit ≥ 6 mEq/L, blood transfusion within the first 12 hours), but without severe head injury (motor Glasgow Coma Scale [GCS] score < 4). MOF temporal trends (Denver MOF score > 3) were adjusted for admission risk factors (age, sex, body max index, Injury Severity Score [ISS], systolic blood pressure, and base deficit) using survival analysis. RESULTS A total of 1,643 patients from four institutions were evaluated. MOF incidence decreased over time (from 17% in 2003–2004 to 9.8% in 2009–2010). MOF-related death rate (33% in 2003–2004 to 36% in 2009–2010), intensive care unit stay, and mechanical ventilation duration did not change over the study period. Adjustment for admission risk factors confirmed the crude trends. MOF patients required much longer ventilation and intensive care unit stay, compared with non-MOF patients. Most of the MOF-related deaths occurred within 2 days of the MOF diagnosis. Lung and cardiac dysfunctions became less frequent (57.6% to 50.8%, 20.9% to 12.5%, respectively), but kidney and liver failure rates did not change (10.1% to 12.5%, 15.2% to 14.1%). CONCLUSION Postinjury MOF remains a resource-intensive, morbid, and lethal condition. Lung injury is an enduring challenge and should be a research

  1. Aerobic intensity and pacing pattern during the six-minute walk test in patients with multiple sclerosis.

    PubMed

    Dalgas, Ulrik; Kjølhede, Tue; Gijbels, Domien; Romberg, Anders; Santoyo, Carme; de Noordhout, Benoit Maertens; Knuts, Kathy; Feys, Peter

    2014-01-01

    To examine the aerobic intensity level and pacing pattern during the 6-min walk test (6MWT) in persons with multiple sclerosis, taking into account time of day, fatigue, disability level and multiple sclerosis subtype. Cross-sectional study. Eighty multiple sclerosis patients (Expanded Disability Status Scale, EDSS ≤ 6.5). Participants performed the 6MWT at 3 different time-points (morning, noon, afternoon) during 1 day. Heart rate and pacing strategy (distance covered every minute) were registered. A sub-group analysis determined the effects of fatigue, disability level and multiple sclerosis subtype. The relative aerobic intensity was constant throughout the day (67 ± 10% of estimated maximal heart rate). In all sub-groups heart rate increased and distance walked declined after the first minute (p < 0.001). The mild EDSS sub-group showed a slightly larger increase throughout the 6MWT in heart rate development, while no differences were seen in sub-groups of fatigue and multiple sclerosis subtype. In most sub-groups walking speed was fastest in the first minute and constant during the final 4 minutes. In patients with multiple sclerosis aerobic intensity is moderate during the 6MWT and unaffected by time of day. Disability may have some influence on aerobic intensity, but not on pacing strategy during the 6MWT, whereas neither fatigue nor multiple sclerosis subtype has any effect.

  2. Intensity distribution based space and time division multiple access technique for hybrid-LOS indoor optical wireless communication

    NASA Astrophysics Data System (ADS)

    Miyamoto, Shinichi; Kawamoto, Kenji; Sampei, Seiichi

    2010-02-01

    This paper proposes a space division and time division multiple access (SD/TDMA) technique based on intensity distributions for hybrid line-of-sight (hybrid-LOS) indoor optical wireless communication system. At first, a novel signal discrimination scheme for the spatially multiplexed optical signals is proposed and it is applied to a space division multiple access (SDMA) in hybrid-LOS system. In the proposed scheme, multiple terminals simultaneously transmit their optical signals to access point (AP) using on-off-keying (OOK) modulation, and the spatially multiplexed optical signals are received by a photodetectors array (PD-array), where multiple PDs are disposed to observe the spatial intensity distribution of optical signals. Because the terminals transmit their data using OOK modulation, the spatial intensity distribution observed by the AP equipped with PD-array is subject to the data transmitted from individual terminals, and the AP can identify the terminals transmitting the optical signal by determining the transmitted intensity distribution. Of course, the transmitted intensity distributions are not orthogonal signal and the discriminability of transmitted intensity distributions is much related to the differences of intensity distributions. This implies that the number of terminals that can simultaneously access to the AP will be limited and it is determined by the differences of the candidate intensity distributions. In order to enhance the discriminability of the transmitted intensity distributions, the proposed signal discrimination scheme is further applied to the SD/TDMA. In the SD/TDMA, the discriminability required to enable SDMA is ensured by introducing a scheduling algorithm in which terminals with higher discriminatory of transmitted intensity distributions are allocated to the same time slot. Numerical results show that SD/TDMA using proposed signal discrimination scheme increases the throughput and the number of terminals that can access to

  3. Frameless single-isocenter intensity modulated stereotactic radiosurgery for simultaneous treatment of multiple intracranial metastases.

    PubMed

    Lau, Steven K M; Zhao, Xiao; Carmona, Ruben; Knipprath, Erik; Simpson, Daniel R; Nath, Sameer K; Kim, Gwe-Ya; Hattangadi, Jona A; Chen, Clark C; Murphy, Kevin T

    2014-08-01

    Stereotactic radiosurgery (SRS) is well accepted treatment for patients with intracranial metastases, but the role of frameless radiosurgery is not well defined. Here, we describe our clinical experience applying a novel single-isocenter technique to frameless intensity modulated stereotactic radiosurgery (IMRS) for simultaneous treatment of multiple intracranial metastases. Between 2006 and 2012, 100 consecutive patients received frameless IMRS for multiple intracranial metastases using a single, centrally-located isocenter. Among these, 29 patients were treated for progressive or recurrent intracranial disease. A total of 465 metastases (median, 4 per patient, range, 2-18) were treated to a median dose of 20 Gy (range, 15-50 Gy). Follow-up including clinical examination and magnetic resonance imaging (MRI) occurred every 3 months. Median follow-up for all patients was 4.3 months (range, 0.2-58.3 months), with 83 patients (83.0%) followed until their death. For the remaining 17 patients alive at the time of analysis, median follow-up was 9.2 months (range, 2.2-58.3 months). Overall survival at 6 months was 49.5% [95% confidence interval (CI), 35.3-63.6%]. Local control at 6 and 12 months was 88.9% (95% CI, 79.1-98.6%) and 81.5% (95% CI, 65.2-97.7%), respectively. Regional failure was observed in 39 patients (39%), and 25 patients (25%) received salvage therapy. Grade 3 or greater treatment-related toxicity was observed in 4 patients (4%) and included intracranial hemorrhage, seizure, and radionecrosis. Median total treatment time was 17.2 minutes (range, 2.8-55.3 minutes). Single-isocenter IMRS for multiple intracranial metastases can produce clinical outcomes comparable to those of conventional radiosurgery techniques.

  4. Frameless single-isocenter intensity modulated stereotactic radiosurgery for simultaneous treatment of multiple intracranial metastases

    PubMed Central

    Lau, Steven K. M.; Zhao, Xiao; Carmona, Ruben; Knipprath, Erik; Simpson, Daniel R.; Nath, Sameer K.; Kim, Gwe-Ya; Hattangadi, Jona A.; Chen, Clark C.; Murphy, Kevin T.

    2015-01-01

    Purpose Stereotactic radiosurgery (SRS) is well accepted treatment for patients with intracranial metastases, but the role of frameless radiosurgery is not well defined. Here, we describe our clinical experience applying a novel single-isocenter technique to frameless intensity modulated stereotactic radiosurgery (IMRS) for simultaneous treatment of multiple intracranial metastases. Methods and materials Between 2006 and 2012, 100 consecutive patients received frameless IMRS for multiple intracranial metastases using a single, centrally-located isocenter. Among these, 29 patients were treated for progressive or recurrent intracranial disease. A total of 465 metastases (median, 4 per patient, range, 2–18) were treated to a median dose of 20 Gy (range, 15–50 Gy). Follow-up including clinical examination and magnetic resonance imaging (MRI) occurred every 3 months. Results Median follow-up for all patients was 4.3 months (range, 0.2–58.3 months), with 83 patients (83.0%) followed until their death. For the remaining 17 patients alive at the time of analysis, median follow-up was 9.2 months (range, 2.2–58.3 months). Overall survival at 6 months was 49.5% [95% confidence interval (CI), 35.3–63.6%]. Local control at 6 and 12 months was 88.9% (95% CI, 79.1–98.6%) and 81.5% (95% CI, 65.2–97.7%), respectively. Regional failure was observed in 39 patients (39%), and 25 patients (25%) received salvage therapy. Grade 3 or greater treatment-related toxicity was observed in 4 patients (4%) and included intracranial hemorrhage, seizure, and radionecrosis. Median total treatment time was 17.2 minutes (range, 2.8–55.3 minutes). Conclusions Single-isocenter IMRS for multiple intracranial metastases can produce clinical outcomes comparable to those of conventional radiosurgery techniques. PMID:25821723

  5. Transpiration-Inspired Fabrication of Opal Capillary with Multiple Heterostructures for Multiplex Aptamer-Based Fluorescent Assays.

    PubMed

    Gao, Bingbing; Tang, Litianyi; Zhang, Dagan; Xie, Zhuoying; Su, Enben; Liu, Hong; Gu, Zhongze

    2017-09-27

    In this work we report a method for the fabrication of opal capillary with multiple heterostructures for aptamer-based assays. The method is inspired by plant transpiration. During the fabrication, monodisperse SiO2 nanoparticles (NPs) self-assemble in a glass capillary, with the solvent gradually evaporating from the top end of the capillary. By a simple change of the colloid solution that wicks through the capillary, multiple heterostructures can be easily prepared inside the capillary. On the surface of the SiO2 NPs, polydopamine is coated for immobilization of aminomethyl-modified aptamers. The aptamers are used for fluorescent detection of adenosine triphosphate (ATP) and thrombin. Owing to fluorescence enhancement effect of the photonic heterstructures, the fluorescent signal for detection is amplified up to 40-fold. The limit of detection is 32 μM for ATP and 8.1 nM for thrombin. Therefore, we believe this method is promising for the fabrication of analytical capillary devices for point-of-care testing.

  6. Differentiating between heat pain intensities: the combined effect of multiple autonomic parameters.

    PubMed

    Treister, Roi; Kliger, Mark; Zuckerman, Galit; Goor Aryeh, Itay; Eisenberg, Elon

    2012-09-01

    Although it is well known that pain induces changes in autonomic parameters, the extent to which these changes correlate with the experience of pain is under debate. The aim of the present study was to compare a combination of multiple autonomic parameters and each parameter alone in their ability to differentiate among 4 categories of pain intensity. Tonic heat stimuli (1minute) were individually adjusted to induce no pain, low, medium, and high pain in 45 healthy volunteers. Electrocardiogram, photoplethysmogram, and galvanic skin response were recorded, and the following parameters were calculated: heart rate; heart rate variability-high frequency (0.15 to 0.4Hz) spectral power; skin conductance level; number of skin conduction fluctuations; and photoplethysmographic pulse wave amplitude. A combination of parameters was created by fitting an ordinal cumulative logit model to the data and using linear coefficients of the model. Friedman test with post-hoc Wilcoxon test were used to compare between pain intensity categories for every parameter alone and for their linear combination. All of the parameters successfully differentiated between no pain and all other pain categories. However, none of the parameters differentiated between all 3 pain categories (i.e., low and medium; medium and high; low and high). In contrast, the linear combination of parameters significantly differentiated not only between pain and no pain, but also between all pain categories (P<.001 to .02). These results suggest that multiparameter approaches should be further investigated to make progress toward reliable autonomic-based pain assessment. Copyright © 2012 International Association for the Study of Pain. Published by Elsevier B.V. All rights reserved.

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

    PubMed

    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.

  8. Probabilistic multiple sclerosis lesion classification based on modeling regional intensity variability and local neighborhood information.

    PubMed

    Harmouche, Rola; Subbanna, Nagesh K; Collins, D Louis; Arnold, Douglas L; Arbel, Tal

    2015-05-01

    In this paper, a fully automatic probabilistic method for multiple sclerosis (MS) lesion classification is presented, whereby the posterior probability density function over healthy tissues and two types of lesions (T1-hypointense and T2-hyperintense) is generated at every voxel. During training, the system explicitly models the spatial variability of the intensity distributions throughout the brain by first segmenting it into distinct anatomical regions and then building regional likelihood distributions for each tissue class based on multimodal magnetic resonance image (MRI) intensities. Local class smoothness is ensured by incorporating neighboring voxel information in the prior probability through Markov random fields. The system is tested on two datasets from real multisite clinical trials consisting of multimodal MRIs from a total of 100 patients with MS. Lesion classification results based on the framework are compared with and without the regional information, as well as with other state-of-the-art methods against the labels from expert manual raters. The metrics for comparison include Dice overlap, sensitivity, and positive predictive rates for both voxel and lesion classifications. Statistically significant improvements in Dice values ( ), for voxel-based and lesion-based sensitivity values ( ), and positive predictive rates ( and respectively) are shown when the proposed method is compared to the method without regional information, and to a widely used method [1]. This holds particularly true in the posterior fossa, an area where classification is very challenging. The proposed method allows us to provide clinicians with accurate tissue labels for T1-hypointense and T2-hyperintense lesions, two types of lesions that differ in appearance and clinical ramifications, and with a confidence level in the classification, which helps clinicians assess the classification results.

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

    PubMed

    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.

  10. Fluorescence lifetime and intensity of terbium-doped dipicolinic acid in water, HCl, and sodium acetate buffer solutions.

    PubMed

    Makoui, Anali; Killinger, Dennis K

    2009-02-01

    The lifetimes of the individual fluorescing lines from the terbium-doped dipicolinic acid (DPA) complex have been measured and reported, for the first time to our knowledge. These lifetimes have been measured as a function of terbium and dipicolinic acid concentration, solvent pH, and solvent composition for water, HCl, and sodium acetate buffer solutions. Fluorescence lifetimes over the range from 0.75 to 1.07 ms were measured. The maximum fluorescence was obtained for distilled water solutions.

  11. [Severe thrombocytopenia on admission to the intensive care unit in patients with multiple organ failure].

    PubMed

    Marco-Schulke, C M; Sánchez-Casado, M; Hortigüela-Martín, V A; Quintana-Díaz, M; Rodríguez-Villar, S; Pérez-Pedrero, M J; Velasco-Ramos, A; Canabal-Berlanga, A; Arrese-Cosculluela, M Á

    2012-04-01

    To evaluate the frequency of severe thrombocytopenia (STCP) (≤ 50,000/μl) in the first 24 hours in patients with multiple organ dysfunction syndrome, and the factors that influence its occurrence. A retrospective, observational study. AREA: Medical-surgical intensive care unit (ICU). Tertiary hospital. Those with failure of at least two organs, according to SOFA criteria, with the exclusion of neurological and traumatologic critical cases. Medical history, regular medication, baseline functional status, demographic variables, severity scores in ICU, multiple-organ failure data, course in ICU and main hospital data. A total of 587 patients were included; 6.3% (37 patients) presented with STCP during the first day of admission; 64.6% were men; SOFA 8 (5-10); APACHE II 18 (13-24); APACHE IV 59 (46-73); 32.5% were surgical patients. A total of 79.9% subsequently needed mechanical ventilation, and 71.4% required vasoactive drugs. Overall stay in ICU: 4 (2-10) days, main hospital stay 18 (9-35) days. A total of 29.2% died in the ICU; 11.7% developed STCP during admission to the ICU. Multivariate analysis found the main determining factors in the occurrence of thrombocytopenia on admission to be: history of hospitalization in the last year, albumin and bilirubin levels, and sepsis. The prevalence of STCP among critical patients was 6.3%. Its occurrence was associated with albumin and bilirubin levels, sepsis, and with patient admittance in the last year. Copyright © 2011 Elsevier España, S.L. and SEMICYUC. All rights reserved.

  12. Carbon Dots: A Modular Activity to Teach Fluorescence and Nanotechnology at Multiple Levels

    ERIC Educational Resources Information Center

    Pham, Susan N.; Kuether, Joshua E.; Gallagher, Miranda J.; Hernandez, Rodrigo Tapia; Williams, Denise N.; Zhi, Bo; Mensch, Arielle C.; Hamers, Robert J.; Rosenweig, Zeev; Fairbrother, Howard; Krause, Miriam O. P.; Feng, Z. Vivian; Haynes, Christy L.

    In recent years, nanomaterials have entered our daily lives via consumer products; thus, it has become increasingly important to implement activities to introduce these novel materials into chemistry curricula. Here we introduce a newly developed fluorescent nanomaterial, carbon dots, as a more environmentally friendly alternative to heavy-metal…

  13. Ultra-Fast Excited State Dynamics in Green Fluorescent Protein: Multiple States and Proton Transfer

    NASA Astrophysics Data System (ADS)

    Chattoraj, Mita; King, Brett A.; Bublitz, Gerold U.; Boxer, Steven G.

    1996-08-01

    The green fluorescent protein (GFP) of the jellyfish Aequorea Victoria has attracted widespread interest since the discovery that its chromophore is generated by the autocatalytic, posttranslational cyclization and oxidation of a hexapeptide unit. This permits fusion of the DNA sequence of GFP with that of any protein whose expression or transport can then be readily monitored by sensitive fluorescence methods without the need to add exogenous fluorescent dyes. The excited state dynamics of GFP were studied following photo-excitation of each of its two strong absorption bands in the visible using fluorescence upconversion spectroscopy (about 100 fs time resolution). It is shown that excitation of the higher energy feature leads very rapidly to a form of the lower energy species, and that the excited state interconversion rate can be markedly slowed by replacing exchangeable protons with deuterons. This observation and others lead to a model in which the two visible absorption bands correspond to GFP in two ground-state conformations. These conformations can be slowly interconverted in the ground state, but the process is much faster in the excited state. The observed isotope effect suggests that the initial excited state process involves a proton transfer reaction that is followed by additional structural changes. These observations may help to rationalize and motivate mutations that alter the absorption properties and improve the photo stability of GFP.

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

    NASA Astrophysics Data System (ADS)

    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.

  15. A Solar-Pumped Fluorescence Model for Line-By-Line Emission Intensities in the B-X, A-X, and X-X Band Systems of 12C14N

    NASA Technical Reports Server (NTRS)

    Paganini, L.; Mumma, M. J.

    2016-01-01

    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(sup 2)Pi(sub I) and B(sup 2)Sum(sup +) electronically excited states followed by cascade to ro-vibrational levels of X(sup 2)Sum(sup +), and direct solar infrared pumping of ro-vibrational levels in the X(sup 2)Sum(sup +) 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(sup 2)Sum(sup +) (1-0) in comet C/2014 Q2 (Lovejoy), thereby identifying multiple emission lines observed at IR wavelengths.

  16. A Solar-Pumped Fluorescence Model for Line-By-Line Emission Intensities in the B-X, A-X, and X-X Band Systems of 12C14N

    NASA Technical Reports Server (NTRS)

    Paganini, L.; Mumma, M. J.

    2016-01-01

    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(sup 2)Pi(sub I) and B(sup 2)Sum(sup +) electronically excited states followed by cascade to ro-vibrational levels of X(sup 2)Sum(sup +), and direct solar infrared pumping of ro-vibrational levels in the X(sup 2)Sum(sup +) 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(sup 2)Sum(sup +) (1-0) in comet C/2014 Q2 (Lovejoy), thereby identifying multiple emission lines observed at IR wavelengths.

  17. Intensity-modulated radiosurgery with rapidarc for multiple brain metastases and comparison with static approach

    SciTech Connect

    Wang Jiazhu; Pawlicki, Todd; Rice, Roger; Mundt, Arno J.; Sandhu, Ajay; Lawson, Joshua; Murphy, Kevin T.

    2012-04-01

    Rotational RapidArc (RA) and static intensity-modulated radiosurgery (IMRS) have been used for brain radiosurgery. This study compares the 2 techniques from beam delivery parameters and dosimetry aspects for multiple brain metastases. Twelve patients with 2-12 brain lesions treated with IMRS were replanned using RA. For each patient, an optimal 2-arc RA plan from several trials was chosen for comparison with IMRS. Homogeneity, conformity, and gradient indexes have been calculated. The mean dose to normal brain and maximal dose to other critical organs were evaluated. It was found that monitor unit (MU) reduction by RA is more pronounced for cases with larger number of brain lesions. The MU-ratio of RA and IMRS is reduced from 104% to 39% when lesions increase from 2 to 12. The dose homogeneities are comparable in both techniques and the conformity and gradient indexes and critical organ doses are higher in RA. Treatment time is greatly reduced by RA in intracranial radiosurgery, because RA uses fewer MUs, fewer beams, and fewer couch angles.

  18. Coupling functions for NM total intensity and different multiplicities: Analytical approach

    NASA Astrophysics Data System (ADS)

    Lev, Dorman

    Coupling functions for NM total intensity and different multiplicities play important role when we by observed data of cosmic ray (CR) variations on the ground based detectors (mostly neutron monitors and muon telescopes) tried to determine the primary variations of CR energy spectrum out of the Earth’s atmosphere and magnetosphere, into interplanetary space. This is especially important for forecasting expected radiation hazards from solar CR, because by determined primary solar CR energy spectrum in the interplanetary space it is possible to determine effective time of solar CR ejection into solar wind, source function and the diffusion coefficient of solar CR propagation in space in dependence of particle energy and distance from the Sun. Coupling functions are important also for investigations of Forbush effect and precursory effects for forecasting dangerous interplanetary shock waves. We check obtained results for coupling functions by latitude expedition experimental data. We found how coupling functions depend from the level of solar activity and pressure on the level of observations. Obtained results are presented in the analytical forms that are convenient to use for any NM at any place on the Earth.

  19. Intensity-modulated radiosurgery with rapidarc for multiple brain metastases and comparison with static approach.

    PubMed

    Wang, Jia-Zhu; Pawlicki, Todd; Rice, Roger; Mundt, Arno J; Sandhu, Ajay; Lawson, Joshua; Murphy, Kevin T

    2012-01-01

    Rotational RapidArc (RA) and static intensity-modulated radiosurgery (IMRS) have been used for brain radiosurgery. This study compares the 2 techniques from beam delivery parameters and dosimetry aspects for multiple brain metastases. Twelve patients with 2-12 brain lesions treated with IMRS were replanned using RA. For each patient, an optimal 2-arc RA plan from several trials was chosen for comparison with IMRS. Homogeneity, conformity, and gradient indexes have been calculated. The mean dose to normal brain and maximal dose to other critical organs were evaluated. It was found that monitor unit (MU) reduction by RA is more pronounced for cases with larger number of brain lesions. The MU-ratio of RA and IMRS is reduced from 104% to 39% when lesions increase from 2 to 12. The dose homogeneities are comparable in both techniques and the conformity and gradient indexes and critical organ doses are higher in RA. Treatment time is greatly reduced by RA in intracranial radiosurgery, because RA uses fewer MUs, fewer beams, and fewer couch angles. Copyright © 2012 American Association of Medical Dosimetrists. Published by Elsevier Inc. All rights reserved.

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

    PubMed

    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.

  1. Fluorescence intensity ratio thermometer methodology of eliminating the "decoupling" effect of a pair of thermally coupled energy levels of rare-earth ions.

    PubMed

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

    2017-04-01

    By separating the thermal and nonradiative relaxation population, the fluorescence intensity ratio (FIR) of a pair of thermally coupled energy levels of rare-earth ion is reformulated. For a pair of thermally coupled levels, if the ratio of the thermal population in the upper level to the total population of the lower level abides by the Boltzmann distribution law, the general FIR would be modulated by the proportion of the total population to the thermal population in the upper level. By defining the reciprocal of the proportion as the thermal population degree (η), the product ηFIR will follow the pure Boltzmann distribution law. Considering the fluorescent transient process, the η values may be obtained from the weights of the fluorescent dynamic components of the upper level. A method to calculate this η factor is presented.

  2. A fluorescently labeled dendronized polymer-enzyme conjugate carrying multiple copies of two different types of active enzymes.

    PubMed

    Grotzky, Andrea; Nauser, Thomas; Erdogan, Huriye; Schlüter, A Dieter; Walde, Peter

    2012-07-18

    A hybrid structure of a synthetic dendronized polymer, two different types of enzymes (superoxide dismutase and horseradish peroxidase), and a fluorescent dye (fluorescein) was synthesized. Thereby, a single polymer chain carried multiple copies of the two enzymes and the fluorescein. The entire attachment chemistry is based on UV/vis-quantifiable bis-aryl hydrazone bond formation that allows direct quantification of bound molecules: 60 superoxide dismutase, 120 horseradish peroxidase, and 20 fluorescein molecules on an average polymer chain of 2000 repeating units. To obtain other enzyme ratios the experimental conditions were altered accordingly. Moreover, it could be shown that both enzymes remained fully active and catalyzed a two-step cascade reaction.

  3. Aptamer induced multicoloured Au NCs-MoS2 "switch on" fluorescence resonance energy transfer biosensor for dual color simultaneous detection of multiple tumor markers by single wavelength excitation.

    PubMed

    Xu, Shenghao; Feng, Xiuying; Gao, Teng; Liu, Gufan; Mao, Yaning; Lin, Jiehua; Yu, Xijuan; Luo, Xiliang

    2017-08-29

    An aptamer induced "switch on" fluorescence resonance energy transfer (FRET) biosensor for the simultaneous detection of multiple tumor markers (e.g., AFP and CEA) combining molybdenum disulfide (MoS2) nanosheets with multicolored Au NCs by a single excitation was developed for the first time. Here, AFP aptamer functionalized green colored Au NCs (510 nm) and CEA aptamer functionalized red colored Au NCs (650 nm) are used as energy donors, while MoS2 is used as energy receptor. On the basis of recording the change of the recovered fluorescence intensity at 510 nm and 650 nm upon the addition of targets CEA and AFP, these two tumor markers can be simultaneously quantitatively detected, with detection limits of 0.16 and 0.21 ng mL(-1) (3σ) for AFP and CEA, respectively. In addition, it is noteworthy that the developed biosensor can not only realize accurate quantitative determination of multiple tumor markers by fluorescent intensity, but also be applied in semi-quantitative determination through photo visualization. More importantly, confocal microscope experiments prove that serums from normal and hepatoma patients can also be visually and qualitatively discriminated by this FRET-based biosensor with a single excitation wavelength, indicating promising potential of this assay for clinical diagnosis. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. An extension to laser-induced fluorescence measures multiple velocity components simultaneously

    NASA Astrophysics Data System (ADS)

    Ross, T. J.; Newsham, D.; Rynn, N.

    1996-09-01

    An extension to laser-induced fluorescence is presented. The new technique, called intermodulated optical tagging, can be used to measure multidimensional velocity distribution functions directly as well as to tag ions in two velocity and three spatial coordinates. The first application makes the technique a possible replacement for tomography, the second makes it useful for heretofore difficult transport measurements. Application to velocity-space transport perpendicular to the magnetic field is discussed for a quiescent plasma.

  5. Palus Somni - Anomalies in the correlation of Al/Si X-ray fluorescence intensity ratios and broad-spectrum visible albedos. [lunar surface mineralogy

    NASA Technical Reports Server (NTRS)

    Clark, P. E.; Andre, C. G.; Adler, I.; Weidner, J.; Podwysocki, M.

    1976-01-01

    The positive correlation between Al/Si X-ray fluorescence intensity ratios determined during the Apollo 15 lunar mission and a broad-spectrum visible albedo of the moon is quantitatively established. Linear regression analysis performed on 246 1 degree geographic cells of X-ray fluorescence intensity and visible albedo data points produced a statistically significant correlation coefficient of .78. Three distinct distributions of data were identified as (1) within one standard deviation of the regression line, (2) greater than one standard deviation below the line, and (3) greater than one standard deviation above the line. The latter two distributions of data were found to occupy distinct geographic areas in the Palus Somni region.

  6. Palus Somni - Anomalies in the correlation of Al/Si X-ray fluorescence intensity ratios and broad-spectrum visible albedos. [lunar surface mineralogy

    NASA Technical Reports Server (NTRS)

    Clark, P. E.; Andre, C. G.; Adler, I.; Weidner, J.; Podwysocki, M.

    1976-01-01

    The positive correlation between Al/Si X-ray fluorescence intensity ratios determined during the Apollo 15 lunar mission and a broad-spectrum visible albedo of the moon is quantitatively established. Linear regression analysis performed on 246 1 degree geographic cells of X-ray fluorescence intensity and visible albedo data points produced a statistically significant correlation coefficient of .78. Three distinct distributions of data were identified as (1) within one standard deviation of the regression line, (2) greater than one standard deviation below the line, and (3) greater than one standard deviation above the line. The latter two distributions of data were found to occupy distinct geographic areas in the Palus Somni region.

  7. Design of Multiple Logic Gates Based on Chemically Triggered Fluorescence Switching of Functionalized Polyethylenimine.

    PubMed

    Pan, Yi; Shi, Yupeng; Chen, Zhihua; Chen, Junying; Hou, Mengfei; Chen, Zhanpeng; Li, Cheuk-Wing; Yi, Changqing

    2016-04-13

    In this study, two new functionalized polyethylenimine (PEI), PEIR and PEIQ, have been synthesized by covalently conjugating rhodamine 6G (R6G) or 8-chloroacetyl-aminoquinoline (CAAQ) and have been investigated for their sensing capabilities toward metal ions and anions basing on fluorescence on-off and off-on mechanisms. When triggered by protons, metal ions, or anions, functionalized PEIs can behave as a fluorescence switch, leading to a multiaddressable system. Inspired by these results, functionalized PEI-based logic systems capable of performing elementary logic operations (YES, NOT, NOR, and INHIBIT) and integrative logic operations (OR + INHIBIT) have been constructed by observing the change in the fluorescence with varying the chemical inputs such as protons, metal ions, and anions. Due to its characteristics, such as high sensitivity and fast response, developing functionalized PEI as a new material to perform logic operations may pave a new avenue to construct the next generation of molecular devices with better applicability for biomedical research.

  8. L subshell fluorescence cross-sections and relative intensity ratios of some elements in the atomic range 72⩽ Z⩽92

    NASA Astrophysics Data System (ADS)

    Gürol, A.; Karabulut, A.

    2003-08-01

    The Lα 1,2, Lγ 1, Lγ 2,3,6,8, Lγ 4,4' and Lγ 5 X-ray fluorescence cross-sections for some elements in the atomic range 72⩽ Z⩽92 were measured at 59.54 keV. L subshell fluorescence cross-sections (σ L1, σ L2 and σ L3) and relative intensity ratios (σ L1/σ L2, σ L1/σ L3 and σ L2/σ L3) were determined by using these experimental results. In addition, these cross-sections and relative intensity ratios were calculated theoretically. These experimental and theoretical relative intensity ratios were compared with each other and a fairly good agreement between these experimental and theoretical results were found. Besides, our results for L subshell fluorescence cross-sections were compared with the values of Şahin et al. (J. Phys. B 33 (2000) 93). Good agreement between these values was found for σ L3, but there is no agreement for σ L1 and σ L2.

  9. From the Inside Looking Out--An Intensive Interaction Group for People with Profound and Multiple Learning Disabilities

    ERIC Educational Resources Information Center

    Leaning, Brian; Watson, Tessa

    2006-01-01

    This paper describes the use of a new project, "from the inside looking out" (FILO) (N. Richardson Unpublished data), to develop communication, interaction and emotional literacy skills with people with profound and multiple learning disabilities. The authors utilized tools derived from Intensive Interaction Therapy [D. Hewett & M. Nind (1994)…

  10. Nanoscale energy-route selector consisting of multiple photo-switchable fluorescence-resonance-energy-transfer structures on DNA

    NASA Astrophysics Data System (ADS)

    Fujii, Ryo; Nishimura, Takahiro; Ogura, Yusuke; Tanida, Jun

    2015-04-01

    We report on a nanoscale energy-route selector consisting of multiple fluorescence resonance energy transfer (FRET) structures switched by external signaling with multiple wavelengths of light. In each FRET structure, a specific activator molecule is incorporated to a FRET pair of a donor and an acceptor to control the activation of the acceptor. Owing to this configuration, the FRET structures are switched independently, and an energy route is selected. Two photo-switchable FRET structures, one consists of Alexa Fluor 568 (donor), Cy5 (acceptor), and Alexa Fluor 405 (activator), and the other consists of Alexa Fluor 568 (donor), Cy5.5 (acceptor), and Cy3 (activator), were constructed using DNA strands modified with fluorescence molecules. Switching rates for the individual FRET structures were measured as 64 and 49 %, respectively. An energy-route selector was then assembled with the FRET structures which share a single donor. Experimental results demonstrate that the energy route can be changed repeatedly by activation control using three wavelengths of light.

  11. A fluorescent microsphere-based method for assay of multiple analytes in plasma.

    PubMed

    Bernhard, Oliver K; Mathias, Rommel A; Barnes, Thomas W; Simpson, Richard J

    2011-01-01

    Measurement of multiple analytes can provide increased sensitivity and specificity for the detection and management of disease. The enzyme-linked immunosorbent assay (ELISA) is currently the "gold standard" for protein quantification; however, individual assays for each analyte must be performed, placing demand on sample volume. On the contrary, multiplex assays using microsphere-based technologies allow for multiple analytes to be simultaneously assayed within a single sample. Here, we present a protocol for the preparation and development of a multiple-analyte assay in human plasma using the BioPlex 200 platform (Bio-Rad), which incorporates xMAP technology (Luminex).

  12. A Statistical Assessment of the Impact of Agricultural Land Use Intensity on Regional Surface Water Quality at Multiple Scales

    PubMed Central

    Zhang, Weiwei; Li, Hong; Sun, Danfeng; Zhou, Liandi

    2012-01-01

    Understanding the effects of intensive agricultural land use activities on water resources is essential for natural resource management and environmental improvement. In this paper, multi-scale nested watersheds were delineated and the relationships between two representative water quality indexes and agricultural land use intensity were assessed and quantified for the year 2000 using multi-scale regression analysis. The results show that the log-transformed nitrate-nitrogen (NO3-N) index exhibited a relationship with chemical fertilizer input intensity and several natural factors, including soil loss, rainfall and sunlight at the first order watershed scale, while permanganate index (CODMn) had a positive relationship with another two input intensities of pesticides and agricultural plastic mulch and organic manure at the fifth order watershed scale. The first order watershed and the fifth order watershed were considered as the watershed adaptive response units for NO3-N and CODMn, respectively. The adjustment of agricultural input and its intensity may be carried out inside the individual watershed adaptive response unit. The multiple linear regression model demonstrated the cause-and-effect relationship between agricultural land use intensity and stream water quality at multiple scales, which is an important factor for the maintenance of stream water quality. PMID:23202839

  13. Chemically Induced Fluorescence Switching of Carbon-Dots and Its Multiple Logic Gate Implementation

    PubMed Central

    Dhenadhayalan, Namasivayam; Lin, King-Chuen

    2015-01-01

    Investigations were carried out on the carbon-dots (C-dots) based fluorescent off - on (Fe3 + - S2O32−) and on - off (Zn2 + - PO43−) sensors for the detection of metal ions and anions. The sensor system exhibits excellent selectivity and sensitivity towards the detection of biologically important Fe3 + , Zn2 +  metal ions and S2O32−, PO43− anions. It was found that the functional group on the C-dots surface plays crucial role in metal ions and anions detection. Inspired by the sensing results, we demonstrate C-dots based molecular logic gates operation using metal ions and anions as the chemical input. Herein, YES, NOT, OR, XOR and IMPLICATION (IMP) logic gates were constructed based on the selection of metal ions and anions as inputs. This carbon-dots sensor can be utilized as various logic gates at the molecular level and it will show better applicability for the next generation of molecular logic gates. Their promising properties of C-dots may open up a new paradigm for establishing the chemical logic gates via fluorescent chemosensors. PMID:25943914

  14. Chemically Induced Fluorescence Switching of Carbon-Dots and Its Multiple Logic Gate Implementation

    NASA Astrophysics Data System (ADS)

    Dhenadhayalan, Namasivayam; Lin, King-Chuen

    2015-05-01

    Investigations were carried out on the carbon-dots (C-dots) based fluorescent off - on (Fe3 + - S2O32-) and on - off (Zn2 + - PO43-) sensors for the detection of metal ions and anions. The sensor system exhibits excellent selectivity and sensitivity towards the detection of biologically important Fe3 + , Zn2 +  metal ions and S2O32-, PO43- anions. It was found that the functional group on the C-dots surface plays crucial role in metal ions and anions detection. Inspired by the sensing results, we demonstrate C-dots based molecular logic gates operation using metal ions and anions as the chemical input. Herein, YES, NOT, OR, XOR and IMPLICATION (IMP) logic gates were constructed based on the selection of metal ions and anions as inputs. This carbon-dots sensor can be utilized as various logic gates at the molecular level and it will show better applicability for the next generation of molecular logic gates. Their promising properties of C-dots may open up a new paradigm for establishing the chemical logic gates via fluorescent chemosensors.

  15. Fluorescent intensity-based differential counting of FITC-doped silica nanoparticles: applications of CD4+ T-cell detection in microchip-type flowcytometers

    NASA Astrophysics Data System (ADS)

    Yun, Hoyoung; Bang, Hyunwoo; Lee, Won Gu; Lim, Hyunchang; Park, Junha; Lee, Joonmo; Riaz, Asif; Cho, Keunchang; Chung, Chanil; Han, Dong-Chul; Chang, Jun Keun

    2007-12-01

    Although CD4+ T-cells are an important target of HIV detection, there have been still major problems in making a diagnosis and monitoring in the third world and the region with few medical facilities. Then, it is necessary to use portable diagnosis devices at low cost when you put an enumeration of CD4+ T-cells. In general, the counting of CD4 below 200cells/uL makes it necessary to initiate antiretroviral treatment in adults (over 13 years old). However, lymphocyte subsets (including CD4 counts) of infants and young children are higher than those of adults. This fact shows the percentage of CD4+ T-cells of blood subsets, i.e., CD4/CD45%, CD4/CD8% or CD4/CD3% means a more reliable indicator of HIV infection than absolute counts in children. To know the percentage of CD4+ T-cell by using two fluorescent dyes of different emission wavelength, at least, one laser and two PMT detectors are in general needed. Then, it is so hard to develop a portable device like a 'toaster size' because this makes such a device more complex including many peripheral modules. In this study, we developed a novel technique to control the intensity of fluorescent dye-doped silica nanoparticles. I synthesized FITC-doped silica nanoparticles conjugated CD4 antibody 10 times brighter than FITC-conjugated CD45 antibody. With the difference of intensity of two fluorescent dyes, we measured two parameters by using only a single detector and laser. Most experiments were achieved with uFACS (microfabricated fluorescence-activated cell sorter) on an inverted microscope (IX71, Olympus). In conclusion, this method enables us to discriminate the difference between CD4 and CD45 in an intensity domain simultaneously. Furthermore, this technique would make it possible develop much cheaper and smaller devices which can count the number of CD4 T-cells.

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

    PubMed

    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.

  17. Optimal patterns for sequentially multiple focusing in high intensity focused ultrasound and their application to thermal dose

    NASA Astrophysics Data System (ADS)

    Shim, Mun-Bo; Lee, Hyoungki; Lee, Hotaik; Park, Junho; Ahn, Minsu

    2012-11-01

    The purpose of this study is to propose a new method for multiple-focus generation to shorten overall treatment time as well as to avoid the formation of high intensity regions outside the target volume. A numerical simulation of the acoustic fields produced by a 1017-element spherical-section ultrasound phased array transducer operating at a frequency of 1.0MHz with 16 cm radius of curvature is performed for the proposed multiple-focus generation. The total foci are partitioned into the several patterns because multiple focusing generally gives rise to the grating lobes outside of the three dimensional region of interest even if applying the optimization of intensity gain in determining the phases and amplitudes of the excitation source vector. The optimization problem is repeatedly formulated in term of the focal points until the multiple-focus patterns cover all the focal points. Genetic algorithm is used for selecting the patterns without the grating lobes. The obtained set of multiple-focus patterns can sequentially be used to necrose a given volume in the short time as well as in the safe treatment. The proposed method might prove useful to improve the speed and safety of focused ultrasound thermal ablation. This strategy will also be effective for any transducers as well as for other cases of multiple-focus generation.

  18. Predicting Distribution and Inter-Annual Variability of Tropical Cyclone Intensity from a Stochastic, Multiple-Linear Regression Model

    NASA Astrophysics Data System (ADS)

    Lee, C. Y.; Tippett, M. K.; Sobel, A. H.; Camargo, S. J.

    2014-12-01

    We are working towards the development of a new statistical-dynamical downscaling system to study the influence of climate on tropical cyclones (TCs). The first step is development of an appropriate model for TC intensity as a function of environmental variables. We approach this issue with a stochastic model consisting of a multiple linear regression model (MLR) for 12-hour intensity forecasts as a deterministic component, and a random error generator as a stochastic component. Similar to the operational Statistical Hurricane Intensity Prediction Scheme (SHIPS), MLR relates the surrounding environment to storm intensity, but with only essential predictors calculated from monthly-mean NCEP reanalysis fields (potential intensity, shear, etc.) and from persistence. The deterministic MLR is developed with data from 1981-1999 and tested with data from 2000-2012 for the Atlantic, Eastern North Pacific, Western North Pacific, Indian Ocean, and Southern Hemisphere basins. While the global MLR's skill is comparable to that of the operational statistical models (e.g., SHIPS), the distribution of the predicted maximum intensity from deterministic results has a systematic low bias compared to observations; the deterministic MLR creates almost no storms with intensities greater than 100 kt. The deterministic MLR can be significantly improved by adding the stochastic component, based on the distribution of random forecasting errors from the deterministic model compared to the training data. This stochastic component may be thought of as representing the component of TC intensification that is not linearly related to the environmental variables. We find that in order for the stochastic model to accurately capture the observed distribution of maximum storm intensities, the stochastic component must be auto-correlated across 12-hour time steps. This presentation also includes a detailed discussion of the distributions of other TC-intensity related quantities, as well as the inter

  19. Reasoning about Intensive Quantities in Whole-Number Multiplication? A Possible Basis for Ratio Understanding

    ERIC Educational Resources Information Center

    Simon, Martin A.; Placa, Nicora

    2012-01-01

    One of the challenges of learning ratio concepts is that it involves intensive quantities, a type of quantity that is more conceptually demanding than those that are evaluated by counting or measuring (extensive quantities). In this paper, we engage in an exploration of the possibility of developing reasoning about intensive quantities during the…

  20. Health care utilization before and after intensive care unit admission in multiple sclerosis.

    PubMed

    Marrie, Ruth Ann; Bernstein, Charles N; Peschken, Christine A; Hitchon, Carol A; Chen, Hui; Fransoo, Randall; Garland, Allan

    2015-07-01

    The incidence of intensive care unit (ICU) admission is elevated in the multiple sclerosis (MS) population but the reasons for this are incompletely understood, as are outcomes post-ICU admission. Among MS patients we examined the association between ICU admission and health care utilization in the year preceding admission, and compared health care utilization following ICU admission among persons with MS and persons from the general population. We used population-based administrative data from Manitoba, Canada to identify 4237 MS cases of which 2547 were incident. We compared the incidence rates of ICU admission in the prevalent MS population according to health care utilization in the year before admission, adjusting for age, sex, comorbidity and socioeconomic status. Among incident cases of MS we compared rates of health care utilization after ICU admission to those in a matched general population cohort. We used generalized linear models adjusting for age, sex, socioeconomic status, region, comorbidity and utilization before admission. Of 4219 prevalent MS cases, 222 (5.3%) were admitted to the ICU. After adjustment, any hospitalization in the prior year conferred an 80% increased incidence, and physician visits in the highest tertile and prescription costs in the highest quartile in the prior year each conferred a more than two-fold increased incidence of admission. Among 2547 incident cases of MS, 109 (4.3%) were admitted to the ICU and 93 survived their admission. Thirty-eight percent of the MS population were re-hospitalized in the year following admission, similar to the matched population (33.8%). Seven percent of both populations were readmitted to the ICU. The MS population had more hospital days after ICU admission than the matched population (adjusted RR 3.11; 95% CI: 1.34-5.90). After adjustment the number of physician visits did not differ between populations. The incidence of ICU admission is higher among persons with MS who have higher prior

  1. Targeting accuracy of single-isocenter intensity-modulated radiosurgery for multiple lesions.

    PubMed

    Calvo-Ortega, J F; Pozo, M; Moragues, S; Casals, J

    2017-01-01

    To investigate the targeting accuracy of intensity-modulated SRS (IMRS) plans designed to simultaneously treat multiple brain metastases with a single isocenter. A home-made acrylic phantom able to support a film (EBT3) in its coronal plane was used. The phantom was CT scanned and three coplanar small targets (a central and two peripheral) were outlined in the Eclipse system. Peripheral targets were 6 cm apart from the central one. A reference IMRS plan was designed to simultaneously treat the three targets, but only a single isocenter located at the center of the central target was used. After positioning the phantom on the linac using the room lasers, a CBCT scan was acquired and the reference plan were mapped on it, by placing the planned isocenter at the intersection of the landmarks used in the film showing the linac isocenter. The mapped plan was then recalculated and delivered. The film dose distribution was derived using a cloud computing application (www.radiochromic.com) that uses a triple-channel dosimetry algorithm. Comparison of dose distributions using the gamma index (5%/1 mm) were performed over a 5 × 5 cm(2) region centered over each target. 2D shifts required to get the best gamma passing rates on the peripheral target regions were compared with the reported ones for the central target. The experiment was repeated ten times in different sessions. Average 2D shifts required to achieve optimal gamma passing rates (99%, 97%, 99%) were 0.7 mm (SD: 0.3 mm), 0.8 mm (SD: 0.4 mm) and 0.8 mm (SD: 0.3 mm), for the central and the two peripheral targets, respectively. No statistical differences (p > 0.05) were found for targeting accuracy between the central and the two peripheral targets. The study revealed a targeting accuracy within 1 mm for off-isocenter targets within 6 cm of the linac isocenter, when a single-isocenter IMRS plan is designed. Copyright © 2017 American Association of Medical Dosimetrists. Published by Elsevier Inc. All rights reserved.

  2. An unusual cause of multiple organ dysfunction syndrome in the pediatric intensive care unit: hemophagocytic lymphohistiocytosis.

    PubMed

    Karapinar, Bülent; Yilmaz, Deniz; Balkan, Can; Akin, Mehmet; Ay, Yilmaz; Kvakli, Kaan

    2009-05-01

    To report our experience in children with primary or secondary hemophagocytic lymphohistiocytosis (HLH) presented with multiple organ dysfunction syndrome (MODS) in pediatric intensive care unit (PICU). The records of patients with a diagnosis of HLH and MODS between January 2005 and January 2008 were reviewed. The patients' characteristics, treatment modalities, and outcomes were assessed. PICU of Ege University Hospital. Twelve children who were hospitalized in the PICU met the diagnostic criteria for HLH, and presented with MODS were entered into the study. The median age of the patients was 3 years (range, 2 months-15.5 years). Six patients had a history of parental consanguinity and two had an affected sibling. Five of the patients were classified as primary HLH. All of the patients had hepatosplenomegaly, elevated ferritin levels, hypofibrinogenemia, anemia, thrombocytopenia, and hemophagocytosis in bone marrow examination at presentation. The median Pediatric Logistic Organ Dysfunction score of the patients at onset was 51 (range, 12-62). Four patients had six, four had five, two had four, and the remaining two had three organ dysfunctions. Organ dysfunction, other than hematologic dysfunction which was present in all patients, was most commonly seen in hepatic (n = 11, 91.7%), respiratory (n = 11, 91.7%), and cardiovascular systems (n = 10, 83.3%). Although nine patients showed neurologic dysfunction including convulsion and coma, renal failure was detected in five patients. Eleven patients were supported with mechanical ventilation and four patients required hemodialysis. Eight patients were treated according to the HLH 2004 treatment protocol, consisting of cyclosporine A, etoposide, and dexamethasone. The remaining four patients received only intravenous immunoglobulin and supportive treatment. Seven of the patients died. HLH is a frequently lethal disease and with a clinical presentation similar to severe sepsis, MODS, disseminated intravascular

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

    NASA Astrophysics Data System (ADS)

    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

  4. [Control of photocoagulation intensity by thermo-induced release of a fluorescent marker encapsulated in liposomes: study of an in vivo vascular model].

    PubMed

    Desmettre, T; Mordon, S; Soulie, S; Devoisselle, J M; Mitchell, V

    1996-01-01

    To evaluate the feasibility of thermal damage assessment of blood vessels by using laser-induced release of liposome-encapsulated dye. A skin flap window model of aluminium was implanted on the loose skin on the back of adults Golden hamsters to expose skin blood vessels in vivo. Thermosensitive liposomes (DSPC) loaded with 5,6-Carboxyfluorescein were injected together with a specific Indocyanine green (ICG) formulation (O/W emulsion) in order to enhance diode laser absorption. Photocoagulations were then performed on the vessels with a diode laser (lambda = 810 nm, P = 0.8W, phi = 1.3 mm, 1 to 6s). Fluorescence measurements were realized with an ultra high sensitivity intensified camera (Hamamatsu Argus 50 imaging system). Two different fluorescence intensity curves corresponding to the variability of absorption of the targets were observed. Variability was related to the amount of ICG. For each curve, 3 zones were identified: (i) for fluences ranging from 60 +/- 20 J/cm2 to 110 +/- 20 J/cm2 a transient intravascular fluorescence was observed only for the loser pulses targeted on the vessels, (ii) for fluences ranging from 110 +/- 20 J/cm2 to 190 +/- 20 J/cm2 a permanent fluorescent spot limited to the vessel was observed for the laser pulses targeted on the vessels; for the laser pulses targeted on the skin a transient low fluorescence circular spot was observed. For this fluence range a selective photocoagulation of a vessel was performed. (iii) for fluences ranging from 190 +/- 20 J/cm2 to 300 +/- 20 J/cm2 persistent intense fluorescence spots were observed on both skin and vessels. This type of fluorescence was related to an overdosage. These results are in fair agreement with the data of the literature about liposomes and with the data we obtained in a previous study on a vascular model. This study demonstrates the interest of a laser-induced release of liposome-encapsulated dye for a real-time quantification of thermal damage. Such a method could be useful

  5. Multiple-wavelength operation of electroabsorption intensity modulator array fabricated using the one-step quantum well intermixing process

    NASA Astrophysics Data System (ADS)

    Ng, S. L.; Lim, H. S.; Lam, Y. L.; Chan, Y. C.; Ooi, B. S.; Aimez, V.; Beauvais, J.; Beerens, J.

    2002-09-01

    Multiple-wavelength selective channel electroabsorption intensity modulators have been fabricated on a single InGaAs/InGaAsP chip using a one-step quantum well intermixing process. This technique was demonstrated for tailoring the intensity modulator operating wavelength by incorporating low-energy (360 keV) phosphorus ions implantation induced disordering process with gray-mask lithography technology. A modulation depth of -15 dB has been measured from these devices with a voltage swing of -4.5 V.

  6. Enhanced intensity variation for multiple-plane phase retrieval using a spatial light modulator as a convenient tunable diffuser.

    PubMed

    Almoro, Percival F; Pham, Quang Duc; Serrano-Garcia, David Ignacio; Hasegawa, Satoshi; Hayasaki, Yoshio; Takeda, Mitsuo; Yatagai, Toyohiko

    2016-05-15

    In the multiple-plane phase retrieval method, a tedious-to-fabricate phase diffuser plate is used to increase the axial intensity variation for a nonstagnating iterative reconstruction of a smooth object wavefront. Here we show that a spatial light modulator (SLM) can be used as an easily controllable diffuser for phase retrieval. The polarization modulation at the SLM facilitates independent formation of orthogonally polarized scattered and specularly reflected beams. Through an analyzer, the polarization states are filtered enabling beam interference, thereby efficiently encoding the phase information in the axially diverse speckle intensity measurements. The technique is described using wave propagation and Jones calculus, and demonstrated experimentally on technical and biological samples.

  7. Effects of ferrofluid and phytoalexin spirobrassinin on thioflavin-T-based fluorescence in cerebrospinal fluid of the elderly and multiple sclerosis patients.

    PubMed

    Kristofikova, Zdena; Gazova, Zuzana; Siposova, Katarina; Bartos, Ales; Ricny, Jan; Kotoucova, Jolana; Sirova, Jana; Ripova, Daniela

    2014-08-01

    It is well known that misfolded peptides/proteins can play a role in processes of normal ageing and in the pathogenesis of many diseases including Alzheimer's disease. Previously, we evaluated samples of cerebrospinal fluid from patients with Alzheimer's disease and multiple sclerosis by means of thioflavin-T-based fluorescence. We observed attenuated effects of magnetite nanoparticles operated via anti-aggregation actions on peptides/proteins from patients with Alzheimer's disease but not from those with multiple sclerosis when compared to age-related controls. In this study, we have evaluated the in vitro effects of anti-aggregation operating ferrofluid and phytoalexin spirobrassinin in the cerebrospinal fluid of patients with multiple sclerosis and Alzheimer's disease. We have found significant differences in native fluorescence (λ excitation = 440 nm, λ emission = 485 nm) of samples among particular groups (young controls < multiple sclerosis, Alzheimer's disease < old controls). Differences among groups were observed also in thioflavin-T-based fluorescence (young controls = multiple sclerosis < Alzheimer's disease < old controls) and the most marked change from native to thioflavin-T-based fluorescence was found in young controls (28-40 years old people). Both ferrofluid and spirobrassinin evoked drops in thioflavin-T-based fluorescence; however, ferrofluid was more efficient in old controls (54-75 years old people) and spirobrassinin in multiple sclerosis patients, both compared to young controls. The results are discussed especially in relation to aggregated peptides/proteins and liposoluble fluorescent products of lipid peroxidation. Based on the significant effect of spirobrassinin in vitro, we suggest that spirobrassinin may be of therapeutic value in multiple sclerosis.

  8. Fluorescence from Multiple Chromophore Hydrogen-Bonding States in the Far-Red Protein TagRFP675.

    PubMed

    Konold, Patrick E; Yoon, Eunjin; Lee, Junghwa; Allen, Samantha L; Chapagain, Prem P; Gerstman, Bernard S; Regmi, Chola K; Piatkevich, Kiryl D; Verkhusha, Vladislav V; Joo, Taiha; Jimenez, Ralph

    2016-08-04

    Far-red fluorescent proteins are critical for in vivo imaging applications, but the relative importance of structure versus dynamics in generating large Stokes-shifted emission is unclear. The unusually red-shifted emission of TagRFP675, a derivative of mKate, has been attributed to the multiple hydrogen bonds with the chromophore N-acylimine carbonyl. We characterized TagRFP675 and point mutants designed to perturb these hydrogen bonds with spectrally resolved transient grating and time-resolved fluorescence (TRF) spectroscopies supported by molecular dynamics simulations. TRF results for TagRFP675 and the mKate/M41Q variant show picosecond time scale red-shifts followed by nanosecond time blue-shifts. Global analysis of the TRF spectra reveals spectrally distinct emitting states that do not interconvert during the S1 lifetime. These dynamics originate from photoexcitation of a mixed ground-state population of acylimine hydrogen bond conformers. Strategically tuning the chromophore environment in TagRFP675 might stabilize the most red-shifted conformation and result in a variant with a larger Stokes shift.

  9. Probing the binding interaction of AKR with human serum albumin by multiple fluorescence spectroscopy and molecular modeling.

    PubMed

    Li, Yan; Chen, Chun; Zhang, Chunping; Duan, Jingyu; Yao, Huankai; Wei, Qunli

    2017-05-01

    Human serum albumin (HSA) is the major transport protein affording endogenous and exogenous substances in plasma. It can affect the behavior and efficacy of chemicals in vivo through the binding interaction. AKR (3-O-α-l-arabinofuranosyl-kaempferol-7-O-α-l-rhamnopyranoside) is a flavonoid diglycoside with modulation of estrogen receptors (ERs). Herein, we investigated the binding interaction between AKR and HSA by multiple fluorescence spectroscopy and molecular modeling. As a result, AKR specifically binds in site I of HSA through hydrogen bonds, van der Waals force, and electrostatic interaction. The formation of AKR-HSA complex in binding process is spontaneously exothermic and leads to the static fluorescence quenching through affecting the microenvironment around the fluorophores. The complex also affects the backbone of HSA and makes AKR access to fluorophores. Molecular modeling gives the visualization of the interaction between AKR and HSA as well as ERs. The affinity of AKR with HSA is higher than the competitive site marker Warfarin. In addition, docking studies reveal the binding interaction of AKR with ERs through hydrogen bonds, van der Waals force, hydrophobic, and electrostatic interactions. And AKR is more favorable to ERβ. These results unravel the binding interaction of AKR with HSA and mechanism as an ERs modulator.

  10. Minimization of self-quenching fluorescence on dyes conjugated to biomolecules with multiple labeling sites via asymmetrically charged NIR fluorophores

    PubMed Central

    Zhegalova, Natalia G.; He, Shawn; Zhou, Haiying; Kim, David M.; Berezin, Mikhail Y.

    2014-01-01

    Self-aggregation of dyes even at low concentrations pose a considerable challenge in preparing sufficiently bright molecular probes for in vivo imaging, particularly in the conjugation of near infrared cyanine dyes to polypeptides with multiple labeling sites. Such self-aggregation leads to a significant energy transfer between the dyes resulting in severe quenching and low brightness of the targeted probe. To address this problem, we designed a novel type of dye with an asymmetrical distribution of charge. Asymmetrical distribution prevents the chromophores from π-stacking thus minimizing the energy transfer and fluorescence quenching. The conjugation of the dye to polypeptides showed only a small presence of an H-aggregate band in the absorption spectra and, hence, a relatively high quantum efficiency. PMID:24764130

  11. Completely distinguishing individual A-genome chromosomes and their karyotyping analysis by multiple bacterial artificial chromosome - fluorescence in situ hybridization.

    PubMed

    Wang, Kai; Guan, Bing; Guo, Wangzhen; Zhou, Baoliang; Hu, Yan; Zhu, Yichao; Zhang, Tianzhen

    2008-02-01

    Twenty bacterial artificial chromosome (BAC) clones that could produce bright signals and no or very low fluorescence in situ hybridization (FISH) background were identified from Gossypium arboreum cv. JLZM, and G. hirsutum accession (acc.) TM-1 and 0-613-2R. Combining with 45S and 5S rDNA, a 22-probe cocktail that could identify all 13 G. arboreum chromosomes simultaneously was developed. According to their homology with tetraploid cotton, the G. arboreum chromosomes were designated as A1-A13, and a standard karyotype analysis of G. arboreum was presented. These results demonstrated an application for multiple BAC-FISH in cotton cytogenetic studies and a technique to overcome the problem of simultaneous chromosome recognition in mitotic cotton cells.

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

    SciTech Connect

    Worley, Christopher G; Soderberg, Constance B; Townsend, Lisa E

    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.

  13. Analysis of multiple scattering suppression using structured laser illumination planar imaging in scattering and fluorescing media.

    PubMed

    Kristensson, E; Araneo, L; Berrocal, E; Manin, J; Richter, M; Aldén, M; Linne, M

    2011-07-04

    The accuracy, precision and limitations of the imaging technique named Structured Laser Illumination Planar Imaging (SLIPI) have been investigated. SLIPI, which allows multiply scattered light to be diminished, has previously demonstrated improvements in image quality and contrast for spray imaging. In the current study the method is applied to a controlled confined environment consisting of a mixture of water and monodisperse polystyrene microspheres. Elastic scattering and fluorescence are studied and the results obtained when probing different particle concentrations and diameters conclusively show the advantages of SLIPI for imaging within moderately turbid media. Although the technique presents both good repeatability and agreement with the Beer-Lambert law, discrepancies in its performance were, however, discovered. Photons undergoing scattering without changing their incident trajectory cannot be discriminated and, owing to differences in scattering phase functions, probing larger particles reduces the suppression of multiply scattered light. However, in terms of visibility such behavior is beneficial as it allows denser media to be probed. It is further demonstrated that the suppression of diffuse light performs equally well regardless of whether photons propagate along the incident direction or towards the camera. In addition, this filtering process acts independently on the spatial distribution of the multiply scattered light but is limited by the finite dynamic range and unavoidable signal noise of the camera.

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

    PubMed

    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.

  15. Potential rainfall-intensity and pH-driven shifts in the apparent fluorescent composition of dissolved organic matter in rainwater.

    PubMed

    Zhou, Yongqiang; Yao, Xiaolong; Zhang, Yibo; Shi, Kun; Zhang, Yunlin; Jeppesen, Erik; Gao, Guang; Zhu, Guangwei; Qin, Boqiang

    2017-05-01

    Perturbations of rainwater chromophoric dissolved organic matter (CDOM) fluorescence induced by changes in rainfall intensity and pH were investigated by field observations and laboratory pH titrations. Microbial humic-like fluorophores dominated the rainwater CDOM pool, followed by tryptophan-like and tyrosine-like substances. Increased rainfall intensity had notable dilution effects on all six fluorescent components (C1-C6) identified using parallel factor (PARAFAC) analysis, the effect being especially pronounced for the microbial humic-like C1, tryptophan-like C3, and tyrosine-like C5. The results also indicated that increasing pH from 7 to 9 led to decreased fluorescence intensity (Fmax) of all the six components, while a pH increase from 5 to 7, resulted in increasing Fmax of terrestrial humic-like C2, tyrosine-like C5, and tryptophan-like C6 and decreasing microbial humic-like C1, tryptophan-like C3, and fulvic-like C4. Two-dimensional correlation spectroscopy (2D-COS) demonstrated that synchronous fluorescence responded first to pH modifications at fulvic-like wavelength (λEx/λEm = ∼316/416 nm), followed by tyrosine-like wavelength (λEx/λEm = ∼204/304 nm), tryptophan-like wavelength (λEx/λEm = ∼226/326 nm), microbial humic-like wavelength (∼295/395 nm), and finally terrestrial humic-like wavelength (∼360/460 nm). Our results suggest that a decrease in areas affected by acid rain in South China occurring at present may possibly result in apparent compositional changes of CDOM fluorescence. The decreased rainfall in South-West China and increased rainfall in North-West China during the past five decades may possibly accordingly result in increased and decreased Fmax of all the six components identified in South-West and North-West China, respectively. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Sensitive fluorescent detection of DNA methyltransferase using nicking endonuclease-mediated multiple primers-like rolling circle amplification.

    PubMed

    Huang, Juan; Li, Xiao-Yu; Du, Yi-Chen; Zhang, Li-Na; Liu, Ke-Ke; Zhu, Li-Na; Kong, De-Ming

    2017-05-15

    Sensitive and reliable detection of DNA methyltransferase (MTase) is of great significance for both early tumor diagnosis and therapy. In this study, a simple, label-free and sensitive DNA MTase-sensing method was developed on the basis of a nicking endonuclease-mediated multiple primers-like rolling circle amplification (RCA) strategy. In this method, a dumbbell RCA template was prepared by blunt-end ligation of two molecules of hairpin DNA. In addition to the primer-binding sequence, the dumbbell template contained another three important parts: 5'-CCGG-3' sequences in double-stranded stems, nicking endonuclease recognition sites and C-rich sequences in single-stranded loops. The introduction of 5'-CCGG-3' sequences allows the dumbbell template to be destroyed by the restriction endonuclease, HpaII, but is not destroyed in the presence of the target MTase-M.SssI MTase. The introduction of nicking endonuclease recognition sites makes the M.SssI MTase-protected dumbbell template-mediated RCA proceed in a multiple primers-like exponential mode, thus providing the RCA with high amplification efficiency. The introduction of C-rich sequences may promote the folding of amplification products into a G-quadruplex structure, which is specifically recognized by the commercially available fluorescent probe thioflavin T. Improved RCA amplification efficiency and specific fluorescent recognition of RCA products provide the M.SssI MTase-sensing platform with high sensitivity. When a dumbbell template containing four nicking endonuclease sites is used, highly specific M.SssI MTase activity detection can be achieved in the range of 0.008-50U/mL with a detection limit as low as 0.0011U/mL. Simple experimental operation and mix-and-detection fluorescent sensing mode ensures that M.SssI MTase quantitation works well in a real-time RCA mode, thus further simplifying the sensing performance and making high throughput detection possible. The proposed MTase-sensing strategy was also

  17. Onsite naked eye determination of cysteine and homocysteine using quencher displacement-induced fluorescence recovery of the dual-emission hybrid probes with desired intensity ratio.

    PubMed

    Wang, Kan; Qian, Jing; Jiang, Ding; Yang, Zhengting; Du, Xiaojiao; Wang, Kun

    2015-03-15

    Simple, inexpensive, portable sensing strategies for those clinically relevant molecules have attained a significant positive impact on the health care system. Herein, we have prepared a dual-emission ratiometric fluorescence probe with desired intensity ratio and demonstrated its efficiency for onsite naked eye determination of cysteine (Cys) and homocysteine (Hcy). The hybrid probe has been designed by hybridizing two differently sized CdTe quantum dots (QDs), in which the red-emitting CdTe QDs (rQDs) entrapped in the silica sphere acting as the reference signal, and the green-emitting CdTe QDs (gQDs) covalently attached on the silica surface serving as the response signal. When 1,10-phenanthroline with strong coordination ability to Cd atoms in gQDs was introduced, the fluorescence of the gQDs was effectively quenched, while the fluorescence of the rQDs stayed constant. Upon exposure to different contents of Cys or Hcy, the fluorescence of gQDs can be recovered gradually due to the displacement of the quencher. Based on the background signal of rQDs, the variations of the sensing system display continuous fluorescence color changes from red to green, which can be easily observed by the naked eye. The assay requires ∼20min and has a detection limit of 2.5 and 1.7μM for Cys and Hcy, respectively. Furthermore, we demonstrate that this sensing scheme can be fully integrated in a filter paper-based assay, thus enabling a potential point-of-care application featuring easy operation, low power consumption, and low fabrication costs.

  18. NIST/ISAC standardization study: variability in assignment of intensity values to fluorescence standard beads and in cross calibration of standard beads to hard dyed beads.

    PubMed

    Hoffman, Robert A; Wang, Lili; Bigos, Martin; Nolan, John P

    2012-09-01

    Results from a standardization study cosponsored by the International Society for Advancement of Cytometry (ISAC) and the US National Institute of Standards and Technology (NIST) are reported. The study evaluated the variability of assigning intensity values to fluorophore standard beads by bead manufacturers and the variability of cross calibrating the standard beads to stained polymer beads (hard-dyed beads) using different flow cytometers. Hard dyed beads are generally not spectrally matched to the fluorophores used to stain cells, and spectral response varies among flow cytometers. Thus if hard dyed beads are used as fluorescence calibrators, one expects calibration for specific fluorophores (e.g., FITC or PE) to vary among different instruments. Using standard beads surface-stained with specific fluorophores (FITC, PE, APC, and Pacific Blue™), the study compared the measured intensity of fluorophore standard beads to that of hard dyed beads through cross calibration on 133 different flow cytometers. Using robust CV as a measure of variability, the variation of cross calibrated values was typically 20% or more for a particular hard dyed bead in a specific detection channel. The variation across different instrument models was often greater than the variation within a particular instrument model. As a separate part of the study, NIST and four bead manufacturers used a NIST supplied protocol and calibrated fluorophore solution standards to assign intensity values to the fluorophore beads. Values assigned to the reference beads by different groups varied by orders of magnitude in most cases, reflecting differences in instrumentation used to perform the calibration. The study concluded that the use of any spectrally unmatched hard dyed bead as a general fluorescence calibrator must be verified and characterized for every particular instrument model. Close interaction between bead manufacturers and NIST is recommended to have reliable and uniformly assigned

  19. Comparison of Pollen Transfer Dynamics by Multiple Floral Visitors: Experiments with Pollen and Fluorescent Dye

    PubMed Central

    ADLER, LYNN S.; IRWIN, REBECCA E.

    2006-01-01

    • Background and Aims Most plant species are visited by a diversity of floral visitors. Pollen transfer of the four most common pollinating bee species and one nectar-robbing bee of the distylous plant Gelsemium sempervirens were compared. • Methods Naturally occurring pollen loads carried by the common floral visitor species of G. sempervirens were compared. In addition, dyed pollen donor flowers and sequences of four emasculated recipient flowers in field cages were used to estimate pollen transfer, and the utility of fluorescent dye powder as an analogue for pollen transfer was determined. • Key Results Xylocopa virginica, Osmia lignaria and Habropoda laboriosa carried the most G. sempervirens pollen on their bodies, followed by Bombus bimaculatus and Apis mellifera. However, B. bimaculatus, O. lignaria and H. laboriosa transferred significantly more pollen than A. mellifera. Nectar-robbing X. virginica transferred the least pollen, even when visiting legitimately. Dye particles were strongly correlated with pollen grains on a stigma, and therefore provide a good analogue for pollen in this system. The ratio of pollen : dye across stigmas was not affected by bee species or interactions between bee species and floral morphology. However, dye transfer was more sensitive than pollen transfer to differences in floral morphology. • Conclusions The results from this study add to a growing body of literature highlighting that floral visitors vary in pollination effectiveness, and that visitors carrying the most pollen on their bodies may not always be the most efficient at depositing pollen on stigmas. Understanding the magnitude of variability in pollinator quality is one important factor for predicting how different pollinator taxa may influence the evolution of floral traits. PMID:16299005

  20. Plum-blossom needling promoted PpIX fluorescence intensity from 5-aminolevulinic acid in porcine skin model and patients with actnic keratosis.

    PubMed

    Chen, Jia; Zhang, Yunfeng; Wang, Peiru; Wang, Bo; Zhang, Guolong; Wang, Xiuli

    2016-09-01

    Plum-blossom needling might enhance transdermal penetration of topically applied drugs by creating vertical channels. The purpose of this study was to evaluate drug delivery assisted by plum-blossom needling comparing with CO2 laser ablative fractional resurfacing (AFR) using 5-aminolevulinic acid (5-ALA), a porphyrin precursor, as a test drug. Ex vivo porcine skin was treated with plum-blossom needle(HWATO, Suzhou medical supplies factory Co., Ltd. China) or CO2 laser AFR before topical application of 20% 5-ALA(Sigma-Aldrich, Co., USA)cream, placebo cream and no cream. ALA-induced porphyrin fluorescence was measured by fluorescence microscopy at skin depths down to 1800μm. Needling was done by tapping the skin vertically from 5cm high above quickly. AFR was performed with a 10.6μm wavelength prototype CO2 laser, using stacked single pulses of 3 millisecond and 91.6mJ per pulse. Plum-blossom needling after ALA application was also done. Fluorescence intensity on lesion surface was examined by curalux spectrum analyzer (Laser Institute of Munich University, Germany) and VAS pain score was recorded in a randomized split-lesion clinical trial including 6 patients, 8 actinic keratosis lesions. AFR created regular cone-shaped channels surrounded by a 70μm thin layer of thermally coagulated dermis, respectively. The cone is approximately 200μm in diameter at the opening and 1850μm in depth. Plum-blossom needle created irregular cone-shaped channels of approximately 180μm in diameter at the opening and it always drags a tail-which was shaped from the closed deeper channels. There was no porphyrin fluorescence in placebo cream or untreated skin sites. Plum-blossom needling followed by ALA application enhanced drug delivery with significantly higher porphyrin fluorescence at the edge of hole (P<0.005) and 100μm far from the hole (P=0.000) versus AFR followed by ALA application at skin depths of 120 and 500μm. Needling after ALA application presented higher porphyrin

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

    SciTech Connect

    Ross, J.B.A.; Schwartz, G.P.; Laws, W.R. ); Wyssbrod, H.R.; Porter, R.A. ); Michaels, C.A. )

    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.

  2. Understanding walking activity in multiple sclerosis: step count, walking intensity and uninterrupted walking activity duration related to degree of disability.

    PubMed

    Neven, An; Vanderstraeten, Annelien; Janssens, Davy; Wets, Geert; Feys, Peter

    2016-09-01

    In multiple sclerosis (MS), physical activity (PA) is most commonly measured as number of steps, while also walking intensity and walking activity duration are keys for a healthy lifestyle. The aim of this study was to investigate (1) the number of steps persons with MS (PwMS) take; (2) the number of steps they take at low and moderate intensity; and (3) their walking activity duration for 2, 3, 6, 10, 12 and 14 uninterrupted minutes; all related to the degree of disability. 64 PwMS participated, distinguished in a mild (n = 31) and moderate MS subgroup (n = 34) based on their ambulatory dysfunction (Disease Steps). Standardized clinical tests were performed, and step data from the StepWatch Activity Monitor were collected for seven consecutive days. The results showed that (1) step count in PwMS was lower than PA recommendations, and is negatively influenced by a higher disability degree. (2) No walking was registered during 77 % of the day. PwMS are making steps for 22 % at low and only 1 % at moderate intensity. (3) Both MS subgroups rarely walk for more than six uninterrupted minutes, especially not at moderate intensity. PwMS need to be encouraged to make steps at moderate intensity, and to make steps for longer periods of time (minimal ten uninterrupted minutes).

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

    SciTech Connect

    Wu, C.Y.R.; Chen, F.Z.; Hung, T.; Judge, D.L.

    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.

  4. Multiple metrics for quantifying the intensity of water consumption of energy production

    NASA Astrophysics Data System (ADS)

    Spang, E. S.; Moomaw, W. R.; Gallagher, K. S.; Kirshen, P. H.; Marks, D. H.

    2014-10-01

    Discussion of the environmental implications of worldwide energy demand is currently dominated by the effects of carbon dioxide (CO2) emissions on global climate. At the regional scale, however, water resource challenges associated with energy systems are a growing concern. This paper, based on an inventory of national energy portfolios, posits an indicator-based framework for characterizing regional energy portfolios’ relative water intensity. These calculations extend upon a previous paper that established a method for calculating the national water consumption of energy production (WCEP) at the global level. Intensity indicators are based on normalizing the WCEP results with a set of additional indicators (including population, gross domestic product, total energy production, and regional water availability). The results show great variability in water consumption across nations, as well as across the various water intensity measures that were applied. Therefore, it is best to apply this full suite of indicators to each country to develop an integrated understanding of the intensity of water use for energy across countries.

  5. Response of intertidal macrofauna to multiple disturbance types and intensities - an experimental approach.

    PubMed

    Whomersley, P; Huxham, M; Bolam, S; Schratzberger, M; Augley, J; Ridland, D

    2010-06-01

    Two of the best-supported theories which describe the effects of disturbance within marine benthic habitats are the organic enrichment 'Successional Model' and the 'Intermediate Disturbance Hypothesis'. Underlying these models, biological mechanisms thought to drive community change include competition, facilitation, inhibition, tolerance and random colonisation. To further examine the effects of disturbance an experimental test of the effects of different types (burial, raking and organic enrichment) and intensities of disturbance on infaunal intertidal communities at two different sites with similar suites of species was carried out. The same type and frequency of disturbance, applied to the two different sites, produced different responses at the species, community and trophic group level. In models that assume a linear relationship between disturbance intensity and effect, knowledge of the intensity of any novel disturbance, combined with the original disturbance regime experienced by a community (i.e. its 'starting point'), should be sufficient to predict final community characteristics. The current results do not conform to such a linear interpretation, as at both sites the intensity of treatments did not always predict the degree of disturbance. Therefore the response to disturbance may depend on site-specific factors such as the history of prior disturbance and the inherent ecological plasticity exhibited by many benthic species. Whilst current models perform well in predicting benthic responses to gross disturbance, detecting subtler effects requires a recognition that community response may depend on the site, the species and the sources of disturbance.

  6. The interactive effects of press/pulse intensity and duration on regime shifts at multiple scales

    USDA-ARS?s Scientific Manuscript database

    Regime shifts are difficult-to-reverse transitions that occur when an ecosystem reorganizes around a new set of self-reinforcing feedbacks. Regime shifts are predicted to occur when the intensity of some exogenous driver variable, such as temperature, annual harvest rate, or nutrient addition rate, ...

  7. A declaration of independence for Mg/Si. [Al/Si intensity ratio predictive usefulness for Mg/Si intensity ratio in lunar X-ray fluorescence

    NASA Technical Reports Server (NTRS)

    Hubbard, N.; Keith, J. E.

    1978-01-01

    The weak covariation that exists between Al/Si and Mg/Si for large areas of the lunar surface is little, if any, stronger than that forced on a random set of numbers that are subject to closure. The Mg and Al variations implied by the Mg/Si and Al/Si intensity ratio data are qualitatively like those seen in lunar soil sample data. Two petrogenetic provinces are suggested for terra materials; one appears to have 50% higher Mg values than the other. Using the improved data, Mg/Si variations can be studied at a signal-to-noise ratio greater than 5/1.

  8. A novel analytical model for intensity modulated and direct detected multiple-input-multiple-output system with time related zero forcing equalization over multimode fiber

    NASA Astrophysics Data System (ADS)

    Fang, Wuliang; Shao, Yufeng; chi, Nan

    2012-07-01

    In this paper, basing on tap delay lines filter model and model spatial coupling theory, we build up a novel analytical model for an intensity modulated and direct detected multiple-input-multiple-output (IM-DDMIMO) system over multimode fiber. At the receiver side, time related zero forcing (ZF) equalization was used to recover signals. With this model, we theoretically and by simulation analyzed a 2 × 2 multimode fiber MIMO system utilizing offset launching scheme. It's found that two received streams can be well recovered by equalization. Compared with traditional single-input-single-output (SISO) system, such 2 × 2MIMO system can provide at least 5 dB Bit error rate (BER) performance improvement.

  9. Application of Peak Intensity Analysis to Measurements of Protein Binding to Lipid Vesicles and Erythrocytes Using Fluorescence Correlation Spectroscopy: Dependence on Particle Size.

    PubMed

    Antonenko, Yuri N; Lapashina, Anna S; Kotova, Elena A; Ramonova, Alla A; Moisenovich, Mikhail M; Agapov, Igor I

    2017-02-01

    Fluorescence correlation spectroscopy (FCS) is a sensitive analytical tool for investigation of processes accompanied by changes in the mobility of molecules and complexes. In the present work, peak intensity analysis (PIA) in combination with the solution stirring using FCS setup was applied to explore the interaction between fluorescently labeled protein ligands and corresponding receptors located on membranes. In the system composed of biotinylated liposomes and fluorescently labeled streptavidin as a ligand, PIA allowed us to determine the optimum receptor concentration and demonstrate the essential dependence of the binding efficacy on the length of the linker between the biotin group and the polar head group of the lipid. The binding was dependent on the size of liposomes which was varied by lipid extrusion through filters of different pore diameters. The sensitivity of the method was higher with the liposomes of larger sizes. The PIA approach can be applied not only to liposomes but also to relatively large objects, e.g., erythrocytes or Sepharose beads derivatized with lactose as a receptor for the binding of viscumin and ricin.

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

    PubMed Central

    Spagnolo, Daniel M.; Gyanchandani, Rekha; Al-Kofahi, Yousef; Stern, Andrew M.; Lezon, Timothy R.; Gough, Albert; Meyer, Dan E.; Ginty, Fiona; Sarachan, Brion; Fine, Jeffrey; Lee, Adrian V.; Taylor, D. Lansing; Chennubhotla, S. Chakra

    2016-01-01

    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 components

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

    NASA Astrophysics Data System (ADS)

    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.

  12. Application of novel low-intensity nonscanning fluorescence lifetime imaging microscopy for monitoring excited state dynamics in individual chloroplasts and living cells of photosynthetic organisms

    NASA Astrophysics Data System (ADS)

    Eckert, Hann-Jörg; Petrášek, Zdeněk; Kemnitz, Klaus

    2006-10-01

    Picosecond fluorescence lifetime imaging microscopy (FLIM) provides a most valuable tool to analyze the primary processes of photosynthesis in individual cells and chloroplasts of living cells. In order to obtain correct lifetimes of the excited states, the peak intensity of the exciting laser pulses as well as the average intensity has to be sufficiently low to avoid distortions of the kinetics by processes such as singlet-singlet annihilation, closing of the reaction centers or photoinhibition. In the present study this requirement is achieved by non-scanning wide-field FLIM based on time- and space-correlated single-photon counting (TSCSPC) using a novel microchannel plate photomultiplier with quadrant anode (QA-MCP) that allows parallel acquisition of time-resolved images under minimally invasive low-excitation conditions. The potential of the wide-field TCSPC method is demonstrated by presenting results obtained from measurements of the fluorescence dynamics in individual chloroplasts of moss leaves and living cells of the chlorophyll d-containing cyanobacterium Acaryochloris marina.

  13. Estimating Dengue Transmission Intensity from Sero-Prevalence Surveys in Multiple Countries

    PubMed Central

    Imai, Natsuko; Dorigatti, Ilaria; Cauchemez, Simon; Ferguson, Neil M.

    2015-01-01

    Background Estimates of dengue transmission intensity remain ambiguous. Since the majority of infections are asymptomatic, surveillance systems substantially underestimate true rates of infection. With advances in the development of novel control measures, obtaining robust estimates of average dengue transmission intensity is key for assessing both the burden of disease from dengue and the likely impact of interventions. Methodology/Principal Findings The force of infection (λ) and corresponding basic reproduction numbers (R0) for dengue were estimated from non-serotype (IgG) and serotype-specific (PRNT) age-stratified seroprevalence surveys identified from the literature. The majority of R0 estimates ranged from 1–4. Assuming that two heterologous infections result in complete immunity produced up to two-fold higher estimates of R0 than when tertiary and quaternary infections were included. λ estimated from IgG data were comparable to the sum of serotype-specific forces of infection derived from PRNT data, particularly when inter-serotype interactions were allowed for. Conclusions/Significance Our analysis highlights the highly heterogeneous nature of dengue transmission. How underlying assumptions about serotype interactions and immunity affect the relationship between the force of infection and R0 will have implications for control planning. While PRNT data provides the maximum information, our study shows that even the much cheaper ELISA-based assays would provide comparable baseline estimates of overall transmission intensity which will be an important consideration in resource-constrained settings. PMID:25881272

  14. Estimating Dengue Transmission Intensity from Case-Notification Data from Multiple Countries

    PubMed Central

    Imai, Natsuko; Dorigatti, Ilaria; Cauchemez, Simon; Ferguson, Neil M.

    2016-01-01

    Background Despite being the most widely distributed mosquito-borne viral infection, estimates of dengue transmission intensity and associated burden remain ambiguous. With advances in the development of novel control measures, obtaining robust estimates of average dengue transmission intensity is key for assessing the burden of disease and the likely impact of interventions. Methodology/Principle Findings We estimated the force of infection (λ) and corresponding basic reproduction numbers (R0) by fitting catalytic models to age-stratified incidence data identified from the literature. We compared estimates derived from incidence and seroprevalence data and assessed the level of under-reporting of dengue disease. In addition, we estimated the relative contribution of primary to quaternary infections to the observed burden of dengue disease incidence. The majority of R0 estimates ranged from one to five and the force of infection estimates from incidence data were consistent with those previously estimated from seroprevalence data. The baseline reporting rate (or the probability of detecting a secondary infection) was generally low (<25%) and varied within and between countries. Conclusions/Significance As expected, estimates varied widely across and within countries, highlighting the spatio-temporally heterogeneous nature of dengue transmission. Although seroprevalence data provide the maximum information, the incidence models presented in this paper provide a method for estimating dengue transmission intensity from age-stratified incidence data, which will be an important consideration in areas where seroprevalence data are not available. PMID:27399793

  15. Modelling and implementation of a fixed-length-extension to measure fluorescent intensity in bioprocesses using an optical sensor

    NASA Astrophysics Data System (ADS)

    Sardesai, Neha; Al-Adhami, Mustafa; Rao, Govind; Kostov, Yordan

    2016-05-01

    Fluorescent proteins are often used as reporters of protein concentration in biology and biomedicine applications. They can be detected using a fluorimeter equipped with fiber optics for ease of access. However, small changes in the path length due to change in the position, or immersion depth of the optical fiber results in large changes in readings. To alleviate the situation, the fiber is equipped with a fixed-length-extension that provides constant path length. The operation of the fiber equipped fluorimeter is theoretically modelled and practically verified in this paper.

  16. Intensive Long Distance Running as a Possible Cause of Multiple Splanchnic Arterial Aneurysms: A Case Report

    PubMed Central

    Jang, Lee Chan; Park, Sung Su

    2016-01-01

    This is a case report that suggests the possible association between multiple splanchnic arterial aneurysms and long-distance running. The clinical features of one patient admitted at Chungbuk National University Hospital for treatment of multiple splanchnic arterial aneurysms were reviewed. A 54-year-old man had a recurrent, intermittent and epigastric pain for 2 months. There was no abnormality in gastroscopy and colonoscopy. An abdominal computed tomography angiography documented calcified superior mesenteric artery (SMA) and splenic artery aneurysms. The patient had a history of recreational long-distance running for over 10 years. His average running time per week was more than 10 hours. There was no evidence of systemic arteritis, connective tissue disorder or infectious process that may have caused the aneurysms. He did not take any drugs. The SMA aneurysm was opened, and the aneurysmal segment of SMA was replaced with a vein graft. The splenic aneurysm was observed. The patient recovered without any sequelae. PMID:27699161

  17. A Technique for Estimating Intensity of Emotional Expressions and Speaking Styles in Speech Based on Multiple-Regression HSMM

    NASA Astrophysics Data System (ADS)

    Nose, Takashi; Kobayashi, Takao

    In this paper, we propose a technique for estimating the degree or intensity of emotional expressions and speaking styles appearing in speech. The key idea is based on a style control technique for speech synthesis using a multiple regression hidden semi-Markov model (MRHSMM), and the proposed technique can be viewed as the inverse of the style control. In the proposed technique, the acoustic features of spectrum, power, fundamental frequency, and duration are simultaneously modeled using the MRHSMM. We derive an algorithm for estimating explanatory variables of the MRHSMM, each of which represents the degree or intensity of emotional expressions and speaking styles appearing in acoustic features of speech, based on a maximum likelihood criterion. We show experimental results to demonstrate the ability of the proposed technique using two types of speech data, simulated emotional speech and spontaneous speech with different speaking styles. It is found that the estimated values have correlation with human perception.

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

    NASA Astrophysics Data System (ADS)

    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.

  19. Duration of an intense laser pulse can determine the breakage of multiple chemical bonds.

    PubMed

    Xie, Xinhua; Lötstedt, Erik; Roither, Stefan; Schöffler, Markus; Kartashov, Daniil; Midorikawa, Katsumi; Baltuška, Andrius; Yamanouchi, Kaoru; Kitzler, Markus

    2015-08-14

    Control over the breakage of a certain chemical bond in a molecule by an ultrashort laser pulse has been considered for decades. With the availability of intense non-resonant laser fields it became possible to pre-determine femtosecond to picosecond molecular bond breakage dynamics by controlled distortions of the electronic molecular system on sub-femtosecond time scales using field-sensitive processes such as strong-field ionization or excitation. So far, all successful demonstrations in this area considered only fragmentation reactions, where only one bond is broken and the molecule is split into merely two moieties. Here, using ethylene (C2H4) as an example, we experimentally investigate whether complex fragmentation reactions that involve the breakage of more than one chemical bond can be influenced by parameters of an ultrashort intense laser pulse. We show that the dynamics of removing three electrons by strong-field ionization determines the ratio of fragmentation of the molecular trication into two respectively three moieties. We observe a relative increase of two-body fragmentations with the laser pulse duration by almost an order of magnitude. Supported by quantum chemical simulations we explain our experimental results by the interplay between the dynamics of electron removal and nuclear motion.

  20. Duration of an intense laser pulse can determine the breakage of multiple chemical bonds

    NASA Astrophysics Data System (ADS)

    Xie, Xinhua; Lötstedt, Erik; Roither, Stefan; Schöffler, Markus; Kartashov, Daniil; Midorikawa, Katsumi; Baltuška, Andrius; Yamanouchi, Kaoru; Kitzler, Markus

    2015-08-01

    Control over the breakage of a certain chemical bond in a molecule by an ultrashort laser pulse has been considered for decades. With the availability of intense non-resonant laser fields it became possible to pre-determine femtosecond to picosecond molecular bond breakage dynamics by controlled distortions of the electronic molecular system on sub-femtosecond time scales using field-sensitive processes such as strong-field ionization or excitation. So far, all successful demonstrations in this area considered only fragmentation reactions, where only one bond is broken and the molecule is split into merely two moieties. Here, using ethylene (C2H4) as an example, we experimentally investigate whether complex fragmentation reactions that involve the breakage of more than one chemical bond can be influenced by parameters of an ultrashort intense laser pulse. We show that the dynamics of removing three electrons by strong-field ionization determines the ratio of fragmentation of the molecular trication into two respectively three moieties. We observe a relative increase of two-body fragmentations with the laser pulse duration by almost an order of magnitude. Supported by quantum chemical simulations we explain our experimental results by the interplay between the dynamics of electron removal and nuclear motion.

  1. Duration of an intense laser pulse can determine the breakage of multiple chemical bonds

    PubMed Central

    Xie, Xinhua; Lötstedt, Erik; Roither, Stefan; Schöffler, Markus; Kartashov, Daniil; Midorikawa, Katsumi; Baltuška, Andrius; Yamanouchi, Kaoru; Kitzler, Markus

    2015-01-01

    Control over the breakage of a certain chemical bond in a molecule by an ultrashort laser pulse has been considered for decades. With the availability of intense non-resonant laser fields it became possible to pre-determine femtosecond to picosecond molecular bond breakage dynamics by controlled distortions of the electronic molecular system on sub-femtosecond time scales using field-sensitive processes such as strong-field ionization or excitation. So far, all successful demonstrations in this area considered only fragmentation reactions, where only one bond is broken and the molecule is split into merely two moieties. Here, using ethylene (C2H4) as an example, we experimentally investigate whether complex fragmentation reactions that involve the breakage of more than one chemical bond can be influenced by parameters of an ultrashort intense laser pulse. We show that the dynamics of removing three electrons by strong-field ionization determines the ratio of fragmentation of the molecular trication into two respectively three moieties. We observe a relative increase of two-body fragmentations with the laser pulse duration by almost an order of magnitude. Supported by quantum chemical simulations we explain our experimental results by the interplay between the dynamics of electron removal and nuclear motion. PMID:26271602

  2. Fiber-optic holography employing multiple beam fringe stabilization and object/reference beam intensity variability.

    PubMed

    Muhs, J D; Leilabady, P A; Corke, M

    1988-09-01

    The use of fiber optics in the field of optical holography is discussed with emphasis on the design of systems used to overcome several inherent shortcomings associated with fiber-optic holographic systems. Specifically, random environmentally induced optical phase changes within the fiber are minimized by employing a Michelson interferometer in conjunction with a closed loop feedback system. Furthermore, by using several passive single-mode couplers, complete object illumination via several illumination fibers is observed. Finally, by implementing a Mach-Zehnder interferometric technique, control of the object and reference beam intensity ratios in a fiber-optic holographic system can be accomplished. The resulting schemes are very stable and highly versatile systems suitable for remote holographic interferometric sensing and other applications where conventional holography techniques are impractical. Experimental results on fringe visibility, fringe stability, and the stabilization of object/reference beam intensity ratios are also given along with a composite summary of the overall system constraints associated with fiber-optic holographic systems.

  3. Sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using DNA/dye conjugate as a fluorescent label.

    PubMed

    Fan, Ziyan; Keum, Young Soo; Li, Qing X; Shelver, Weilin L; Guo, Liang-Hong

    2012-05-01

    Indirect competitive immunoassays were developed on protein microarrays for the sensitive and simultaneous detection of multiple environmental chemicals in one sample. In this assay, a DNA/SYTOX Orange conjugate was employed as an antibody label to increase the fluorescence signal and sensitivity of the immunoassays. Epoxy-modified glass slides were selected as the substrate for the production of 4 × 4 coating antigen microarrays. With this signal-enhancing system, competition curves for 17β-estradiol (E2), benzo[a]pyrene (BaP) and 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) were obtained individually on the protein microarray. The IC(50) and calculated limit of detection (LOD) are 0.32 μg L(-1) and 0.022 μg L(-1) for E2, 37.2 μg L(-1) and 24.5 μg L(-1) for BaP, and 31.6 μg L(-1) and 2.8 μg L(-1) for BDE-47, respectively. LOD of E2 is 14-fold lower than the value reported in a previous study using Cy3 labeled antibody (Du et al., Clin. Chem, 2005, 51, 368-375). The results of the microarray immunoassay were within 15% of chromatographic analysis for all three pollutants in spiked river water samples, thus verifying the immunoassay. Simultaneous detection of E2, BaP and BDE-47 in one sample was demonstrated. There was no cross-reaction in the immunoassay between these three environmental chemicals. These results suggest that microarray-based immunoassays with DNA/dye conjugate labels are useful tools for the rapid, sensitive, and high throughput screening of multiple environmental contaminants.

  4. Robust Bayesian Fluorescence Lifetime Estimation, Decay Model Selection and Instrument Response Determination for Low-Intensity FLIM Imaging.

    PubMed

    Rowley, Mark I; Coolen, Anthonius C C; Vojnovic, Borivoj; Barber, Paul R

    2016-01-01

    We present novel Bayesian methods for the analysis of exponential decay data that exploit the evidence carried by every detected decay event and enables robust extension to advanced processing. Our algorithms are presented in the context of fluorescence lifetime imaging microscopy (FLIM) and particular attention has been paid to model the time-domain system (based on time-correlated single photon counting) with unprecedented accuracy. We present estimates of decay parameters for mono- and bi-exponential systems, offering up to a factor of two improvement in accuracy compared to previous popular techniques. Results of the analysis of synthetic and experimental data are presented, and areas where the superior precision of our techniques can be exploited in Förster Resonance Energy Transfer (FRET) experiments are described. Furthermore, we demonstrate two advanced processing methods: decay model selection to choose between differing models such as mono- and bi-exponential, and the simultaneous estimation of instrument and decay parameters.

  5. Robust Bayesian Fluorescence Lifetime Estimation, Decay Model Selection and Instrument Response Determination for Low-Intensity FLIM Imaging

    PubMed Central

    Rowley, Mark I.; Coolen, Anthonius C. C.; Vojnovic, Borivoj; Barber, Paul R.

    2016-01-01

    We present novel Bayesian methods for the analysis of exponential decay data that exploit the evidence carried by every detected decay event and enables robust extension to advanced processing. Our algorithms are presented in the context of fluorescence lifetime imaging microscopy (FLIM) and particular attention has been paid to model the time-domain system (based on time-correlated single photon counting) with unprecedented accuracy. We present estimates of decay parameters for mono- and bi-exponential systems, offering up to a factor of two improvement in accuracy compared to previous popular techniques. Results of the analysis of synthetic and experimental data are presented, and areas where the superior precision of our techniques can be exploited in Förster Resonance Energy Transfer (FRET) experiments are described. Furthermore, we demonstrate two advanced processing methods: decay model selection to choose between differing models such as mono- and bi-exponential, and the simultaneous estimation of instrument and decay parameters. PMID:27355322

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

    PubMed

    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.

  7. Centralized triage for multiple intensive care units: the central intensivist physician.

    PubMed

    Romig, Mark; Latif, Asad; Pronovost, Peter; Sapirstein, Adam

    2010-01-01

    Subspecialization of critical care units and overall increasing demand for critical care services has led to inefficiencies in allocation of critical care resources with potential impacts on hospital economics and patient outcomes. Centralized management of critical care resource allocation within an institution may improve use while simultaneously ensuring quality of patient care. The authors' institution has implemented a Central Intensivist Physician (CIP) program to oversee resource allocation within the adult surgical intensive care units (ICUs). The result has been an improvement in patient flow throughout the surgical ICUs manifested by steady case cancellation rates despite increasing acuity and length of stay. Additionally, triage duties have been shifted from the individual unit physician to the CIP, resulting in improved provider satisfaction from improved continuity of rounds. The authors conclude that the CIP program may improve overall critical care resource use while maintaining unit specialization within a large tertiary care hospital setting.

  8. A methodology to study multiple sclerosis (MS) based on distributions of standardized intensities in segmented tissue regions

    NASA Astrophysics Data System (ADS)

    Lei, T.; Udupa, J. K.; Odhner, D.; Mishra, S.; Wu, G.; Schwartz, E.; Ying, G.-S.; Iwanaga, T.; Desiderio, L.; Balcer, L.

    2006-03-01

    This paper presents (1) an improved hierarchical method for segmenting the component tissue regions in fast spin echo T2 and PD images of the brain of Multiple Sclerosis (MS) patients, and (2) a methodology to characterize the disease utilizing the distributions of standardized T2 and PD intensities in the segmented tissue regions. First, the background intensity inhomogeneities are corrected and the intensity scales are standardized for all acquired images. The segmentation method imposes a feedback-like procedure on our previously developed hierarchical brain tissue segmentation method. With gradually simplified patterns in images and stronger evidences, pathological objects are recognized and segmented in an interplay fashion. After the brain parenchymal (BP) mask is generated, an under-estimated gray matter mask (uGM) and an over-estimated white matter mask (oWM) are created. Pure WM (PWM) and lesion (LS) masks are extracted from the all-inclusive oWM mask. By feedback, accurate GM and WM masks are subsequently formed. Finally, partial volume regions of GM and WM as well as Dirty WM (DWM) masks are generated. Intensity histograms and their parameters (peak height, peak location, and 25th, 50th and 75th percentile values) are computed for both T2 and PD images within each tissue region. Tissue volumes are also estimated. Spearman correlation coefficient rank test is then utilized to assess if there exists a trend between clinical states and the image-based parameters. This image analysis method has been applied to a data set consisting of 60 patients with MS and 20 normal controls. LS related parameters and clinical Extended Disability Status Scale (EDSS) scores demonstrate modest correlations. Almost every intensity-based parameter shows statistical difference between normal control and patient groups with a level better than 5%. These results can be utilized to monitor disease progression in MS.

  9. Correlation of Neonatal Intensive Care Unit Performance Across Multiple Measures of Quality of Care

    PubMed Central

    Profit, J; Zupancic, JAF; Gould, JB; Pietz, K; Kowalkowski, MA; Draper, D; Hysong, SJ; Petersen, LA

    2014-01-01

    Objectives To examine whether high performance on one measure of quality is associated with high performance on others and to develop a data-driven explanatory model of neonatal intensive care unit (NICU) performance. Design We conducted a cross-sectional data analysis of a statewide perinatal care database. Risk-adjusted NICU ranks were computed for each of 8 measures of quality selected based on expert input. Correlations across measures were tested using the Pearson correlation coefficient. Exploratory factor analysis was used to determine whether underlying factors were driving the correlations. Setting Twenty-two regional NICUs in California. Patients In total, 5445 very low-birth-weight infants cared for between January 1, 2004, and December 31, 2007. Main Outcomes Measures Pneumothorax, growth velocity, health care–associated infection, antenatal corticosteroid use, hypothermia during the first hour of life, chronic lung disease, mortality in the NICU, and discharge on any human breast milk. Results The NICUs varied substantially in their clinical performance across measures of quality. Of 28 unit-level correlations only 6 were significant (P < .05). Correlations between pairs of quality measures were strong (ρ > .5) for 1 pair, moderate (.3 < |ρ| < .5) for 8 pairs, weak (.1 < |ρ| < .3) for 5 pairs and negligible (|ρ| < .1) for 14 pairs. Exploratory factor analysis revealed 4 underlying factors of quality in this sample. Pneumothorax, mortality in the NICU, and antenatal corticosteroid use loaded on factor 1; growth velocity and health care–associated infection loaded on factor 2; chronic lung disease loaded on factor 3; and discharge on any human breast milk loaded on factor 4. Conclusion In this sample, the ability of individual measures of quality to explain overall quality of neonatal intensive care was modest. PMID:23403539

  10. Fluorescent optical position sensor

    DOEpatents

    Weiss, Jonathan D.

    2005-11-15

    A fluorescent optical position sensor and method of operation. A small excitation source side-pumps a localized region of fluorescence at an unknown position along a fluorescent waveguide. As the fluorescent light travels down the waveguide, the intensity of fluorescent light decreases due to absorption. By measuring with one (or two) photodetectors the attenuated intensity of fluorescent light emitted from one (or both) ends of the waveguide, the position of the excitation source relative to the waveguide can be determined by comparing the measured light intensity to a calibrated response curve or mathematical model. Alternatively, excitation light can be pumped into an end of the waveguide, which generates an exponentially-decaying continuous source of fluorescent light along the length of the waveguide. The position of a photodetector oriented to view the side of the waveguide can be uniquely determined by measuring the intensity of the fluorescent light emitted radially at that location.

  11. Endocrine, metabolic, nutritional and body composition abnormalities are common in advanced intensively-treated (transplanted) multiple myeloma.

    PubMed

    Greenfield, D M; Boland, E; Ezaydi, Y; Ross, R J M; Ahmedzai, S H; Snowden, J A

    2014-07-01

    Modern treatment strategies have increased life expectancy in multiple myeloma, but little is known about the endocrine, metabolic and nutritional status of long-term survivors. We performed endocrine, metabolic, bone, body composition and nutritional evaluations in 32 patients with intensively-treated, advanced but stable, myeloma a median duration of 6 years from diagnosis and three lines of intensive treatment, including at least one haematopoietic SCT procedure. All patients were off active treatment. There was a high prevalence of endocrine dysfunction: hypothyroidism (9%), hypogonadism (65% males) and elevated prolactin (19%). Adrenocortical function was preserved despite large cumulative corticosteroid pretreatment. Biochemical markers were consistent with postmenopausal status in all females and infertility in males. Nutritionally, 59% were vitamin D insufficient/deficient, reduced serum folate in 25% and vitamin B12 in 6%. Total body DEXA scanning confirmed 'sarcopenic-obesity' in 65%, but reduced bone density was seen in a minority. We conclude that potentially correctable endocrine, metabolic and nutritional abnormalities are prevalent in heavily-treated patients with stable multiple myeloma. Preservation of bone supports the efficacy of bisphosphonate treatment from diagnosis, but sarcopenic-obesity may contribute to frailty. Ultimately, multi-system screening and appropriate interventions may optimise quality of long-term survival and further studies are warranted.

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

    PubMed

    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.

  13. Step-rate cut-points for physical activity intensity in patients with multiple sclerosis: The effect of disability status.

    PubMed

    Agiovlasitis, Stamatis; Sandroff, Brian M; Motl, Robert W

    2016-02-15

    Evaluating the relationship between step-rate and rate of oxygen uptake (VO2) may allow for practical physical activity assessment in patients with multiple sclerosis (MS) of differing disability levels. To examine whether the VO2 to step-rate relationship during over-ground walking differs across varying disability levels among patients with MS and to develop step-rate thresholds for moderate- and vigorous-intensity physical activity. Adults with MS (N=58; age: 51 ± 9 years; 48 women) completed one over-ground walking trial at comfortable speed, one at 0.22 m · s(-1) slower, and one at 0.22 m · s(-1) faster. Each trial lasted 6 min. VO2 was measured with portable spirometry and steps with hand-tally. Disability status was classified as mild, moderate, or severe based on Expanded Disability Status Scale scores. Multi-level regression indicated that step-rate, disability status, and height significantly predicted VO2 (p<0.05). Based on this model, we developed step-rate thresholds for activity intensity that vary by disability status and height. A separate regression without height allowed for development of step-rate thresholds that vary only by disability status. The VO2 during over-ground walking differs among ambulatory patients with MS based on disability level and height, yielding different step-rate thresholds for physical activity intensity. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Effect of multiple time delays on intensity fluctuation dynamics in fiber ring lasers.

    PubMed

    Franz, Anthony L; Roy, Rajarshi; Shaw, Leah B; Schwartz, Ira B

    2008-07-01

    The effect of time delay on nonlinear oscillators is an important problem in the study of dynamical systems. The dynamics of an erbium-doped fiber ring laser with an extra loop providing time-delayed feedback is studied experimentally by measuring the intensity of the laser. The delay time for the feedback is varied from approximately 0.3 to approximately 900 times the cavity round-trip time, over four orders of magnitude, by changing the length of fiber in the delay line. Depending on the delay, we observe either regular oscillations or complex dynamics. The size of the fluctuations increases for delays long compared with the round-trip time of the laser cavity. The complexity of the fluctuations is quantified by creating spatiotemporal representations of the time series and performing a Karhunen-Loève decomposition. The complexity increases with increasing delay time. The experiment is extended by mutually coupling two fiber ring lasers together. The delay time for the mutual coupling is varied from approximately 0.2 to approximately 600 times the cavity round-trip time, over four orders of magnitude again. In this case the fluctuations are generally larger than the single laser case. The complexity of the dynamics for the mutually coupled system is less at short delays and larger at long delays when compared to the uncoupled case. The width of the optical spectra of the coupled lasers also narrows.

  15. A single versus multiple bouts of moderate-intensity exercise for fat metabolism.

    PubMed

    Goto, Kazushige; Tanaka, Kiyoji; Ishii, Naokata; Uchida, Sunao; Takamatsu, Kaoru

    2011-05-01

    This study compared the fat metabolism between 'a single bout of 30-min exercise' and 'three bouts of 10-min exercise' of the same intensity (60% maximal oxygen uptake) and total exercise duration (30 min). Nine healthy men participated in three trials: (1) a single 30-min bout of exercise (Single), (2) three 10-min bouts of exercise, separated by a 10-min rest (Repeated) and (3) rest (Rest). Each exercise was performed with a cycle ergometer at 60% of maximal oxygen uptake, followed by 180-min rest. Blood lactate concentration increased significantly after exercise in the Single and Repeated trials (P < 0.05), but the Single trial showed a significantly higher value during the recovery period (P < 0.05). No significant difference was observed in the responses of plasma glycerol concentration. The Repeated trial produced a smaller increase in the ratings of perceived exertion during the exercise (P < 0.01). During the exercise, no significant difference was observed in respiratory exchange ratio (RER) between the Single and Repeated trials. However, the RER values during the recovery period were significantly lower in the Repeated trial than in the Single and Rest trials (P < 0.05), indicating higher relative contribution of fat oxidation in the Repeated trial (P < 0.05). These results suggest that the repetition of 10-min of moderate exercise can contribute to greater exercise-induced fat oxidation compared with a single 30-min bout of continuous exercise.

  16. Environmental controls on storm intensity and charge structure in multiple regions of the continental United States

    NASA Astrophysics Data System (ADS)

    Fuchs, Brody R.; Rutledge, Steven A.; Bruning, Eric C.; Pierce, Jeffrey R.; Kodros, John K.; Lang, Timothy J.; MacGorman, Donald R.; Krehbiel, Paul R.; Rison, William

    2015-07-01

    A database consisting of approximately 4000 storm observations has been objectively analyzed to determine environmental characteristics that produce high radar reflectivities above the freezing level, large total lightning flash rates on the order of 10 flashes per minute, and anomalous vertical charge structures (most notably, dominant midlevel positive charge). The storm database is drawn from four regions of the United States featuring distinct environments, each with coinciding Lightning Mapping Array (LMA) network data. LMAs are able to infer total lightning flash rates using flash clustering algorithms, such as the one implemented in this study. Results show that anomalous charge structures inferred from LMA data, significant lightning flash rates, and increased radar reflectivities above the freezing level tend to be associated with environments that have high cloud base heights (approximately 3 km above ground level) and large atmospheric instability, quantified by normalized convective available potential energy (NCAPE) near 0.2 m s-2. Additionally, we infer that aerosols may affect storm intensity. Maximum flash rates were observed in storms with attributed aerosol concentrations near 1000 cm-3, while total flash rates decrease when aerosol concentrations exceed 1500 cm-3, consistent with previous studies. However, this effect is more pronounced in regions where the NCAPE and cloud base height are low. The dearth of storms with estimated aerosol concentrations less than 700 cm-3 (approximately 1% of total sample) does not provide a complete depiction of aerosol invigoration.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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.

  19. Fluorescence melting curve analysis using self-quenching dual-labeled peptide nucleic acid probes for simultaneously identifying multiple DNA sequences.

    PubMed

    Ahn, Jeong Jin; Kim, Youngjoo; Lee, Seung Yong; Hong, Ji Young; Kim, Gi Won; Hwang, Seung Yong

    2015-09-01

    Previous fluorescence melting curve analysis (FMCA) used intercalating dyes, and this method has restricted application. Therefore, FMCA methods such as probe-based FMCA and molecular beacons were studied. However, the usual dual-labeled probes do not possess adequate fluorescence quenching ability and sufficient specificity, and molecular beacons with the necessary stem structures are hard to design. Therefore, we have developed a peptide nucleic acid (PNA)-based FMCA method. PNA oligonucleotide can have a much higher melting temperature (Tm) value than DNA. Therefore, short PNA probes can have adequate Tm values for FMCA, and short probes can have higher specificity and accuracy in FMCA. Moreover, dual-labeled PNA probes have self-quenching ability via single-strand base stacking, which makes PNA more favorable. In addition, this method can facilitate simultaneous identification of multiple DNA templates. In conventional real-time polymerase chain reaction (PCR), one fluorescence channel can identify only one DNA template. However, this method uses two fluorescence channels to detect three types of DNA. Experiments were performed with one to three different DNA sequences mixed in a single tube. This method can be used to identify multiple DNA sequences in a single tube with high specificity and high clarity. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Highly Efficient Multiple-Anchored Fluorescent Probe for the Detection of Aniline Vapor Based on Synergistic Effect: Chemical Reaction and PET.

    PubMed

    Jiao, Zinuo; Zhang, Yu; Xu, Wei; Zhang, Xiangtao; Jiang, Haibo; Wu, Pengcheng; Fu, Yanyan; He, Qingguo; Cao, Huimin; Cheng, Jiangong

    2017-05-26

    A multiple-anchored fluorescent probe ((((hexane-1,6-diylbis(2,7-bis(4-formyl)-phenyl)-9H-fluorine-9,9-diyl))-bis(hexane-6,1-diyl))-bis(9H-carbazole-9,3,6-triyl))-tetrakis(benzene-4,1-diyl))-tetraformyl-(8FP-2F) with eight aldehyde groups was designed and synthesized. The molecule has four branches and highly twisted structure. Furthermore, it tends to self-assemble into nanospheres, which is beneficial for gaseous analyte penetration and high fluorescence quantum efficiency. Among gaseous analytes, detection of aniline vapor is extraordinarily important in the control of environmental issues and human diseases. Herein, 8FP-2F was introduced to detect aniline vapor with distinguished sensitivity and selectivity via simple Schiff base reaction at room temperature. After exposure to saturate aniline vapor, the 89% fluorescence of 8FP-2F was quenched in 50 s and the detection limit was as low as 3 ppb. Further study showed the suitable HOMO/LUMO energy levels and matched orbital symmetry between probe and aniline molecules ensured chemical reaction and PET process work together. The synergistic effect resulted in a significant sensing performance and fluorescence quenching toward aniline vapor. Moreover, the multiple active sites structure of 8FP-2F means it could be applied for constructing many interesting structures and highly efficient organic optoelectronic functional materials.

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

    PubMed

    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.

  2. A simple but efficient strategy to enhance hydrostability of intensely fluorescent Mg-based coordination polymer (CP) via forming a composite of CP with hydrophobic PVDF.

    PubMed

    Zhai, Lu; Zhang, Wen-Wei; Zuo, Jing-Lin; Ren, Xiao-Ming

    2016-02-28

    A coordination polymer (CP) of Mg(2+) with 1,3,5-benzenetricarboxylate (BTC(3-)) was synthesized using a solvothermal method. The Mg-CP, with a formula of Mg3(BTC)(HCOO)3(DMF)3, crystallizes in the trigonal space group P3[combining macron], with cell parameters of a = b = 13.972(5) Å, c = 8.090(5) Å and V = 1367.6(11) Å(3), and shows a lamella structure built from planar rosette-type hexanuclear architectures. The Mg-CP emits intense blue fluorescence arising from π* → π transition of intra-ligand of BTC(3-) with 21.69% quantum yield, yet it exhibits poor stability to water. The composites of Mg-CP with hydrophobic polyvinylidene fluoride (PVDF) were sequentially prepared by mechanically mixed, tableted and annealed processes, which showed good compatibility between Mg-CP and PVDF, high hydrostability, and intense blue emission. This study suggests a simple but efficient method to solve the drawbacks of some functional CPs unstable to water and to promote them as practical applications in the field of functional materials.

  3. A prospective Phase II clinical trial of 5-aminolevulinic acid to assess the correlation of intraoperative fluorescence intensity and degree of histologic cellularity during resection of high-grade gliomas.

    PubMed

    Lau, Darryl; Hervey-Jumper, Shawn L; Chang, Susan; Molinaro, Annette M; McDermott, Michael W; Phillips, Joanna J; Berger, Mitchel S

    2016-05-01

    OBJECT There is evidence that 5-aminolevulinic acid (ALA) facilitates greater extent of resection and improves 6-month progression-free survival in patients with high-grade gliomas. But there remains a paucity of studies that have examined whether the intensity of ALA fluorescence correlates with tumor cellularity. Therefore, a Phase II clinical trial was undertaken to examine the correlation of intensity of ALA fluorescence with the degree of tumor cellularity. METHODS A single-center, prospective, single-arm, open-label Phase II clinical trial of ALA fluorescence-guided resection of high-grade gliomas (Grade III and IV) was held over a 43-month period (August 2010 to February 2014). ALA was administered at a dose of 20 mg/kg body weight. Intraoperative biopsies from resection cavities were collected. The biopsies were graded on a 4-point scale (0 to 3) based on ALA fluorescence intensity by the surgeon and independently based on tumor cellularity by a neuropathologist. The primary outcome of interest was the correlation of ALA fluorescence intensity to tumor cellularity. The secondary outcome of interest was ALA adverse events. Sensitivities, specificities, positive predictive values (PPVs), negative predictive values (NPVs), and Spearman correlation coefficients were calculated. RESULTS A total of 211 biopsies from 59 patients were included. Mean age was 53.3 years and 59.5% were male. The majority of biopsies were glioblastoma (GBM) (79.7%). Slightly more than half (52.5%) of all tumors were recurrent. ALA intensity of 3 correlated with presence of tumor 97.4% (PPV) of the time. However, absence of ALA fluorescence (intensity 0) correlated with the absence of tumor only 37.7% (NPV) of the time. For all tumor types, GBM, Grade III gliomas, and recurrent tumors, ALA intensity 3 correlated strongly with cellularity Grade 3; Spearman correlation coefficients (r) were 0.65, 0.66, 0.65, and 0.62, respectively. The specificity and PPV of ALA intensity 3 correlating

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

    NASA Technical Reports Server (NTRS)

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    NASA Technical Reports Server (NTRS)

    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.

  7. Assessment of curing behavior of light-activated dental composites using intensity correlation based multiple reference optical coherence tomography.

    PubMed

    Dsouza, Roshan; Subhash, Hrebesh; Neuhaus, Kai; Kantamneni, Ramakrishna; McNamara, Paul M; Hogan, Josh; Wilson, Carol; Leahy, Martin

    2016-01-01

    Monitoring the curing kinetics of light-activated resin is a key area of research. These resins are used in restorative applications and particularly in dental applications. They can undergo volumetric shrinkage due to poor control of the depth dependent curing process, modulated by the intensity and duration of the curing light source. This often results in the formation of marginal gaps, causing pain and damage to the restoration site. In this study, we demonstrate the capabilities of a correlation method applied using a multiple references optical coherence tomography (MR-OCT) architecture to monitor the curing of the resin. A MR-OCT system is used in this study to monitor the curing of the resin. The system operates at the center wavelength of 1310 nm with an A-scan rate of 1200 A-scans per second. The axial and lateral resolution of the system is ∼13 μm and ∼27 μm. The method to determine the intensity correlation between adjacent B-frames is based on the Pearson correlation coefficient for a region of interest. Calculating the correlation coefficient for multiple B-frames related to the first B-frame at regular spaced time points, shows for a noncured resin a reduction of the correlation coefficient over time due to Brownian motion. The time constant of the reduction of the correlation value is a measure for the progress of the polymerization during LED light irradiation of the resin. The proposed approach is potentially a low-cost, powerful and unique optical imaging modality for measuring the curing behavior of dental resin and other resins, coatings, and adhesives in medical and industrial applications. To demonstrate the proposed method to monitor the curing process, a light-activated resin composite from GRADIA DIRECT ANTERIOR (GC Corporation, Japan) is studied. The curing time of resin was measured and monitored as a function of depth. The correlation coefficient method is highly sensitive to Brownian motion. The process of curing results in

  8. Fluorescence lifetime excitation cytometry by kinetic dithering.

    PubMed

    Li, Wenyan; Vacca, Giacomo; Castillo, Maryann; Houston, Kevin D; Houston, Jessica P

    2014-07-01

    Flow cytometers are powerful high-throughput devices that capture spectroscopic information from individual particles or cells. These instruments provide a means of multi-parametric analyses for various cellular biomarkers or labeled organelles and cellular proteins. However, the spectral overlap of fluorophores limits the number of fluorophores that can be used simultaneously during experimentation. Time-resolved parameters enable the quantification of fluorescence decay kinetics, thus circumventing common issues associated with intensity-based measurements. This contribution introduces fluorescence lifetime excitation cytometry by kinetic dithering (FLECKD) as a method to capture multiple fluorescence lifetimes using a hybrid time-domain approach. The FLECKD approach excites fluorophores by delivering short pulses of light to cells or particles by rapid dithering and facilitates measurement of complex fluorescence decay kinetics by flow cytometry. Our simulations demonstrated a resolvable fluorescence lifetime value as low as 1.8 ns (±0.3 ns) with less than 20% absolute error. Using the FLECKD instrument, we measured the shortest average fluorescence lifetime value of 2.4 ns and found the system measurement error to be ±0.3 ns (SEM), from hundreds of monodisperse and chemically stable fluorescent microspheres. Additionally, we demonstrate the ability to detect two distinct excited state lifetimes from fluorophores in single cells using FLECKD. This approach presents a new ability to resolve multiple fluorescence lifetimes while retaining the fluidic throughput of a cytometry system. The ability to discriminate more than one average fluorescence lifetime expands the current capabilities of high-throughput and intensity-based cytometry assays as the need to tag one single cell with multiple fluorophores is now widespread.

  9. Fluorescence lifetime excitation cytometry by kinetic dithering

    PubMed Central

    Li, Wenyan; Vacca, Giacomo; Castillo, Maryann; Houston, Kevin D; Houston, Jessica P

    2014-01-01

    Flow cytometers are powerful high-throughput devices that capture spectroscopic information from individual particles or cells. These instruments provide a means of multi-parametric analyses for various cellular biomarkers or labeled organelles and cellular proteins. However, the spectral overlap of fluorophores limits the number of fluorophores that can be used simultaneously during experimentation. Time-resolved parameters enable the quantification of fluorescence decay kinetics, thus circumventing common issues associated with intensity-based measurements. This contribution introduces fluorescence lifetime excitation cytometry by kinetic dithering (FLECKD) as a method to capture multiple fluorescence lifetimes using a hybrid time-domain approach. The FLECKD approach excites fluorophores by delivering short pulses of light to cells or particles by rapid dithering and facilitates measurement of complex fluorescence decay kinetics by flow cytometry. Our simulations demonstrated a resolvable fluorescence lifetime value as low as 1.8 ns (±0.3 ns) with less than 20% absolute error. Using the FLECKD instrument, we measured the shortest average fluorescence lifetime value of 2.4 ns and found the system measurement error to be ±0.3 ns (SEM), from hundreds of monodisperse and chemically stable fluorescent microspheres. Additionally, we demonstrate the ability to detect two distinct excited state lifetimes from fluorophores in single cells using FLECKD. This approach presents a new ability to resolve multiple fluorescence lifetimes while retaining the fluidic throughput of a cytometry system. The ability to discriminate more than one average fluorescence lifetime expands the current capabilities of high-throughput and intensity-based cytometry assays as the need to tag one single cell with multiple fluorophores is now widespread. PMID:24668857

  10. SU-E-J-70: Evaluation of Multiple Isocentric Intensity Modulated and Volumetric Modulated Arc Therapy Techniques Using Portal Dosimetry

    SciTech Connect

    Muralidhar, K Raja; Pangam, S; Kolla, J; Ponaganti, S; Ali, M; Vuba, S; Mariyappan, P; Babaiah, M; Komanduri, K

    2015-06-15

    Purpose: To develop a method for verification of dose distribution in a patient during treatment using multiple isocentric Intensity modulated and volumetric modulated arc therapy techniques with portal dosimetry. Methods: Varian True Beam accelerator, equipped with an aS1000 megavoltage electronic portal imaging device (EPID) has an integrated image mode for portal dosimetry (PD). The source-to-imager distance was taken at 150 cm to avoid collision to the table. Fourteen fractions were analyzed for this study. During shift in a single plan from one isocenter to another isocenter, EPID also shifted longitudinally for each field by taking the extent of divergence of beam into the consideration for EPID distance of 150cm. Patients were given treatment everyday with EPID placed in proper position for each field. Several parameters were obtained by comparing the dose distribution between fractions to fraction. The impact of the intra-fraction and inter-fraction of the patient in combination with isocenter shift of the beams were observed. Results: During treatment, measurements were performed by EPID and were evaluated by the gamma method. Analysis was done between fractions for multiple isocenter treatments. The pass rates of the gamma analysis with a criterion of 3% and 3 mm for the 14 fractions were over 97.8% with good consistency. Whereas maximum gamma exceeded the criteria in few fractions (in<1 cc vol). Average gamma was observed in the criteria of 0.5%. Maximum dose difference and average dose differences were less than 0.22 CU and 0.01 CU for maximum tolerance of 1.0 CU and 0.2 CU respectively. Conclusion: EPID with extended distance is ideal method to verify the multiple isocentric dose distribution in patient during treatment, especially cold and hot spots in junction dose. Verification of shifts as well as the dose differences between each fraction due to inter-fraction and intra-fraction of the patient can be derived.

  11. Combined genetic algorithm and multiple linear regression (GA-MLR) optimizer: Application to multi-exponential fluorescence decay surface.

    PubMed

    Fisz, Jacek J

    2006-12-07

    The optimization approach based on the genetic algorithm (GA) combined with multiple linear regression (MLR) method, is discussed. The GA-MLR optimizer is designed for the nonlinear least-squares problems in which the model functions are linear combinations of nonlinear functions. GA optimizes the nonlinear parameters, and the linear parameters are calculated from MLR. GA-MLR is an intuitive optimization approach and it exploits all advantages of the genetic algorithm technique. This optimization method results from an appropriate combination of two well-known optimization methods. The MLR method is embedded in the GA optimizer and linear and nonlinear model parameters are optimized in parallel. The MLR method is the only one strictly mathematical "tool" involved in GA-MLR. The GA-MLR approach simplifies and accelerates considerably the optimization process because the linear parameters are not the fitted ones. Its properties are exemplified by the analysis of the kinetic biexponential fluorescence decay surface corresponding to a two-excited-state interconversion process. A short discussion of the variable projection (VP) algorithm, designed for the same class of the optimization problems, is presented. VP is a very advanced mathematical formalism that involves the methods of nonlinear functionals, algebra of linear projectors, and the formalism of Fréchet derivatives and pseudo-inverses. Additional explanatory comments are added on the application of recently introduced the GA-NR optimizer to simultaneous recovery of linear and weakly nonlinear parameters occurring in the same optimization problem together with nonlinear parameters. The GA-NR optimizer combines the GA method with the NR method, in which the minimum-value condition for the quadratic approximation to chi(2), obtained from the Taylor series expansion of chi(2), is recovered by means of the Newton-Raphson algorithm. The application of the GA-NR optimizer to model functions which are multi

  12. Fluorescence characteristics of 5-carboxytetramethylrhodamine linked covalently to the 5' end of oligonucleotides: multiple conformers of single-stranded and double-stranded dye-DNA complexes.

    PubMed Central

    Vámosi, G; Gohlke, C; Clegg, R M

    1996-01-01

    Fluorescence steady-state and lifetime experiments have been carried out on duplex and single-stranded DNA molecules labeled at the 5' ends with 5-carboxytetramethylrhodamine (TMRh). The temperature and ionic strength of the solutions were varied over large ranges. The results reveal at least three well-defined states of the TMRh-DNA molecules for the single-stranded as well as for the double-stranded DNA molecules. Two states are fluorescent, with lifetimes in the range of 0.5-1 ns and 2.5-3 ns. A third state of TMRh-DNA does not fluoresce (a dark species of TMRh-DNA). The distribution of the TMRh-DNA molecules among these three states is strongly temperature and ionic strength dependent. Estimates are made of some reaction parameters of the multistate model. The results are discussed in terms of the photophysics of TMRh, and consequences of the multiple conformers of TMRh-DNA for studies involving fluorescence studies with TMRh-labeled DNA are considered. PMID:8842236

  13. Fluorescence study of sugars

    NASA Astrophysics Data System (ADS)

    Thongjamroon, Sunida; Pattanaporkratana, Apichart

    2015-07-01

    We studied photoemission of monosaccharides and disaccharides using laser-induced fluorescence spectroscopy. A 532- nm, 10 mW, laser was used to excite the samples and back-scattering signals were collected by a spectrometer. We found that most sugars show weak fluorescence in solid phase but do not fluoresce when dissolved in water solutions. The emission spectra show similar peak intensity at 590 nm, but they are different in emission intensities. We suggest that the fluorescence spectra may be used to differentiate sugar type, even though the origin of the fluorescence is unclear and needed further study.

  14. Assessment of crop productivity over intensively managed agriculture regions in India and Australia using solar-induced fluorescence remote sensing data

    NASA Astrophysics Data System (ADS)

    Devadas, R.; Huete, A. R.; Patel, N. R.; Padalia, H.; Restrepo-Coupe, N.; Kuruvilla, A.

    2015-12-01

    Satellite based estimation of solar-induced terrestrial fluorescence (SIF) is considered to be a direct measure of photosynthetic functional status of the vegetation. Prior studies have shown SIF to more accurately retrieve the productivity of intensively managed croplands, as in the U.S. corn belt. In this study, we assessed and compared agricultural productivity over two intensive crop production regions in Australia and India using SIF data, traditional spectral measures, and crop yield data. Regional level wheat yield data were obtained for the Indo-Gangetic Plains (IGP) in India and the Murray Darling Basin (MDB) in Australia for analyses with GOME-2 SIF satellite and MODIS VI measurements, and gross primary productivity from flux towers. We investigated the importance of integrating traditional meteorological parameters and ground based data with time-series vegetation indices for scaling of SIF to obtain robust yield prediction models for application across years and continents. This study further explored the relationship of inter annual variations in crop phenology metrics through SIF retrievals and its relationship with crop yields. The IGP study region showed systematic cycles of double cropping. MDB region on the other hand showed cycles of pronounced winter cropping and a weaker and variable second cropping over the analysis period. For various winter wheat crop seasons in IGP, from 2007 to 2012, SIF explained and accounted between 48 to 74 per cent of the variations in regional wheat yields. Similar results were obtained in the case of MDB also, however, the relationship between SIF and yield estimates was weaker (R2 = 0.44). SIF measurements, as a surrogate of crop productivity, were considerably higher over the highly productive IGP region in almost all the years considered. The SIF data shows immense potential for modelling agricultural productivity, particularly as the resolution of SIF retrievals continues to improve.

  15. Uranium enrichment measurements using the intensity ratios of self-fluorescence X-rays to 92* keV gamma ray in UXK alpha spectral region.

    PubMed

    Yücel, H; Dikmen, H

    2009-04-30

    In this paper, the known multigroup gamma-ray analysis method for uranium (MGAU) as one of the non-destructive gamma-ray spectrometry methods has been applied to certified reference nuclear materials (depleted, natural and enriched uranium) containing (235)U isotope in the range of 0.32-4.51% atom (235)U. Its analysis gives incorrect results for the low component (235)U in depleted and natural uranium samples where the build-up of the decay products begins to interfere with the analysis. The results reveal that the build-up of decay products seems to be significant and thus the algorithms for the presence of decay products should be improved to resulting in the correct enrichment value. For instance, for the case of (235)U analysis in depleted uranium or natural ore samples, self-induced X-rays such as 94.6 keV and 98.4 keV lying in UXK(alpha) spectral region used by MGAU can be excluded from the calculation. Because the significant increases have been observed in the intensities of uranium self-induced X-rays due to gamma-ray emissions with above 100 keV energy arising from decay products of (238)U and (235)U and these parents. Instead, the use of calibration curve to be made between the intensity ratios of self-fluorescence X-rays to 92(*)keV gamma-ray and the certified (235)U abundances is suggested for the determination of (235)U when higher amounts of decay products are detected in the gamma-ray spectrum acquired for the MGAU analysis.

  16. Temporal and spatial changes in dissolved organic carbon concentration and fluorescence intensity of fulvic acid like materials in mountainous headwater catchments

    NASA Astrophysics Data System (ADS)

    Terajima, Tomomi; Moriizumi, Mihoko

    2013-02-01

    SummaryDissolved organic carbon (DOC) such as humic substances are key to understanding the aquatic environment in catchments, because they, containing a large number of phenolic and carboxylic acid groups, adsorb many kinds of inorganic materials and also affect nutrition and carbon transport in catchments. To understand the detailed DOC dynamics, we conducted hydrological observations at mountainous headwater catchments dominated by different vegetation types (planted evergreen coniferous forest of 1.29 ha and natural deciduous broadleaf forest of 1.28 ha). The relationship between DOC concentrations and fluorescence intensity of fulvic acid-like materials (F-FAM) were positively correlated in both catchments but different between soil extracts, baseflow, and near surface flow represented by biomat flow. The ratios of change in F-FAM to that in DOC concentration (F-FAM/DOC) were higher in the baseflow (about 6 in both catchments) and lower in the soil extracts (about 4.5 in both catchments, respectively). However, the relationship in stormflow was distributed between the trends of baseflow and soil extracts. The higher F-FAM/DOC in baseflow may thus indicate that DOC (and FAM) in groundwater discharge mainly contributed to the stream flow, and the stormflow mainly reflect subsurface flow through soil during most rainstorms. In contrast, a high F-FAM/DOC ratio (>6) appeared in the stormflow of both catchments especially during large storms of short duration and high intensity following a dry antecedent period. The F-FAM/DOC in biomat flow developing distinctly in the coniferous catchment was high (about 6.5). Thus, rapid shallow subsurface flow through the biomat or near-surface of slopes might explain the unique transport dynamics of DOC and FAM in stormflows with the high F-FAM/DOC ratio. These results imply that the DOC and FAM relationship responds variably depending on both the distribution of soil organic matter and rainwater flow paths in steep slopes as

  17. High Intensity Training May Reverse the Fiber Type Specific Decline in Myogenic Stem Cells in Multiple Sclerosis Patients

    PubMed Central

    Farup, Jean; Dalgas, Ulrik; Keytsman, Charly; Eijnde, Bert O.; Wens, Inez

    2016-01-01

    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 and 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) was quantified using ATPase histochemistry. At baseline the SCs per fiber was lower in type II compared to type I fibers 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 per fCSA in MS patients increased by 165% (p < 0.05) and 135% (p < 0.05), respectively. Furthermore, the type II fiber MN content tended (p = 0.06) to be increased by 35% 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. PMID:27303309

  18. Autologous haematopoietic stem cell transplantation with an intermediate intensity conditioning regimen in multiple sclerosis: the Italian multi-centre experience.

    PubMed

    Mancardi, G L; Sormani, M P; Di Gioia, M; Vuolo, L; Gualandi, F; Amato, M P; Capello, E; Currò, D; Uccelli, A; Bertolotto, A; Gasperini, C; Lugaresi, A; Merelli, E; Meucci, G; Motti, L; Tola, M R; Scarpini, E; Repice, A M; Massacesi, L; Saccardi, R

    2012-06-01

    Over recent years numerous patients with severe forms of multiple sclerosis (MS) refractory to conventional therapies have been treated with intense immunosuppression followed by autologous haematopoietic stem cell transplantation (AHSCT). The clinical outcome and the toxicity of AHSCT can be diverse, depending on the various types of conditioning protocols and on the disease phase. To report the Italian experience on all the consecutive patients with MS treated with AHSCT with an intermediate intensity conditioning regimen, named BEAM/ATG, in the period from 1996 to 2008. Clinical and magnetic resonance imaging outcomes of 74 patients were collected after a median follow-up period of 48.3 (range = 0.8-126) months. Two patients (2.7%) died from transplant-related causes. After 5 years, 66% of patients remained stable or improved. Among patients with a follow-up longer than 1 year, eight out of 25 subjects with a relapsing-remitting course (31%) had a 6-12 months confirmed Expanded Disability Status Scale improvement > 1 point after AHSCT as compared with one out of 36 (3%) patients with a secondary progressive disease course (p = 0.009). Among the 18 cases with a follow-up longer than 7 years, eight (44%) remained stable or had a sustained improvement while 10 (56%), after an initial period of stabilization or improvement with median duration of 3.5 years, showed a slow disability progression. This study shows that AHSCT with a BEAM/ATG conditioning regimen has a sustained effect in suppressing disease progression in aggressive MS cases unresponsive to conventional therapies. It can also cause a sustained clinical improvement, especially if treated subjects are still in the relapsing-remitting phase of the disease.

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

    NASA Astrophysics Data System (ADS)

    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.

  20. Multi-wavelength fluorescence tomography

    NASA Astrophysics Data System (ADS)

    Kwong, Tiffany C.; Lo, Pei-An; Cho, Jaedu; Nouizi, Farouk; Chiang, Huihua K.; Kim, Chang-Seok; Gulsen, Gultekin

    2016-03-01

    The strong scattering and absorption of light in biological tissue makes it challenging to model the propagation of light, especially in deep tissue. This is especially true in fluorescent tomography, which aims to recover the internal fluorescence source distribution from the measured light intensities on the surface of the tissue. The inherently ill-posed and underdetermined nature of the inverse problem along with strong tissue scattering makes Fluorescence Tomography (FT) extremely challenging. Previously, multispectral detection fluorescent tomography (FT) has been shown to improve the image quality of FT by incorporating the spectral filtering of biological tissue to provide depth information to overcome the inherent absorption and scattering limitations. We investigate whether multi-wavelength fluorescent tomography can be used to distinguish the signals from multiple fluorophores with overlapping fluorescence spectrums using a unique near-infrared (NIR) swept laser. In this work, a small feasibility study was performed to see whether multi-wavelength FT can be used to detect subtle shifts in the absorption spectrum due to differences in fluorophore microenvironment.

  1. Advantages of estimating parameters of photosynthesis model by fitting A-Ci curves at multiple subsaturating light intensities

    NASA Astrophysics Data System (ADS)

    Fu, W.; Gu, L.; Hoffman, F. M.

    2013-12-01

    The photosynthesis model of Farquhar, von Caemmerer & Berry (1980) is an important tool for predicting the response of plants to climate change. So far, the critical parameters required by the model have been obtained from the leaf-level measurements of gas exchange, namely the net assimilation of CO2 against intercellular CO2 concentration (A-Ci) curves, made at saturating light conditions. With such measurements, most points are likely in the Rubisco-limited state for which the model is structurally overparameterized (the model is also overparameterized in the TPU-limited state). In order to reliably estimate photosynthetic parameters, there must be sufficient number of points in the RuBP regeneration-limited state, which has no structural over-parameterization. To improve the accuracy of A-Ci data analysis, we investigate the potential of using multiple A-Ci curves at subsaturating light intensities to generate some important parameter estimates more accurately. Using subsaturating light intensities allow more RuBp regeneration-limited points to be obtained. In this study, simulated examples are used to demonstrate how this method can eliminate the errors of conventional A-Ci curve fitting methods. Some fitted parameters like the photocompensation point and day respiration impose a significant limitation on modeling leaf CO2 exchange. The multiple A-Ci curves fitting can also improve over the so-called Laisk (1977) method, which was shown by some recent publication to produce incorrect estimates of photocompensation point and day respiration. We also test the approach with actual measurements, along with suggested measurement conditions to constrain measured A-Ci points to maximize the occurrence of RuBP regeneration-limited photosynthesis. Finally, we use our measured gas exchange datasets to quantify the magnitude of resistance of chloroplast and cell wall-plasmalemma and explore the effect of variable mesophyll conductance. The variable mesophyll conductance

  2. Ultrapure Blue Thermally Activated Delayed Fluorescence Molecules: Efficient HOMO-LUMO Separation by the Multiple Resonance Effect.

    PubMed

    Hatakeyama, Takuji; Shiren, Kazushi; Nakajima, Kiichi; Nomura, Shintaro; Nakatsuka, Soichiro; Kinoshita, Keisuke; Ni, Jingping; Ono, Yohei; Ikuta, Toshiaki

    2016-04-13

    Ultrapure blue-fluorescent molecules based on thermally activated delayed fluorescence are developed. Organic light-emitting diode (OLED) devices employing the new emitters exhibit a deep blue emission at 467 nm with a full-width at half-maximum of 28 nm, CIE coordinates of (0.12, 0.13), and an internal quantum efficiency of ≈100%, which represent record-setting performance for blue OLED devices. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Influence of hydrological and climatic parameters on spatial-temporal variability of fluorescence intensity and DOC of karst percolation waters in the Santana Cave System, Southeastern Brazil

    NASA Astrophysics Data System (ADS)

    Cruz, F. W.; Karmann, I.; Magdaleno, G. B.; Coichev, N.; Viana, O.

    2005-02-01

    Fluorescence intensity (FI) and organic carbon concentration of groundwater percolating through soil and rock into the Santana Cave were monitored at eight different cave sites between 2000 and 2002 to investigate their relationships to climatic parameters, stalactite discharge and thickness of rock overlying the cave. FI values, compared among sampling sites, are inversely proportional to depth and directly proportional to discharge; in contrast, dissolved organic matter (DOC) shows no significant spatial variability. Time-series analysis demonstrated similarities in DOC trends of different waters, but no correlation was observed with FI trends. Combined evaluation of DOC of infiltration waters, rainfall data and chemical parameters of Fe, O 2, pH, Eh in soil solution indicate that peaks in DOC content coincide with more reduced conditions in the soil and have a lag time of 2-3 months after heavy showers. Variation of FI throughout the year occurs at all sampling sites but only higher drip discharge and rimstone pool waters were correlatable to rainfall events. FI of lower discharge sampling sites shows similar trends, but no relationship between drip discharge and rainfall variation was observed. Ranges and means of FI for all drip waters were significantly higher in the 2001-2002 period than in the preceding 2000-2001 period, which correlates with a 5.5 °C increase in mean austral winter temperatures in 2001. Hence, FI variations of karst waters that form carbonate speleothems under a humid subtropical climate may provide a useful proxy in paleoenvironmental reconstruction.

  4. [The changing process of population probability and fluorescence intensity in ErP5O14 noncrystalline from 0 second to stabilization].

    PubMed

    Chen, Xiao-Bo; Chen, Ying; Li, Song; Yao, Wen-Ting; Wu, Zheng-Long

    2012-06-01

    The results of numerical simulation were compared respectively with and without considering the improved coefficient for the energy transfer process in ErP5O14 noncrystalline under 521.8, 450.0, 405.5 and 378.5 nm lights excitation. The results showed that it is essential to take the coefficient into calculation where energy transfer plays a key role. The relative fluorescence intensity ratios of 4I13/2-->4I15/2 to 4I11/2-->4I15/2 under 523.8, 450.0 and 378.5 nm lights excitation were 2.11, 2.82 and 2.99 times larger than that under 979.3 nm light excitation respectively. There are obviously effective near-infrared (NIR) quantum cutting of 4I13/2-->4I15/2 transition. The result indicates that ErP5O14 noncrystalline has potential application in the enhancement of conversion efficiency of germanium solar cells.

  5. Single-isocenter frameless intensity-modulated stereotactic radiosurgery for simultaneous treatment of multiple brain metastases: clinical experience.

    PubMed

    Nath, Sameer K; Lawson, Joshua D; Simpson, Daniel R; Vanderspek, Lauren; Wang, Jia-Zhu; Alksne, John F; Ciacci, Joseph; Mundt, Arno J; Murphy, Kevin T

    2010-09-01

    To describe our clinical experience using a unique single-isocenter technique for frameless intensity-modulated stereotactic radiosurgery (IM-SRS) to treat multiple brain metastases. Twenty-six patients with a median of 5 metastases (range, 2-13) underwent optically guided frameless IM-SRS using a single, centrally located isocenter. Median prescription dose was 18 Gy (range, 14-25). Follow-up magnetic resonance imaging (MRI) and clinical examination occurred every 2-4 months. Median follow-up for all patients was 3.3 months (range, 0.2-21.3), with 20 of 26 patients (77%) followed up until their death. For the remaining 6 patients alive at the time of analysis, median follow-up was 14.6 months (range, 9.3-18.0). Total treatment time ranged from 9.0 to 38.9 minutes (median, 21.0). Actuarial 6- and 12-month overall survivals were 50% (95% confidence interval [C.I.], 31-70%) and 38% (95% C.I., 19-56%), respectively. Actuarial 6- and 12-month local control (LC) rates were 97% (95% C.I., 93-100%) and 83% (95% C.I., 71-96%), respectively. Tumors 1.5 cm (98% vs. 90%, p = 0.008). New intracranial metastatic disease occurring outside of the treatment volume was observed in 7 patients. Grade >or=3 toxicity occurred in 2 patients (8%). Frameless IM-SRS using a single-isocenter approach for treating multiple intracranial metastases can produce clinical outcomes that compare favorably with those of conventional SRS in a much shorter treatment time (<40 minutes). Given its faster treatment time, this technique is appealing to both patients and personnel in busy clinics. Copyright (c) 2010 Elsevier Inc. All rights reserved.

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

    PubMed

    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. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. SPECIAL ISSUE DEVOTED TO MULTIPLE RADIATION SCATTERING IN RANDOM MEDIA: Path method for reconstructing images in fluorescence optical tomography

    NASA Astrophysics Data System (ADS)

    Kravtsenyuk, Olga V.; Lyubimov, Vladimir V.; Kalintseva, Natalie A.

    2006-11-01

    A reconstruction method elaborated for the optical diffusion tomography of the internal structure of objects containing absorbing and scattering inhomogeneities is considered. The method is developed for studying objects with fluorescing inhomogeneities and can be used for imaging of distributions of artificial fluorophores whose aggregations indicate the presence of various diseases or pathological deviations.

  8. Influence of phase transitions on green fluorescence intensity ratio in Er3+ doped K0.5Na0.5NbO3 ceramic.

    PubMed

    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 2H11/2 and 4S3/2 states of Er3+ doped K0.5Na0.5NbO3 (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 (TO-T) point and 13 K at the Curie point (TC). 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.

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

    PubMed Central

    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.

    PubMed

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

    2009-03-09

    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. To report whole-body and organ-specific fluorescence imaging to characterize the transgenic cyan fluorescent protein mouse. 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). 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. Determination of fluorescence intensity of different organs. 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. The cyan fluorescent protein mouse should lead to a deeper understanding of pancreatic function and pathology, including cancer.

  11. Multiple temperature effects on up-conversion fluorescences of Er{sup 3+}-Y b{sup 3+}-Mo{sup 6+} codoped TiO{sub 2} and high thermal sensitivity

    SciTech Connect

    Cao, B. S.; Wu, J. L.; Wang, X. H.; He, Y. Y.; Feng, Z. Q.; Dong, B. E-mail: bscao@dlnu.edu.cn; Rino, L.

    2015-08-15

    We report multiple temperature effects on green and red up-conversion emissions in Er{sup 3+}-Y b{sup 3+}-Mo{sup 6+} codoped TiO{sub 2} phosphors. With increasing temperature, the decrease of the red emission from {sup 4}F{sub 9/2}→{sup 4}I{sub 15/2}, the increase of green emission from {sup 2}H{sub 11/2}→{sup 4}I{sub 15/2} and another unchanged green emission from {sup 4}S{sub 3/2}→{sup 4}I{sub 15/2} 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 (S{sub max}) and optimum operating temperature (T{sub max}) 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.

  12. Development and primary application of a fluorescent liquid bead array for the simultaneous identification of multiple genetically modified maize.

    PubMed

    Han, Xueqing; Wang, Huiyu; Chen, Hongjun; Mei, Lin; Wu, Shaoqiang; Jia, Guangle; Cheng, Tao; Zhu, Shuifang; Lin, Xiangmei

    2013-11-15

    An integration event-specific fluorescent liquid bead array was developed for the simultaneous identification of 10 genetically modified (GM) maize, including Bt176, Bt11, MON810, NK603, GA21, MON88017, MON89034, MIR604, T25 and MIR162, as well as one non-GM maize. The system comprised 11 specific oligonucleotide probes labeled with an amino group and coupled to fluorescence-encoded microspheres. To enable fluorescence detection, 11 pairs of primers labeled with biotin at the 5' ends were used. The hybridization signal of biotinylated PCR product to the probe-coupled microspheres was then detected. The limit of detection of this assay was 0.1% for GM maize, which is lower than the current labeling threshold levels enforced in the EU (0.9%). The results of the positive and negative controls were consistent with their expected situation, which showed that the method was highly specific. We detected GM maize in 20 of the 1370 commercial food samples tested, which were labeled as containing maize. The overall sensitivity, specificity, rapidity and high throughput capacity of this liquid chip system suggest that it could provide a significant improvement over current methods, and potentially offer an improved platform for further research into the detection of other GM plants.

  13. Fluorescent fiber diagnostics

    DOEpatents

    Toeppen, John S.

    1994-10-04

    A fluorescent fiber (13) having a doped core (16) is pumped (11) by light (18) of a relatively short wavelength to produce fluorescence at a longer wavelength that is detected by detector (24). The level of fluorescence is monitored (26) and evaluated to provide information as to the excitation of the fiber (13) or the environment thereof. In particular, the level of intensity of the detected fluorescence may be used to measure the intensity of a light beam (18) passing axially through an optical fiber system (12) (FIG. 1 ), or the intensity of a light beam (46) passing radially through a fluorescent fiber (13) (FIG. 2 ), or the level of a fluid (32) in a tank (31) (FIG. 3 ), or a scintillation event (37) in a fluorescent fiber (13) pumped to produce amplification of the scintillation event (FIG. 4 ).

  14. Fluorescent fiber diagnostics

    DOEpatents

    Toeppen, John S.

    1994-01-01

    A fluorescent fiber (13) having a doped core (16) is pumped (11) by light (18) of a relatively short wavelength to produce fluorescence at a longer wavelength that is detected by detector (24). The level of fluorescence is monitored (26) and evaluated to provide information as to the excitation of the fiber (13) or the environment thereof. In particular, the level of intensity of the detected fluorescence may be used to measure the intensity of a light beam (18) passing axially through an optical fiber system (12) (FIG. 1 ), or the intensity of a light beam (46) passing radially through a fluorescent fiber (13) (FIG. 2 ), or the level of a fluid (32) in a tank (31) (FIG. 3 ), or a scintillation event (37) in a fluorescent fiber (13) pumped to produce amplification of the scintillation event (FIG. 4 ).

  15. Frequency upconversion and fluorescence intensity ratio method in Yb3+-ion-sensitized Gd2O3:Er3+-Eu3+ phosphors for display and temperature sensing

    NASA Astrophysics Data System (ADS)

    Ranjan, Sushil Kumar; Soni, Abhishek Kumar; Rai, Vineet Kumar

    2017-09-01

    Near infrared (NIR) to visible frequency upconversion emission studies in Er3+-Eu3+/Er3+-Eu3+-Yb3+ co-doped/tri-doped Gd2O3 phosphors prepared by the co-precipitation technique have been explored under 980 nm laser diode radiation. The developed phosphors were characterized with the help of XRD, FE-SEM and FTIR analysis. No upconversion (UC) emission was found in the Eu3+-doped Gd2O3 phosphor. UC emission from Eu3+ ions along with Er3+ ions was observed in Er3+-Eu3+ and Er3+-Eu3+-Yb3+ co-doped/tri-doped phosphors. The UC emission arising from the Er3+ and Eu3+ ions was enhanced several times due to the incorporation of Yb3+ ions. The processes involved in the UC emission were obtained on the basis of the effect of energy transfer/sensitization through the Yb3+ → Er3+ → Eu3+ process. The red/green intensity ratio was improved from 0.16 to 1.50 and 1.01 to 1.50 for Er3+-Eu3+-Yb3+ tri-doped phosphors as compared to the Er3+-doped and Er3+-Yb3+ co-doped phosphors, respectively, at a fixed pump power density. A UC fluorescence intensity ratio (FIR)-based temperature sensing study was performed in the prepared Er3+-Eu3+-Yb3+ tri-doped Gd2O3 phosphors for green upconversion emission bands in the 300 K–443 K temperature range. A maximum sensor sensitivity of about ∼0.0043 K‑1 at 300 K was achieved for the synthesized tri-doped phosphors upon excitation with a 980 nm laser diode. The colour coordinates lying in the green–yellow region are invariant, with variation in pump power density and temperature. The observed results support the utility of the prepared tri-doped phosphors in optical temperature sensing, display devices and NIR to visible upconverters.

  16. Robust effects of cloud superparameterization on simulated daily rainfall intensity statistics across multiple versions of the Community Earth System Model

    DOE PAGES

    Kooperman, Gabriel J.; Pritchard, Michael S.; Burt, Melissa A.; ...

    2016-02-01

    This study evaluates several important statistics of daily rainfall based on frequency and amount distributions as simulated by a global climate model whose precipitation does not depend on convective parameterization—Super-Parameterized Community Atmosphere Model (SPCAM). Three superparameterized and conventional versions of CAM, coupled within the Community Earth System Model (CESM1 and CCSM4), are compared against two modern rainfall products (GPCP 1DD and TRMM 3B42) to discriminate robust effects of superparameterization that emerge across multiple versions. The geographic pattern of annual-mean rainfall is mostly insensitive to superparameterization, with only slight improvements in the double-ITCZ bias. However, unfolding intensity distributions reveal several improvementsmore » in the character of rainfall simulated by SPCAM. The rainfall rate that delivers the most accumulated rain (i.e., amount mode) is systematically too weak in all versions of CAM relative to TRMM 3B42 and does not improve with horizontal resolution. It is improved by superparameterization though, with higher modes in regions of tropical wave, Madden-Julian Oscillation, and monsoon activity. Superparameterization produces better representations of extreme rates compared to TRMM 3B42, without sensitivity to horizontal resolution seen in CAM. SPCAM produces more dry days over land and fewer over the ocean. Updates to CAM’s low cloud parameterizations have narrowed the frequency peak of light rain, converging toward SPCAM. Poleward of 50°, where more rainfall is produced by resolved-scale processes in CAM, few differences discriminate the rainfall properties of the two models. Lastly, these results are discussed in light of their implication for future rainfall changes in response to climate forcing.« less

  17. Robust effects of cloud superparameterization on simulated daily rainfall intensity statistics across multiple versions of the Community Earth System Model

    NASA Astrophysics Data System (ADS)

    Kooperman, Gabriel J.; Pritchard, Michael S.; Burt, Melissa A.; Branson, Mark D.; Randall, David A.

    2016-03-01

    This study evaluates several important statistics of daily rainfall based on frequency and amount distributions as simulated by a global climate model whose precipitation does not depend on convective parameterization—Super-Parameterized Community Atmosphere Model (SPCAM). Three superparameterized and conventional versions of CAM, coupled within the Community Earth System Model (CESM1 and CCSM4), are compared against two modern rainfall products (GPCP 1DD and TRMM 3B42) to discriminate robust effects of superparameterization that emerge across multiple versions. The geographic pattern of annual-mean rainfall is mostly insensitive to superparameterization, with only slight improvements in the double-ITCZ bias. However, unfolding intensity distributions reveal several improvements in the character of rainfall simulated by SPCAM. The rainfall rate that delivers the most accumulated rain (i.e., amount mode) is systematically too weak in all versions of CAM relative to TRMM 3B42 and does not improve with horizontal resolution. It is improved by superparameterization though, with higher modes in regions of tropical wave, Madden-Julian Oscillation, and monsoon activity. Superparameterization produces better representations of extreme rates compared to TRMM 3B42, without sensitivity to horizontal resolution seen in CAM. SPCAM produces more dry days over land and fewer over the ocean. Updates to CAM's low cloud parameterizations have narrowed the frequency peak of light rain, converging toward SPCAM. Poleward of 50°, where more rainfall is produced by resolved-scale processes in CAM, few differences discriminate the rainfall properties of the two models. These results are discussed in light of their implication for future rainfall changes in response to climate forcing.

  18. 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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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.

  20. Intensity and Breadth of Participation in Organized Activities during the Adolescent Years: Multiple Associations with Youth Outcomes

    ERIC Educational Resources Information Center

    Denault, Anne-Sophie; Poulin, Francois

    2009-01-01

    The goal of this study was to examine initial levels and rates of change in the intensity and breadth of participation in organized activities during the adolescent years, and how these participation practices were related to youth outcomes in later adolescence. The main objectives were (a) to examine growth curves of intensity and breadth of…

  1. Signal to Noise Ratio in Digital Lock-in Detection for Multiple Intensity-Modulated Signals in CO2 Laser Absorption Spectrometer

    NASA Astrophysics Data System (ADS)

    CHEN, S.; Lin, B.; Harrison, F. W.; Nehrir, A. R.; Campbell, J. F.; Refaat, T.; Abedin, N. M.; Obland, M. D.; Ismail, S.; Meadows, B. L.

    2013-12-01

    NASA Langley Research Center is investigating Intensity-Modulated, Continuous-Wave Laser Absorption Spectrometers (LASs) for the measurement of atmospheric carbon dioxide (CO2) column mixing ratio from both air- and space-borne platforms. The LAS system uses high-power fiber lasers/amplifiers in the 1.57-um CO2 absorption band and the 1.26-um O2 absorption band in the transmitters and simultaneous digital lock-in detection for the multiple intensity-modulated signals with different modulation waveforms , such as simple sinusoidal waves at different frequencies, associated with different wavelengths in the receivers. The Signal to Noise Ratio (SNR) of the simultaneous digital lock-in detection in the system is of interest for the system designs and the performance prediction of airborne and space-borne implementations in the future. This paper will discuss the properties of the signals and various noises in the LAS system, especially for the simultaneous digital lock-in detection with a single detector for the multiple intensity-modulated signals at different frequencies. The numerical simulation of the SNR for the simultaneous digital lock-in detection in terms of relative intensity of the multiple modulated signals and the integration time, and an initial experimental verification will be presented.

  2. An algorithm for the realization of three-dimensional laser imaging system's intensity image and range image based on multiple-slit streak tube

    NASA Astrophysics Data System (ADS)

    Han, Shaokun; Yu, Xiaojiang; Jiang, Ming; Liu, Xunliang

    2008-03-01

    The technology of three-dimensional laser imaging is applied widely in the field of military use and civilian use etc. There are mainly two methods for three-dimensional laser imaging. One of them is based on APD arrays, and the other is based on streak tube. The latter represents relatively mature technology for providing high-resolution 3D laser radar images. In both of them, the realization of intensity image and range image is the foundation and key of three-dimensional laser imaging. It presents the method for three-dimensional laser imaging using multiple-slit streak tube to get clear, exact intensity image and range image. The multiple-slit streak tube imaging lidar (MS-STIL) approach uses several slits instead of the usual single slit to provide a number of additional capabilities over conventional laser radar systems. And it researches into the algorithm for the realization of intensity image and range image and processes the simulative streak tube image with it via the analysis of multiple-slit streak tube's imaging theory, and finally carries through the simulation of intensity image and range image.

  3. The fluorescence intensity of the lipophilic probe N-phenyl-1-naphthylamine responds to the oxidation-reduction state of the respiratory chain in everted membrane vesicles of Escherichia coli.

    PubMed

    Sedgwick, E G; Bragg, P D

    1987-06-22

    N-Phenyl-1-naphthylamine (NPN), a reagent which has been used previously to probe the fluidity or microviscosity of the membrane lipids of intact cells of Escherichia coli, was found to respond to metabolic changes in everted inner membrane vesicles from this organism. NPN was bound to the vesicles to produce a steady-state level of fluorescence intensity. Addition of substrate or ATP did not alter the fluorescence. However, following complete removal of oxygen from the medium by oxidation of substrate through the respiratory chain, there was an increase in the fluorescence of NPN. Reoxidation of the components of the respiratory chain by the addition of oxygen, ferricyanide, fumarate or nitrate decreased fluorescence to the steady-state level until the oxidant had been completely reduced. The fluorescence changes were insensitive to the state of energization of the membrane. It is proposed that NPN responds to the state of reduction of components of the respiratory chain either directly by reacting with a component of the chain or indirectly through an effect transmitted to the membrane by a change in the conformation of respiratory chain components.

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

    SciTech Connect

    Ma, L; Dong, P; Larson, D; Keeling, V; Hossain, S; Ahmad, S; Sahgal, A

    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

  5. High-intensity interval training combined with resistance training improves physiological capacities, strength and quality of life in multiple sclerosis patients: a pilot study.

    PubMed

    Zaenker, Pierre; Favret, Fabrice; Lonsdorfer, Evelyne; Muff, Guillaume; DE Seze, Jérôme; Isner-Horobeti, Marie-Eve

    2017-07-05

    Numerous studies have shown that mild-to-moderate intensity or resistance exercise training improves physical capacities such as, peak oxygen consumption, maximal tolerated power and strength in multiple sclerosis patients. However, few studies have evaluated the effects of high intensity interval training associated to with resistance training. Only few studies have analysed difference between men and women before and after combined training. Moreover, the evaluation of exercise between ambulatory multiple sclerosis patients without disability (EDSS 0- 3) and patients with disabilities (EDSS 3.5-5) was not largely published. The main objective of our study was to determine if high intensity interval training combined with resistance training improved aerobic and strength capacities as well as quality of life in multiple sclerosis patients and if gender and disabilities play a role in these changes. This study was an open-label uncontrolled study. The study was performed outside from conventional care facilities and including homebased training. 26 multiple sclerosis patients have completed the program (19 women/7men, Age 44.6+7.9years, Expanded Disability Status Score 2 (0-5)). We conducted a 12-week programme of high intensity interval training combined with resistance training at body weight. Peak oxygen consumption, maximal tolerated power, lactates, isokinetic strength of quadriceps and hamstrings (at 90°/sec, 180°/sec and 240°/sec) and quality of life were evaluated before and after the programme. Peak oxygen consumption and maximum tolerated power improved by 13.5% and 9.4%, respectively. Isokinetic muscle strength increased in both quadriceps and hamstrings at each speed, with a rebalancing of strength between the two legs in quadriceps. Quality of life was also enhanced in three domains. Women showed better improvements than men in VO2peak, maximal tolerated power, lactates at the end of test, and heart rate peak, strength in both quadriceps and

  6. Feasibility of transferring fluorescent in situ hybridization probes to an 18S rRNA gene phylochip and mapping of signal intensities.

    PubMed

    Metfies, Katja; Medlin, Linda K

    2008-05-01

    DNA microarray technology offers the possibility to analyze microbial communities without cultivation, thus benefiting biodiversity studies. We developed a DNA phylochip to assess phytoplankton diversity and transferred 18S rRNA probes from dot blot or fluorescent in situ hybridization (FISH) analyses to a microarray format. Similar studies with 16S rRNA probes have been done determined that in order to achieve a signal on the microarray, the 16S rRNA molecule had to be fragmented, or PCR amplicons had to be <150 bp in length to minimize the formation of a secondary structure in the molecule so that the probe could bind to the target site. We found different results with the 18S rRNA molecule. Four out of 12 FISH probes exhibited false-negative signals on the microarray; eight exhibited strong but variable signals using full-length 18S RNA molecules. A systematic investigation of the probe's accessibility to the 18S rRNA gene was made using Prymenisum parvum as the target. Fourteen additional probes identical to this target covered the regions not tested with existing FISH probes. Probes with a binding site in the first 900 bp of the gene generated positive signals. Six out of nine probes binding in the last 900 bp of the gene produced no signal. Our results suggest that although secondary structure affected probe binding, the effect is not the same for the 18S rRNA gene and the 16S rRNA gene. For the 16S rRNA gene, the secondary structure is stronger in the first half of the molecule, whereas in the 18S rRNA gene, the last half of the molecule is critical. Probe-binding sites within 18S rRNA gene molecules are important for the probe design for DNA phylochips because signal intensity appears to be correlated with the secondary structure at the binding site in this molecule. If probes are designed from the first half of the 18S rRNA molecule, then full-length 18S rRNA molecules can be used in the hybridization on the chip, avoiding the fragmentation and the

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

    DOE PAGES

    Holliday, K. S.; Kohlgruber, T. A.; Tran, I. C.; ...

    2016-08-28

    Development of next generation red phosphors for commercial lighting requires understanding of how increased luminescence is achieved by various treatment strategies. In our 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. Furthermore, the oxidation state of themore » Pr was probed by NEXAFS and revealed that both Nb5+ co-doping and NH3 treatment reduced the number of non-fluorescing Pr4+ centers. We performed calculations in order 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.« less

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

    NASA Astrophysics Data System (ADS)

    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.

  9. Multiple versus single injections of fluorescent microspheres for the determination of regional organ blood flow in septic sheep.

    PubMed

    Lange, Matthias; Hamahata, Atsumori; Traber, Daniel L; Nakano, Yoshimitsu; Traber, Lillian D; Enkhbaatar, Perenlei

    2013-07-01

    Determination of regional blood flow by the injection of microspheres in sepsis models is crucial for the experimental evaluation of the influence of experimental treatment strategies on organ perfusion. However, multiple injections may critically increase the total quantity of microspheres, thereby restricting regional microcirculation and altering the results of blood flow measurements. This study was designed to compare the results of multiple versus single injections of microspheres in an established ovine sepsis model. Injury was induced by smoke inhalation and instillation of Pseudomonas aeruginosa into the lungs. Twenty sheep were studied for 4, 8, 12, 18, or 24 h, respectively. Microspheres were injected at the end of the study period and the animals were euthanized and organ tissues were harvested. Another four sheep were studied for 24 h and multiple microsphere injections were performed at the above indicated time points in the same animals. Tracheal blood flow significantly increased and blood flow to the pancreas and ileum significantly decreased versus baseline in both groups (P < 0.05 each). Blood flow to the ileum, renal cortex and skin did not significantly change versus baseline in both groups (P > 0.05). Blood flow was higher to the trachea in the multiple injection group at 18 h (P = 0.048) and to the ileum at 12 h (P = 0.049), and lower to the skin at 18 h (P = 0.015). In conclusion, the results indicate that multiple versus single microsphere injections induced no or negligible alterations during ovine sepsis. This finding may help reduce the quantity of animals needed in future experiments.

  10. Chair-side detection of Prevotella Intermedia in mature dental plaque by its fluorescence.

    PubMed

    Nomura, Yoshiaki; Takeuchi, Hiroaki; Okamoto, Masaaki; Sogabe, Kaoru; Okada, Ayako; Hanada, Nobuhiro

    2017-06-01

    Prevotella intermedia/nigrescens is one of the well-known pathogens causing periodontal diseases, and the red florescence excited by the visible blue light caused by the protoporphyrin IX in the bacterial cells could be useful for the chair-side detection. The aim of this study was to evaluated levels of periodontal pathogen, especially P. intermedia in clinical samples of red fluorescent dental plaque. Thirty two supra gingival plaque samples from six individuals were measured its fluorescence at 640nm wavelength excited by 409nm. Periodontopathic bacteria were counted by the Invader PLUS PCR assay. Co-relations the fluorescence intensity and bacterial counts were analyzed by Person's correlation coefficient and simple and multiple regression analysis. Positive and negative predictive values of the fluorescence intensities for with or without P. intermedia in supragingival plaque was calculated. When relative fluorescence unit (RFU) were logarithmic transformed, statistically significant linear relations between RFU and bacterial counts were obtained for P. intermedia, Porphyromonas gingivalis and Tannerella forsythia. By the multiple regression analysis, only P. intermedia had statistically significant co-relation with fluorescence intensities. All of the fluorescent dental plaque contained P. intermedia m. In contrast, 28% of non-fluorescent plaques contained P. intermedia. To check the fluorescence dental plaque in the oral cavity could be the simple chair-side screening of the mature dental plaque before examining the periodontal pathogens especially P. intermedia by the PCR method. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. 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).

    PubMed

    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) (p<0.05). Of the bacteria 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.

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

    PubMed

    Hu, Houchun H; Pokorney, Amber; Towbin, Richard B; Miller, Jeffrey H

    2016-10-01

    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.

  13. The on-line each hour and each minute automatically correction data of total NM intensity and different multiplicities on snow effect

    NASA Astrophysics Data System (ADS)

    Lev, Dorman; Zukerman, Igor; Pustilnik, Lev; Dai, Uri; Shternlib, Abracham; Shai Applbaum, David; Kazantsev, Vasilii; Kozliner, Lev; Ben Israel, Isaac

    In our report Dorman et al. “Snow effect for total NM intensity and different multiplicities on Mt. Hermon during 1998 - 2014”, we described the method to determine the snow effect in the total NM intensity and different multiplicities. By using regression coefficients obtained for the long period of observations, obtained in this paper, we developed method of automatically correction on-line each hour and each minute data of total NM intensity and different multiplicities on snow effect. We show that expected average errors in this method for one hour observation is about 0.2%, what is comparable with the statistical error. We show also how to correct on-line automatically one-minute data on snow effect. Corrected on-line one minute data can be now used for the forecasting of great radiation hazards from solar flares and estimation of expected total fluency and radiation hazards for satellites electronics and astronauts health, as well as for people and electronics on regular airlines at altitudes about 10 km. Corrected on-line one hour data can be now used for the forecasting of great magnetic storms, dangerous for satellites, technologies, and people health. https://www.cospar-assembly.org/user/download2.php?id=29566&type=preview

  14. Multiplicity

    DTIC Science & Technology

    1991-04-01

    practice as a "[descent] into that inner circle of the Inferno where the damned endlessly degate multiplicity for sentencing." United States v. Barnard...select the charges to be brought in a particular case"). 19 Brown v. Ohio, 432 U.S. 161, 165 (1977). 20 Whalen v. United States, 445 U.S. at 689. 21...parte Lange, 8-5 U.S. (19 Wall.) 163 (1874). Cf. Brown v. Ohio, 432 U.S. at 165 ("once the legislature has acted courts may not impose more than one

  15. Characterization of the T 1 state of porphyrin molecules on the basis of numerical modeling of decrease and increase of fluorescence intensity kinetics

    NASA Astrophysics Data System (ADS)

    Stanishevsky, I. V.; Arabei, S. M.; Chernyavskii, V. A.; Solovyev, K. N.

    2016-11-01

    For a number of porphyrin molecules, it was shown that their excitation by light pulses of a rectangular step shape and a certain duration led to experimentally observed dynamical decrease and increase of fluorescence kinetics due to changes in the population of the lower (metastable) triplet T 1 state. On the basis of exact analytical expressions for a three-energy-level model, a simple analytical relationships between the rate constants of intramolecular processes and experimentally measured parameters of the fluorescence kinetics were obtained. The three-dimensional isotropic orientation of the molecules in the framework of the chosen model was taken into account by numerical methods and allowed to simulate adequately the experimentally observed fluorescence kinetics. The T 1 state lifetimes of the studied porphyrin molecules in polymer matrices were determined from experimental curves using numerical methods for solving inverse problems. The obtained values correlated with literature data. Features and advantages of this approach were discussed.

  16. Multiple-overtone resonance Raman scattering and fluorescence from I{sub 2} species adsorbed on silver surfaces

    SciTech Connect

    Sibbald, M.S.; Chumanov, G.; Small, G.; Cotton, T.M.

    1998-07-01

    A detailed excitation profile of a Raman progression consisting of up to six overtones and a fundamental band at 123 cm{sup {minus}1} observed from iodide adsorbed on an electrochemically roughened silver surface at 20 K is analyzed. The excitation profile was constructed from 77 spectra obtained by tuning the laser wavelength in {approximately}0.25 nm steps through the spectral range 409 nm{endash}433 nm. The shift between resonances in the excitation profile, corresponding to the spacing between vibronic levels in the excited state, is also equal to 123 cm{sup {minus}1} indicating that the ground state and excited state potential energy surfaces have the same shape. Only two distinct resonances spaced three vibrational quanta apart were evident in the profile for each band in the progression. Curve fitting of the Raman band shapes indicates that each overtone is composed of one sharp and one broad band, whereas the fundamental contains only one sharp component. The measured width of the fundamental was less than 2.5 cm{sup {minus}1} FWHM, limited by the instrument function. It is proposed that the sharp Raman bands represent a normal vibrational mode of a surface-adsorbed I{sub 2}-like species with the width determined by the intrinsic vibrational dephasing in the ground state. On the other hand, the broad Raman bands reflect vibronic coupling between different I{sub 2}-like species adsorbed on the same Ag cluster. The broad bandwidths result from both dephasing associated with the vibronic coupling and the intrinsic vibrational dephasing. Other weak emission bands are attributed to resonance fluorescence corresponding to direct transitions from higher vibronic levels of the excited state to the ground state. An emission at 429.9 nm is assigned to exciton recombination in small silver iodide clusters which are formed after spontaneous oxidation of the iodide-modified silver surface. {copyright} {ital 1998 American Institute of Physics.}

  17. Simultaneous multiple wavelength fluorescence video microscopy shows Ca2+ regulation of pH in living cells

    NASA Astrophysics Data System (ADS)

    Morris, Stephen J.; Beatty, Diane M.; Chronwall, Bibie M.

    1994-08-01

    We have designed an epifluorescence video microscope for simultaneous dual excitation of indo 1 (for [Ca2+]i) at 350 nm and SNARF 1 (for pHi) at 540 nm. The microscope will simultaneously capture all four emission images at 405, 475, 575, and 640 nm from the two ratio dyes at video frame or field rates. Popular dyes for measuring [Ca2+]i, such as indo 1 and fura 2, have pH- dependent Kd's; thus changes in pHi can be misinterpreted as changes in [Ca2+]i. For any pixel (or region of interest), we use the pH value to generate the appropriate Kd. The corrected Kd is then used to calculate a corrected calcium value. Using the imaging system, we show that, for the peptide secreting melanotropes of the pituitary intermediate lobe, changes in intracellular calcium ([Ca2PLU)]i) produce changes in intracellular pH (pHi). Melanotropes grown in primary explant culture and double-loaded with indo-1 acetoxy methyl ester (AM) and SNARF-1 AM, were examined for Ca2+/pH interactions. Following experimentation, cells were positively identified by (beta) -endorphin fluorescence immunohistochemistry. K-induced depolarization of melanotropes produced increases in [Ca2PLU)]i due to activation of L-type Ca-channels. Ca2+ entry was closely coupled to reductions in pHi. Effects were dependent upon entry of extracellular Ca2+ rather than release from intracellular stores. The close association between increases in intracellular Ca2+ and H+ suggest that the pHi changes are due to release of H+ upon binding of Ca2+ to intracellular buffers. Although the pH drop is `passive,' it will be sensed by all cytoplasmic components. Thus it represents a second messenger pathway, akin to the generation of cAMP or inositol trisphosphate, which cannot fail to influence numerous pH-dependent cell activities.

  18. Prevalence of multiple organ dysfunction in the pediatric intensive care unit: Pediatric Risk of Mortality III versus Pediatric Logistic Organ Dysfunction scores for mortality prediction

    PubMed Central

    Hamshary, Azza Abd Elkader El; Sherbini, Seham Awad El; Elgebaly, HebatAllah Fadel; Amin, Samah Abdelkrim

    2017-01-01

    Objectives To assess the frequency of primary multiple organ failure and the role of sepsis as a causative agent in critically ill pediatric patients; and calculate and evaluate the accuracy of the Pediatric Risk of Mortality III (PRISM III) and Pediatric Logistic Organ Dysfunction (PELOD) scores to predict the outcomes of critically ill children. Methods Retrospective study, which evaluated data from patients admitted from January to December 2011 in the pediatric intensive care unit of the Children's Hospital of the University of Cairo. Results Out of 237 patients in the study, 72% had multiple organ dysfunctions, and 45% had sepsis with multiple organ dysfunctions. The mortality rate in patients with multiple organ dysfunction was 73%. Independent risk factors for death were mechanical ventilation and neurological failure [OR: 36 and 3.3, respectively]. The PRISM III score was more accurate than the PELOD score in predicting death, with a Hosmer-Lemeshow X2 (Chi-square value) of 7.3 (df = 8, p = 0.5). The area under the curve was 0.723 for PRISM III and 0.78 for PELOD. Conclusion A multiple organ dysfunctions was associated with high mortality. Sepsis was the major cause. Pneumonia, diarrhea and central nervous system infections were the major causes of sepsis. PRISM III had a better calibration than the PELOD for prognosis of the patients, despite the high frequency of the multiple organ dysfunction syndrome.

  19. Fluorescent zinc indicators for neurobiology.

    PubMed

    Thompson, R B; Peterson, Dwight; Mahoney, William; Cramer, Michele; Maliwal, Badri P; Suh, Sang Won; Frederickson, Chris; Fierke, Carol; Herman, Petr

    2002-07-30

    Mounting evidence indicates that zinc has multiple roles in cell biology, viz. as a part of metalloenzyme catalytic sites, as a structural component of gene regulatory proteins, and (like calcium) as a free signal ion, particularly in the cortex of the brain. While most Zn(II) in the brain is tightly bound, such that free Zn(II) levels extracellularly and intracellularly are likely to be picomolar, a subset of glutamatergic neurons possess weakly bound zinc in presynaptic boutons which is released at micromolar levels in response to a variety of stimuli. Key to further progress in understanding the multiple roles of zinc will be the availability of fluorescent indicator systems that will permit quantitative determination and imaging of zinc fluxes and levels over a broad concentration range both intracellularly and extracellularly using fluorescence microscopy. Towards that end, we have compared a variety of fluorescent indicators for their sensitivity to Zn(II) and Cu(II), selectivity for Zn(II) in the presence of potential interferents such as Ca(II) or Mg(II), and potential for quantitative imaging. The commercially available probes Fura-2, Mag-Fura-5, Newport Green DCF, and FuraZin-1 were compared with the carbonic anhydrase-based indicator systems for selectivity and sensitivity. In addition, intracellular levels of Zn following excitotoxic insult were determined by single pixel fluorescence lifetime microscopy of Newport Green DCF, and extracellular levels of free zinc following stimulus of rat hippocampal slices were determined ratiometrically with a carbonic anhydrase-based indicator system. These results suggest that zinc ion at high nM to microM levels can be accurately quantitated by FuraZin-1 ratiometrically or by Newport Green DCF by fluorescence lifetime; and at levels down to pM by intensity ratio, lifetime, or polarization using carbonic anhydrase-based systems. Copyright 2002 Elsevier Science B.V.

  20. Stochastic extreme downscaling model for an assessment of changes in rainfall intensity-duration-frequency curves over South Korea using multiple regional climate models

    NASA Astrophysics Data System (ADS)

    So, Byung-Jin; Kim, Jin-Young; Kwon, Hyun-Han; Lima, Carlos H. R.

    2017-10-01

    A conditional copula function based downscaling model in a fully Bayesian framework is developed in this study to evaluate future changes in intensity-duration frequency (IDF) curves in South Korea. The model incorporates a quantile mapping approach for bias correction while integrated Bayesian inference allows accounting for parameter uncertainties. The proposed approach is used to temporally downscale expected changes in daily rainfall, inferred from multiple CORDEX-RCMs based on Representative Concentration Pathways (RCPs) 4.5 and 8.5 scenarios, into sub-daily temporal scales. Among the CORDEX-RCMs, a noticeable increase in rainfall intensity is observed in the HadGem3-RA (9%), RegCM (28%), and SNU_WRF (13%) on average, whereas no noticeable changes are observed in the GRIMs (-2%) for the period 2020-2050. More specifically, a 5-30% increase in rainfall intensity is expected in all of the CORDEX-RCMs for 50-year return values under the RCP 8.5 scenario. Uncertainty in simulated rainfall intensity gradually decreases toward the longer durations, which is largely associated with the enhanced strength of the relationship with the 24-h annual maximum rainfalls (AMRs). A primary advantage of the proposed model is that projected changes in future rainfall intensities are well preserved.

  1. COMPARISON OF RANDOM SITE SELECTION AT MULTIPLE INTENSITIES FOR THE ASSESSMENT OF THE OHIO RIVER FISH COMMUNITY

    EPA Science Inventory

    The Ohio River Valley Sanitation Commission (ORSANCO) is a compact of eight states representing interests in the Ohio River basin that has been instrumental in the development of biological monitoring of the Ohio River. In the past, ORSANCO has conducted intensive surveys by samp...

  2. Development of a hybrid modeling approach for predicting intensively managed Douglas-fir growth at multiple scales.

    Treesearch

    A. Weiskittel; D. Maguire; R. Monserud

    2007-01-01

    Hybrid models offer the opportunity to improve future growth projections by combining advantages of both empirical and process-based modeling approaches. Hybrid models have been constructed in several regions and their performance relative to a purely empirical approach has varied. A hybrid model was constructed for intensively managed Douglas-fir plantations in the...

  3. COMPARISON OF RANDOM SITE SELECTION AT MULTIPLE INTENSITIES FOR THE ASSESSMENT OF THE OHIO RIVER FISH COMMUNITY

    EPA Science Inventory

    The Ohio River Valley Sanitation Commission (ORSANCO) is a compact of eight states representing interests in the Ohio River basin that has been instrumental in the development of biological monitoring of the Ohio River. In the past, ORSANCO has conducted intensive surveys by samp...

  4. Investigating State Restriction in Fluorescent Protein FRET Using Time-Resolved Fluorescence and Anisotropy

    PubMed Central

    2016-01-01

    Most fluorescent proteins exhibit multiexponential fluorescence decays, indicating a heterogeneous excited state population. FRET between fluorescent proteins should therefore involve multiple energy transfer pathways. We recently demonstrated the FRET pathways between EGFP and mCherry (mC), upon the dimerization of 3-phosphoinositide dependent protein kinase 1 (PDK1), to be highly restricted. A mechanism for FRET restriction based on a highly unfavorable κ2 orientation factor arising from differences in donor–acceptor transition dipole moment angles in a far from coplanar and near static interaction geometry was proposed. Here this is tested via FRET to mC arising from the association of glutathione (GSH) and glutathione S-transferase (GST) with an intrinsically homogeneous and more mobile donor Oregon Green 488 (OG). A new analysis of the acceptor window intensity, based on the turnover point of the sensitized fluorescence, is combined with donor window intensity and anisotropy measurements which show that unrestricted FRET to mC takes place. However, a long-lived anisotropy decay component in the donor window reveals a GST-GSH population in which FRET does not occur, explaining previous discrepancies between quantitative FRET measurements of GST-GSH association and their accepted values. This reinforces the importance of the local donor–acceptor environment in mediating energy transfer and the need to perform spectrally resolved intensity and anisotropy decay measurements in the accurate quantification of fluorescent protein FRET. PMID:28217242

  5. UV radiation at the fluorescence excitation maxima produces significant changes in the fluorescence of skin

    NASA Astrophysics Data System (ADS)

    Kollias, Nikiforos; Tian, W. D.; Zonios, George I.; Brancaleon, Lorenzo; Gillies, Robert

    1999-06-01

    Fluorescence excitation spectra of skin have been determined to be stable and reproducible. Three major bands dominate the wavelength range 280-400 nm. The major epidermal band due to tryptophan moieties appears at 295+/-5 nm and the major dermal bands due to collagen cross links appear at 335+/-5 nm and at 370+/-5 nm. The tryptophan fluorescence intensity has been found to increase with exposure to UV radiation; the UVB wavelengths are more effective than the UVA wavelengths. The PDCCL fluorescence intensity has been found to decrease dramatically with exposure to UVA in a wavelength specific way. The maximum of the action spectrum for this process is centered at the maximum of the excitation spectrum. The fluorescence of the skin recovers within 24 hours following exposure to UVA from single exposures. Multiple exposures produce permanent changes, in a follow-up of 8 weeks. the changes in the tryptophan fluorescence are probably due to changes in the molecular environment brought about by changes in the electrolyte balance in the epidermis following exposure. The changes in the dermis following UVA exposure appear to be associated with change in the collagen cross links, either through their association with other dermal species leading to quenching of the fluorescence or by the formation of stronger cross links with a smaller quantum efficiency. As these changes are immediate both alterations may provide the means for in vivo UV dosimetry.

  6. Monte Carlo fluorescence microtomography

    NASA Astrophysics Data System (ADS)

    Cong, Alexander X.; Hofmann, Matthias C.; Cong, Wenxiang; Xu, Yong; Wang, Ge

    2011-07-01

    Fluorescence microscopy allows real-time monitoring of optical molecular probes for disease characterization, drug development, and tissue regeneration. However, when a biological sample is thicker than 1 mm, intense scattering of light would significantly degrade the spatial resolution of fluorescence microscopy. In this paper, we develop a fluorescence microtomography technique that utilizes the Monte Carlo method to image fluorescence reporters in thick biological samples. This approach is based on an l0-regularized tomography model and provides an excellent solution. Our studies on biomimetic tissue scaffolds have demonstrated that the proposed approach is capable of localizing and quantifying the distribution of optical molecular probe accurately and reliably.

  7. Fluorescence lifetime imaging ophthalmoscopy.

    PubMed

    Dysli, Chantal; Wolf, Sebastian; Berezin, Mikhail Y; Sauer, Lydia; Hammer, Martin; Zinkernagel, Martin S

    2017-09-01

    Imaging techniques based on retinal autofluorescence have found broad applications in ophthalmology because they are extremely sensitive and noninvasive. Conventional fundus autofluorescence imaging measures fluorescence intensity of endogenous retinal fluorophores. It mainly derives its signal from lipofuscin at the level of the retinal pigment epithelium. Fundus autofluorescence, however, can not only be characterized by the spatial distribution of the fluorescence intensity or emission spectrum, but also by a characteristic fluorescence lifetime function. The fluorescence lifetime is the average amount of time a fluorophore remains in the excited state following excitation. Fluorescence lifetime imaging ophthalmoscopy (FLIO) is an emerging imaging modality for in vivo measurement of lifetimes of endogenous retinal fluorophores. Recent reports in this field have contributed to our understanding of the pathophysiology of various macular and retinal diseases. Within this review, the basic concept of fluorescence lifetime imaging is provided. It includes technical background information and correlation with in vitro measurements of individual retinal metabolites. In a second part, clinical applications of fluorescence lifetime imaging and fluorescence lifetime features of selected retinal diseases such as Stargardt disease, age-related macular degeneration, choroideremia, central serous chorioretinopathy, macular holes, diabetic retinopathy, and retinal artery occlusion are discussed. Potential areas of use for fluorescence lifetime imaging ophthalmoscopy will be outlined at the end of this review. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  8. Fluorescence lidar method for remote monitoring of effects on vegetation

    NASA Astrophysics Data System (ADS)

    Matvienko, Gennady; Timofeev, Valery; Grishin, Anatoly; Fateyeva, Natalia

    2006-09-01

    Plants constantly interact with environment, mainly, by means of photosynthesis and soil nutrition. The state of plant photosynthetic apparatus that reflects the general physiological state of a plant, can be analyzed remotely on a basis of laser-induced fluorescence using a fluorescence lidar. In this respect, a fluorescence lidar can be a technical means of remote sensing of the effects on vegetation including chemical soil pollution. Among a series of applications, of interest is development of a lidar technique for detecting the effects of oil products and mechanical disturbances. This paper is devoted to the application of the fluorescence lidar technique to monitoring mechanical and chemical impacts on the woody vegetation typical of Siberia. A physical basis of this technique is the red fluorescence of chlorophyll of green plants excited by the second harmonic (532 nm) of Nd:YAG laser. Red fluorescence of plants consists of two bands centered at 685 and 740 nm which is conditioned by functioning of two photosystems. As in situ experiments show, the indicated photosystems and, respectively, the fluorescence on these bands respond differently to feeding disturbances and mechanical impacts, making the increase in the fluorescence intensity informative. Time criteria of fluorescence characteristics were obtained at single and multiple effects on the vegetation. The paper describes a lidar system that meets the requirements for detecting the effects on vegetation.

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

    PubMed

    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.

  10. Balanced crystalloids versus saline in the intensive care unit: study protocol for a cluster-randomized, multiple-crossover trial.

    PubMed

    Semler, Matthew W; Self, Wesley H; Wang, Li; Byrne, Daniel W; Wanderer, Jonathan P; Ehrenfeld, Jesse M; Stollings, Joanna L; Kumar, Avinash B; Hernandez, Antonio; Guillamondegui, Oscar D; May, Addison K; Siew, Edward D; Shaw, Andrew D; Bernard, Gordon R; Rice, Todd W

    2017-03-16

    Saline, the intravenous fluid most commonly administered to critically ill adults, contains a high chloride content, which may be associated with acute kidney injury and death. Whether using balanced crystalloids rather than saline decreases the risk of acute kidney injury and death among critically ill adults remains unknown. The Isotonic Solutions and Major Adverse Renal Events Trial (SMART) is a pragmatic, cluster-level allocation, cluster-level crossover trial being conducted between 1 June 2015 and 30 April 2017 in five intensive care units at Vanderbilt University Medical Center in Nashville, TN, USA. SMART compares saline (0.9% sodium chloride) with balanced crystalloids (clinician's choice of lactated Ringer's solution or Plasma-Lyte A®). Each intensive care unit is assigned to provide either saline or balanced crystalloids each month, with the assigned crystalloid alternating monthly over the course of the trial. All adults admitted to participating intensive care units during the study period are enrolled and followed until hospital discharge or 30 days after enrollment. The anticipated enrollment is approximately 14,000 patients. The primary outcome is Major Adverse Kidney Events within 30 days-the composite of in-hospital death, receipt of new renal replacement therapy, or persistent renal dysfunction (discharge creatinine ≥200% of baseline creatinine). Secondary clinical outcomes include in-hospital mortality, intensive care unit-free days, ventilator-free days, vasopressor-free days, and renal replacement therapy-free days. Secondary renal outcomes include new renal replacement therapy receipt, persistent renal dysfunction, and incidence of stage 2 or higher acute kidney injury. This ongoing pragmatic trial will provide the largest and most comprehensive comparison to date of clinical outcomes with saline versus balanced crystalloids among critically ill adults. For logistical reasons, SMART was prospectively registered separately for the medical

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

    PubMed

    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.

  12. Effects of self-hypnosis training and cognitive restructuring on daily pain intensity and catastrophizing in individuals with multiple sclerosis and chronic pain.

    PubMed

    Jensen, Mark P; Ehde, Dawn M; Gertz, Kevin J; Stoelb, Brenda L; Dillworth, Tiara M; Hirsh, Adam T; Molton, Ivan R; Kraft, George H

    2011-01-01

    Fifteen adults with multiple sclerosis were given 16 sessions of treatment for chronic pain that included 4 sessions each of 4 different treatment modules: (a) an education control intervention; (b) self-hypnosis training (HYP); (c) cognitive restructuring (CR); and (d) a combined hypnosis-cognitive restructuring intervention (CR-HYP). The findings supported the greater beneficial effects of HYP, relative to CR, on average pain intensity. The CR-HYP treatment appeared to have beneficial effects greater than the effects of CR and HYP alone. Future research examining the efficacy of an intervention that combines CR and HYP is warranted.

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

    NASA Astrophysics Data System (ADS)

    McMillan, A. M. S.; Harvey, M. J.; Martin, R. J.; Bromley, A. M.; Evans, M. J.; Mukherjee, S.; Laubach, J.

    2013-10-01

    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 m and 1.0 m. The gradient resolution achieved corresponded to the ability to detect an inter-plot N2O flux difference of 26.4 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.

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

    NASA Astrophysics Data System (ADS)

    McMillan, A. M. S.; Harvey, M. J.; Martin, R. J.; Bromley, A. M.; Evans, M. J.; Mukherjee, S.; Laubach, J.

    2014-05-01

    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.

  15. Polar plot representation of time-resolved fluorescence.

    PubMed

    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.

  16. Impacts of cloud superparameterization on projected daily rainfall intensity climate changes in multiple versions of the Community Earth System Model

    DOE PAGES

    Kooperman, Gabriel J.; Pritchard, Michael S.; Burt, Melissa A.; ...

    2016-09-26

    Changes in the character of rainfall are assessed using a holistic set of statistics based on rainfall frequency and amount distributions in climate change experiments with three conventional and superparameterized versions of the Community Atmosphere Model (CAM and SPCAM). Previous work has shown that high-order statistics of present-day rainfall intensity are significantly improved with superparameterization, especially in regions of tropical convection. Globally, the two modeling approaches project a similar future increase in mean rainfall, especially across the Inter-Tropical Convergence Zone (ITCZ) and at high latitudes, but over land, SPCAM predicts a smaller mean change than CAM. Changes in high-order statisticsmore » are similar at high latitudes in the two models but diverge at lower latitudes. In the tropics, SPCAM projects a large intensification of moderate and extreme rain rates in regions of organized convection associated with the Madden Julian Oscillation, ITCZ, monsoons, and tropical waves. In contrast, this signal is missing in all versions of CAM, which are found to be prone to predicting increases in the amount but not intensity of moderate rates. Predictions from SPCAM exhibit a scale-insensitive behavior with little dependence on horizontal resolution for extreme rates, while lower resolution (~2°) versions of CAM are not able to capture the response simulated with higher resolution (~1°). Furthermore, moderate rain rates analyzed by the “amount mode” and “amount median” are found to be especially telling as a diagnostic for evaluating climate model performance and tracing future changes in rainfall statistics to tropical wave modes in SPCAM.« less

  17. Long-term dynamics of tropical walking sticks in response to multiple large-scale and intense disturbances.

    PubMed

    Willig, Michael R; Presley, Steven J; Bloch, Christopher P

    2011-02-01

    Understanding the effects of disturbance and secondary succession on spatio-temporal patterns in the abundance of species is stymied by a lack of long-term demographic data, especially in response to infrequent and high intensity disturbances, such as hurricanes. Moreover, resistance and resilience to hurricane-induced disturbance may be mediated by legacies of previous land use, although such interactive effects are poorly understood, especially in tropical environments. We address these central issues in disturbance ecology by analyzing an extensive dataset, spanning the impacts of Hurricanes Hugo and Georges, on the abundance of a Neotropical walking stick, Lamponius portoricensis, in tabonuco rainforest of Puerto Rico during the wet and dry seasons from 1991 to 2007. By synthesizing data from two proximate sites in tabonuco forest, we show that resistance to Hurricane Hugo (97% reduction in abundance) was much less than resistance to Hurricane Georges (21% reduction in abundance). Based on a powerful statistical approach (generalized linear mixed-effects models with Poisson error terms), we documented that the temporal trajectories of abundance during secondary succession (i.e., patterns of resilience) differed between hurricanes and among historical land use categories, but that the effects of hurricanes and land use histories were independent of each other. These complex results likely arise because of differences in the intensities of the two hurricanes with respect to microclimatic effects (temperature and moisture) in the forest understory, as well as to time-lags in the response of L. portoricensis to changes in the abundance and distribution of preferred food plants (Piper) in post-hurricane environments.

  18. Multiple Escherichia coli RecQ helicase monomers cooperate to unwind long DNA substrates: a fluorescence cross-correlation spectroscopy study.

    PubMed

    Li, Na; Henry, Etienne; Guiot, Elvire; Rigolet, Pascal; Brochon, Jean-Claude; Xi, Xu-Guang; Deprez, Eric

    2010-03-05

    The RecQ family helicases catalyze the DNA unwinding reaction in an ATP hydrolysis-dependent manner. We investigated the mechanism of DNA unwinding by the Escherichia coli RecQ helicase using a new sensitive helicase assay based on fluorescence cross-correlation spectroscopy (FCCS) with two-photon excitation. The FCCS-based assay can be used to measure the unwinding activity under both single and multiple turnover conditions with no limitation related to the size of the DNA strands constituting the DNA substrate. We found that the monomeric helicase was sufficient to perform the unwinding of short DNA substrates. However, a significant increase in the activity was observed using longer DNA substrates, under single turnover conditions, originating from the simultaneous binding of multiple helicase monomers to the same DNA molecule. This functional cooperativity was strongly dependent on several factors, including DNA substrate length, the number and size of single-stranded 3'-tails, and the temperature. Regarding the latter parameter, a strong cooperativity was observed at 37 degrees C, whereas only modest or no cooperativity was observed at 25 degrees C regardless of the nature of the DNA substrate. Consistently, the functional cooperativity was found to be tightly associated with a cooperative DNA binding mode. We also showed that the cooperative binding of helicase to the DNA substrate indirectly accounts for the sigmoidal dependence of unwinding activity on ATP concentration, which also occurs only at 37 degrees C but not at 25 degrees C. Finally, we further examined the influences of spontaneous DNA rehybridization (after helicase translocation) and the single-stranded DNA binding property of helicase on the unwinding activity as detected in the FCCS assay.

  19. CO2, H2O, and chlorophyll fluorescence retrieved from OCO-2 measurements using a fast radiative transfer model approximating multiple scattering effects

    NASA Astrophysics Data System (ADS)

    Reuter, Maximilian; Bovensmann, Heinrich; Buchwitz, Michael; Burrows, John P.; Heymann, Jens; Noël, Stefan; Rozanov, Vladimir; Schneising, Oliver

    2017-04-01

    Carbon dioxide is the most important anthropogenic greenhouse gas. Its global increasing concentration in the Earth's atmosphere is the main driver for global climate change. In spite of its importance, there are still large uncertainties on its global sources and sinks. Satellite measurements have the potential to reduce these surface flux uncertainties. However, the demanding accuracy requirements usually involve the need for precise radiative transfer calculations in a scattering atmosphere. These can be computationally so expensive that hundreds or thousands of CPU cores are need to keep up with the data stream of an instrument like OCO-2. Future instruments will further increase the amount of soundings at least by an order of magnitude. A radiative transfer model has been developed approximating scattering effects by multiple scattering at an optically thin scattering layer reducing the computational costs by several orders of magnitude. The model can be used to simulate the radiance in all three OCO-2 spectral bands allowing the simultaneous retrieval of CO2, H2O, and chlorophyll fluorescence. First retrieval results for OCO-2 data will be presented.

  20. Identification of peanut (Arachis hypogaea) chromosomes using a fluorescence in situ hybridization system reveals multiple hybridization events during tetraploid peanut formation.

    PubMed

    Zhang, Laining; Yang, Xiaoyu; Tian, Li; Chen, Lei; Yu, Weichang

    2016-09-01

    The cultivated peanut Arachis hypogaea (AABB) is thought to have originated from the hybridization of Arachis duranensis (AA) and Arachis ipaënsis (BB) followed by spontaneous chromosome doubling. In this study, we cloned and analyzed chromosome markers from cultivated peanut and its wild relatives. A fluorescence in situ hybridization (FISH)-based karyotyping cocktail was developed with which to study the karyotypes and chromosome evolution of peanut and its wild relatives. Karyotypes were constructed in cultivated peanut and its two putative progenitors using our FISH-based karyotyping system. Comparative karyotyping analysis revealed that chromosome organization was highly conserved in cultivated peanut and its two putative progenitors, especially in the B genome chromosomes. However, variations existed between A. duranensis and the A genome chromosomes in cultivated peanut, especially for the distribution of the interstitial telomere repeats (ITRs). A search of additional A. duranensis varieties from different geographic regions revealed both numeric and positional variations of ITRs, which were similar to the variations in tetraploid peanut varieties. The results provide evidence for the origin of cultivated peanut from the two diploid ancestors, and also suggest that multiple hybridization events of A. ipaënsis with different varieties of A. duranensis may have occurred during the origination of peanut. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

  1. Introducing Amylo-Glo, a novel fluorescent amyloid specific histochemical tracer especially suited for multiple labeling and large scale quantification studies.

    PubMed

    Schmued, Larry; Raymick, James; Tolleson, William; Sarkar, Sumit; Zhang, Yi-Hong; Bell-Cohn, Ashlee

    2012-07-30

    One of the hallmark pathologies associated with Alzheimer's disease (AD) is the conspicuous deposition of extracellular amyloid plaques within the forebrain. These plaques are primarily composed of fibrular aggregates of the A-beta peptide. Traditional methods for the histological localization of these plaques typically rely on the use of the tracers Congo Red or Thioflavin S. This study describes the characterization of a novel fluorescent histochemical probe, Amylo-Glo, for the high resolution and contrast localization of amyloid plaques in brain tissue sections. Potential advantages over conventional amyloid plaque stains such as Congo Red or Thioflavin S can be attributed to its unique chemical and spectral properties. Specifically, it results in a very bright blue UV excitable stain under physiological conditions that will not bleed through when illuminated with other filters. Its brightness makes it ideal for low magnification quantification studies, while its unique excitation/emission profile and mild staining conditions makes it ideal for combination with multiple immunofluorescent labeling studies. Published by Elsevier B.V.

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

    PubMed

    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.

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

    PubMed Central

    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 (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. PMID:26623308

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

    PubMed

    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.

  5. Low intensity lympho-ablative regimen followed by autologous hematopoietic stem cell transplantation in severe forms of multiple sclerosis: A MRI-based clinical study.

    PubMed

    Curro', Daniela; Vuolo, Luisa; Gualandi, Francesca; Bacigalupo, Andrea; Roccatagliata, Luca; Capello, Elisabetta; Uccelli, Antonio; Saccardi, Riccardo; Sormani, Maria Pia; Mancardi, Gianluigi

    2015-10-01

    Autologous hematopoietic stem cell transplantation (AHSCT) has been successfully used to treat aggressive forms of multiple sclerosis (MS) that are unresponsive to approved therapies. In the last years, in view of the risk of mortality related to the procedure, the utilization of low-intensity conditioning regimens has been considered. To report magnetic resonance imaging (MRI) and clinical data in a small cohort of patients treated with a low-intensity lympho-ablative regimen, followed by AHSCT. Seven patients affected by relapsing-remitting MS (RRMS) underwent AHSCT, with cyclophosphamide 120 mg/kg in 2 days as the conditioning regimen; and were then followed with serial MRI evaluations until 36 months, with clinical evaluations until 60 months. The mean number of gadolinium (Gd)-enhancing lesions significantly decreased after treatment, but a complete suppression of inflammatory activity was not obtained. No deaths occurred, but every patient developed adverse events, although not severe. After 5 years of follow-up, two patients remained stable, one patient markedly improved and four patients had a mild progression of the disease. Only one patient experienced a relapse after treatment. A low-intensity conditioning regimen with AHSCT has a profound effect on MRI inflammation and relapses, but is not able to completely abrogate MRI activity and disease progression of aggressive RRMS. © The Author(s), 2015.

  6. The CREATE Strategy for Intensive Analysis of Primary Literature Can Be Used Effectively by Newly Trained Faculty to Produce Multiple Gains in Diverse Students

    PubMed Central

    Stevens, Leslie M.

    2014-01-01

    The CREATE (Consider Read, Elucidate the hypotheses, Analyze and interpret the data, and Think of the next Experiment) strategy aims to demystify scientific research and scientists while building critical thinking, reading/analytical skills, and improved science attitudes through intensive analysis of primary literature. CREATE was developed and piloted at the City College of New York (CCNY), a 4-yr, minority-serving institution, with both upper-level biology majors and first-year students interested in science, technology, engineering, and mathematics. To test the extent to which CREATE strategies are broadly applicable to students at private, public, research-intensive, and/or primarily undergraduate colleges/universities, we trained a cohort of faculty from the New York/New Jersey/Pennsylvania area in CREATE pedagogies, then followed a subset, the CREATE implementers (CIs), as they taught all or part of an existing course on their home campuses using CREATE approaches. Evaluation of the workshops, the CIs, and their students was carried out both by the principal investigators and by an outside evaluator working independently. Our data indicate that: intensive workshops change aspects of faculty attitudes about teaching/learning; workshop-trained faculty can effectively design and teach CREATE courses; and students taught by such faculty on multiple campuses make significant cognitive and affective gains that parallel the changes documented previously at CCNY. PMID:26086655

  7. Quantification of fluorescent reporters in plant cells.

    PubMed

    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.

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

    NASA Astrophysics Data System (ADS)

    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., <5 GPa) shock events. Aqueous alteration was enhanced by those impacts that were of sufficient intensity to open high permeability fracture networks that could connect to fluid reservoirs.

  9. Amobarbital treatment of multiple personality. Use of structured video tape interviews as a basis for intensive psychotherapy.

    PubMed

    Hall, R C; LeCann, A F; Schoolar, J C

    1978-09-01

    The case of a 30-year-old woman with five distinct personalities is presented. The patient was treated, using a system of structured video taped sodium amobarbital interviews, in which areas to be explored were developed in psychotherapy. Tapes were played for the patient after each session. The taped material was used as the basis for psychotherapeutic investigation. The patient evidenced many of the features previously reported in cases of multiple personality, specifically: being the product of an unwanted pregnancy in a repressively rigid family; emotional distancing by one parent; strong sibling rivalry with an adopted sib; family history of mental illness; a traumatic first sexual experience (rape); a marriage to a maladjusted individual in an attempt to escape the parental home; a high internalized standard of performance and an inability to display anger or negative feelings toward the parents. In the course of treatment, the patient's personalties fused and she was able to accept each component as part of herself. No further fragmentation has occurred during the year following discharge. The therapy technique minimized dependency, and the possiblity of addiction to amobarbital interviews permitted more active patient therapy involvement, and set clear-cut goals and expectations for improvement before further amobarbital interviews could be conducted.

  10. 3D multiple optical trapping of Au-nanoparticles and prokaryote E. coli using intra-cavity generated non-circular beam of inhomogeneous intensity

    NASA Astrophysics Data System (ADS)

    Kumar, R.; Shakher, C.; Mehta, D. S.

    2010-06-01

    We report 3D multiple trapping of dielectric polystyrene (PS) beads and gold nano-particles (GNPs) in single beam optical tweezers system using an asymmetric beam of inhomogeneous intensity distribution. This special kind of beam of quasi-TEM11 profile was generated from intra-cavity CW-laser source operating at 532 nm. Multiple trapping of both the low refractive index rod-like Escherichia coli bacteria and 253 nm plasmonic GNPs dispersed in 1.025 μm PS beads which were homogenized in de-ionized water was realized utilizing this spatial beam. Laser-GNPs interaction rendered the enhancement of local surface plasmon resonance field around GNPs causing long-range aggregation of PS beads. The multiple trapping of plasmonic GNPs by the present simple method might find applications for micro- and nano-connectors, underlying physical processes in light-matter interaction assays for inter-particle force analysis, cancer diagnostic and photothermolysis, surface-enhanced Raman scattering (SERS) spectroscopy, and surface plasmon based biological and chemical sensors.

  11. Efficacy and safety of reduced-intensity induction therapy with a bortezomib-based regimen in elderly patients with multiple myeloma

    PubMed Central

    Sopeña, María; Clavero, Estela Martín; Villa, Paula

    2012-01-01

    The therapeutic effects of bortezomib in untreated and refractory/relapsed multiple myeloma have been demonstrated in several clinical trials, displaying superiority to the conventional treatments. However, many treatment-related toxicities, such as bone marrow suppression, infections and peripheral neuropathy, are well known and lead to treatment discontinuation and dose modification, especially in elderly patients. The purpose of this review is to summarize the published literature concerning the efficacy and safety of reduced-intensity induction therapy with bortezomib-based regimens in elderly patients with multiple myeloma. We used the VISTA trial as a reference and compared it with the seven trials identified in a systematic search. The data suggest that low-dose bortezomib significantly reduces therapy-related toxicities, especially neuropathy, and decreases the rate of discontinuation compared with the twice-weekly regimen, without losing efficacy. In light of this review, we suggest that once-weekly infusion of bortezomib in addition to melphalan–prednisone may be considered as a new standard of care in frontline treatment of elderly patients with symptomatic multiple myeloma. PMID:23556121

  12. Use of Tunable, Pulsed Dye Laser for Quantitative Fluorescence in Syphilis Serology (FTA-ABS Test)

    PubMed Central

    Kasatiya, S. S.; Lambert, N. G.; Laurence, R. A.

    1974-01-01

    A pulsed dye laser was used as an excitation source in a fluorescent treponemal antibody absorption (FTA-ABS) test. A high precision in quantitative fluorescence was obtained with this high-power excitation source coupled to an electronic detection system and a storage oscilloscope by standardization of fluorescence evaluation and through elimination of human error. One 0.4-μs pulse exposure was sufficient to record fluorescence intensity data on the oscilloscope. Absence of fading of fluorescence after repeated excitation permitted multiple readings of the same microscope field. Almost 100% reproducible results were obtained for the FTA-ABS test with 40 samples. Electronic detection of fluorescence and the high sensitivity obtained with laser excitation raise doubts about the relative value of quantitative immunofluorescence in the FTA-ABS test. PMID:4598221

  13. Fluorescent Applications to Crystallization

    NASA Technical Reports Server (NTRS)

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

    2006-01-01

    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 tests with model proteins have shown that labeling u to 5 percent of the protein molecules does not affect the X-ray data quality obtained . The presence of the trace fluorescent label gives a number of advantages. Since the label is covalently attached to the protein molecules, it "tracks" the protein s response to the crystallization conditions. 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 darker background. Non-protein structures, such as salt crystals, do not show up under fluorescent illumination. Crystals have the highest protein concentration and are readily observed 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. Preliminary tests, using model proteins, indicates that we can use 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 more rapid amorphous precipitation kinetics may overwhelm and trap more slowly formed ordered assemblies, which subsequently show up as regions of brighter fluorescence intensity. Experiments are now being carried out to test this approach using a wider range, of proteins. The trace fluorescently labeled crystals will also

  14. Intense Inflammation and Nerve Damage in Early Multiple Sclerosis Subsides at Older Age: A Reflection by Cerebrospinal Fluid Biomarkers

    PubMed Central

    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

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

    PubMed

    Danielson, Eric; Lee, Sang H

    2014-01-01

    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.

  16. Absolute brightness of fluorescent microspheres.

    PubMed

    Finger, Isaac; Phillips, Scott; Mobley, Elizabeth; Tucker, Robert; Hess, Henry

    2009-02-07

    The absolute brightness of fluorescent particles, such as dye-containing nano- and microspheres or quantum dots, is a critical design parameter for many applications relying on fluorescence detection. The absolute brightness, defined as the ratio of radiant intensity of emission to illumination intensity of excitation, of nile-red fluorescent microspheres with a 1 micrometre diameter is measured to be 4.2 +/- 1 x 10(-16) m(2)/sr, and the implications for the design of kinesin motor protein-powered "smart dust" devices and the remote detection of fluorescence are discussed.

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

    PubMed

    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.

  18. The evaluation of minimal residual disease in multiple myeloma by fluorescent molecular beacons in real time PCR of IgH gene rearrangements and correlation with flow cytometry.

    PubMed

    Kara, I O; Duman, B B; Afsar, C U

    2013-01-01

    Multiple myeloma (MM) patients relapse after a period of time despite longer disease-free survival due to novel treatment options. In this study we aimed to assess the value of real-time polymerase chain reaction (RT-PCR) for detecting the immunoglobulin heavy chain (IgH) gene rearrangement using allele-specific molecular beacons as fluorescence probes to quantify minimal residual disease (MRD) and also to correlate post-treatment flow cytometric detection of plasma cells' (PCs) expression of CD19, CD38, CD45, CD56 and CD138 in MM. After diagnosis of 17 MM patients, the CDR1, CDR2 and CDR3 regions of the IgH gene were analysed and sequenced to identify IgH's clonal nature. Unique sequences of the clonal IgH rearrangement were used to design specific molecular beacon probes for each MM patient. Examined were also the co-expression of CD19, CD38, CD45, CD56, and CD138 molecules in bone marrow aspirates of patients with MM by flow cytometry. Detection of MRD was positive in 13 (76%) of 17 patients by RT-PCR. The infiltration ratio was significantly correlated with CD138 expression (p=0.009). Significant correlation was also found between RT-PCR detection of MRD and CD138 expression (p=0.006). Nevertheless, no correlation was observed among other surface antigens (CD38, CD45, CD56). Our results indicated that RT-PCR with specific molecular beacons provide a feasible, accurate and reproducible method for the determination of MRD in MM. Flow cytometry detection of CD138 expression may be used as a disease marker in addition to RT-PCR.

  19. The ROCK Inhibitor Y-27632 Improves Recovery of Human Embryonic Stem Cells after Fluorescence-Activated Cell Sorting with Multiple Cell Surface Markers

    PubMed Central

    Emre, Nil; Vidal, Jason G.; Elia, Jeanne; O'Connor, Eric D.; Paramban, Rosanto I.; Hefferan, Michael P.; Navarro, Roman; Goldberg, Danielle S.; Varki, Nissi M.; Marsala, Martin; Carson, Christian T.

    2010-01-01

    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, identification and

  20. The Rate Constant for Fluorescence Quenching

    ERIC Educational Resources Information Center

    Legenza, Michael W.; Marzzacco, Charles J.

    1977-01-01

    Describes an experiment that utilizes fluorescence intensity measurements from a Spectronic 20 to determine the rate constant for the fluorescence quenching of various aromatic hydrocarbons by carbon tetrachloride in an ethanol solvent. (MLH)

  1. The Rate Constant for Fluorescence Quenching

    ERIC Educational Resources Information Center

    Legenza, Michael W.; Marzzacco, Charles J.

    1977-01-01

    Describes an experiment that utilizes fluorescence intensity measurements from a Spectronic 20 to determine the rate constant for the fluorescence quenching of various aromatic hydrocarbons by carbon tetrachloride in an ethanol solvent. (MLH)

  2. Dexamethasone, cyclophosphamide, idarubicin and etoposide (DC-IE): a novel, intensive induction chemotherapy regimen for patients with high-risk multiple myeloma.

    PubMed

    Ballester, O F; Moscinski, L C; Fields, K K; Hiemenz, J W; Zorsky, P E; Goldstein, S C; Saba, H I; Spiers, A S; Kronish, L; Sullivan, P; Elfenbein, G J

    1997-03-01

    We evaluated toxicities and responses to a novel, dose intensive and time sequenced, chemotherapy programme (DC-IE) in 45 patients with high-risk myeloma. DC-IE consisted of: dexamethasone (days 1-4); cyclophosphamide (day 5); idarubicin and etoposide (days 8-10). Complete response (CR) was achieved in four patients, six patients achieved near complete responses (nCR) and 21 patients achieved a partial remission (PR). Overall response rate was 76% (CI 56-94%) for newly diagnosed patients (n = 21) and 62% (CI 36-81%) for relapsed/refractory patients (n = 24). Toxicities were limited to myelosuppression; two patients died of sepsis during neutropenia (4%). DC-IE is active and tolerable for high-risk multiple myeloma, including patients with relapsed or refractory disease to anthracycline containing regimens.

  3. Dose-dense and less dose-intense Total Therapy 5 for gene expression profiling-defined high-risk multiple myeloma

    PubMed Central

    Jethava, Y; Mitchell, A; Zangari, M; Waheed, S; Schinke, C; Thanendrarajan, S; Sawyer, J; Alapat, D; Tian, E; Stein, C; Khan, R; Heuck, C J; Petty, N; Avery, D; Steward, D; Smith, R; Bailey, C; Epstein, J; Yaccoby, S; Hoering, A; Crowley, J; Morgan, G; Barlogie, B; van Rhee, F

    2016-01-01

    Multiple myeloma (MM) is a heterogeneous disease with high-risk patients progressing rapidly despite treatment. Various definitions of high-risk MM are used and we reported that gene expression profile (GEP)-defined high risk was a major predictor of relapse. In spite of our best efforts, the majority of GEP70 high-risk patients relapse and we have noted higher relapse rates during drug-free intervals. This prompted us to explore the concept of less intense drug dosing with shorter intervals between courses with the aim of preventing inter-course relapse. Here we report the outcome of the Total Therapy 5 trial, where this concept was tested. This regimen effectively reduced early mortality and relapse but failed to improve progression-free survival and overall survival due to relapse early during maintenance. PMID:27471869

  4. 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).

    PubMed

    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 <7.5%. Secondary outcomes included body weight, 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

  5. Noninvasive tongue stimulation combined with intensive cognitive and physical rehabilitation induces neuroplastic changes in patients with multiple sclerosis: A multimodal neuroimaging study.

    PubMed

    Leonard, Gabriel; Lapierre, Yves; Chen, Jen-Kai; Wardini, Rima; Crane, Joelle; Ptito, Alain

    2017-01-01

    Multiple sclerosis (MS) patients have central nervous system (CNS) lesions that may impede cognitive and sensorimotor function. Few rehabilitative therapies are available. The objective of this paper is to study effects of noninvasive tongue stimulation using the Portable Neuromodulation Stimulator (PoNS™) combined with intensive cognitive and physical rehabilitation on working memory, gait, balance and concomitant changes in the brain. Fourteen MS patients, seven each in an active and a sham stimulation group, participated. Participants received intensive physical therapy and working memory training for 14 weeks. Functional magnetic resonance imaging (fMRI) using motor imagery and working-memory tasks were completed prior to and following therapy, as were sensory organization tests (SOT), motor performance measures, and neuropsychological assessment. On the SOT, the active group showed significant improvement from baseline. fMRI revealed significant blood oxygen level-dependent signal changes in the left primary motor cortex for the Active Group, while the sham group had increased activity in bilateral premotor cortices. All individuals improved on working-memory tasks, but only the active group showed increased dorsolateral prefrontal cortex activity. In this cohort of MS patients, the results suggest that PoNS stimulation can enhance motor performance and working memory while also driving neuroplasticity. Further studies are warranted to explore these findings.

  6. Noninvasive tongue stimulation combined with intensive cognitive and physical rehabilitation induces neuroplastic changes in patients with multiple sclerosis: A multimodal neuroimaging study

    PubMed Central

    Lapierre, Yves; Chen, Jen-Kai; Wardini, Rima; Crane, Joelle; Ptito, Alain

    2017-01-01

    Background Multiple sclerosis (MS) patients have central nervous system (CNS) lesions that may impede cognitive and sensorimotor function. Few rehabilitative therapies are available. Objectives The objective of this paper is to study effects of noninvasive tongue stimulation using the Portable Neuromodulation Stimulator (PoNS™) combined with intensive cognitive and physical rehabilitation on working memory, gait, balance and concomitant changes in the brain. Methods Fourteen MS patients, seven each in an active and a sham stimulation group, participated. Participants received intensive physical therapy and working memory training for 14 weeks. Functional magnetic resonance imaging (fMRI) using motor imagery and working-memory tasks were completed prior to and following therapy, as were sensory organization tests (SOT), motor performance measures, and neuropsychological assessment. Results On the SOT, the active group showed significant improvement from baseline. fMRI revealed significant blood oxygen level-dependent signal changes in the left primary motor cortex for the Active Group, while the sham group had increased activity in bilateral premotor cortices. All individuals improved on working-memory tasks, but only the active group showed increased dorsolateral prefrontal cortex activity. Conclusions In this cohort of MS patients, the results suggest that PoNS stimulation can enhance motor performance and working memory while also driving neuroplasticity. Further studies are warranted to explore these findings. PMID:28607750

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

    PubMed

    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.

  8. Clinical practice and risk factors for immediate complications of endotracheal intubation in the intensive care unit: a prospective, multiple-center study.

    PubMed

    Jaber, Samir; Amraoui, Jibba; Lefrant, Jean-Yves; Arich, Charles; Cohendy, Robert; Landreau, Liliane; Calvet, Yves; Capdevila, Xavier; Mahamat, Aba; Eledjam, Jean-Jacques

    2006-09-01

    To describe the current practice of physicians, to report complications associated with endotracheal intubation (ETI) performed in THE intensive care unit (ICU), and to isolate predictive factors of immediate life-threatening complications. Multiple-center observational study. Seven intensive care units of two university hospitals. : We evaluated 253 occurrences of ETI in 220 patients. From January 1 to June 30, 2003, data related to all ETI performed in ICU were collected. Information regarding patient descriptors, procedures, and immediate complications were analyzed. The main indications to intubate the trachea were acute respiratory failure, shock, and coma. Some 148 ETIs (59%) were performed by residents. At least one severe complication occurred in 71 ETIs (28%): severe hypoxemia (26%), hemodynamic collapse (25%), and cardiac arrest (2%). The other complications were difficult intubation (12%), cardiac arrhythmia (10%), esophageal intubation (5%), and aspiration (2%). Presence of acute respiratory failure and the presence of shock as an indication for ETI were identified as independent risk factors for occurrence of complications, and ETI performed by a junior physician supervised by a senior (i.e., two operators) was identified as a protective factor for the occurrence of complications. ETI in ICU patients is associated with a high rate of immediate and severe life-threatening complications. Independent risk factors of complication occurrence were presence of acute respiratory failure and presence of shock as an indication for ETI. Further studies should aim to better define protocols for intubation in critically ill patients to make this procedure safer.

  9. Common fluorescent proteins for single-molecule localization microscopy

    NASA Astrophysics Data System (ADS)

    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.

  10. Integrated Fluorescence

    NASA Technical Reports Server (NTRS)

    Tuma, Margaret (Inventor); Gruhlke, Russell W. (Inventor)

    1998-01-01

    A detection method is integrated with a filtering method and an enhancement method to create a fluorescence sensor that can be miniaturized. The fluorescence sensor comprises a thin film geometry including a waveguide layer, a metal film layer and sensor layer. The thin film geometry of the fluorescence sensor allows the detection of fluorescent radiation over a narrow wavelength interval. This enables wavelength discrimination and eliminates the detection of unwanted light from unknown or spurious sources.

  11. 7,8-benzoflavone binding to human cytochrome P450 3A4 reveals complex fluorescence quenching, suggesting binding at multiple protein sites.

    PubMed

    Marsch, Glenn A; Carlson, Benjamin T; Guengerich, F Peter

    2017-03-20

    Human cytochrome P450 (P450) 3A4 is involved in the metabolism of one-half of marketed drugs and shows cooperative interactions with some substrates and other ligands. The interaction between P450 3A4 and the known allosteric effector 7,8-benzoflavone (α-naphthoflavone, αNF) was characterized using steady-state fluorescence spectroscopy. The binding interaction of P450 3A4 and αNF effectively quenched the fluorescence of both the enzyme and ligand. The Hill Equation and Stern-Volmer fluorescence quenching models were used to evaluate binding of ligand to enzyme. P450 3A4 fluorescence was quenched by titration with αNF; at the relatively higher [αNF]/[P450 3A4] ratios in this experiment, two weaker quenching interactions were revealed (Kd 1.8-2.5 and 6.5 μM). A range is given for the stronger interaction since αNF quenching of P450 3A4 fluorescence changed the protein spectral profile: quenching of 315 nm emission was slightly more efficient (Kd 1.8 μM) than the quenching of protein fluorescence at 335 and 355 nm (Kd 2.5 and 2.1 μM, respectively). In the reverse titration, αNF fluorescence was quenched by P450 3A4; at the lower [αNF]/[P450 3A4] ratios here, two strong quenching interactions were revealed (Kd 0.048 and 1.0 μM). Thus, four binding interactions of αNF to P450 3A4 are suggested by this study, one of which may be newly recognized and which could affect studies of drug oxidations by this important enzyme.

  12. Self-interference fluorescence microscopy with three-phase detection for depth-resolved confocal epi-fluorescence imaging.

    PubMed

    Braaf, Boy; de Boer, Johannes F

    2017-03-20

    Three-dimensional confocal fluorescence imaging of in vivo tissues is challenging due to sample motion and limited imaging speeds. In this paper a novel method is therefore presented for scanning confocal epi-fluorescence microscopy with instantaneous depth-sensing based on self-interference fluorescence microscopy (SIFM). A tabletop epi-fluorescence SIFM setup was constructed with an annular phase plate in the emission path to create a spectral self-interference signal that is phase-dependent on the axial position of a fluorescent sample. A Mach-Zehnder interferometer based on a 3 × 3 fiber-coupler was developed for a sensitive phase analysis of the SIFM signal with three photon-counter detectors instead of a spectrometer. The Mach-Zehnder interferometer created three intensity signals that alternately oscillated as a function of the SIFM spectral phase and therefore encoded directly for the axial sample position. Controlled axial translation of fluorescent microsphere layers showed a linear dependence of the SIFM spectral phase with sample depth over axial image ranges of 500 µm and 80 µm (3.9 × Rayleigh range) for 4 × and 10 × microscope objectives respectively. In addition, SIFM was in good agreement with optical coherence tomography depth measurements on a sample with indocyanine green dye filled capillaries placed at multiple depths. High-resolution SIFM imaging applications are demonstrated for fluorescence angiography on a dye-filled capillary blood vessel phantom and for autofluorescence imaging on an ex vivo fly eye.

  13. Cell-based and in vivo spectral analysis of fluorescent proteins for multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Salomonnson, Emma; Mihalko, Laura Anne; Verkhusha, Vladislav V.; Luker, Kathryn E.; Luker, Gary D.

    2012-09-01

    Multiphoton microscopy of cells and subcellular structures labeled with fluorescent proteins is the state-of-the-art technology for longitudinal imaging studies in tissues and living animals. Successful analysis of separate cell populations or signaling events by intravital microscopy requires optimal pairing of multiphoton excitation wavelengths with spectrally distinct fluorescent proteins. While prior studies have analyzed two photon absorption properties of isolated fluorescent proteins, there is limited information about two photon excitation and fluorescence emission profiles of fluorescent proteins expressed in living cells and intact tissues. Multiphoton microscopy was used to analyze fluorescence outputs of multiple blue, green, and red fluorescent proteins in cultured cells and orthotopic tumor xenografts of human breast cancer cells. It is shown that commonly used orange and red fluorescent proteins are excited efficiently by 750 to 760 nm laser light in living cells, enabling dual color imaging studies with blue or cyan proteins without changing excitation wavelength. It is also shown that small incremental changes in excitation wavelength significantly affect emission intensities from fluorescent proteins, which can be used to optimize multi-color imaging using a single laser wavelength. These data will direct optimal selection of fluorescent proteins for multispectral two photon microscopy.

  14. Improved self-efficacy in persons with relapsing remitting multiple sclerosis after an intensive social cognitive wellness program with participation of support partners: a 6-months observational study

    PubMed Central

    2014-01-01

    Background For persons with multiple sclerosis (MS) it is important to preserve their autonomy, in spite of increasing disability. A major factor mediating autonomy is self-efficacy. According to the social cognitive theory stressors are crucial determinants of self-efficacy, as well as the interaction with partners. Methods In an explorative observational study we assessed in 47 persons with MS (PwMS) the effect of an intense, multidisciplinary, 3-day, social cognitive wellness program with the participation of support partners, after 1, 3 and 6 months. Primary outcomes: self-efficacy-control and -function (Multiple Sclerosis Self-Efficacy Scale [MSSES]),limitations to and problems with participation and autonomy (Impact on Participation and Autonomy [IPA] scale). Secondary outcomes: health-related quality of life (HRQoL) (MS Quality of Life-54 Items [MSQoL-54] questionnaire), anxiety, depression (Hospital Anxiety and Depression Scale [HADS]), and fatigue (Modified Fatigue Impact Scale-5 Items [MFIS-5]). Disability was measured with the Expanded Disability Status Scale (EDSS). Percentage changes from baseline were tested with T-tests, level of significance 0.05. Results In the whole group the MSQoL-54 Mental score was increased at 1, 3 and 6 months (+16.0%, +13.2%, +12.2%), and the MSQoL-54 Physical (+10.2%) at 6 months, with no changes in other outcomes. The relapsing remitting (RR) subgroup (n = 20) had at 6 months an increase in the MSSES-Control score (+24.8%) and in the MSQoL54 Mental and Physical scores (+22.3%, +17.6%). Progressive patients (n = 22) only showed an increase in the MSQoL-54 Mental score (+11.5%) at 1 month. In the low-disability (EDSS < 4.0) subgroup the MSSES-Control score was increased (+23.8%) at 6 months, and the IPA-Limitations and -Problems scores decreased at 3 months (−6.1%, −8.8%); the MSQoL-54 Mental score had increased at 1, 3 and 6 months (+19.3%, +21.5%, +19.3%). In the high-disability (EDSS > =4

  15. Ethynyl-linked (pyreno)pyrrole-naphthyridine and aniline-naphthyridine molecules as fluorescent sensors of guanine via multiple hydrogen bondings.

    PubMed

    Lu, Shao-Hung; Selvi, Srinivasan; Fang, Jim-Min

    2007-01-05

    New fluorescent molecular sensors for 9-alkylguanines were constructed by conjugation of 2-acetamido-1,8-naphthyridine with N-Boc-pyrrole, N-Boc-pyreno[2,1-b]pyrrole, or acetanilide moieties via an ethynyl bridge. In combination with the triple hydrogen-bonding motif of 2-acetamidonaphthyridine toward alkylguanine, an additional binding site was provided by the substituent properly located on the pyrrole or aniline ring to enhance the affinity of these receptor molecules. Besides the ESI-MS analyses, the binding events were readily monitored by the absorption and fluorescence changes in the visible region.

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

    PubMed

    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.

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

    PubMed Central

    2011-01-01

    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. PMID:21801450

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

    PubMed

    Bak, Kate; Dobrow, Mark J; Hodgson, David; Whitton, Anthony

    2011-07-31

    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. 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. 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. 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.

  19. A prospective PETHEMA study of tandem autologous transplantation versus autograft followed by reduced-intensity conditioning allogeneic transplantation in newly diagnosed multiple myeloma.

    PubMed

    Rosiñol, Laura; Pérez-Simón, José Antonio; Sureda, Anna; de la Rubia, Javier; de Arriba, Felipe; Lahuerta, Juan José; González, José David; Díaz-Mediavilla, Joaquín; Hernández, Belén; García-Frade, Javier; Carrera, Dolores; León, Angel; Hernández, Miguel; Abellán, Pascual Fernández; Bergua, Juan Miguel; San Miguel, Jesús; Bladé, Joan

    2008-11-01

    One hundred ten patients with multiple myeloma (MM) failing to achieve at least near-complete remission (nCR) after a first autologous stem cell transplantation (ASCT) were scheduled to receive a second ASCT (85 patients) or a reduced-intensity-conditioning allograft (allo-RIC; 25 patients), depending on the human leukocyte antigen (HLA)-identical sibling donor availability. There was a higher increase in complete remission (CR) rate (40% vs 11%, P = .001) and a trend toward a longer progression-free survival (PFS; median, 31 months vs not reached, P = .08) in favor of allo-RIC. In contrast, it was associated with a trend toward a higher transplantation-related mortality (16% vs 5%, P = .07), a 66% chance of chronic graft-versus-host disease and no statistical difference in event-free survival and overall survival. Although the PFS plateau observed with allo-RIC is very encouraging, this procedure is associated with high morbidity and mortality, and therefore it should still be considered investigational and restricted to well-designed prospective clinical trials. This trial is registered at ClinicalTrials.gov ID number NCT00560053.

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

    PubMed

    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. 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.

    PubMed

    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

  2. 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.

    PubMed

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

  4. Acclimation of tobacco leaves to high light intensity drives the plastoquinone oxidation system--relationship among the fraction of open PSII centers, non-photochemical quenching of Chl fluorescence and the maximum quantum yield of PSII in the dark.

    PubMed

    Miyake, Chikahiro; Amako, Katsumi; Shiraishi, Naomasa; Sugimoto, Toshio

    2009-04-01

    Responses of the reduction-oxidation level of plastoquinone (PQ) in the photosynthetic electron transport (PET) system of chloroplasts to growth light intensity were evaluated in tobacco plants. Plants grown in low light (150 micromol photons m-2 s-1) (LL plants) were exposed to a high light intensity (1,100 micromol photons m-2 s-1) for 1 d. Subsequently, the plants exposed to high light (LH plants) were returned back again to the low light condition: these plants were designated as LHL plants. Both LH and LHL plants showed higher values of non-photochemical quenching of Chl fluorescence (NPQ) and the fraction of open PSII centers (qL), and lower values of the maximum quantum yield of PSII in the dark (Fv/Fm), compared with LL plants. The dependence of qL on the quantum yield of PSII [Phi(PSII)] in LH and LHL plants was higher than that in LL plants. To evaluate the effect of an increase in NPQ and decrease in Fv/Fm on qL, we derived an equation expressing qL in relation to both NPQ and Fv/Fm, according to the lake model of photoexcitation of the PSII reaction center. As a result, the heat dissipation process, shown as NPQ, did not contribute greatly to the increase in qL. On the other hand, decreased Fv/Fm did contribute to the increase in qL, i.e. the enhanced oxidation of PQ under photosynthesis-limited conditions. Thylakoid membranes isolated from LH plants, having high qL, showed a higher tolerance against photoinhibition of PSII, compared with those from LL plants. We propose a 'plastoquinone oxidation system (POS)', which keeps PQ in an oxidized state by suppressing the accumulation of electrons in the PET system in such a way as to regulate the maximum quantum yield of PSII.

  5. Long-term outcomes of autologous hematopoietic stem cell transplantation with reduced-intensity conditioning in multiple sclerosis: physician's and patient's perspectives.

    PubMed

    Shevchenko, Jury L; Kuznetsov, Alexey N; Ionova, Tatyana I; Melnichenko, Vladimir Y; Fedorenko, Denis A; Kurbatova, Kira A; Gorodokin, Gary I; Novik, Andrei A

    2015-07-01

    High-dose immunosuppressive therapy (HDIT) with autologous hematopoietic stem cell transplantation (AHSCT) is a promising approach to treatment of multiple sclerosis (MS) patients. In this paper, we present the long-term outcomes of a prospective single-center study with the analysis of the safety and efficacy of HDIT + AHSCT with reduced-intensity BEAM-like conditioning regimen in 99 MS patients: mean age-35 years old; male/female-39/60; median Expanded Disability Status Scale (EDSS) = 3.5; 43 relapsing/remitting MS, 56 progressive MS. No transplant-related deaths were observed. The mobilization and transplantation procedures were well tolerated. At 6 months post-transplant, neurological improvement or stabilization was observed in all the patients except one. Cumulative incidence of disease progression was 16.7 % at 8 years after HDIT + AHSCT. Estimated event-free survival at median follow-up of 48.9 months was 80 %: 83.3 % in relapsing/remitting MS vs 75.5 % in progressive MS. Sixty-four patients who did not progress during the first 3 years post-transplant and were monitored for more than 3 years were included in long-term outcome analysis. At the median long-term follow-up of 62 months, 47 % of patients improved by at least 0.5 points on the EDSS scale as compared to baseline and exhibited improvement during the entire period of follow-up; 45 % of patients were stable. No active, new, or enlarging lesions on magnetic resonance imaging were registered in patients without disease progression. AHSCT was accompanied by a significant improvement in patient's quality of life. Due to the fact that patient selection was quite different to the other studies and that the information about disease activity prior in the disease course and its treatment was inhomogeneous, comparison with the results in the literature should be done with caution. Thus, the risk/benefit ratio of HDIT + AHSCT with reduced-intensity BEAM-like conditioning regimen in our population

  6. Determination of barometric coefficients for total neutron intensity and neutron multiplicities on the base of Emilio Segre' Observatory data corrected for primary variations according to Rome neutron monitor data

    NASA Astrophysics Data System (ADS)

    Dorman, L. I.

    By the barometric coefficients determined in a first approximation in the way from sea level to the place of stationary operation, we corrected for barometric effect the total neutron intensity and intensities of neutron multiplicities detected by a 6NM-64 neutron monitor installed inside the Emilio Segre' Israelo-Italian moving laboratory (Mt. Hermon, Israel, 2020 m a.s.l.). The period June-December 1998 was analysed. We compared the obtained results with the Rome 17NM-64 neutron monitor data and corrected the Emilio Segre' Observatory data for primary variations. We determined with high accuracy barometric coefficients for the total neutron monitor counting rate and for the intensities of detected neutron multiplicities m=1, m=2, m=3, m=4, m=5, m=6, m=7 and m≥8.

  7. Epi-Fluorescence Microscopy

    PubMed Central

    Webb, Donna J.; Brown, Claire M.

    2012-01-01

    Epi-fluorescence microscopy is available in most life sciences research laboratories, and when optimized can be a central laboratory tool. In this chapter, the epi-fluorescence light path is introduced and the various components are discussed in detail. Recommendations are made for incident lamp light sources, excitation and emission filters, dichroic mirrors, objective lenses, and charge-coupled device (CCD) cameras in order to obtain the most sensitive epi-fluorescence microscope. The even illumination of metal-halide lamps combined with new “hard” coated filters and mirrors, a high resolution monochrome CCD camera, and a high NA objective lens are all recommended for high resolution and high sensitivity fluorescence imaging. Recommendations are also made for multicolor imaging with the use of monochrome cameras, motorized filter turrets, individual filter cubes, and corresponding dyes that are the best choice for sensitive, high resolution multicolor imaging. Images should be collected using Nyquist sampling and should be corrected for background intensity contributions and nonuniform illumination across the field of view. Photostable fluorescent probes and proteins that absorb a lot of light (i.e., high extinction co-efficients) and generate a lot of fluorescence signal (i.e., high quantum yields) are optimal. A neuronal immune-fluorescence labeling protocol is also presented. Finally, in order to maximize the utility of sensitive wide-field microscopes and generate the highest resolution images with high signal-to-noise, advice for combining wide-field epi-fluorescence imaging with restorative image deconvolution is presented. PMID:23026996

  8. Fluorescent image tracking velocimeter

    DOEpatents

    Shaffer, Franklin D.

    1994-01-01

    A multiple-exposure fluorescent image tracking velocimeter (FITV) detects and measures the motion (trajectory, direction and velocity) of small particles close to light scattering surfaces. The small particles may follow the motion of a carrier medium such as a liquid, gas or multi-phase mixture, allowing the motion of the carrier medium to be observed, measured and recorded. The main components of the FITV include: (1) fluorescent particles; (2) a pulsed fluorescent excitation laser source; (3) an imaging camera; and (4) an image analyzer. FITV uses fluorescing particles excited by visible laser light to enhance particle image detectability near light scattering surfaces. The excitation laser light is filtered out before reaching the imaging camera allowing the fluoresced wavelengths emitted by the particles to be detected and recorded by the camera. FITV employs multiple exposures of a single camera image by pulsing the excitation laser light for producing a series of images of each particle along its trajectory. The time-lapsed image may be used to determine trajectory and velocity and the exposures may be coded to derive directional information.

  9. Synthesis, characterization, and plasmonic enhancement of fluorescent carbon nanodots

    NASA Astrophysics Data System (ADS)

    Schmitz, Rachel Danielle

    Carbon nanodots are fluorescent nanoparticles that have unique photophysical properties that make them ideal candidates as luminescent probes for various fields of study. Although the synthetic routes of carbon nanodots have been extensively investigated, many of them produce carbon nanodots with low fluorescence quantum yields and/or require surface modifications to obtain fluorescent nanoparticles. This research discusses the development of a combustion-based method to synthesize carbon nanodots along with the characterizations and modifications of the nanodots photophysical properties. First, the synthetic aspect of this research investigated two combustion-based synthetic pathways of the carbon nanodots. The first pathway (candle-based) utilized the oxidation of candle soot with nitric acid followed by multiple neutralization and separation steps that resulted in carbon nanodots with low quantum yields ( 2%). The second pathway (methane-based), utilizing methane gas, produced, and collected the carbon nanodots directly from a flame, which resulted in significantly less experimental time and carbon nanodots with higher quantum yields ( 30%). The photophysical characterizations of the synthesized carbon nanodots revealed an excitation wavelength dependent fluorescence, a broad absorption, good photostability, and complex intensity decays. The photophysical properties of the methane-based nanodots were further investigated for a more in-depth understanding of the fluorescence-based structural architecture, utilizing fluorescence quenching methods. These experiments revealed that the fluorescence quenching of the nanodots can occur through both dynamic and static quenching mechanisms depending on the type of quencher utilized. Also, these experiments helped to elucidate the origin of the luminescence of carbon nanodots. While carrying out the quenching experiments, a novel temperature dependent fluorescence property was observed and analyzed, which showed an

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

    PubMed

    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.

  11. Flow cytometric fluorescence lifetime analysis of DNA binding fluorochromes

    SciTech Connect

    Crissman, Harry A.; Cui, H. H.; 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

  12. Nine New Fluorescent Probes

    NASA Astrophysics Data System (ADS)

    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

  13. Representational overlap of adjacent fingers in multiple areas of human primary somatosensory cortex depends on electrical stimulus intensity: an fMRI study.

    PubMed

    Krause, T; Kurth, R; Ruben, J; Schwiemann, J; Villringer, K; Deuchert, M; Moosmann, M; Brandt, S; Wolf, K; Curio, G; Villringer, A

    2001-04-27

    Functional magnetic resonance imaging (fMRI) was used to examine the influence of non-painful electrical stimulus intensity on the BOLD response in human primary somatosensory cortex (SI). In ten healthy subjects, index and middle finger of the right hand were stimulated separately at two different stimulus intensities. The activated volume of single finger representations as well as the volume of representational overlap of the two activations increased following an increase in stimulus intensity. This effect was seen in two different subdivisions of SI, one in the depth of the central sulcus, presumably corresponding to Brodmann area (BA) 3b, and one on the crown of the postcentral gyrus, presumably corresponding to BA 1/2. Relative overlap (ratio of overlap volume to volume of individual finger representation) was larger in BA 1/2 than in BA 3b. Additionally, in both areas relative overlap increased significantly from low to high stimulus intensity. Relative overlap did not change when different correlation thresholds were employed arguing against an unspecific 'spillover effect'. Analysis of signal intensity time courses indicated that the response difference to high versus low stimulus strength was not present during the initial seconds of stimulation, during which both led to a similar signal intensity increase. Only during the following maintenance level of the response did the response to high stimulus intensity reach a significantly higher plateau level than the one due to low intensity stimulation, an effect which was present in both areas, BA 3b and BA 1/2, respectively.

  14. Prediction of soil organic carbon in an intensively managed reclamation zone of eastern China: A comparison of multiple linear regressions and the random forest model.

    PubMed

    Zhang, Huan; Wu, Pengbao; Yin, Aijing; Yang, Xiaohui; Zhang, Ming; Gao, Chao

    2017-08-15

    Organic carbon is a key component of soils and plays a fundamental role in soil fertility and climate change. Determining the importance of potential drivers of soil organic carbon (SOC) and thus predicting the distribution of SOC are important for measuring carbon sequestration or emissions. Coastal wetlands are precious land resources that are currently undergoing rapid reclamation in China. The alternations in soil physicochemical conditions caused by reclamation can strongly impact the cycle of organic carbon. However, identification of the important drivers of SOC dynamics and prediction of SOC using the potential drivers remain largely unclear. In this study, we used classification and regression tree (CART) to identify the importance of the potential drivers of SOC at 241 sites from an intensively managed reclamation zone of eastern China. Multiple linear regressions (MLR) and random forest (RF) models were applied to predict the distribution of SOC using continuous variables, such as the contents of Cl, CaO, Fe2O3, Al2O3, SiO2, clay, silt, and sand as well as the soil pH, along with categorical variables, such as land use and reclamation duration. The results indicate that the soil/sediment pH was the most important variable impacting SOC, followed by the Cl and silt contents. The RF and MLR involving all predictor variables produced much higher R(2) and lower error indices than the RF and MLR models involving independent variables (pH and CaO). RF performed much better than MLR as it revealed much lower error indices (ME, MSE, and RMSE) and a higher R(2) than MLR. The superiority of RF in predicting SOC is related to its capability to deal with non-linear and hierarchical relationships between SOC and predictors. Analyses of land use effects on SOC dynamics indicated that paddy soils were superior in sequestering SOC than other land use types, which is likely ascribed to the rapid desalination and dealkalization of paddy field management. Therefore, paddy

  15. The photon counting histogram in fluorescence fluctuation spectroscopy.

    PubMed Central

    Chen, Y; Müller, J D; So, P T; Gratton, E

    1999-01-01

    Fluorescence correlation spectroscopy (FCS) is generally used to obtain information about the number of fluorescent particles in a small volume and the diffusion coefficient from the autocorrelation function of the fluorescence signal. Here we demonstrate that photon counting histogram (PCH) analysis constitutes a novel tool for extracting quantities from fluorescence fluctuation data, i.e., the measured photon counts per molecule and the average number of molecules within the observation volume. The photon counting histogram of fluorescence fluctuation experiments, in which few molecules are present in the excitation volume, exhibits a super-Poissonian behavior. The additional broadening of the PCH compared to a Poisson distribution is due to fluorescence intensity fluctuations. For diffusing particles these intensity fluctuations are caused by an inhomogeneous excitation profile and the fluctuations in the number of particles in the observation volume. The quantitative relationship between the detected photon counts and the fluorescence intensity reaching the detector is given by Mandel's formula. Based on this equation and considering the fluorescence intensity distribution in the two-photon excitation volume, a theoretical expression for the PCH as a function of the number of molecules in the excitation volume is derived. For a single molecular species two parameters are sufficient to characterize the histogram completely, namely the average number of molecules within the observation volume and the detected photon counts per molecule per sampling time epsilon. The PCH for multiple molecular species, on the other hand, is generated by successively convoluting the photon counting distribution of each species with the others. The influence of the excitation profile upon the photon counting statistics for two relevant point spread functions (PSFs), the three-dimensional Gaussian PSF conventionally employed in confocal detection and the square of the Gaussian

  16. Plasmonic fluorescent quantum dots.

    PubMed

    Jin, Yongdong; Gao, Xiaohu

    2009-09-01

    Combining multiple discrete components into a single multifunctional nanoparticle could be useful in a variety of applications. Retaining the unique optical and electrical properties of each component after nanoscale integration is, however, a long-standing problem. It is particularly difficult when trying to combine fluorophores such as semiconductor quantum dots with plasmonic materials such as gold, because gold and other metals can quench the fluorescence. So far, the combination of quantum dot fluorescence with plasmonically active gold has only been demonstrated on flat surfaces. Here, we combine fluorescent and plasmonic activities in a single nanoparticle by controlling the spacing between a quantum dot core and an ultrathin gold shell with nanometre precision through layer-by-layer assembly. Our wet-chemistry approach provides a general route for the deposition of ultrathin gold layers onto virtually any discrete nanostructure or continuous surface, and should prove useful for multimodal bioimaging, interfacing with biological systems, reducing nanotoxicity, modulating electromagnetic fields and contacting nanostructures.

  17. Chromosome characterization using single fluorescent dye

    DOEpatents

    Crissman, Harry A.; Hirons, Gregory T.

    1995-01-01

    Chromosomes are characterized by fluorescent emissions from a single fluorescent dye that is excited over two different wavelengths. A mixture containing chromosomes is stained with a single dye selected from the group consisting of TOTO and YOYO and the stained chromosomes are placed in a flow cytometer. The fluorescent dye is excited sequentially by a first light having a wavelength in the ultraviolet range to excite the TOTO or YOYO to fluoresce at a first intensity and by a second light having a wavelength effective to excite the TOTO or YOYO dye to fluoresce at a second intensity. Specific chromosomes may be identified and sorted by intensity relationships between the first and second fluorescence emissions.

  18. Fluorescent refrigeration

    DOEpatents

    Epstein, Richard I.; Edwards, Bradley C.; Buchwald, Melvin I.; Gosnell, Timothy R.

    1995-01-01

    Fluorescent refrigeration is based on selective radiative pumping, using substantially monochromatic radiation, of quantum excitations which are then endothermically redistributed to higher energies. Ultimately, the populated energy levels radiatively deexcite emitting, on the average, more radiant energy than was initially absorbed. The material utilized to accomplish the cooling must have dimensions such that the exciting radiation is strongly absorbed, but the fluorescence may exit the material through a significantly smaller optical pathlength. Optical fibers and mirrored glasses and crystals provide this requirement.

  19. Fluorescence of Melanin.

    NASA Astrophysics Data System (ADS)

    Gallas, James Martin

    1981-06-01

    Optical fluorescence in aqueous suspensions of synthetic dopa melanin has been detected and investigated. A fluorescence lifetime of 8.3 ns was measured in pulsed laser experiments. Fluorescence was observed for excitation between 290 and 420 nm. The emission as a function of wavelength displayed a single broad maximum falling between 440 and 480 nm, depending on excitation wavelength. The dependence of the quantum efficiency for fluorescence on parameters such as solution temperature, viscosity, pH, and concentration of metal ion, such as copper, has been investigated. The emission intensity decreased at both high and low values of pH, with increasing temperatures, and with increasing metal ion concentration, and decreasing viscosity. The pH dependence of the fluorescence can be related to changes in the ionization state of the various ionizable groups attached to the fluorophores. A self consistent model was developed in which proton transfer from these groups was fast compared with fluorescence rates for one type of fluorophore and slow compared with those for another type. The fluorescence dependence on copper ion concentration is explained in terms of a model invoking a reaction between melanin fluorophores and copper ions leading to the formation of a melanin-copper complex which is itself fluorescent. A model incorporating diffusion controlled chemical reactions between fluorescents groups and quencher groups on the melanin polymer is developed and used to explain the observed dependence of melanin fluorescence on solvent viscosity. Over the temperature range 20(DEGREES)C-70(DEGREES)C, an Arrhenius type behavior was found with an activation enthalpy of 1.3 Kcal/mole. Using the temperature dependence of the viscous quenching model as well as the temperature dependence of the fluorophore-radical equilibrium concentration leads to a temperature dependence which is in reasonable agreement with the observed behavior. Many aspects of the experimental results

  20. Correlation fluorescence method of amine detection

    NASA Astrophysics Data System (ADS)

    Myslitsky, Valentin F.; Tkachuk, Svetlana S.; Rudeichuk, Volodimir M.; Strinadko, Miroslav T.; Slyotov, Mikhail M.; Strinadko, Marina M.

    1997-12-01

    The amines fluorescence spectra stimulated by UV laser radiation are investigated in this paper. The fluorescence is stimulated by the coherent laser beam with the wavelength 0.337 micrometers . At the sufficient energy of laser stimulation the narrow peaks of the fluorescence spectra are detected besides the wide maximum. The relationship between the fluorescence intensity and the concentration of amines solutions are investigated. The fluorescence intensity temporal dependence on wavelength 0.363 micrometers of the norepinephrine solution preliminarily radiated by UV laser with wavelength 0.337 micrometers was found. The computer stimulated and experimental investigations of adrenaline and norepinephrine mixtures fluorescence spectra were done. The correlation fluorescent method of amines detection is proposed.

  1. Method of determining the optimal dilution ratio for fluorescence fingerprint of food constituents.

    PubMed

    Trivittayasil, Vipavee; Tsuta, Mizuki; Kokawa, Mito; Yoshimura, Masatoshi; Sugiyama, Junichi; Fujita, Kaori; Shibata, Mario

    2015-01-01

    Quantitative determination by fluorescence spectroscopy is possible because of the linear relationship between the intensity of emitted fluorescence and the fluorophore concentration. However, concentration quenching may cause the relationship to become nonlinear, and thus, the optimal dilution ratio has to be determined. In the case of fluorescence fingerprint (FF) measurement, fluorescence is measured under multiple wavelength conditions and a method of determining the optimal dilution ratio for multivariate data such as FFs has not been reported. In this study, the FFs of mixed solutions of tryptophan and epicatechin of different concentrations and composition ratios were measured. Principal component analysis was applied, and the resulting loading plots were found to contain useful information about each constituent. The optimal concentration ranges could be determined by identifying the linear region of the PC score plotted against total concentration.

  2. A simple model for understanding the fluorescence behavior of Au25 nanoclusters

    NASA Astrophysics Data System (ADS)

    Wang, Shuxin; Zhu, Xiuyi; Cao, Tiantian; Zhu, Manzhou

    2014-05-01

    In this work, we synthesized Au25 nanoclusters protected by 2-(naphthalen-2-yl)ethanethiolate. Our experiments revealed that the luminescence of this nanocluster consists of two bands, namely, band I centered at 740 nm and band II centered at 680 nm. Compared with 2-phenylethanethiolate protected Au25 nanoclusters, this new nanocluster has a much higher QY (quantum yield) value (6.5 times higher). Fluorescence lifetime measurements showed multiple components, i.e. 0.15 ns, ~20 ns and ~150 ns. With an increase in the electropositivity of the nanocluster, the fluorescence intensity of the nanocluster exhibits a significant enhancement. Since the 2-(naphthalen-2-yl)ethanethiolate protected Au25 nanocluster shares the same Au13/Au12 core-shell structure as the 2-phenylethanethiolate protected nanocluster, the band II fluorescence implies that the surface ligands play a major role in the origin of the fluorescence.

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

    NASA Astrophysics Data System (ADS)

    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.

  4. Analysis of Stroma Labeling During Multiple Passage of a Sarcoma Imageable Patient-derived Orthotopic Xenograft (iPDOX) in Red Fluorescent Protein Transgenic Nude Mice.

    PubMed

    Kiyuna, Tasuku; Murakami, Takashi; Tome, Yasunori; Kawaguchi, Kei; Igarashi, Kentaro; Miyake, Kentaro; Kanaya, Fuminori; Singh, Arun; Eilber, Fritz C; Hoffman, Robert M

    2017-03-16

    A patient-derived orthotopic xenograft (PDOX) model of undifferentiated pleomorphic sarcoma (UPS) was previously established that acquired red fluorescent protein (RFP)-expressing stroma by growth in an RFP transgenic nude mouse. In the present study, an imageable PDOX model (iPDOX) of UPS was established by orthotopic implantation in the biceps femoris of transgenic RFP nude mice. After the tumors grew to a diameter of 10 mm, they were harvested and the brightest portion of the tumors were subsequently orthotopically transplanted to both RFP and non-colored nude mice. The UPS PDOX tumor was again transplanted to RFP transgenic and non-colored nude mice and finally a 3(rd) passage was made in the same manner. Five UPS tumors from each passage in both RFP and non-colored mouse models were harvested. The FV1000 confocal microscope was used to visualize and quantitate the RFP area of the resected tumors. The average percent fluorescent area in the first passage of RFP mice was 34 ± 22%; in the second passage, 34 ± 20%; and 36 ± 11% in the third passage of RFP transgenic nude mice. The average tumor RFP area in the first passage from RFP mice to non-colored mice was 20 ± 7%; in the second passage, 28 ± 11%; in the third passage was 27 ± 13%. The present results demonstrate the extensive and stable acquisition of stroma by the UPS-tumor growing orthotopically in transgenic RFP nude mice (iPDOX). This model can be used for screening for effective drugs for individual patients and drug discovery. This article is protected by copyright. All rights reserved.

  5. Novel full‐spectral flow cytometry with multiple spectrally‐adjacent fluorescent proteins and fluorochromes and visualization of in vivo cellular movement

    PubMed Central

    Futamura, Koji; Sekino, Masashi; Hata, Akihiro; Ikebuchi, Ryoyo; Nakanishi, Yasutaka; Egawa, Gyohei; Kabashima, Kenji; Watanabe, Takeshi; Furuki, Motohiro

    2015-01-01

    Abstract Flow cytometric analysis with multicolor fluoroprobes is an essential method for detecting biological signatures of cells. Here, we present a new full‐spectral flow cytometer (spectral‐FCM). Unlike conventional flow cytometer, this spectral‐FCM acquires the emitted fluorescence for all probes across the full‐spectrum from each cell with 32 channels sequential PMT unit after dispersion with prism, and extracts the signals of each fluoroprobe based on the spectral shape of each fluoroprobe using unique algorithm in high speed, high sensitive, accurate, automatic and real‐time. The spectral‐FCM detects the continuous changes in emission spectra from green to red of the photoconvertible protein, KikGR with high‐spectral resolution and separates spectrally‐adjacent fluoroprobes, such as FITC (Emission peak (Em) 519 nm) and EGFP (Em 507 nm). Moreover, the spectral‐FCM can measure and subtract autofluorescence of each cell providing increased signal‐to‐noise ratios and improved resolution of dim samples, which leads to a transformative technology for investigation of single cell state and function. These advances make it possible to perform 11‐color fluorescence analysis to visualize movement of multilinage immune cells by using KikGR‐expressing mice. Thus, the novel spectral flow cytometry improves the combinational use of spectrally‐adjacent various FPs and multicolor fluorochromes in metabolically active cell for the investigation of not only the immune system but also other research and clinical fields of use. © 2015 The Authors. Cytometry Part A Published by Wiley Periodicals, Inc. on behalf of ISAC PMID:26217952

  6. High-intensity resistance training in multiple sclerosis - An exploratory study of effects on immune markers in blood and cerebrospinal fluid, and on mood, fatigue, health-related quality of life, muscle strength, walking and cognition.

    PubMed

    Kierkegaard, Marie; Lundberg, Ingrid E; Olsson, Tomas; Johansson, Sverker; Ygberg, Sofia; Opava, Christina; Holmqvist, Lotta Widén; Piehl, Fredrik

    2016-03-15

    High-intensity resistance training is unexplored in people with multiple sclerosis. To evaluate effects of high-intensity resistance training on immune markers and on measures of mood, fatigue, health-related quality of life, muscle strength, walking and cognition. Further, to describe participants' opinion and perceived changes of the training. Twenty patients with relapsing-remitting multiple sclerosis performed high-intensity resistance training at an intensity of 80% of one-repetition maximum, twice a week for 12 weeks. Blood and optional cerebrospinal fluid samples, and data on secondary outcome measures were collected before and after intervention. A study-specific questionnaire was used for capturing participants' opinion. Seventeen participants completed the study. Plasma cytokine levels of tumor necrosis factor were significantly decreased post-intervention (p=0.001). Exploratory cytokine analyses in cerebrospinal fluid (n=8) did not reveal major changes. Significant and clinically important improvements were found in fatigue (p=0.001) and health-related quality of life (p=0.004). Measures of mood (p=0.002), muscle strength (p ≤ 0.001), walking speed (p=0.013) and cognition (p=0.04) were also improved. A majority of participants evaluated the training as very good and perceived changes to the better. High-intensity resistance training in persons with relapsing remitting multiple sclerosis with low disability had positive effects on peripheral pro-inflammatory cytokine levels, led to clinically relevant improvements in measures of fatigue and health-related quality of life, and was well tolerated. These results provide a basis for a larger randomized trial. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Treatment planning strategy for whole-brain radiotherapy with hippocampal sparing and simultaneous integrated boost for multiple brain metastases using intensity-modulated arc therapy.

    PubMed

    Pokhrel, Damodar; Sood, Sumit; McClinton, Christopher; Shen, Xinglei; Lominska, Christopher; Saleh, Habeeb; Badkul, Rajeev; Jiang, Hongyu; Mitchell, Melissa; Wang, Fen

    2016-01-01

    To retrospectively evaluate the accuracy, plan quality and efficiency of intensity-modulated arc therapy (IMAT) for hippocampal sparing whole-brain radiotherapy (HS-WBRT) with simultaneous integrated boost (SIB) in patients with multiple brain metastases (m-BM). A total of 5 patients with m-BM were retrospectively replanned for HS-WBRT with SIB using IMAT treatment planning. The hippocampus was contoured on diagnostic T1-weighted magnetic resonance imaging (MRI) which had been fused with the planning CT image set. The hippocampal avoidance zone (HAZ) was generated using a 5-mm uniform margin around the paired hippocampi. The m-BM planning target volumes (PTVs) were contoured on T1/T2-weighted MRI registered with the 3D planning computed tomography (CT). The whole-brain planning target volume (WB-PTV) was defined as the whole-brain tissue volume minus HAZ and m-BM PTVs. Highly conformal IMAT plans were generated in the Eclipse treatment planning system for Novalis-TX linear accelerator consisting of high-definition multileaf collimators (HD-MLCs: 2.5-mm leaf width at isocenter) and 6-MV beam. Prescription dose was 30Gy for WB-PTV and 45Gy for each m-BM in 10 fractions. Three full coplanar arcs with orbit avoidance sectors were used. Treatment plans were evaluated using homogeneity (HI) and conformity indices (CI) for target coverage and dose to organs at risk (OAR). Dose delivery efficiency and accuracy of each IMAT plan was assessed via quality assurance (QA) with a MapCHECK device. Actual beam-on time was recorded and a gamma index was used to compare dose agreement between the planned and measured doses. All 5 HS-WBRT with SIB plans met WB-PTV D2%, D98%, and V30Gy NRG-CC001 requirements. The plans demonstrated highly conformal and homogenous coverage of the WB-PTV with mean HI and CI values of 0.33 ± 0.04 (range: 0.27 to 0.36), and 0.96 ± 0.01 (range: 0.95 to 0.97), respectively. All 5 hippocampal sparing patients met protocol guidelines with maximum dose and

  8. Laser-excited fluorescence for measuring atmospheric pollution

    NASA Technical Reports Server (NTRS)

    Menzies, R. T.

    1975-01-01

    System measures amount of given pollutant at specific location. Infrared laser aimed at location has wavelength that will cause molecules of pollutant to fluoresce. Detector separates fluorescence from other radiation and measures its intensity to indicate concentration of pollutant.

  9. Temperature-dependent fluorescence in nanodiamonds

    NASA Astrophysics Data System (ADS)

    Su, Li-Xia; Lou, Qing; Zang, Jin-Hao; Shan, Chong-Xin; Gao, Yuan-Fei

    2017-02-01

    Here, we report that nanodiamonds (NDs) exhibit blue fluorescence with an emission peak at around 400 nm. With increasing temperature, the peak energy of fluorescence was found to demonstrate a blue shift, possibly due to excited excitons populating higher-energy states, such as oxidation defect states. The intensity evolution of the fluorescence was attributed to a thermally activated process. Moreover, the bandwidth of fluorescence also increased because of exciton–phonon interactions and ionized impurity scattering. The above results indicate that the fluorescence of NDs could originate from radiative recombination through intrinsic transitions between highly localized π states.

  10. Fluorescence lifetime image of a single halobacterium

    NASA Astrophysics Data System (ADS)

    Wang, Hui-Ping; Nakabayashi, Takakazu; Tsujimoto, Kazuo; Miyauchi, Seiji; Kamo, Naoki; Ohta, Nobuhiro

    2007-07-01

    Fluorescence lifetime imaging (FLIM) has been reported for a single halobacterium loaded with 2',7'-bis-(carboxyethyl)-5(6)-carboxyfluorescein (BCECF). FLIM of halobacteria exhibits two distinct fluorescence lifetimes, representing different environments from each other in different halobacteria. The observed difference in the fluorescence lifetime appears to be due to the difference in electric field inside a cell. Fluorescence intensity of BCECF in polyvinyl alcohol film is quenched by an external electric field, indicating that the fluorescence lifetime of BCECF is affected by an electric field.

  11. 5-color multiplexed microwave-accelerated metal-enhanced fluorescence: detection and analysis of multiple DNA sequences from within one sample well within a few seconds.

    PubMed

    Dragan, Anatoliy; Geddes, Chris D

    2014-11-01

    We present a potentially highly sensitive and selective bio-assay for the potential detection of any five different DNA sequences from one sample in one well. The assay is based on a DNA "rapid catch and signal" (DNA-RCS) technology developed for the detection of different DNA sequences from a sample well area. Our signal amplification utilizes the metal-enhanced fluorescence (MEF) of dyes attached to the probe-DNAs, which hybridizes with the pre-formed mixture of anchor-DNA scaffolds on silver island films (SiFs). Low-power microwave irradiation accelerates both the formation of the anchor-DNA scaffold on the SiF-surface and anchor/probe DNA hybridization, i.e. "rapid catch" of target DNAs from a bulk solution, decreasing the assay run time from hours to only a few seconds. Localization of signaling dye-labels close to the SiFs make them extremely photostable, which allows for collecting/integrating the signal over a long time period. To demonstrate a 5 color DNA assay (5-plex) we have used a range of readily available Alexa™ dyes. Advantages and perspectives of the RCS-technologies ability to detect 5 different DNA sequences from within one plate-well are discussed.

  12. Near infrared fluorescent dual ligand functionalized Au NCs based multidimensional sensor array for pattern recognition of multiple proteins and serum discrimination.

    PubMed

    Xu, Shenghao; Gao, Teng; Feng, Xiuying; Fan, Xiaojian; Liu, Gufan; Mao, Yaning; Yu, Xijuan; Lin, Jiehua; Luo, Xiliang

    2017-11-15

    Here, a multidimensional sensor array capable of analyzing various proteins and discriminating between serums from different stages of breast cancer patients were developed based on six kinds of near infrared fluorescent dual ligand functionalized Au NCs (functionalized with different amino acids) as sensing receptors. These six kinds of different amino acids functionalized Au NCs were synthesized for the first time within 2h due to the direct donation of delocalized electrons of electron-rich atoms or groups of the ligands to the Au core. Based on this, ten proteins could be simultaneously and effectively discriminated by this "chemical nose/tongue" sensor array. Linear discrimination analysis (LDA) of the response patterns showed successful differentiation of the analytes at concentrations as low as 10nM with high identification accuracy. Isothermal titration calorimetry (ITC) experiment illustrates that Au NCs interacted with proteins mainly by hydrogen bonding and van der Waals forces. Furthermore, the greatest highlight of this sensor array is demonstrated by successfully discriminating between serums from different stages of breast cancer patients (early, middle and late) and healthy people, suggesting great potential for auxiliary diagnosis. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Fluorescent refrigeration

    DOEpatents

    Epstein, R.I.; Edwards, B.C.; Buchwald, M.I.; Gosnell, T.R.

    1995-09-05

    Fluorescent refrigeration is based on selective radiative pumping, using substantially monochromatic radiation, of quantum excitations which are then endothermically redistributed to higher energies. Ultimately, the populated energy levels radiatively deexcite emitting, on the average, more radiant energy than was initially absorbed. The material utilized to accomplish the cooling must have dimensions such that the exciting radiation is strongly absorbed, but the fluorescence may exit the material through a significantly smaller optical pathlength. Optical fibers and mirrored glasses and crystals provide this requirement. 6 figs.

  14. Dual time-resolved temperature-jump fluorescence and infrared spectroscopy for the study of fast protein dynamics

    NASA Astrophysics Data System (ADS)

    Davis, Caitlin M.; Reddish, Michael J.; Dyer, R. Brian

    2017-05-01

    Time-resolved temperature-jump (T-jump) coupled with fluorescence and infrared (IR) spectroscopy is a powerful technique for monitoring protein dynamics. Although IR spectroscopy of the polypeptide amide I mode is more technically challenging, it offers complementary information because it directly probes changes in the protein backbone, whereas, fluorescence spectroscopy is sensitive to the environment of specific side chains. With the advent of widely tunable quantum cascade lasers (QCL) it is possible to efficiently probe multiple IR frequencies with high sensitivity and reproducibility. Here we describe a dual time-resolved T-jump fluorescence and IR spectrometer and its application to study protein folding dynamics. A Q-switched Ho:YAG laser provides the T-jump source for both time-resolved IR and fluorescence spectroscopy, which are probed by a QCL and Ti:Sapphire laser, respectively. The Ho:YAG laser simultaneously pumps the time-resolved IR and fluorescence spectrometers. The instrument has high sensitivity, with an IR absorbance detection limit of < 0.2 mOD and a fluorescence sensitivity of 2% of the overall fluorescence intensity. Using a computer controlled QCL to rapidly tune the IR frequency it is possible to create a T-jump induced difference spectrum from 50 ns to 0.5 ms. This study demonstrates the power of the dual time-resolved T-jump fluorescence and IR spectroscopy to resolve complex folding mechanisms by complementary IR absorbance and fluorescence measurements of protein dynamics.

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

    SciTech Connect

    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.

    2016-08-28

    Development of next generation red phosphors for commercial lighting requires understanding of how increased luminescence is achieved by various treatment strategies. In our 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. Furthermore, 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. We performed calculations in order 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.

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

    SciTech Connect

    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.

    2016-08-28

    Development of next generation red phosphors for commercial lighting requires understanding of how increased luminescence is achieved by various treatment strategies. In our 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. Furthermore, 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. We performed calculations in order 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.

  17. The CREATE Strategy for Intensive Analysis of Primary Literature Can Be Used Effectively by Newly Trained Faculty to Produce Multiple Gains in Diverse Students

    ERIC Educational Resources Information Center

    Stevens, Leslie M.; Hoskins, Sally G.

    2014-01-01

    The CREATE (Consider Read, Elucidate the hypotheses, Analyze and interpret the data, and Think of the next Experiment) strategy aims to demystify scientific research and scientists while building critical thinking, reading/analytical skills, and improved science attitudes through intensive analysis of primary literature. CREATE was developed and…

  18. The CREATE Strategy for Intensive Analysis of Primary Literature Can Be Used Effectively by Newly Trained Faculty to Produce Multiple Gains in Diverse Students

    ERIC Educational Resources Information Center

    Stevens, Leslie M.; Hoskins, Sally G.

    2014-01-01

    The CREATE (Consider Read, Elucidate the hypotheses, Analyze and interpret the data, and Think of the next Experiment) strategy aims to demystify scientific research and scientists while building critical thinking, reading/analytical skills, and improved science attitudes through intensive analysis of primary literature. CREATE was developed and…

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

    PubMed

    Anderson, Cheryl B; Hughes, Sheryl O; Fuemmeler, Bernard F

    2009-07-01

    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 children (mean age 9.9 years) reported attitudes on importance of vigorous and moderate intensity team and individually performed sports/activities, as well as household chores. Separate structural models (LISREL 8.7) for girls and boys tested whether parental attitudes were related to child TV and computer via child attitudes, sport team participation, and physical activity, controlling for demographic factors. Child 7-day physical activity, sport teams, weekly TV, computer. Parent-child attitude congruence was more prevalent among boys, and attitudes varied by ethnicity, parent education, and number of children. Positive parent-child attitudes for vigorous team sports were related to increased team participation and physical activity, as well as reduced TV and computer in boys and girls. Value of moderate intensity household chores, such as cleaning house and doing laundry, was related to decreased team participation and increased TV in boys. Only organized team sports, not general physical activity, was related to reduced TV and computer. Results support parents' role in socializing children's achievement task values, affecting child activity by transferring specific attitudes. Value of vigorous intensity sports provided the most benefits to activity and reduction of sedentary behavior, while valuing household chores had unexpected negative effects.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    PubMed Central

    Klitgaard, Kirstine; Boye, Mette; Capion, Nynne; Jensen, Tim K.

    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 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 rRNA-directed oligonucleotide probes. Two phylotypes, phylotype 1 (PT1) and PT2, were not closely related to any characterized treponemal species. PT7 was 99.3% identical to Treponema denticola, while PT9 resembled T. vincentii by 96%. The remaining phylotypes, PT3, PT4, PT5, PT6, and PT8, and Treponema brennaborense had previously been isolated from DD lesions. Forty DD biopsy specimens were examined for Treponema by FISH. With one exception, all of the biopsy specimens revealed epidermotropic, intermingled infection with three or more different phylotypes (mean, 4.7). The most prevalent species were PT1 (95%), PT6 (93%), and PT3 (85%). While colonization by PT3 was confined to the surface of the epidermis, both PT1 and PT6 invaded deep into the stratum spinosum and were seen in ulcerated dermal papillae. In two cases, all 10 phylotypes were demonstrated. Furthermore, FISH with a Treponema group-specific probe showed 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. PMID:18562583

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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.

  4. Fluorescent microspheres

    NASA Technical Reports Server (NTRS)

    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.

  5. Vibronic model of PTCDA stacks: fluorescence and relaxation energies

    NASA Astrophysics Data System (ADS)

    Hennessy, M. H.; Pascal, R. A., Jr.; Soos, Zoltan G.

    1999-12-01

    Face-to-face stacks of PTCDA molecules in films and multiple quantum wells are modeled using Frenkel and charge-transfer excitons that are coupled to a characteristic molecular vibration. The intensity and vibronic structure of film fluorescence are extensions of the mixed Frenkel-CT model for absorption and electroabsorption. Ab initio relaxation energies of PTCDA excited states and molecular ions provide independent estimates of model parameters, while the (pi) - (pi) * transition dipole gives the hopping integral J. PTCDA's close connections to conjugated hydrocarbons and its structural and electronic simplicity are emphasized.

  6. Integrated fluorescence analysis system

    DOEpatents

    Buican, Tudor N.; Yoshida, Thomas M.

    1992-01-01

    An integrated fluorescence analysis system enables a component part of a sample to be virtually sorted within a sample volume after a spectrum of the component part has been identified from a fluorescence spectrum of the entire sample in a flow cytometer. Birefringent optics enables the entire spectrum to be resolved into a set of numbers representing the intensity of spectral components of the spectrum. One or more spectral components are selected to program a scanning laser microscope, preferably a confocal microscope, whereby the spectrum from individual pixels or voxels in the sample can be compared. Individual pixels or voxels containing the selected spectral components are identified and an image may be formed to show the morphology of the sample with respect to only those components having the selected spectral components. There is no need for any physical sorting of the sample components to obtain the morphological information.

  7. Delayed fluorescence in photosynthesis.

    PubMed

    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 fluorescenc