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Sample records for fluorescence intensity multiple

  1. Fluorescence detection by intensity changes for high-performance thin-layer chromatography separation of lipids using automated multiple development.

    PubMed

    Cebolla, Vicente L; Jarne, Carmen; Domingo, Pilar; Domínguez, Andrés; Delgado-Camón, Aránzazu; Garriga, Rosa; Galbán, Javier; Membrado, Luis; Gálvez, Eva M; Cossío, Fernando P

    2011-05-13

    Changes in emission of berberine cation, induced by non-covalent interactions with lipids on silica gel plates, can be used for detecting and quantifying lipids using fluorescence scanning densitometry in HPTLC analysis. This procedure, referred to as fluorescence detection by intensity changes (FDIC) has been used here in combination with automated multiple development (HPTLC/AMD), a gradient-based separation HPTLC technique, for separating, detecting and quantifying lipids from different families. Three different HPTLC/AMD gradient schemes have been developed for separating: neutral lipid families and steryl glycosides; different sphingolipids; and sphingosine-sphinganine mixtures. Fluorescent molar responses of studied lipids, and differences in response among different lipid families have been rationalized in the light of a previously proposed model of FDIC response, which is based on ion-induced dipole interactions between the fluorophore and the analyte. Likewise, computational calculations using molecular mechanics have also been a complementary useful tool to explain high FDIC responses of cholesteryl and steryl-derivatives, and moderate responses of sphingolipids. An explanation for the high FDIC response of cholesterol, whose limit of detection (LOD) is 5 ng, has been proposed. Advantages and limitations of FDIC application have also been discussed. PMID:21145556

  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. Anomalous fluorescence line intensity in megavoltage bremsstrahlung

    NASA Astrophysics Data System (ADS)

    Pereira, Nino; Litz, Marc; Merkel, George; Schumer, Joseph; Seely, John; Carroll, Jeff

    2009-11-01

    A Cauchois transmission crystal spectrometer intended for laser plasma diagnostics has measured an anomalous ratio between the fluorescence lines in megavoltage bremsstrahlung. When observed in reflection, Kα1 fluorescence is twice as strong as the Kβ line, as is usual. However, in forward-directed bremsstrahlung from a 2 MV end point linear accelerator with a tungsten converter, the Kα1 and Kβ fluorescence are approximately equal. The anomalous fluorescence line ratio, unity, reflects the large amount of fluorescence generated on the side of the converter where the electrons enter, and the differential attenuation of the fluorescence photons as they pass through the converter to opposite side. Understanding of fluorescence in megavoltage bremsstrahlung is relevant to the explanation of anomalous line ratios in spectra produced by high-energy electrons generated by intense femtosecond laser irradiation.

  4. The Development of Fluorescence Intensity Standards

    PubMed Central

    Gaigalas, A. K.; Li, Li; Henderson, O.; Vogt, R.; Barr, J.; Marti, G.; Weaver, J.; Schwartz, A.

    2001-01-01

    The use of fluorescence as an analytical technique has been growing over the last 20 years. A major factor in inhibiting more rapid growth has been the inability to make comparable fluorescence intensity measurements across laboratories. NIST recognizes the need to develop and provide primary fluorescence intensity standard (FIS) reference materials to the scientific and technical communities involved in these assays. The critical component of the effort will be the cooperation between the Federal laboratories, the manufacturers, and the technical personnel who will use the fluorescence intensity standards. We realize that the development and use of FIS will have to overcome many difficulties. However, as we outline in this article, the development of FIS is feasible.

  5. Manipulating fluorescence intensity with mechanical strains

    NASA Astrophysics Data System (ADS)

    Zhao, Weiwei; Bi, Kedong; Zhang, Hongze; Guo, Xitao; Ni, Zhenhua; Chen, Yunfei

    2015-01-01

    In this paper we show that the fluorescence of rhodamine 6G (R6G) can be manipulated by applying mechanical strains to gold nanoparticles (GNs) sandwiched in graphene/GNs/polydimethylsiloxane (graphene/GNs/PDMS) structure. The fluorescence intensity of R6G on the graphene/GNs/PDMS sample shows a gradual increase with the mechanical strain. However, the fluorescence intensity of R6G on the graphene/PDMS structure without the GNs buried in between is almost unchanged under the action of the external mechanical strain. Experiment results indicate that the gap distance change between the GNs is the main cause of the fluorescence intensity increase and graphene, as a passivation layer, does not block the energy transfer from R6G to GNs. Compared with that tuning the gap distance between GNs by preparing various GNs samples, applying macroscopic mechanical strain on GNs is a simple way to manipulate the fluorescence intensity of a specific material and brings a new perspective for optoelectronic applications.

  6. Highly intense fluorescence of novel carbon nanocrystals combined with a DNAzyme-assisted autocatalytic multiple amplification strategy for sensitive detection of thrombin.

    PubMed

    Wang, Xiaochun; Lu, Zhengkun; Tan, Lu; Jie, Guifen

    2016-05-10

    In this work, novel water-soluble carbon nanocrystals (CNCs) with excellent fluorescence were prepared, and successfully applied to sensitive fluorescence detection of thrombin by using an enzyme-assisted autocatalytic DNA recycling amplification strategy. PMID:27079442

  7. 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

  8. An optical microsensor to measure fluorescent light intensity in biofilms.

    PubMed

    Beyenal, Haluk; Yakymyshyn, Chris; Hyungnak, Jeon; Davis, Catherine C; Lewandowski, Zbigniew

    2004-09-01

    We have developed an optical microsensor to quantify fluorescent light intensity distribution in biofilms. The optical system consisted of a beam splitter, light couplers, filters and a spectrophotometer able to accept the fiberoptic cable to measure fluorescent light intensity. The emitted light, fluorescence from the biofilm, was collected at the tip of the optical microsensor and was transferred to a spectrophotometer via a fiberoptic cable. The total fluorescent light intensity was evaluated from the emission spectrum by numerical integration. The newly developed fiberoptic microsensor was tested using a Staphylococcus aureus strain producing yellow fluorescent protein (YFP) grown as biofilm. We used a 405-nm violet laser diode for excitation, and measured the emission intensity between 480 nm and 540 nm. The optical microsensor that quantifies fluorescent light intensity is a promising tool in biofilm research which often requires detection and quantification of fluorescent light intensity distribution generated by various fluorescent proteins. PMID:15279941

  9. Three Dimensional Fluorescence Imaging Using Multiple Light-Sheet Microscopy

    PubMed Central

    Mohan, Kavya; Purnapatra, Subhajit B.; Mondal, Partha Pratim

    2014-01-01

    We developed a multiple light-sheet microscopy (MLSM) system capable of 3D fluorescence imaging. Employing spatial filter in the excitation arm of a SPIM system, we successfully generated multiple light-sheets. This improves upon the existing SPIM system and is capable of 3D volume imaging by simultaneously illuminating multiple planes in the sample. Theta detection geometry is employed for data acquisition from multiple specimen layers. This detection scheme inherits many advantages including, background reduction, cross-talk free fluorescence detection and high-resolution at long working distance. Using this technique, we generated equi-intense light-sheets of thickness approximately with an inter-sheet separation of . Moreover, the light-sheets generated by MLSM is found to be 2 times thinner than the state-of-art SPIM system. Imaging of fluorescently coated yeast cells of size (encaged in Agarose gel-matrix) is achieved. Proposed imaging technique may accelerate the field of fluorescence microscopy, cell biology and biophotonics. PMID:24911061

  10. 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. PMID:26826684

  11. Phycobiliprotein fusion proteins: versatile intensely fluorescent constructs

    NASA Astrophysics Data System (ADS)

    Glazer, Alexander N.; Cai, Yuping A.; Tooley, Aaron J.

    2004-06-01

    Since 1982, phycobiliproteins have served as fluorescent labels in a wide variety of cell and molecule analyses. The exceptional spectroscopic properties of these labels include very high absorbance coefficients and quantum yields, and large Stokes shifts. The spectroscopic diversity of these reagents is restricted to a subset of naturally occurring phycobiliproteins with stable assembly states in vitro, whose target specificity is generated by chemical conjugation to proteins or small molecules. The latter step generates heterogeneity. These limitations have been overcome by expressing various recombinant phycobiliprotein constructs in the cyanobacterium Anabaena sp. PCC7120. Modular recombinant phycobiliprotein-based labels were constructed with some or all of the following features (a) an affinity purification tag; (b) a stable oligomerization domain (to maintain stable higher order assemblies of the phycobiliprotein monomers at very low protein concentration); (c) a biospecific recognition domain. Such phycobiliprotein constructs are readily purified from crude cell extracts by affinity chromatography and used directly as fluorescent labels. To generate constructs for intracellular in vivo labeling, the entire pathways for the biosynthesis of the His-tagged holo- α (phycocyanobilin-bearing) subunit of phycocyanin (emission max. 641 nm) and of the His-tagged holo-α (phycobiliviolin-bearing) subunit of phycoerythrocyanin (emission max. 582 nm) were reconstituted in Escherichia coli.

  12. 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.

  13. Dipyrrolylquinoxaline difluoroborates with intense red solid-state fluorescence.

    PubMed

    Yu, Changjiang; Hao, Erhong; Li, Tingting; Wang, Jun; Sheng, Wanle; Wei, Yun; Mu, Xiaolong; Jiao, Lijuan

    2015-08-21

    A set of organic fluorescent dyes of dipyrrolylquinoxalines (PQs ) and their BF2 complexes (BPQs ) were synthesized from commercial reagents, and were characterized by their X-ray structural analysis, and optical and electrochemical properties. BPQs showed intense broad absorption in the visible region in the solution-state. In comparison with that of PQs , there is an over 110 nm red-shift of the absorption maximum in the BPQs (up to 583 nm). Interestingly, dyes all exhibit red solid-state fluorescence with moderate to high fluorescence quantum yields except for PQ which showed bright yellow solid-state fluorescence. X-ray structures of BPQs showed the planar structure of quinoxaline with one pyrrole unit via the BF2 chelation and the almost perpendicular orientation of the uncoordinated pyrrole to the NBN core plane (the dihedral angle of 70-73°). The extended π-conjugation was in good agreement with the observed red-shift of the spectra. These dyes formed well-ordered intermolecular packing structures via the intermolecular hydrogen bonding between the N atoms of quinoxaline moieties and the NH units of adjacent pyrroles. The lack of π-π stacking in their crystal packing structures may explain the interestingly intense solid-state fluorescence of these dyes. PMID:26152609

  14. 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.

  15. 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. PMID:24692055

  16. A supramolecular cross-linked conjugated polymer network for multiple fluorescent sensing.

    PubMed

    Ji, Xiaofan; Yao, Yong; Li, Jinying; Yan, Xuzhou; Huang, Feihe

    2013-01-01

    A supramolecular cross-linked network was fabricated and demonstrated to act as a multiple fluorescent sensor. It was constructed from a fluorescent conjugated polymer and a bisammonium salt cross-linker driven by dibenzo[24]crown-8/secondary ammonium salt host-guest interactions. Compared with the conjugated polymer, the network has weak fluorescence due to the aggregation of polymer chains. Thanks to the multiple stimuli-responsiveness of host-guest interactions, the fluorescence intensity of the system can be enhanced by four types of signals, including potassium cation, chloride anion, pH increase, and heating. Hence, the network can serve as a cation sensor, an anion sensor, a pH sensor, and a temperature sensor. It can be used in both solution and thin film. Interestingly, exposure of a film made from this supramolecular cross-linked network to ammonia leads to an increase of fluorescence, making it a good candidate for gas detection. PMID:23259828

  17. 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.

  18. A triple-color fluorescent probe for multiple nuclease assays.

    PubMed

    Xu, Qinfeng; Zhang, Yihong; Zhang, Chun-yang

    2015-06-01

    We develop a triple-color fluorescent probe which may function as a lab-on-a-DNA-molecule for simultaneous detection of multiple exonucleases/restriction endonucleases. This triple-color fluorescent probe can be further applied for the discrimination of seven exonucleases and four cell lines as well as the screening of various nuclease inhibitors. PMID:25940190

  19. The instantaneous light-intensity function of a fluorescent lamp

    NASA Astrophysics Data System (ADS)

    Gluskin, Emanuel; Topalis, Frangiskos V.; Kateri, Ifigenia; Bisketzis, Nikolas

    2006-05-01

    Using some simple physics and “system” considerations, the instantaneous light intensity function ψ(t) of a fluorescent lamp fed via a regular ballast from the 50 60 Hz line is argued to be ψ(t)=ψ+bp(t), where p(t) is the instantaneous power function of the lamp, and b is a constant, and experiment confirms this formula well. The main frequency of ψ(t), the very significant singularity of its waveform, and the relative intensity of the ripple, i.e., the depth of the modulation, are the focus. The results are important for research into the vision problem that some humans (autistic, but others, too) experience regarding fluorescent light. The inertia of the processes in the lamp which are responsible for the light emission, provides some nonzero emission at the instants when p(t) has zeros. The smaller the volume of the tube and the mass of the gas are, the more weakly the inertia of the processes is expressed, and the relatively smaller is ψ. However, it should be very difficult to theoretically obtain ψ(t), in particular ψ, from the very complicated physics of the low-pressure discharge in the tube. We conclude that ψ has to be connected with the (also easily measured) lamp's inductance. The work should attract more attention of the physicists to the properties of the common fluorescent lamps.

  20. Unmixing multiple adjacent fluorescent targets with multispectral excited fluorescence molecular tomography.

    PubMed

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

    2016-06-20

    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 unmix multiple adjacent fluorescent targets that have different functional features but are labeled with the same fluorochrome. A method based on independent component analysis for multispectral excited FMT was proposed in our previous study. It showed that double fluorescent targets with certain edge-to-edge distance (EED) could be unmixed by the method. In this study, the situation is promoted to unmix multiple adjacent fluorescent targets (i.e., more than two fluorescent targets and EED=0). Phantom experiments on the resolving ability of the proposed algorithm demonstrate that the algorithm performs well in unmixing multiple adjacent fluorescent targets in both lateral and axial directions. And also, we recovered the locational information of each independent fluorescent target and described the variable trends of the corresponding fluorescent targets under the excitation spectrum. This method is capable of unmixing multiple fluorescent targets with small EED but labeled with the same fluorochrome, and may be used in imaging of nonspecific probe targeting and metabolism of drugs. PMID:27409108

  1. 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.

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

    PubMed Central

    Hur, Kwang-Ho; Mueller, Joachim D.

    2015-01-01

    The brightness measured by fluorescence fluctuation spectroscopy specifies the average stoichiometry of a labeled protein in a sample. Here we extended brightness analysis, which has been mainly applied in eukaryotic cells, to prokaryotic cells with E. coli serving as a model system. The small size of the E. coli cell introduces unique challenges for applying brightness analysis that are addressed in this work. Photobleaching leads to a depletion of fluorophores and a reduction of the brightness of protein complexes. In addition, the E. coli cell and the point spread function of the instrument only partially overlap, which influences intensity fluctuations. To address these challenges we developed MSQ analysis, which is based on the mean Q-value of segmented photon count data, and combined it with the analysis of axial scans through the E. coli cell. The MSQ method recovers brightness, concentration, and diffusion time of soluble proteins in E. coli. We applied MSQ to measure the brightness of EGFP in E. coli and compared it to solution measurements. We further used MSQ analysis to determine the oligomeric state of nuclear transport factor 2 labeled with EGFP expressed in E. coli cells. The results obtained demonstrate the feasibility of quantifying the stoichiometry of proteins by brightness analysis in a prokaryotic cell. PMID:26099032

  3. 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.

  4. Quantitating Fluorescence Intensity From Fluorophore: Assignment of MESF Values

    PubMed Central

    Gaigalas, A. K.; Wang, Lili; Schwartz, Abe; Marti, Gerald E.; Vogt, Robert F.

    2005-01-01

    A procedure is presented to convert the comparison of measured fluorescence signals into a comparison of fluorescence yields (FY). The fluorescence yield, which is a property of a solution or a suspension, is defined as the product of the fluorophore concentration and the molecular quantum yield. The paper revises the measurement model which relates the measured fluorescence signal to the FY. The equality of FY of two solutions provides an equivalence between the concentrations of fluorophore in the two solutions. The equivalence is the basis for quantitation in terms of molecules of equivalent soluble fluorophore (MESF). The quantitation procedure starts with the measurement of fluorescence signals from a serial dilution of fluorescein solutions to obtain a calibration of a fluorometer. The fluorometer is used to measure the fluorescence signal of a suspension of microspheres with immobilized fluorescein isothiocyanate (FITC). The calibration is used to obtain the concentration of soluble fluorophores which gives the same fluorescence signal as the microsphere suspension. The number concentration of microspheres is measured and the equality of fluorescence yields is used to obtain the number of soluble fluorescein molecules equivalent to a single microsphere. PMID:27308107

  5. 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.

  6. Ambiguous dependence of fluorescence intensity of trees on chlorophyll concentration

    NASA Astrophysics Data System (ADS)

    Zavoruev, Valeriy V.; Zavorueva, Elena N.

    2014-11-01

    Using fluorimetry Junior PAM (Heinz Walz GmbH, Germany) fluorescence parameters of leaves Prinsepia sinensis, Crataegus chlorocarca M, Acer negúndo, Bétula péndula are studied. It was found that the dependence of maximum fluorescence (Fm) plants on the concentration of chlorophyll depends on the sampling method during of vegetation. The correctness of sampling proves during vegetation is substantiated.

  7. 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.

  8. INTER-LABORATORY STUDY OF CELLULAR FLUORESCENCE INTENSITY MEASUREMENTS WITH FLUORESCEIN-LABELED MICROBEAD STANDARDS

    EPA Science Inventory

    To determine the precision of cellular fluorescence intensity (FI) measurements derived from labeled microbead standards, FI results were compared from 43 different flow cytometers in 34 laboratories. ll laboratories analyzed prepared aliquots of fluoresceinated calf thymocyte nu...

  9. Theory of light quenching: effects of fluorescence polarization, intensity, and anisotropy decays.

    PubMed Central

    Kuśba, J; Bogdanov, V; Gryczynski, I; Lakowicz, J R

    1994-01-01

    Experimental studies have recently demonstrated that fluorescence emission can be quenched by laser light pulses from modern high repetition rate lasers, a phenomenon we call "light quenching." We now describe the theory of light quenching and some of its effects on the steady-state and time-resolved intensity and anisotropy decays of fluorophores. Light quenching can decrease or increase the steady-state or time-zero anisotropy. Remarkably, the light quenching can break the usual z axis symmetry of the excited-state population, and the emission polarization can range from -1 to +1 under selected conditions. The measured anisotropy (or polarization) depends upon whether the observation axis is parallel or perpendicular to the propagation direction of the light quenching beam. The effects of light quenching are different for a single pulse, which results in both excitation and quenching, as compared with a time-delayed quenching pulse. Time-delayed light quenching pulses can result in step-like changes in the time-dependent intensity or anisotropy and are predicted to cause oscillations in the frequency-domain intensity and anisotropy decays. The increasing availability of pulsed laser sources offers the opportunity for a new class of two-pulse or multiple-pulse experiments where the sample is prepared by an excitation pulse, the excited state population is modified by the quenching pulse(s), followed by time- or frequency-domain measurements of the resulting emission. PMID:7858140

  10. High-intensity xenon pulse light source for fluorescence excitation

    NASA Astrophysics Data System (ADS)

    Miyamoto, Makoto; Ueno, Kazuo

    1997-05-01

    A newly developed 60W xenon flash lamp, L6604 and L6605, achieves the goals of longer operating life, higher output, and improved light stability. It operates at 2 Joules per flash input energy with approximately a 4 microsecond flash duration. The stability achieved is 2-3 percent peak-to-peak during a lifetime of 5 X 10e7 flashes, which is almost double that of conventional xenon flash lamps. This newly developed xenon flashlamp should serve as an excellent light source for analytical cytology and other fluorescence instruments. It can function as a high output, stable excitation light source for conventional fluorescence or delayed luminescence with a CCD. Besides providing powerful and stable illumination for absorption analysis of cells on slides, this lamp eliminates the optical artifacts associated with vibration of the stage which often limit throughput. This paper will describe in detail performance improvements obtained from this newly developed xenon flash lamp.

  11. 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.

  12. 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.

  13. Determination of Some Non-sedating Antihistamines via Their Native Fluorescence and Derivation of Some Quantitative Fluorescence Intensity - Structure Relationships.

    PubMed

    El-Kommos, Michael E; El-Gizawy, Samia M; Atia, Noha N; Hosny, Noha M

    2015-11-01

    A validated simple, novel, and rapid spectrofluorimetric method was developed for the determination of some non-sedating antihistamines (NSAs); namely cetirizine (CTZ), ebastine (EBS), fexofenadine (FXD), and loratadine (LOR). The method is based on measuring the native fluorescence of the cited drugs after protonation in acidic media and studying their quantitative fluorescence intensity - structure relationships. There was a linear relationship between the relative fluorescence intensity and the concentration of the investigated drug. Under the optimal conditions, the linear ranges of calibration curves for the determination of the studied NSAs were 0.10-2.0, 0.20-6.0, and 0.02-1.0 [Formula: see text] for (CTZ, FXD), (EBS), and (LOR); respectively. The factors affecting the protonation of the studied drugs were carefully studied and optimized. The method was validated according to ICH guidelines. The suggested method is applicable for the determination of the four investigated drugs in bulk and pharmaceutical dosage forms with excellent recoveries (97.67-103.80%). Quantitative relationships were found between the relative fluorescence intensities of the protonated drugs and their physicochemical parameters namely: the pKa, log P, connectivity indexes (χ(v)) and their squares. Regression equations (76) were obtained and not previously reported. Six of these equations were highly significant and used for the prediction of RFI of the studied NSAs. PMID:26439930

  14. Diagnostic potential of fluorescence of formalin-fixed paraffin-embedded malignant melanoma and pigmented skin lesions: quantitative study of fluorescence intensity using fluorescence microscope and digital imaging.

    PubMed

    Chwirot, B W; Sypniewska, N; Swiatlak, J

    2001-12-01

    The background for this study was reports in the literature of stronger fluorescence observed visually for melanomas compared with benign naevi in formalin-fixed paraffin-embedded sections. Our objective was to carry out a quantitative study of the phenomenon and to investigate if such an approach could be used in the detection of melanomas. Microscopic digital imaging was used to measure quantitatively the fluorescence intensity in specimens from 50 malignant melanomas, four basal cell carcinomas and 58 benign lesions. The mean fluorescence intensity of the melanomas was considerably higher than of the other lesions. For melanomas, the intensity depended both on the distance from the skin surface and the distance from the centre of the lesion. A simple algorithm based on the intensity threshold correctly classified the melanomas with a sensitivity of 74% and a specificity of 59%. Quantitative measurements of the fluorescence of the pigmented skin lesions fixed with formalin and embedded in paraffin can be a useful auxiliary tool for differentiating melanoma from other pigmented lesions histopathologically. PMID:11725203

  15. Quantitating Fluorescence Intensity From Fluorophores: Practical Use of MESF Values

    PubMed Central

    Wang, Lili; Gaigalas, Adolfas K.; Abbasi, Fatima; Marti, Gerald E.; Vogt, Robert F.; Schwartz, Abe

    2002-01-01

    The present work uses fluorescein as the model fluorophore and points out critical steps in the use of MESF (Molecules of Equivalent Soluble Fluorophores) values for quantitative flow cytometric measurements. It has been found that emission spectrum matching between a reference solution and an analyte and normalization by the corresponding extinction coefficient are required for quantifying fluorescence signals using flow cytometers. Because of the use of fluorescein, the pH value of the medium is also critical for accurate MESF assignments. Given that the emission spectrum shapes of microbead suspensions and stained biological cells are not significantly different, the percentage of error due to spectrum mismatch is estimated. We have also found that the emission spectrum of a microbead with a seven-methylene linker between the fluorescein and the bead surface (bead7) provides the best match with the spectra from biological cells. Therefore, bead7 is potentially a better calibration standard for flow cytometers than the existing one that is commercially available and used in the present study.

  16. 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).

  17. Multiple vibration intensities and frequencies for bone mineral density improvement.

    PubMed

    Ezenwa, Bertram; Burns, Edith; Wilson, Charles

    2008-01-01

    Devices that deliver controlled quantum vibration intensities at multiple frequencies (QVIMF) provide optimal stress to the musculoskeletal system for improved bone mineral density and muscle strength. This paper presents development of a QVIMF system and pilot study to determine device performance. Development is centered on specially-designed actuators that comprise multiple nodes of controlled and smooth, but variable rates of contact on a telescoping platform through sets of damping subsystems. The combination of specially-designed actuators and damping subsystems, powered by a DC controlled motor, delivers quantum busts of vibration at multiple frequencies resulting in whole body vibration. An initial feasibility study involved a 79 year old adult male. After IRB approval from both the University of Wisconsin-Milwaukee (UWM) and the Zablocki VA Medical Center, Milwaukee, the subject's bone mineral density (BMD) was measured by dual x-ray absorptimetry (DXA) at baseline. The subject then visited the UWM laboratory for two fifteen-minute vibration sessions per visit, three times a week for a total of 60 visits. Post-vibration BMD was again measured by DXA. Comparison pre- and post-vibration test results showed increases in BMD at the femoral neck, trochanter, total hip, forearm and lower lumbar spine (L1-4). PMID:19163635

  18. Narrow-Band Emitting Solid Fluorescence Reference Standard with Certified Intensity Pattern.

    PubMed

    Hoffmann, Katrin; Spieles, Monika; Bremser, Wolfram; Resch-Genger, Ute

    2015-07-21

    The development of a lanthanum-phosphate glass doped with several rare-earth-ions for use as solid fluorescence standard is described. The cuvette-shaped reference material which shows a characteristic emission intensity pattern upon excitation at 365 nm consisting of a multitude of relatively narrow emission bands in the wavelength region between 450 and 700 nm is intended for the day-to-day performance validation of fluorescence measuring devices. Evaluation of the fluorescent glass includes the determination of all properties which can affect its relative emission intensity profile or contribute to the uncertainty of the certified values like absorption spectra, fluorescence anisotropy, excitation wavelength, and temperature dependence of the spectroscopic features, homogeneity of fluorophore distribution, and photo- and long-term stability. Moreover, a certification procedure was developed including the normalization of the intensity profile consisting of several narrow emission bands and the calculation of wavelength-dependent uncertainties. Criteria for the design, characterization, and working principle of the new reference material BAM-F012 are presented, and possible applications of this ready-to-use fluorescence standard are discussed. PMID:26077510

  19. 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.

  20. Multiphoton fluorescence microscopy: behavior of biological specimens under high-intensity illumination

    NASA Astrophysics Data System (ADS)

    Cheng, Ping C.; Lin, Bai-Ling; Kao, Fu-Jen; Sun, Chi-Kuang

    2000-07-01

    Recent development in multi-photon fluorescence microscopy, second and third harmonic generation microscopy (SHG and THG) and CARS open new dimensions in biological studies. Not only the technologies allow probing the biological specimen both functionally and structurally with increasing spatial and temporal resolution, but also raise the interest in how biological specimens respond to high intensity illumination commonly 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 high intensity illumination. It was found that cells can be seriously damaged by high intensity NIR irradiation even the linear absorption coefficient in low in these wavelengths. Micro-spectroscopy of single chloroplast also allows us to gain insight on the possible photo-damage mechanism. In addition to fluorescence emission, second harmonic generation was observed in the maize protoplasts.

  1. 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. PMID:26322591

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

    PubMed

    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. PMID:23208294

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

    PubMed Central

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

    2012-01-01

    Abstract. 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. PMID:23208294

  4. Precise intensity correlation measurement for atomic resonance fluorescence from optical molasses.

    PubMed

    Nakayama, Kazuyuki; Yoshikawa, Yutaka; Matsumoto, Hisatoshi; Torii, Yoshio; Kuga, Takahiro

    2010-03-29

    We measured the intensity correlation of true thermal light scattered from cold atoms in an optical molasses. Using a single-mode fiber as a transverse mode filter, measurement with maximally high spatial coherence was realized, allowing us to observe ideal photon bunching with unprecedented precision. The measured intensity correlation functions showed a definite bimodal structure with fast damped oscillation from the maximum value of 2.02(3) and slow monotonic decay toward unity. The oscillation can be understood as an interference between elastic and inelastic scattering fields in resonance fluorescence. PMID:20389684

  5. The use of simultaneous confidence bands for comparison of single parameter fluorescent intensity data.

    PubMed

    Kim, Dongha; Donnenberg, Vera S; Wilson, John W; Donnenberg, Albert D

    2016-01-01

    Despite the utility of multiparameter flow cytometry for a wide variety of biological applications, comparing single parameter histograms of fluorescence intensity remains a mainstay of flow cytometric analysis. Even comparisons requiring multiparameter gating strategies often end with single parameter histograms as the final readout. When histograms overlap, analysis relies on comparison of mean or median fluorescence intensities, or determination of percent positive based on an arbitrary cutoff. Earlier attempts to address this problem utilized either simple channel-by-channel subtraction without statistical evaluation, or the Kolmogorov-Smirnov (KS) or Chi-square test statistics, both of which proved to be overly sensitive to small and biologically insignificant differences. Here we present a method for the comparison of two single-parameter histograms based on difference curves and their simultaneous confidence bands generated by bootstrapping raw channel data. Bootstrapping is a nonparametric statistical approach that can be used to generate confidence intervals without distributional assumptions about the data. We have constructed simultaneous confidence bands and show them to be superior to KS and Cox methods. The method constructs 95% confidence bands about the difference curves, provides a P value for the comparison and calculates the area under the difference curve (AUC) as an estimate of percent positive and the area under the confidence band (AUCSCB95), providing a lower estimate of the percent positive. To demonstrate the utility of this new approach we have examined single-color fluorescence intensity data taken from a cell surface proteomic survey of a lung cancer cell line (A549) and a published fluorescence intensity data from a rhodamine efflux assay of P-glycoprotein activity, comparing rhodamine 123 loading and efflux in CD4 and CD8 T-cell populations. SAS source code is provided as supplementary material. PMID:26407241

  6. Two-photon microscopy of living cells by simultaneously exciting multiple endogenous fluorophores and fluorescent proteins

    NASA Astrophysics Data System (ADS)

    Zheng, Wei; Li, Dong; Qu, Jianan Y.

    2010-02-01

    Endogenous fluorophores, such as reduced nicotinamide adenine dinucleotide (NADH), keratin, and tryptophan, have been used as contrast agents for imaging metabolism and morphology of living cells and tissues. Multilabeling which maps the distribution of different targets is an indispensable technique in many biomedical and biochemical studies. Therefore, two-photon excitation fluorescence (TPEF) microscopy of endogenous fluorophores combining with in vivo fluorescence labeling techniques such as genetically encoded fluorescent protein could be a powerful tool for imaging living cells and tissues. However, the challenge is that the excitation and emission wavelengths of these endogenous fluorophores and fluorescence labels are very different. A multi-color ultrafast source is required for the excitation of multiple fluorescence molecules. In this study, we developed a two-photon imaging system with excitations from the pump femtosecond laser and the selected Supercontinuum generated from a photonic crystal fiber (PCF). Multiple endogenous fluorophores and fluorescent proteins such as NADH, tryptophan, green fluorescent protein (GFP), and yellow fluorescent protein (YFP) were excited in their optimal wavelengths alternately or simultaneously. A time- and spectral-resolved detection system was used to record the TPEF signals. This detection technique separated the TPEF signals from multiple sources in time and spectral domains. Cellular organelles such as nucleus, mitochondria, microtubule and Endoplasmic Reticulum (ER), were clearly revealed in the TPEF images.

  7. 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

  8. 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.

  9. 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

  10. Dependence of X-Ray fluorescence intensity on sample specific gravity

    SciTech Connect

    Az'muko, A.A.; Butuina, L.F.; Smagunova, A.N.; Tarasenko, S.V.

    1986-06-01

    In order to understand the physical nature of the experimental relation I=f(n), the authors set up tests that study the dependence of intensity on packing density. Tests were carried out on samples of the minerals galena cassiterite, and zircon, and cassiterite-calcite and zircon-quartz mixtures. The intensities of the Pb L Sn K /SUB alpha/ , and Zr K /SUB alpha/ lines were measured in a KRFS-5 spectrometer with BKhV-6 x-ray tube with tungsten anode. The experimental and theoretical functions I /SUB rel/ =fIn) for galena are shown and the values of the effect understudy are given. It is shown that the observed dependence of fluorescence intensity on sample density is due to the effect of sample surface quality on the value of I /SUB A/ .

  11. Homogeneous immunoassays based on fluorescence emission intensity variations of zinc selenide quantum dot sensors.

    PubMed

    Wang, Jun; Mountziaris, T J

    2013-03-15

    The fluorescence emission intensity of ZnSe quantum dots (QDs) conjugated to proteins to form QD-based biomolecular sensors increases significantly upon binding of the sensors to target proteins in solution. This phenomenon enables the development of homogeneous, separation-free immunoassays for rapid quantitative detection of proteins in solution. Proof-of-principle assays were developed by dosing a solution containing a biomolecular target with a solution containing the corresponding QD-based sensor and monitoring the changes in the peak fluorescence emission intensity of the QDs. Direct immunoassays for detecting basic fibroblast growth factor (bFGF) and prostate-specific antigen (PSA) in solution were demonstrated using QD-anti-bFGF and QD-anti-PSA sensors. A competitive immunoassay for detecting human serum albumin (HSA) was also demonstrated by dosing samples containing HSA with QD-HSA sensors and free anti-HSA antibodies. The QD-HSA sensors were tested in 1000× diluted human serum and found to be unaffected by interference from other proteins. The lower limit of detection of the assays was equal to the lowest sensor concentration in the solution that can be unambiguously detected, typically less than 1 nM. The dynamic range of the assays was determined by identifying the sensor concentration above which optical interference between QDs affected adversely the observed fluorescence emission intensity. The upper limit of this concentration was 2.5 μM for 4 nm QDs. The ZnSe QD-based sensors were stable and preserved ~80% of their initial peak emission intensity after two months in refrigerated storage. These biosensors have potential applications in rapid sensing of target proteins for emergency and point-of-care diagnostic applications. PMID:22960008

  12. 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.

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

    PubMed

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

    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. PMID:27423111

  14. Fe II fluorescence and anomalous C IV doublet intensities in symbiotic novae

    NASA Technical Reports Server (NTRS)

    Michalitsianos, A. G.; Kafatos, M.; Meier, S. R.

    1992-01-01

    The variation of absolute intensities of Bowen-excited Fe II emission in the symbiotic stars RR Tel, RX Pup, and AG Peg is examined. The C IV doublet intensity ratios in RR Tel were not anomalous between 1979 and 1989, and the ratio had typical values within the optically thin range. The intensity of individual Fe II Bowen-excited lines is correlated with the C IV 1548.2 A flux, suggesting the presence of a foreground Fe II region in which fluorescent-excited material responds to flux variations of C IV 1548.2 A. In RX Pup the combined fluxes of Fe II Bowen-pumped lines can account for an appreciable fraction of the flux deficit in the C IV 1548.2 A line when the C IV doublet ratio is less than the optically thick limit of unity. The Fe II Bowen lines in RX Pup exhibit a velocity range from 0 to 80 km/s, where several strong Fe II emission lines correspond to deep absorption structure in the C IV 1548.2 A line profile. In AG Peg and C IV 1548.2 A flux deficit cannot be explained by Fe II fluorescent absorption alone when the C IV doublet ratio anomaly is at an extreme.

  15. Improved phase imaging from intensity measurements in multiple planes

    SciTech Connect

    Soto, Marcos; Acosta, Eva

    2007-11-20

    Problems stemming from quantitative phase imaging from intensity measurements play a key role in many fields of physics. Techniques based on the transport of intensity equation require an estimate of the axial derivative of the intensity to invert the problem. Derivation formulas in two adjacent planes are commonly used to experimentally compute the derivative of the irradiance. Here we propose a formula that improves the estimate of the derivative by using a higher number of planes and taking the noisy nature of the measurements into account. We also establish an upper and lower limit for the estimate error and provide the distance between planes that optimizes the estimate of the derivative.

  16. Fluorescence Intensity- and Lifetime-Based Glucose Sensing Using Glucose/Galactose-Binding Protein

    PubMed Central

    Pickup, John C.; Khan, Faaizah; Zhi, Zheng-Liang; Coulter, Jonathan; Birch, David J. S.

    2013-01-01

    We review progress in our laboratories toward developing in vivo glucose sensors for diabetes that are based on fluorescence labeling of glucose/galactose-binding protein. Measurement strategies have included both monitoring glucose-induced changes in fluorescence resonance energy transfer and labeling with the environmentally sensitive fluorophore, badan. Measuring fluorescence lifetime rather than intensity has particular potential advantages for in vivo sensing. A prototype fiber-optic-based glucose sensor using this technology is being tested.Fluorescence technique is one of the major solutions for achieving the continuous and noninvasive glucose sensor for diabetes. In this article, a highly sensitive nanostructured sensor is developed to detect extremely small amounts of aqueous glucose by applying fluorescence energy transfer (FRET). A one-pot method is applied to produce the dextran-fluorescein isothiocyanate (FITC)-conjugating mesoporous silica nanoparticles (MSNs), which afterward interact with the tetramethylrhodamine isothiocyanate (TRITC)-labeled concanavalin A (Con A) to form the FRET nanoparticles (FITC-dextran-Con A-TRITC@MSNs). The nanostructured glucose sensor is then formed via the self-assembly of the FRET nanoparticles on a transparent, flexible, and biocompatible substrate, e.g., poly(dimethylsiloxane). Our results indicate the diameter of the MSNs is 60 ± 5 nm. The difference in the images before and after adding 20 μl of glucose (0.10 mmol/liter) on the FRET sensor can be detected in less than 2 min by the laser confocal laser scanning microscope. The correlation between the ratio of fluorescence intensity, I(donor)/I(acceptor), of the FRET sensor and the concentration of aqueous glucose in the range of 0.04–4 mmol/liter has been investigated; a linear relationship is found. Furthermore, the durability of the nanostructured FRET sensor is evaluated for 5 days. In addition, the recorded images can be converted to digital images by

  17. 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.

  18. 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.

  19. A direct method for the correction of pressure-induced scrambling of polarized fluorescence intensities.

    PubMed

    Targowski, P; Davenport, L

    1999-10-15

    A simple and direct method for the simultaneous correction of steady-state polarized fluorescence intensities, depolarized (or scrambled) by the effects of applied hydrostatic pressure, is described. In the method discussed here, it is not necessary to first determine the scrambling factors from a separate experiment with a dye immobilized in a rigid medium. Rather correction for depolarizing effects of the high-pressure spectroscopy cell windows is achieved by direct recalculation of the measured polarized data obtained for the sample of interest at the time of data collection. This method of correction is tested for common fluorescent dyes 1, 6-diphenyl-1,3,5-hexatriene (DPH) and 9,10-diphenylanthracene in glycerol where their rotational behavior is well understood. In addition, the pressure-induced "melt" profile for the more complicated biologically relevant system of DPH imbedded within dipalmitoylphosphatidylcholine small unilamellar vesicles has been reexamined. While the method discussed here is used for the correction of steady-state polarized data, it may be easily adapted for use in time-resolved polarized fluorescence measurements. Advantages and limitations of the new correction method are discussed. PMID:10527523

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

    PubMed

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

    2009-12-01

    In this paper the relationship between the chemical structure and fluorescence characteristics of 30 phenylquinolinylethyne (PhQE), and phenylisoquinolinylethyne (PhIE) derivatives compounds employing ab initio calculations have been elucidated. Quantum chemical calculations (6-31G) were carried out to obtain: the optimized geometry, energy levels, charges and dipole moments of these compounds, in the singlet (steady and excited states) and triplet states. The relationship between quantum chemical descriptors, and wavelength of maximum excitation and emission indicated that these two parameters have the most correlation with quantum chemical hardness (eta). Also, stokes shift has the most correlation with the square of difference between the maximum of positive charges in the singlet steady and singlet excited states. The quantitative structure-property relationship (QSPR) of PhQE and PhIE was studied for relative fluorescence intensity (RFI). The genetic algorithm (GA) was applied to select the variables that resulted in the best-fit models. After the variable selection, multiple linear regression (MLR) and support vector machine (SVM) were both utilized to construct linear and non-linear QSPR models, respectively. The SVM model demonstrated a better performance than that of the MLR model. The route mean square error (RMSE) in the training and the test sets for the SVM model was 0.195 and 0.324, and the correlation coefficients were 0.965 and 0.960, respectively, thus revealing the reliability of this model. The resulting data indicated that SVM could be used as a powerful modeling tool for QSPR studies. According to the best of our knowledge, this is the first research on QSPR studies to predict RFI for a series of PhQE and PhIE derivative compounds using SVM. PMID:19854100

  1. Photoactivatable aggregation-induced emission fluorophores with multiple-color fluorescence and wavelength-selective activation.

    PubMed

    Peng, Lu; Zheng, Yue; Wang, Xiaoyan; Tong, Aijun; Xiang, Yu

    2015-03-01

    Photoactivatable (caged) fluorophores are widely used in chemistry, materials, and biology. However, the development of such molecules exhibiting photoactivable solid-state fluorescence is still challenging due to the aggregation-caused quenching (ACQ) effect of most fluorophores in their aggregate or solid states. In this work, we developed caged salicylaldehyde hydrazone derivatives, which are of aggregation-induced emission (AIE) characteristics upon light irradiation, as efficient photoactivatable solid-state fluorophores. These compounds displayed multiple-color emissions and ratiometric (photochromic) fluorescence switches upon wavelength-selective photoactivation, and were successfully applied for photopatterning and photoactivatable cell imaging in a multiple-color and stepwise manner. PMID:25644036

  2. Mapping tillage intensity by integrating multiple remote sensing data

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  3. Fluorescent biosensor for the detection of hyaluronidase: intensity-based ratiometric sensing and fluorescence lifetime-based sensing using a long lifetime azadioxatriangulenium (ADOTA) fluorophore.

    PubMed

    Chib, Rahul; Mummert, Mark; Bora, Ilkay; Laursen, Bo W; Shah, Sunil; Pendry, Robert; Gryczynski, Ignacy; Borejdo, Julian; Gryczynski, Zygmunt; Fudala, Rafal

    2016-05-01

    In this report, we have designed a rapid and sensitive, intensity-based ratiometric sensing as well as lifetime-based sensing probe for the detection of hyaluronidase activity. Hyaluronidase expression is known to be upregulated in various pathological conditions. We have developed a fluorescent probe by heavy labeling of hyaluronic acid with a new orange/red-emitting organic azadioxatriangulenium (ADOTA) fluorophore, which exhibits a long fluorescence lifetime (∼20 ns). The ADOTA fluorophore in water has a peak fluorescence lifetime of ∼20 ns and emission spectra centered at 560 nm. The heavily ADOTA-labeled hyaluronic acid (HA-ADOTA) shows a red shift in the peak emission wavelength (605 nm), a weak fluorescence signal, and a shorter fluorescence lifetime (∼4 ns) due to efficient self-quenching and formation of aggregates. In the presence of hyaluronidase, the brightness and fluorescence lifetime of the sample increase with a blue shift in the peak emission to its original wavelength at 560 nm. The ratio of the fluorescence intensity of the HA-ADOTA probe at 560 and 605 nm can be used as the sensing method for the detection of hyaluronidase. The cleavage of the hyaluronic acid macromolecule reduces the energy migration between ADOTA molecules, as well as the degree of self-quenching and aggregation. This probe can be efficiently used for both intensity-based ratiometric sensing as well as fluorescence lifetime-based sensing of hyaluronidase. The proposed method makes it a rapid and sensitive assay, useful for analyzing levels of hyaluronidase in relevant clinical samples like urine or plasma. Graphical Abstract Scheme showing cleavage of HA-ADOTA probe by hyaluronidase and the change in the emission spectrum of HA-ADOTA probe before and after cleavage by hyaluronidase. PMID:26993308

  4. 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.

  5. 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.

  6. A handheld laser-induced fluorescence detector for multiple applications.

    PubMed

    Fang, Xiao-Xia; Li, Han-Yang; Fang, Pan; Pan, Jian-Zhang; Fang, Qun

    2016-04-01

    In this paper, we present a compact handheld laser-induced fluorescence (LIF) detector based on a 450 nm laser diode and quasi-confocal optical configuration with a total size of 9.1 × 6.2 × 4.1 cm(3). Since there are few reports on the use of 450 nm laser diode in LIF detection, especially in miniaturized LIF detector, we systematically investigated various optical arrangements suitable for the requirements of 450 nm laser diode and system miniaturization, including focusing lens, filter combination, and pinhole, as well as Raman effect of water at 450 nm excitation wavelength. As the result, the handheld LIF detector integrates the light source (450 nm laser diode), optical circuit module (including a 450 nm band-pass filter, a dichroic mirror, a collimating lens, a 525 nm band-pass filter, and a 1.0mm aperture), optical detector (miniaturized photomultiplier tube), as well as electronic module (including signal recording, processing and displaying units). This detector is capable of working independently with a cost of ca. $2000 for the whole instrument. The detection limit of the instrument for sodium fluorescein solution is 0.42 nM (S/N=3). The broad applicability of the present system was demonstrated in capillary electrophoresis separation of fluorescein isothiocyanate (FITC) labeled amino acids and in flow cytometry of tumor cells as an on-line LIF detector, as well as in droplet array chip analysis as a LIF scanner. We expect such a compact LIF detector could be applied in flow analysis systems as an on-line detector, and in field analysis and biosensor analysis as a portable universal LIF detector. PMID:26838391

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

    PubMed Central

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

    2015-01-01

    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. PMID:26080403

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

    PubMed

    Li, Jiayao; Henry, Etienne; Wang, Lanmei; Delelis, Olivier; Wang, Huan; Simon, Françoise; Tauc, Patrick; Brochon, Jean-Claude; Zhao, Yunlong; Deprez, Eric

    2012-01-01

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

  9. Comparative Study of the Fatty Acid Binding Process of a New FABP from Cherax quadricarinatus by Fluorescence Intensity, Lifetime and Anisotropy

    PubMed Central

    Li, Jiayao; Henry, Etienne; Wang, Lanmei; Delelis, Olivier; Wang, Huan; Simon, Françoise; Tauc, Patrick; Brochon, Jean-Claude; Zhao, Yunlong; Deprez, Eric

    2012-01-01

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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

  11. 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. PMID:25416508

  12. 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

  13. 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.

  14. 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.

  15. 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

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

    PubMed

    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

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

    PubMed Central

    Hsieh, Meng-Chang; Chiu, Yi-Hsin; Lin, Sheng-Fu; Chang, Jenq-Yang; Chang, Chia-Ou; Chiang, Huihua Kenny

    2015-01-01

    Fluorescent biosensors have been widely used in biomedical applications. To amplify the intensity of fluorescence signals, this study developed a novel structure for an evanescent wave fiber-optic biosensor by using a Fabry-Perot resonator structure. An excitation light was coupled into the optical fiber through a laser-drilled hole on the proximal end of the resonator. After entering the resonator, the excitation light was reflected back and forth inside the resonator, thereby amplifying the intensity of the light in the fiber. Subsequently, the light was used to excite the fluorescent molecules in the reactive region of the sensor. The experimental results showed that the biosensor signal was amplified eight-fold when the resonator reflector was formed using a 92% reflective coating. Furthermore, in a simulation, the biosensor signal could be amplified 20-fold by using a 99% reflector. PMID:25690548

  18. Limited-projection-angle hybrid fluorescence molecular tomography of multiple molecules.

    PubMed

    Radrich, Karin; Mohajerani, Pouyan; Bussemer, Johanna; Schwaiger, Markus; Beer, Ambros J; Ntziachristos, Vasilis

    2014-04-01

    An advantage of fluorescence methods over other imaging modalities is the ability to concurrently resolve multiple moieties using fluorochromes emitting at different spectral regions. Simultaneous imaging of spectrally separated agents is helpful in interrogating multiple functions or establishing internal controls for accurate measurements. Herein, we investigated multimoiety imaging in the context of a limited-projection-angle hybrid fluorescence molecular tomography (FMT), and x-ray computed tomography implementation and the further registration with positron emission tomography (PET) data. Multichannel FMT systems may image fluorescent probes of varying distribution patterns. Therefore, it is possible that different channels may require different use of priors and regularization parameters. We examined the performance of automatically estimating regularization factors implementing priors, using data-driven regularization specific for limited-projection-angle schemes. We were particularly interested in identifying the implementation variations between hybrid-FMT channels due to probe distribution variation. For this reason, initial validation of the data-driven algorithm on a phantom was followed by imaging different agent distributions in animals, assuming superficial and deep seated activity. We further demonstrate the benefits of combining hybrid FMT with PET to gain multiple readings on the molecular composition of disease. PMID:24770661

  19. Interphase fluorescence in situ hybridization signal detection by computing intensity variance along the optical axis

    NASA Astrophysics Data System (ADS)

    Li, Zheng; Zheng, Bin; Ren, Liqiang; Liu, Hong

    2014-02-01

    Fluorescence in situ Hybridization technology is a commonly used tool to detect chromosome aberrations, which are often pathologically significant. Since manual FISH analysis is a tedious and time-consuming procedure, reliable and robust automated image acquisition and analysis are in demand. Under high magnification objective lenses such as 60x and 100x, the depth of field will often be too small and the FISH probes may not always lie in the same focal plane. A statistical variance based automated FISH analysis method is developed in order to address this problem. On a stack of slices at consecutive image planes with a step size d, the statistical variance alone the z-axis is calculated to form a 2-D matrix. Since pixels shift dramatically to high intensity at FISH probe location, the probes will manifest high peak values in the matrix. A computer-aided detection scheme based on top-hat transform is applied to the matrix to detect FISH probe signals. This study demonstrates a simple and robust method for FISH probe detection as well as a way of 2- D representation of 3-D data.

  20. 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.

  1. Toward the measurement of multiple fluorescence lifetimes in flow cytometry: maximizing multi-harmonic content from cells and microspheres.

    PubMed

    Jenkins, Patrick; Naivar, Mark A; Houston, Jessica P

    2015-11-01

    Flow cytometry is a powerful means for in vitro cellular analyses where multi-fluorescence and multi-angle light scattering can indicate unique biochemical or morphological features of single cells. Yet, to date, flow cytometry systems have lacked the ability to capture complex fluorescence dynamics due to the transient nature of flowing cells. In this contribution we introduce a simple approach for measuring multiple fluorescence lifetimes from a single cytometric event. We leverage square wave modulation, Fourier analysis, and high frequency digitization and show the ability to resolve more than one fluorescence lifetime from fluorescently-labelled cells and microspheres. Illustration of a flow cytometer capable of capturing multiple fluorescence lifetime measurements; creating potential for multi-parametric, time-resolved signals to be captured for every color channel. PMID:25727072

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

    PubMed Central

    Barry, David J.; Durkin, Charlotte H.; Abella, Jasmine V.

    2015-01-01

    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. PMID:25847537

  3. Stress Intensity Factor Solutions for Multiple Edge Cracks in Ceramic Matrix Composites

    NASA Technical Reports Server (NTRS)

    Ghosn, Louis

    1997-01-01

    NASA Lewis Research Center conducted a study to determine the stress intensity factor solutions for periodic arrays of bridged cracks for various crack spacings and crack lengths. Initially, the stress intensity factor of an array of unbridged multiple edge cracks was determined under constant global displacement as well as at a point load along the crack wake. These solutions are expected to contribute toward the development of a damage-based life-prediction methodology for CMC engine components.

  4. 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. PMID:24995295

  5. Numerical evaluation of droplet sizing based on the ratio of fluorescent and scattered light intensities (LIF/Mie technique)

    SciTech Connect

    Charalampous, Georgios; Hardalupas, Yannis

    2011-03-20

    The dependence of fluorescent and scattered light intensities from spherical droplets on droplet diameter was evaluated using Mie theory. The emphasis is on the evaluation of droplet sizing, based on the ratio of laser-induced fluorescence and scattered light intensities (LIF/Mie technique). A parametric study is presented, which includes the effects of scattering angle, the real part of the refractive index and the dye concentration in the liquid (determining the imaginary part of the refractive index). The assumption that the fluorescent and scattered light intensities are proportional to the volume and surface area of the droplets for accurate sizing measurements is not generally valid. More accurate sizing measurements can be performed with minimal dye concentration in the liquid and by collecting light at a scattering angle of 60 deg. rather than the commonly used angle of 90 deg. Unfavorable to the sizing accuracy are oscillations of the scattered light intensity with droplet diameter that are profound at the sidescatter direction (90 deg.) and for droplets with refractive indices around 1.4.

  6. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2016-03-01

    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.

  8. 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

  9. 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

  10. Mixed-Dye-Based Label-Free and Sensitive Dual Fluorescence for the Product Detection of Nucleic Acid Isothermal Multiple-Self-Matching-Initiated Amplification.

    PubMed

    Ding, Xiong; Wu, Wenshuai; Zhu, Qiangyuan; Zhang, Tao; Jin, Wei; Mu, Ying

    2015-10-20

    Visual detections based on fluorescence and the color changes under natural light are two promising product detections for isothermal nucleic acid amplifications (INAAs) such as the isothermal multiple-self-matching-initiated amplification (IMSA) as point-of-care testing techniques. However, the currently used approaches have shortcomings in application. For the former, fluorescence changes recognized by naked eye may be indistinguishable because of single fluorescence emitted and strong background noise, which requires empirical preset of cutoff intensity values. For the latter, visual detection sensitivity under natural light is not comparable to that based on fluorescence. Herein, hydroxyl naphthol blue (HNB) and SYBR Green I (SG) were coupled to acquire a label-free dual fluorescence for the visual product detection of IMSA. The mixed-dye-loaded off-chip (tube-based) and on-chip (microfluidic chip-based) IMSAs for the detection of hepatitis B virus were conducted. The results demonstrated that this dual fluorescence could realize distinguishable fluorescent color changes to improve visual detection sensitivity and avoid the preset of cutoff values. Moreover, the mixed dye is stable when kept at room temperature and compatible with the IMSA's reagents without a contamination-prone step of opening tubes after amplification. Also, this coupled dye inherits the advantages of achieving color changes under natural light from HNB and real-time detection from SG. In conclusion, the mixed-dye-based dual fluorescence has a potential in the point-of-care testing application for realizing off-chip and on-chip product detection of IMSA, loop-mediated isothermal amplification (LAMP), or other INAAs. PMID:26383158

  11. 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.

  12. 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…

  13. Fluorescence molecular tomography on animal model by means of multiple views structured light illumination

    NASA Astrophysics Data System (ADS)

    Ducros, N.; Bassi, A.; Valentini, G.; Canti, G.; Arridge, S.; D'Andrea, C.

    2013-03-01

    Fluorescence molecular tomography (FMT) is quite demanding in terms of acquisition/computational times due to the huge amount of data. Different research groups have proposed compression approaches regarding both illumination (wide field structured light instead of raster point scanning) and detection (compression of the acquired images). The authors have previously proposed a fast FMT reconstruction method based on the combination of a multiple-view approach with a full compression scheme. This method had been successfully tested on a cylindrical phantom and is being generalized in this paper to samples of arbitrary shape. The devised procedure and algorithms have been tested on an ex-vivo mouse.

  14. New fluorescence markers to distinguish co-infecting Trypanosoma brucei strains in experimental multiple infections.

    PubMed

    Balmer, Oliver; Tostado, Cristóbal

    2006-01-01

    Multiple-genotype infections are increasingly recognized as important factors in disease evolution, parasite transmission dynamics, and the evolution of drug resistance. However, the distinction of co-infecting parasite genotypes and the tracking of their dynamics have been difficult with traditional methods based on various genotyping techniques, leaving most questions unaddressed. Here we report new fluorescence markers of various colours that are inserted into the genome of Trypanosoma brucei to phenotypically label live parasites of all life cycle stages. If different parasite strains are labelled with different colours they can be easily distinguished from each other in experimental studies. A total of 10 T. brucei strains were successfully transfected with different fluorescence markers and were monitored in culture, tsetse flies and mice, to demonstrate stability of marker expression. The use of fluorescence activated cell sorting (FACS) allowed rapid and accurate identification of parasite strains labelled with different markers. Cell counts by FACS were virtually identical to counts by traditional microscopy (n=75, Spearman's rho: 0.91, p<0.0001) but were considerably faster and had a significantly lower sampling error (66% lower, d.f.=73, t=-17.1, p<0.0001). Co-infecting strains transfected with fluorescence genes of different colour were easily distinguished by eye and their relative and absolute densities were reliably counted by FACS in experimental multiple infections in mice. Since the FACS can simultaneously determine the population sizes of differently labelled T. brucei strains or subspecies it allows detailed and efficient tracking of multiple-genotype infections within a single host or vector individual, enabling more powerful studies on parasite dynamics. In addition, it also provides a simple way to separate genotypes after experimental mixed infections, to measure responses of the single strains to an applied treatment, thus eliminating the

  15. 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…

  16. 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. PMID:25764015

  17. 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.

  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. PMID:22714463

  19. Short communication: Changes in fluorescence intensity induced by soybean soluble polysaccharide-milk protein interactions during acidification.

    PubMed

    Li, Y H; Wang, W J; Xu, X J; Meng, Y C; Zhang, L W; Chen, J; Qiu, R

    2015-12-01

    Interactions between stabilizer and milk protein are believed to influence the stabilizing behavior of the milk system. We investigated changes in fluorescence intensity induced by interactions of soybean soluble polysaccharide (SSPS) and milk protein (Mp) during acidification. The fluorescence intensity (If) of Mp increased as pH decreased from 6.8 to 5.2. Compared with Mp alone, If of SSPS-Mp mixtures increased as the pH decreased from 6.8 to 5.2. We found that the If of the SSPS-Mp mixture decreased in a pH range from 5.2 to 3.6, which indicated a change in the polarity microenvironment around the Trp residues. We also found that the maximum emission wavelength (λmax) shifted from 337 to 330nm as pH decreased from 6.8 to 3.6, in further support of SSPS interacting with the polar portion of Mp during acidification. Furthermore, an excited monomeric molecule (pyrene exciplex) was found as a ground-state pyrene formed and a broad band was shown at about 450nm. The intensity ratio of the first peak to the third peak (I1:I3) of Mp increased slightly, and the ratio of intensity of pyrene exciplex to monomer (Ie:Im) decreased because pyrene molecules were located in a less hydrophobic microenvironment during acidification. However, the ratio of I1:I3 decreased clearly at pH below 5.6 and the ratio of Ie:Im showed the opposite trend in the SSPS-Mp mixture. Changes in intrinsic and exogenous fluorescence intensity confirmed that interactions of SSPS and Mp could change the polarity of the microenvironment and that SSPS probably interacted with the polar portion of Mp. These results could give insight into the behavior of stabilizers in acid milk products. PMID:26476946

  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. PMID:27161580

  1. Multiple reflection high-energy electron diffraction beam intensity measurement system

    NASA Technical Reports Server (NTRS)

    Resh, J. S.; Jamison, K. D.; Strozier, J.; Ignatiev, A.

    1990-01-01

    A video-based analysis system for reflection high-energy electron diffraction (RHEED) is described which simultaneously measures the intensities and profiles of multiple diffraction beams. This system is used to record real-time RHEED intensity oscillations for layer-by-layer epitaxial growth. Fast Fourier transform analysis of the oscillation data is used to directly determine the growth rate and to accurately obtain phase information about the oscillations. This system is demonstrated and compared to other methods of recording RHEED oscillation data.

  2. 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. PMID:26773794

  3. Titration of a CD45-FITC conjugate to determine the linearity and dynamic range of fluorescence intensity measurements on lymphocytes.

    PubMed

    Powell, M K; Whitfield, W; Redelman, D; Henderson, L O; Vogt, R F

    1998-10-01

    To produce biologic calibrators for relative fluorescence intensity (RFI) measurements, we stained leukocytes with serial dilutions of CD45-FITC conjugate and processed them using our regular whole blood lysis procedure. Cells were stained with conjugate concentrations ranging from twice recommended to a million-fold lower. At the highest concentrations of conjugate, the RFI reached a plateau near the top of the third decade, indicating saturation of CD45 binding sites. As the concentration decreased, the RFI declined in a highly linear relationship between the dilution factor and the histogram channel number. For channel numbers corresponding to the lowest percentiles of the RFI distribution, linearity persisted down to the first half decade. The slope of this relationship revealed a true dynamic range of 4.5 decades, which was comparable to the value obtained with microbead standards calibrated in molecules of equivalent soluble fluorochrome (MESF). Our results suggest that the lower limit of linearity for fluorescence intensity from fluorescein isothiocyanate (FITC)-stained lymphocytes is below 500 MESF and that cellular autofluorescence is the major limiting factor in detecting and quantifying FITC-specific staining. This procedure provides an adroit way of characterizing the linearity and dynamic range of measurements for quantitative fluorescence cytometry using exactly the same matrix, stains, and preparation methods as those used for cellular analytes. PMID:9773883

  4. 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

  5. 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

  6. Determination of sulfur in biodiesel microemulsions using the summation of the intensities of multiple emission lines.

    PubMed

    Young, Carl G; Amais, Renata S; Schiavo, Daniela; Garcia, Edivaldo E; Nóbrega, Joaquim A; Jones, Bradley T

    2011-05-15

    A method for the determination of sulfur in biodiesel samples by inductively coupled plasma optical emission spectrometry which uses microemulsion for sample preparation and the summation of the intensities of multiple emission lines has been developed. Microemulsions were prepared using 0.5 mL of 20% v/v HNO(3), 0.5 mL of Triton X-100, 2-3 mL of biodiesel sample, and diluted with n-propanol to a final volume of 10 mL. Summation of the emission intensities of multiple sulfur lines allowed for increased accuracy and sensitivity. The amounts of sulfur determined experimentally were between 2 and 7 mg L(-1), well below legislative standards for many countries. Recoveries obtained ranged from 72 to 119%, and recoveries obtained for the 182.562 nm line were slightly lower. This is most likely due to its lower sensitivity. Using microemulsion for sample preparation and the summation of the intensities of multiple emission lines for the successful determination of sulfur in biodiesel has been demonstrated. PMID:21482315

  7. 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

  8. One-step fabrication of intense red fluorescent gold nanoclusters and their application in cancer cell imaging

    NASA Astrophysics Data System (ADS)

    Bian, Pingping; Zhou, Jing; Liu, Yueying; Ma, Zhanfang

    2013-06-01

    A one-step method for successfully fabrication of water-soluble and alkanethiol-stabilized Au nanoclusters (NCs) was demonstrated. The novel and facile method was based on simply placing histidine (His), HAuCl4 and 11-mercaptoundcanoic acid (MUA) together at room temperature. The resulting Au NCs were exclusively composed of Au17MUA4His22 (AMH), as demonstrated by the photoluminescence, UV-Vis absorption, electrospray ionization mass and X-ray photoelectron spectroscopy. AMH exhibited intense red fluorescence (λem = 600 nm), a long fluorescence lifetime (7.11 μs), considerable stability, and a large Stoke's shift (320 nm). Based on the excellent properties of the AMH, cell experiments were conducted. Cytotoxicity studies showed that the Au NCs exhibited negligible effects in altering cell proliferation or triggering apoptosis. Cancer cell imaging of HeLa cell lines indicated that the obtained AMH could serve as a promising fluorescent bioprobe for bioimaging. This strategy, based on the one-step method, may offer a novel approach to fabricate other water-soluble and alkanethiol-stabilized metal nanoclusters for application in biolabelling and bioimaging.A one-step method for successfully fabrication of water-soluble and alkanethiol-stabilized Au nanoclusters (NCs) was demonstrated. The novel and facile method was based on simply placing histidine (His), HAuCl4 and 11-mercaptoundcanoic acid (MUA) together at room temperature. The resulting Au NCs were exclusively composed of Au17MUA4His22 (AMH), as demonstrated by the photoluminescence, UV-Vis absorption, electrospray ionization mass and X-ray photoelectron spectroscopy. AMH exhibited intense red fluorescence (λem = 600 nm), a long fluorescence lifetime (7.11 μs), considerable stability, and a large Stoke's shift (320 nm). Based on the excellent properties of the AMH, cell experiments were conducted. Cytotoxicity studies showed that the Au NCs exhibited negligible effects in altering cell proliferation or

  9. 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.

  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. PMID:20018337

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

    NASA Astrophysics Data System (ADS)

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

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

  13. Photoactive yellow protein-based protein labeling system with turn-on fluorescence intensity.

    PubMed

    Hori, Yuichiro; Ueno, Hideki; Mizukami, Shin; Kikuchi, Kazuya

    2009-11-25

    Protein labeling provides significant information about protein function. In this research, we developed a novel protein labeling technique by utilizing photoactive yellow protein (PYP). PYP is a small protein (14 kDa) derived from purple bacteria and binds to 7-hydroxycoumarin-3-carboxylic acid as well as to a natural ligand, 4-hydroxycinnamic acid, through a thioester bond with Cys69. Based on the structure and fluorescence property of this coumarin derivative, we designed two fluorescent probes that bind to PYP. One has an azido moiety, which allows stepwise labeling by click chemistry, and the other is a fluorogenic probe. The live-cell imaging and specific labeling of PYP were achieved by using both probes. The flexibility of the probe design and the small size of the tag protein are great advantages of this system against the existing methods. This novel labeling technique can be used in a wide variety of applications for biological research. PMID:19877615

  14. Monitoring plasmid replication in live mammalian cells over multiple generations by fluorescence microscopy.

    PubMed

    Norby, Kathryn; Chiu, Ya-Fang; Sugden, Bill

    2012-01-01

    Few naturally-occurring plasmids are maintained in mammalian cells. Among these are genomes of gamma-herpesviruses, including Epstein-Barr virus (EBV) and Kaposi's Sarcoma-associated herpesvirus (KSHV), which cause multiple human malignancies (1-3). These two genomes are replicated in a licensed manner, each using a single viral protein and cellular replication machinery, and are passed to daughter cells during cell division despite their lacking traditional centromeres (4-8). Much work has been done to characterize the replications of these plasmid genomes using methods such as Southern blotting and fluorescence in situ hybridization (FISH). These methods are limited, though. Quantitative PCR and Southern blots provide information about the average number of plasmids per cell in a population of cells. FISH is a single-cell assay that reveals both the average number and the distribution of plasmids per cell in the population of cells but is static, allowing no information about the parent or progeny of the examined cell. Here, we describe a method for visualizing plasmids in live cells. This method is based on the binding of a fluorescently tagged lactose repressor protein to multiple sites in the plasmid of interest (9). The DNA of interest is engineered to include approximately 250 tandem repeats of the lactose operator (LacO) sequence. LacO is specifically bound by the lactose repressor protein (LacI), which can be fused to a fluorescent protein. The fusion protein can either be expressed from the engineered plasmid or introduced by a retroviral vector. In this way, the DNA molecules are fluorescently tagged and therefore become visible via fluorescence microscopy. The fusion protein is blocked from binding the plasmid DNA by culturing cells in the presence of IPTG until the plasmids are ready to be viewed. This system allows the plasmids to be monitored in living cells through several generations, revealing properties of their synthesis and partitioning to

  15. 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.

  16. Padé-Laplace method for analysis of fluorescence intensity decay.

    PubMed Central

    Bajzer, Z; Myers, A C; Sedarous, S S; Prendergast, F G

    1989-01-01

    This novel approach to the analysis of multiexponential functions is based on the combined use of the Laplace transform and Padé approximants (Yeramian, E., and P. Claverie. 1987. Nature (Lond.). 326:169-174). It is similar in principle to the well-known Isenberg method of moments (Isenberg, I. 1983. Biophys. J. 43:141-148) traditionally applied to the analysis of fluorescence decay. The advantage of the Padé-Laplace method lies in its ability to detect the number of components in a multiexponential function as well as their parameters. In this paper we modified the original method so that it can be applied to the analysis of multifrequency phase/modulation measurements of fluorescence decay. The method was tested first on simulated data. It afforded recovery up to four distinct lifetime components (and their fractional contributions). In the case of simulated data corresponding to continuous lifetime distributions (nonexponential decay), the results of the analysis by the Padé-Laplace method indicated the absence of discrete exponential components. The method was also applied to real phase/modulation data gathered on known fluorophores and their mixtures and on tryptophan fluorescence in phospholipase A2. The lifetime and fraction recoveries were consistent with those obtained from standard methods involving nonlinear least-square fitting. PMID:2752091

  17. 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.

  18. 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

  19. Supramolecular Fluorescent Nanoparticles Constructed via Multiple Non-Covalent Interactions for the Detection of Hydrogen Peroxide in Cancer Cells.

    PubMed

    Wei, Xuan; Dong, Ruijiao; Wang, Dali; Zhao, Tianyu; Gao, Yongsheng; Duffy, Patrick; Zhu, Xinyuan; Wang, Wenxin

    2015-08-01

    Overabundance of hydrogen peroxide originating from environmental stress and/or genetic mutation can lead to pathological conditions. Thus, the highly sensitive detection of H2 O2 is important. Herein, supramolecular fluorescent nanoparticles self-assembled from fluorescein isothiocyanate modified β-cyclodextrin (FITC-β-CD)/rhodamine B modified ferrocene (Fc-RB) amphiphile were prepared through host-guest interaction between FITC-β-CD host and Fc-RB guest for H2 O2 detection in cancer cells. The self-assembled nanoparticles based on a combination of multiple non-covalent interactions in aqueous medium showed high sensitivity to H2 O2 while maintaining stability under physiological condition. Owing to the fluorescence resonance energy transfer (FRET) effect, addition of H2 O2 led to obvious fluorescence change of nanoparticles from red (RB) to green (FITC) in fluorescent experiments. In vitro study showed the fluorescent nanoparticles could be efficiently internalized by cancer cells and then disrupted by endogenous H2 O2 , accompanying with FRET from "on" to "off". These supramolecular fluorescent nanoparticles constructed via multiple non-covalent interactions are expected to have potential applications in diagnosis and imaging of diseases caused by oxidative stresses. PMID:26133314

  20. 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.

  1. Atmospheric electric field effect for total NM intensity and different multiplicities on Mt Hermon

    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

    Cosmic rays (CR) are an important element of space weather and instrument of space weather forecasting. From this point of view, it is necessary to take into account all factors influencing CR intensity. One of these important factors is the influence of atmospheric electric fields (AEF) during thunderstorms on CR intensity. This is caused by local acceleration (or deceleration, depending on the direction of the AEF and the sign of charged particles) of secondary CR particles (mostly muons and electrons, for CR observations in the low atmosphere or underground). We analysed one minute data on AEF obtained by the ESF-1000 sensor in our observatory on Mt. Hermon, and one minute neutron monitor data corrected on barometric effects and on the effect of snow. While AEF does not influence neutrons, we found significant effects in the observed total neutron intensity and in the intensities of different multiplicities. This is caused mostly by soft negative muons, captured by nuclei of lead (instead of the atom’s electrons) with the formation of mesoatoms. While the cross section of muons relative to strong interactions is very small (the same order as for neutrino), because the captured muon moves about inside the nucleus with very high density, the probability of muon interaction with nucleus is higher than the decay of muon. As result of this interaction the total energy of the rest muon about 100 MeV goes to the excitation of lead nuclei, with emanation of a few neutrons which are detected by the neutron monitor. Therefore, a neutron monitor is an ideal detector for separating positive and negative soft muons (without using a big magnetic system). We obtained results for positively and negatively directed AEF and show existing significant AEF influence on CR intensithttps://www.cospar-assembly.org/user/download2.php?id=29566&type=previewy, biggest for small multiplicities. We give a theoretical explanation of obtained results.

  2. Photoelectron Holography: Exploration of the Multiphoton Ionization and Multiple Rescattering in Intense Laser Fields

    NASA Astrophysics Data System (ADS)

    Chu, Chon-Teng; Li, Peng-Cheng; Chu, Shih-I.

    2015-05-01

    We perform a fully ab initio investigation of the multiphoton ionization (MPI) and electron multiple rescattering dynamics of atomic H driven by intense ultrashort mid-IR laser fields. The time-dependent Schrödinger equation is solved accurately and efficiently by means of the time-dependent generalized pseudospectral method (TDGPS) in the Kramers-Henneberger (KH) frame. We use the semiclassical approach to analyze and visualize all the trajectories during the atom-laser interaction, unveiling the multiple e-parent ion rescattering processes. In this way, we can identify the dominant behaviors of different parts of photoelectron holography to a particular number of times of the electron's revisits to its parent ion. This work was partially supported by DOE.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 antibody labels to increase the fluorescence signal and sensitivity. Ep...

  4. Spatial dynamics of laser-induced fluorescence in an intense laser beam: An experimental and theoretical study with alkali-metal atoms

    NASA Astrophysics Data System (ADS)

    Auzinsh, M.; Berzins, A.; Ferber, R.; Gahbauer, F.; Kalnins, U.

    2016-03-01

    We show that it is possible to model accurately optical phenomena in intense laser fields by taking into account the intensity distribution over the laser beam. We present an extension of an earlier theoretical model that divides an intense laser beam into concentric regions, each with a Rabi frequency that corresponds to the intensity in that region, and solve a set of coupled optical Bloch equations for the density matrix in each region. Experimentally obtained magneto-optical resonance curves for the Fg=2 ⟶Fe=1 transition of the D1 line of 87Rb agree very well with the theoretical model up to a laser intensity of around 200 mW/cm2 for a transition whose saturation intensity is around 4.5 mW/cm2. We examine the spatial dependence of the fluorescence intensity in an intense laser beam experimentally and theoretically. We present and discuss the results of an experiment in which a broad, intense pump laser excites the Fg=4 ⟶Fe=4 transition of the D2 line of cesium while a narrow probe beam scans the atoms within the pump beam and excites the D1 line of cesium, whose fluorescence is recorded as a function of probe beam position. Experimentally obtained spatial profiles of the fluorescence intensity agree qualitatively with the predictions of the model.

  5. The fluorescence intermittency for quantum dots is not power-law distributed: a luminescence intensity resolved approach.

    PubMed

    Schmidt, Robert; Krasselt, Cornelius; Göhler, Clemens; von Borczyskowski, Christian

    2014-04-22

    The photoluminescence (PL) of single emitters like semiconductor quantum dots (QDs) shows PL intermittency, often called blinking. We explore the PL intensities of single CdSe/ZnS QDs in polystyrene (PS), on polyvenylalcohol (PVA), and on silicon oxide (SiOx) by the change-point analysis (CPA). By this, we relate results from the macrotime (sub-ms to 1000 s) and the microtime (0.1-100 ns) range to discrete PL intensities. We conclude that the intensity selected "on"-times in the ms range correspond to only a few (discrete) switching times, while the PL decays in the ns range are multiexponential even with respect to the same selected PL intensity. Both types of relaxation processes depend systematically on the PL intensity in course of a blinking time trace. The overall distribution of on-times does not follow a power law contrary to what has often been reported but can be compiled into 3-4 characteristic on-times. The results can be explained by the recently suggested multiple recombination centers model. Additionally, we can identify a well-defined QD state with a very low PL intensity above the noise level, which we assign to the strongly quenched exciton state. We describe our findings by a model of a hierarchical sequence of hole and electron trapping. Blinking events are the consequence of slow switching processes among these states and depend on the physicochemical properties of the heterogeneous nanointerface of the QDs. PMID:24580107

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

    PubMed Central

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

    2015-01-01

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

  7. Cytogenetic abnormality in patients with multiple myeloma analyzed by fluorescent in situ hybridization

    PubMed Central

    Hu, Ying; Chen, Wenming; Chen, Shilun; Huang, Zhongxia

    2016-01-01

    Objective To analyze the fluorescent in situ hybridization (FISH) data and the association with clinical characteristics, therapy response, and survival time in patients with multiple myeloma. Method We performed a retrospective review of patients with multiple myeloma from November 2010 to April 2014. Results Cytogenetic abnormalities by FISH were detectable in 66% of patients. One cytogenetic abnormality, two cytogenetic abnormalities, and complex abnormalities were detectable in 21.2%, 51.5%, and 27.3% of cases, respectively. 1q21 amplification, t(4p16.3/14q32), and 17p deletion were observed in 69.7%, 30.3%, and 21.2% of cases, respectively. Total response rates (complete response [CR] + near CR + partial response) were 93.8% and 82.1%, respectively, in cytogenetic normality group and abnormality group. CR rates were 50% and 32.1%, respectively. Median overall survival (OS) time was 51 months and 24 months, respectively, in cytogenetic normality group and abnormality group (P<0.05). Median OS time was not significantly different between 1q21 amplification group and no 1q21 amplification group in patients with FISH abnormalities (P>0.05). Median OS time was not significantly different between t(4;14) group and no t(4;14) group in patients with FISH abnormalities (P>0.05). Seven patients of 17p deletion died in 2 years. Conclusion Multiple myeloma is characterized by a high occurrence of chromosomal aberrations. 1q21 amplification and t(4;14) are the most common abnormalities. Multiple cytogenetic abnormalities are frequently observed in the same one patient. The total response rate, CR rate, and OS time are worse in cytogenetic abnormal patients compared with cytogenetic normal patients. Patients with 17p deletion have a very poor prognosis. Future goals of therapy will be to achieve minimal residual disease, biomarkers, and genomic data, which might provide a better estimate of the depth of response to therapy and OS. PMID:27042105

  8. 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.

  9. 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.

  10. When R  >  0.8R 0: fluorescence anisotropy, non-additive intensity, and cluster size

    NASA Astrophysics Data System (ADS)

    Zolmajd-Haghighi, Z.; Hanley, Q. S.

    2016-06-01

    Assembly and clustering feature in many biological processes and homo-FRET and fluorescence anisotropy can assist in estimating the aggregation state of a system. The distance dependence of resonance energy transfer is well described and tested. Similarly, assessment of cluster size using steady state anisotropy is well described for non-oriented systems when R  <  0.8R 0, however, these methods break down when R  >  0.8R 0. Fused trimeric DNA clusters labelled with fluorescein were engineered to provide inter-fluorophore distances from 0.7 to 1.6 R/R 0 and intensity and anisotropy were measured. These constructs cover a range where anisotropy effects depend on distance. Analytical expressions were derived for fully labelled and fractionally labelled clusters and the experimental results analysed. The experimental results showed that: (1) the system underwent distance dependent quenching; (2) when incompletely labelled both doubly and triply labelled forms could be assessed to obtain distance dependent intensity factors; (3) the anisotropy behaviour of a multiply labelled cluster of a particular size depends on the behaviour of the fluorophores and their distance in a cluster. This work establishes that when emission intensity data are available the analytically useful range for investigating clusters does not have to be restricted to R  <  0.8R 0 and is applicable to cases where the anisotropy of a cluster of N fluorophores is not well approximated by r 1/N.

  11. Two-photon imaging of multiple fluorescent proteins by phase-shaping and linear unmixing with a single broadband laser

    PubMed Central

    Brenner, Meredith H.; Cai, Dawen; Swanson, Joel A.; Ogilvie, Jennifer P.

    2013-01-01

    Imaging multiple fluorescent proteins (FPs) by two-photon microscopy has numerous applications for studying biological processes in thick and live samples. Here we demonstrate a setup utilizing a single broadband laser and a phase-only pulse-shaper to achieve imaging of three FPs (mAmetrine, TagRFPt, and mKate2) in live mammalian cells. Phase-shaping to achieve selective excitation of the FPs in combination with post-imaging linear unmixing enables clean separation of the fluorescence signal of each FP. This setup also benefits from low overall cost and simple optical alignment, enabling easy adaptation in a regular biomedical research laboratory. PMID:23938572

  12. High Mean Fluorescence Intensity Donor-Specific Anti-HLA Antibodies Associated With Chronic Rejection Postliver Transplant

    PubMed Central

    O’Leary, J. G.; Kaneku, H.; Susskind, B. M.; Jennings, L. W.; Neri, M. A.; Davis, G. L.; Klintmalm, G. B.; Terasaki, P. I.

    2015-01-01

    In contrast to kidney transplantation where donorspecific anti-HLA antibodies (DSA) negatively impact graft survival, correlation of DSA with clinical outcomes in patients after orthotopic liver transplantation (OLT) has not been clearly established. We hypothesized that DSA are present in patients who develop chronic rejection after OLT. Prospectively collected serial serum samples on 39 primary OLT patients with biopsy-proven chronic rejection and 39 comparator patients were blinded and analyzed for DSA using LABScreen single antigen beads test, where a 1000 mean fluorescence value was considered positive. In study patients, the median graft survival was 15 months, 74% received ≥ one retransplant, 20% remain alive and 87% had ≥ one episode of acute rejection. This is in contrast to comparator patients where 69% remain alive, and no patient needed retransplant or experienced rejection. Thirty-six chronic rejection patients (92%) and 24 (61%) comparator patients had DSA (p = 0.003). Chronic rejection versus comparator patients had higher mean fluorescence intensity (MFI) DSA. Although a further study with larger numbers of patients is needed to identify clinically significant thresholds, there is an association of high-MFI DSA with chronic rejection after OLT. PMID:21672151

  13. 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.

  14. 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.

  15. 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.

  16. Concurrent fluorescence macro-imaging across multiple spectral regions in the visible and the near infrared

    NASA Astrophysics Data System (ADS)

    Kazemzadeh, Farnoud; Haider, Shahid; Jin, Chao; Clausi, David A.; Wong, Alexander

    2015-09-01

    Fluorescent imaging, often synonymous with microscopic imaging, is an imaging modality whereby various features of a target are observed based on assignment of chemical labels. These labels are in most cases indirect tracers of specific structures or chemical compounds which cannot be otherwise identified. The tracers are excited by an illuminating source and they in turn emit light at specific wavelengths. This light is then captured by an imaging device and represented as an indirect observation of the specific feature in the sample. The process of excitation and imaging of the emitted light is performed sequentially and is proportional to the number of tracers or fluorescence species present in the sample. We present an imaging system that can image fluorescent tracers, in the visible and the near Infra-red, simultaneously. This system is capable of illuminating the target with different excitation light sources and capture the corresponding fluorescence images in one snapshot using a series of mirrors to capture different views of the sample. The simultaneously captured image are fused using a computational reconstruction process to present a coherent multispectral fluorescence image. The system is proposed for use in applications where the rapid enumeration of fluorescent species in a large field of view is paramount as opposed to their microscopic image in a narrow field of view. The system was tested using a controlled cocktail solution of four different types fluorescent microspheres and was able to enumerate the microspheres based on their different fluorescent signatures as captured by the system.

  17. 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.

  18. Simultaneous Visualization of Multiple mRNAs and Matrix Metalloproteinases in Living Cells Using a Fluorescence Nanoprobe.

    PubMed

    Pan, Wei; Yang, Huijun; Li, Na; Yang, Limin; Tang, Bo

    2015-04-13

    Simultaneous monitoring of multiple tumour markers is of great significance for improving the accuracy of early cancer detection. In this study, a fluorescence nanoprobe has been prepared that can simultaneously monitor and visualize multiple mRNAs and matrix metalloproteinases (MMPs) in living cells. Confocal fluorescence imaging results indicate that the nanoprobe could effectively distinguish between cancer cells and normal cells even if one tumour maker of normal cells was overexpressed. Furthermore, it can detect changes in the expression levels of mRNAs and MMPs in living cells. The current approach could provide new tools for early cancer detection and monitoring the changes in expression levels of biomarkers during tumour progression. PMID:25752514

  19. A reversible dual-response fluorescence switch for the detection of multiple analytes.

    PubMed

    Geng, Junlong; Liu, Ping; Liu, Bianhua; Guan, Guijian; Zhang, Zhongping; Han, Ming-Yong

    2010-03-22

    This paper reports a reversible dual fluorescence switch for the detection of a proton target and 2,4,6-trinitrotoluene (TNT) with opposite-response results, based on fluorophore derivatization of silica nanoparticles. Fluorescent silica nanoparticles were synthesized through modification of the surface with a nitrobenzoxadiazole (NBD) fluorophore and an organic amine to form a hybrid monolayer of fluorophores and amino ligands; the resultant nanoparticles showed different fluorescence responses to the proton target and TNT. Protonation of the amino ligands leads to fluorescence enhancement due to inhibition of photoinduced electron transfer (PET) between the amine and fluorophore. By contrast, addition of TNT results in fluorescence quenching because a fluorescence resonance energy transfer (FRET) happens between the NBD fluorophore and the formed TNT-amine complex. The fluorescence signal is reversible through washing with the proper solvents and the nanoparticles can be reused after centrifugal separation. Furthermore, these nanoparticles were assembled into chips on an etched silicon wafer for the detection of TNT and the proton target. The assembled chip can be used as a convenient indicator of herbicide (2,4-dichlorophenoxyacetic acid) and TNT residues with the use of only 10 microL of sample. The simple NBD-grafted silica nanoparticles reported here show a reversible signal and good assembly flexibility; thus, they can be applied in multianalyte detection. PMID:20151433

  20. 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

  1. 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

  2. Diurnal Variability in Chlorophyll-a, Carotenoids, CDOM and SO₄(2-) Intensity of Offshore Seawater Detected by an Underwater Fluorescence-Raman Spectral System.

    PubMed

    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 SO₄(2-) 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 SO₄(2-), 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 SO₄(2-). To avoid such interference, the backup Raman spectroscopy system with λex = 785 nm was employed to detect SO₄(2-) 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 SO₄(2-) in the ocean. PMID:27420071

  3. Resolution of fluorescence intensity decays of the two tryptophan residues in glutamine-binding protein from Escherichia coli using single tryptophan mutants.

    PubMed Central

    Axelsen, P H; Bajzer, Z; Prendergast, F G; Cottam, P F; Ho, C

    1991-01-01

    Time correlated single photon counting measurements of tryptophan (Trp) fluorescence intensity decay and other spectroscopic studies were performed on glutamine-binding protein (GlnBP) from Escherichia coli. Using site-specifically mutated forms of the protein in which tyrosine (Tyr) and phenylalanine (Phe) substitute for the Trp residues at positions 32 and 220, we have examined whether wild-type (Wtyp) intensity decay components may be assigned to specific Trp residues. Results indicate that: (a) two exponential intensity decay components are recovered from the Wtyp protein (6.16 ns, 0.46 ns); (b) the long decay component arises from Trp-220 and comprises greater than 90% of the total fluorescence emission; (c) the short component arises from Trp-32 and is highly quenched; (d) all four single-Trp mutants exhibit multiexponential intensity decays, yet equimolar mixtures of two single-Trp mutants yield only two decay components which are virtually indistinguishable from the Wtyp protein; (e) the recovery of additional components in protein mixtures is obscured by statistical noise inherent in the technique of photon counting; (f) various spectroscopic measurements suggest that Trp-Trp interactions occur in the Wtyp protein, but the Wtyp intensity decay may be closely approximated by a linear combination of intensity decays from single-Trp mutants; and (g) inferences derived independently from fluorescence and NMR spectroscopy which pertain to the presence of Trp-Trp interactions and the relative solvent exposure of the two Trp residues are in agreement. PMID:1932553

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

    PubMed

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

    2016-04-28

    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. PMID:27064427

  5. 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.

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

    PubMed Central

    Dewhirst, Mark W.; Shan, S.; Cao, Yiting; Moeller, Benjamin; Yuan, Fan; Li, Chuan-Yuan

    2002-01-01

    The purpose of this report is to present an overview of the use of fluorescence imaging in vivo, with particular emphasis on oncology. It is important to note, however, that many of the methods described herein have been applied to the study of non-malignant tissues as well. Modern medicine and biology research has benefited greatly from an ever-expanding assortment of fluorescent markers and labels. These markers and labels have allowed investigators to observe the behavior and properties of cell and molecular entities of interest in the context of complicated biological systems such as a mammalian cell or a whole mouse. Methods developed to image fluorescence in whole mice have been valuable in studying patterns of tumor growth and metastases. Alternatively, more detailed information and a wide variety of endpoints can be obtained using “intravital” preparations. This review focuses on use of fluorescence imaging for intravital preparations. For detail on fluorescence imaging of whole animals, refer to reviews on this subject [1,2]. For oncologic applications, studies have focused primarily on window chamber preparations that allow for real-time visualization of tumor growth, vascularity, vascular responses to stimulation, vascular permeability, vascular orientation, flow instability, and the like. These endpoints have been used to show that there are functional differences between tumor and normal tissues with respect to these functions under baseline conditions and after therapeutic manipulation. Examples of some of these differences are provided in this review as a means to illustrate how they can be used. PMID:14646042

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

    PubMed

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

    2015-11-01

    A chemosensor consisting of two terpyridines covalently linked to a dithienosilole unit (1) has been synthesized, and its optical and metal sensing properties have been investigated. Due to the metal-organic coordination function, 1 can bind with many transition metal ions and display different fluorescence responses that cause it to function as a "turn-off" fluorescent chemosensor. A significant bathochromic shift in the fluorescence spectra is observed in the presence of Zn(2+). Meanwhile, the emission of 1 is weakened upon exposure to Ag(+) and Fe(2+) and completely quenched by Ni(2+), Co(2+), and Cu(2+). Based on the observed results, several logic gates, such as XNOR, INHIBIT, and IMPLICATION, have been achieved by controlling the chemical inputs. PMID:26099823

  8. Side-entry laser-beam zigzag irradiation of multiple channels in a microchip for simultaneous and highly sensitive detection of fluorescent analytes.

    PubMed

    Anazawa, Takashi; Yokoi, Takahide; Uchiho, Yuichi

    2015-09-01

    A simple and highly sensitive technique for laser-induced fluorescence detection on multiple channels in a plastic microchip was developed, and its effectiveness was demonstrated by laser-beam ray-trace simulations and experiments. In the microchip, with refractive index nC, A channels and B channels are arrayed alternately and respectively filled with materials with refractive indexes nA for electrophoresis analysis and nB for laser-beam control. It was shown that a laser beam entering from the side of the channel array traveled straight and irradiated all A channels simultaneously and effectively because the refractive actions by the A and B channels were counterbalanced according to the condition nA < nC < nB. This technique is thus called "side-entry laser-beam zigzag irradiation". As a demonstration of the technique, when nC = 1.53, nA = 1.41, nB = 1.66, and the cross sections of both eight A channels and seven B channels were the same isosceles trapezoids with 97° base angle, laser-beam irradiation efficiency on the eight A channels by the simulations was 89% on average and coefficient of variation was 4.4%. These results are far superior to those achieved by other conventional methods such as laser-beam expansion and scanning. Furthermore, fluorescence intensity on the eight A channels determined by the experiments agreed well with that determined by the simulations. Therefore, highly sensitive and uniform fluorescence detection on eight A channels was achieved. It is also possible to fabricate the microchips at low cost by plastic-injection molding and to make a simple and compact detection system, thereby promoting actual use of the proposed side-entry laser-beam zigzag irradiation in various fields. PMID:26296140

  9. [Single-donor protocol: Transfusion practices and multiple transfusion risk factors in neonatal intensive care unit].

    PubMed

    Dollat, C; Pierron, C; Keslick, A; Billoir, E; François, A; Jarreau, P-H

    2016-09-01

    In France since 2002, the single-donor transfusion protocol, using four pediatric units from the same adult donor's packed red blood cells (PRBCs) in multiply transfused newborns, is recommended in preterm neonates to reduce the risks of infection and alloimmunization. This protocol is controversial, however, because it causes the transfusion of stored blood, which could have adverse consequences. Before the new recommendations of the French Haute Autorité de santé (National authority for health) in 2015, we conducted a national practice survey in 63 neonatal intensive care units (NICU) and a retrospective study of the characteristics of 103 children transfused within our unit, to better target beneficiaries. The practice survey showed that 30 % of French NICUs no longer used the protocol in 2014, due to logistical or financial problems, or concerns about the transfusion of stored blood. The practices were heterogeneous. Few NICUs used a written protocol. In our NICU, the use of single-donor protocol involved the use of units stored for more than 20 days in half of the cases beginning with the third unit used. Six-term newborns were mainly transfused once, which does not seem to warrant the single-donor transfusion protocol. The use of this protocol caused the loss of 50 % of the manufactured units, which go unused. In multivariate analysis, two factors were predictive of multiple transfusion within our population of 95 premature neonates undergoing transfusion: low-term and a high Clinical Risk Index for Babies (CRIB) score. The risk of multiple transfusions would be reduced by about 15 % for each additional week of gestation and approximately 16 % per point within the CRIB score. These variables integrated into a statistical model predict the risk of multiplying transfusions. According to the ROC curve, a calculated risk higher than 50 % is the appropriate cut-off value to transfuse with the single-donor transfusion protocol. This would limit its

  10. 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)…

  11. 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.

  12. Characterization of Fluorescence of ANS–Tear Lipocalin Complex: Evidence for Multiple-Binding Modes

    PubMed Central

    Gasymov, Oktay K.; Abduragimov, Adil R.; Glasgow, Ben J.

    2010-01-01

    ANS is widely used as a probe for locating binding sites of proteins and studying structural changes under various external conditions. However, the nature of ANS-binding sites in proteins and the accompanying changes in fluorescence properties are controversial. We examined the steady-state and time-resolved fluorescence of the ANS–protein complexes for tear lipocalin (TL) and its mutants in order to discern the origin of lifetime components via analysis that included the multiexponential decay and the model-free maximum entropy methods. Fluorescence lifetimes of ANS–TL complexes can be grouped into two species, 14.01–17.42 ns and 2.72–4.37 ns. The log-normal analyses of fluorescence spectral shapes reveal the heterogeneous nature of both long- and short-lifetime species. The constructed time-resolved emission, amplitude (TRES) and area normalized (TRANES), and decay-associated spectra are consistent with a model that includes heterogeneous modes of ANS binding with two separate lifetime components. The two lifetime components are not derived from solvent relaxation, but rather may represent different binding modes. PMID:18028215

  13. Characterization of fluorescence of ANS-tear lipocalin complex: evidence for multiple-binding modes.

    PubMed

    Gasymov, Oktay K; Abduragimov, Adil R; Glasgow, Ben J

    2007-01-01

    ANS is widely used as a probe for locating binding sites of proteins and studying structural changes under various external conditions. However, the nature of ANS-binding sites in proteins and the accompanying changes in fluorescence properties are controversial. We examined the steady-state and time-resolved fluorescence of the ANS-protein complexes for tear lipocalin (TL) and its mutants in order to discern the origin of lifetime components via analysis that included the multiexponential decay and the model-free maximum entropy methods. Fluorescence lifetimes of ANS-TL complexes can be grouped into two species, 14.01-17.42 ns and 2.72-4.37 ns. The log-normal analyses of fluorescence spectral shapes reveal the heterogeneous nature of both long- and short-lifetime species. The constructed time-resolved emission, amplitude (TRES) and area normalized (TRANES), and decay-associated spectra are consistent with a model that includes heterogeneous modes of ANS binding with two separate lifetime components. The two lifetime components are not derived from solvent relaxation, but rather may represent different binding modes. PMID:18028215

  14. Optimization of the design of a multiple-photon excitation laser scanning fluorescence imaging system

    NASA Astrophysics Data System (ADS)

    Wokosin, David L.; White, John G.

    1997-04-01

    Multi-photon (two or more photon) excitation imaging offers three significant advantages compared to laser-scanning confocal fluorescence microscopy for 3-D and 4-D fluorescence microscopy: considerable reduction in total sample excitation, increased depth penetration, and increased detection sensitivity. All-solid-state ultra-fast lasers offer tremendous potential for affordable, reliable, 'turn-key' multi-photon excitation sources. We have been developing a multi-photon system that utilizes an all-solid- state Nd:YLF excitation source. We have been evaluating the potential of this source for biological microscopy and have been optimizing system parameters for this application area. We have found that the 1047 nm radiation from these lasers can excite by two-photon fluorescence many commonly used fluorophores that are normally excited from blue to yellow light. In addition, we have found that this wavelength readily excites several normally UV excited fluorophores by the mechanism of three-photon excitation. The Nd:YLF laser has proven reliable in operation with nearly 6000 hours logged without significant loss of power. However, the original system produced rather long pulses for multi-photon excitation (300 fs) and a beam shape that was not ideal. We have recently commissioned the development of an improved pulse compressor from the manufacturers that gives narrower pulses (120 fs), improved beam shape, and a smaller insertion loss. This optimized excitation system has 6 times more potential two-photon excited fluorescence and 22 times more potential three-photon excited fluorescence than the prototype system. In addition, by optimizing coatings in the excitation and signal paths, we have improved the descanned detection sensitivity by 20% for two-photon excited fluorescence and 315% for three-photon excited fluorescence. The excitation optical transfer efficiency (1047 nm) of our imaging system is currently 60% to the back aperture of the objective. The

  15. 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

  16. A statistical assessment of the impact of agricultural land use intensity on regional surface water quality at multiple scales.

    PubMed

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

    2012-11-01

    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 (NO(3)-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 (COD(Mn)) 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 NO(3)-N and COD(Mn), 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

  17. Automatic Tumor-Stroma Separation in Fluorescence TMAs Enables the Quantitative High-Throughput Analysis of Multiple Cancer Biomarkers

    PubMed Central

    Lahrmann, Bernd; Halama, Niels; Sinn, Hans-Peter; Schirmacher, Peter; Jaeger, Dirk; Grabe, Niels

    2011-01-01

    The upcoming quantification and automation in biomarker based histological tumor evaluation will require computational methods capable of automatically identifying tumor areas and differentiating them from the stroma. As no single generally applicable tumor biomarker is available, pathology routinely uses morphological criteria as a spatial reference system. We here present and evaluate a method capable of performing the classification in immunofluorescence histological slides solely using a DAPI background stain. Due to the restriction to a single color channel this is inherently challenging. We formed cell graphs based on the topological distribution of the tissue cell nuclei and extracted the corresponding graph features. By using topological, morphological and intensity based features we could systematically quantify and compare the discrimination capability individual features contribute to the overall algorithm. We here show that when classifying fluorescence tissue slides in the DAPI channel, morphological and intensity based features clearly outpace topological ones which have been used exclusively in related previous approaches. We assembled the 15 best features to train a support vector machine based on Keratin stained tumor areas. On a test set of TMAs with 210 cores of triple negative breast cancers our classifier was able to distinguish between tumor and stroma tissue with a total overall accuracy of 88%. Our method yields first results on the discrimination capability of features groups which is essential for an automated tumor diagnostics. Also, it provides an objective spatial reference system for the multiplex analysis of biomarkers in fluorescence immunohistochemistry. PMID:22164226

  18. Meditating metal coenhanced fluorescence and SERS around gold nanoaggregates in nanosphere as bifunctional biosensor for multiple DNA targets.

    PubMed

    Liu, Yong; Wu, Peiyi

    2013-06-26

    Gold nanoparticles (Au NPs) are very attractive candidate nanoparticles in biological assay because of their high chemical stabilities, high homogeneities, good biocompatibilities, and low toxicities. However, molecular beacon assays via encapsulating the combined fluorescence or surface-enhanced Raman scattering (SERS) signals of reporters and Au NPs in nanobarcodes particles usually suffer from fluorescence quenching or weak Raman enhancement when Au NPs are employed (especially with size smaller than 15 nm). Herein, we present a new design of simultaneously realizing metal-enhanced fluorescence and coenhanced surface-enhanced Raman scattering by facilely embedding Ag nanoparticle into the shell of two kinds of Au nanoaggregate (5 and 10 nm), meanwhile, fluorophore is located between the silver core and gold nanoparticle layers and the distance among them is adjusted by SiO2 spacer (Ag@first SiO2 spacer@FiTC+SiO2@second SiO2 spacer@Au nanoaggregate). In this architecture, Ag nanoparticle not only is utilized as an efficient fluorescence enhancer to overcome the common fluorescence quenching around Au nanoaggregates but also behaves like a mirror. Thus, incident light that passes through the SERS-active Au nanoaggregate and the intervening dielectric layer of SiO2 could be reflected multiply from the surface of Ag nanoparticle and coupled with the light at the nanogap between the Au nanoaggregates to further amplify Raman intensity. This results in enhancement factors for fluorescence and SERS ~1.6-fold and more than 300-fold higher than the control samples without silver core under identical experimental conditions, respectively. Moreover, fluorophore and SERS reporters are assembled onto different layers of the concentric hybrid microsphere, resulting in a feasible fabrication protocol when a large number of agents need to be involved into the dual-mode nanobarcodes. A proof-of-concept chip-based DNA sandwich hybridization assay using genetically modified

  19. 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. PMID:27007856

  20. 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.

  1. 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

  2. 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

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

    PubMed

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

    2003-01-21

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

  4. 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…

  5. 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.

  6. Multi-color quantum dot-based fluorescence immunoassay array for simultaneous visual detection of multiple antibiotic residues in milk.

    PubMed

    Song, Erqun; Yu, Mengqun; Wang, Yunyun; Hu, Weihua; Cheng, Dan; Swihart, Mark T; Song, Yang

    2015-10-15

    Antibiotic residues, which are among the most common contaminants in animal-based food products such as milk, have become a significant public health concern. Here, we combine a multicolor quantum dot (QD)-based immunofluorescence assay and an array analysis method to achieve simultaneous, sensitive and visual detection of streptomycin (SM), tetracycline (TC), and penicillin G (PC-G) in milk. Antibodies (Abs) for SM, TC and PC-G were conjugated to QDs with different emission wavelengths (QD 520 nm, QD 565 nm and QD 610 nm) to serve as detection probes (QD-Ab). Then a direct competitive fluorescent immunoassay was performed in antigen-coated microtiter plate wells for simultaneous qualitative and quantitative detection of SM, TC, and PC-G residues, based on fluorescence of the QD-Ab probes. The linear ranges for SM, TC and PC-G were 0.01-25 ng/mL, 0.01-25 ng/mL and 0.01-10 ng/mL, respectively, with detection limit of 5 pg/mL for each of them. Based on fluorescence of the QD-Ab probes, residues of the three antibiotics were determined visually and simultaneously. Compared with a commercial enzyme-linked immunosorbent assay kit, our method could achieve simultaneous analysis of multiple target antibiotics in multiple samples in a single run (high-throughput analysis) and improved accuracy and sensitivity for analysis of residues of the three antibiotics in authentic milk samples. This new analytical tool can play an important role in ameliorating the negative impact of the residual antibiotics on human health and the ecosystem. PMID:26002016

  7. A Record of the Sequence and Intensity of Multiple Impacts in the NWA 7298 H Chondrite

    NASA Astrophysics Data System (ADS)

    Friedrich, J. M.; Weisberg, M. K.; Rivers, M. L.

    2013-09-01

    We demonstrate evidence for the sequence and intensity of at least three distinct impact events affecting NWA 7298. These observations yield new opportunities for investigating the dynamic collisional evolution of asteroids.

  8. 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

  9. Two-photon-like microscopy with orders-of-magnitude lower illumination intensity via two-step fluorescence

    PubMed Central

    Ingaramo, Maria; York, Andrew G.; Andrade, Eric J.; Rainey, Kristin; Patterson, George H.

    2015-01-01

    We describe two-step fluorescence microscopy, a new approach to non-linear imaging based on positive reversible photoswitchable fluorescent probes. The protein Padron approximates ideal two-step fluorescent behaviour: it equilibrates to an inactive state, converts to an active state under blue light, and blue light also excites this active state to fluoresce. Both activation and excitation are linear processes, but the total fluorescent signal is quadratic, proportional to the square of the illumination dose. Here, we use Padron's quadratic non-linearity to demonstrate the principle of two-step microscopy, similar in principle to two-photon microscopy but with orders-of-magnitude better cross-section. As with two-photon, quadratic non-linearity from two-step fluorescence improves resolution and reduces unwanted out-of-focus excitation, and is compatible with structured illumination microscopy. We also show two-step and two-photon imaging can be combined to give quartic non-linearity, further improving imaging in challenging samples. With further improvements, two-step fluorophores could replace conventional fluorophores for many imaging applications. PMID:26333365

  10. Two-photon-like microscopy with orders-of-magnitude lower illumination intensity via two-step fluorescence

    NASA Astrophysics Data System (ADS)

    Ingaramo, Maria; York, Andrew G.; Andrade, Eric J.; Rainey, Kristin; Patterson, George H.

    2015-09-01

    We describe two-step fluorescence microscopy, a new approach to non-linear imaging based on positive reversible photoswitchable fluorescent probes. The protein Padron approximates ideal two-step fluorescent behaviour: it equilibrates to an inactive state, converts to an active state under blue light, and blue light also excites this active state to fluoresce. Both activation and excitation are linear processes, but the total fluorescent signal is quadratic, proportional to the square of the illumination dose. Here, we use Padron's quadratic non-linearity to demonstrate the principle of two-step microscopy, similar in principle to two-photon microscopy but with orders-of-magnitude better cross-section. As with two-photon, quadratic non-linearity from two-step fluorescence improves resolution and reduces unwanted out-of-focus excitation, and is compatible with structured illumination microscopy. We also show two-step and two-photon imaging can be combined to give quartic non-linearity, further improving imaging in challenging samples. With further improvements, two-step fluorophores could replace conventional fluorophores for many imaging applications.

  11. Additive transgene expression and genetic introgression in multiple green-fluorescent protein transgenic crop x weed hybrid generations.

    PubMed

    Halfhill, M D; Millwood, R J; Weissinger, A K; Warwick, S I; Stewart, C N

    2003-11-01

    The level of transgene expression in crop x weed hybrids and the degree to which crop-specific genes are integrated into hybrid populations are important factors in assessing the potential ecological and agricultural risks of gene flow associated with genetic engineering. The average transgene zygosity and genetic structure of transgenic hybrid populations change with the progression of generations, and the green fluorescent protein (GFP) transgene is an ideal marker to quantify transgene expression in advancing populations. The homozygous T(1) single-locus insert GFP/ Bacillus thuringiensis (Bt) transgenic canola ( Brassica napus, cv Westar) with two copies of the transgene fluoresced twice as much as hemizygous individuals with only one copy of the transgene. These data indicate that the expression of the GFP gene was additive, and fluorescence could be used to determine zygosity status. Several hybrid generations (BC(1)F(1), BC(2)F(1)) were produced by backcrossing various GFP/Bt transgenic canola ( B. napus, cv Westar) and birdseed rape ( Brassica rapa) hybrid generations onto B. rapa. Intercrossed generations (BC(2)F(2) Bulk) were generated by crossing BC(2)F(1) individuals in the presence of a pollinating insect ( Musca domestica L.). The ploidy of plants in the BC(2)F(2) Bulk hybrid generation was identical to the weedy parental species, B. rapa. AFLP analysis was used to quantify the degree of B. napus introgression into multiple backcross hybrid generations with B. rapa. The F(1) hybrid generations contained 95-97% of the B. napus-specific AFLP markers, and each successive backcross generation demonstrated a reduction of markers resulting in the 15-29% presence in the BC(2)F(2) Bulk population. Average fluorescence of each successive hybrid generation was analyzed, and homozygous canola lines and hybrid populations that contained individuals homozygous for GFP (BC(2)F(2) Bulk) demonstrated significantly higher fluorescence than hemizygous hybrid

  12. [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. PMID:27228758

  13. 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.

  14. Determination of plume temperature distribution based on the ratios of the radiation intensities of multiple CO2 lines

    NASA Astrophysics Data System (ADS)

    Cieszczyk, S.

    2015-05-01

    New inversion scheme for gas temperature distribution retrieval utilized CO2 spectrum between 2350 cm-1 and 2400 cm-1 is proposed. Inversion model is build base on neural networks. Considered spectral remote sensing method is commonly used for industrial and environmental monitoring. It is a passive single-ended sensor technique in which radiation intensity emerging from a studied object is analyzed. Quantitative investigation of heated gas radiation emission to determine temperature and gas mixture by infrared spectroscopy requires two components apart from optical radiation sensor. First appropriate spectral database and second efficient inversion techniques. In this study calculation of one-dimensional radiative transfer equation have been used for simulation of spectral radiation intensity. To increase quality of retrieval a spectrum preprocessing and feature extraction method is applied. Simulated spectra were parameterized and expressed as ratios of intensities of multiple rotational lines. Each neural network estimates temperature (NN response) at one point on studied path basing on given spectrum (NN input).

  15. Intensity distribution of Fizeau fringes in transmission with the real path of the interfered multiple-beams

    NASA Astrophysics Data System (ADS)

    Ramadan, W. A.

    2014-07-01

    In this paper, a theory is presented to estimate the intensity distribution of Fizeau fringes in transmission. In this theory the real path of the interfered light beams, through the wedge interferometer, has been considered. Interference of multiple beams has been estimated up to 12 beams considering the phase and amplitude for each beam. The summation of these waves has been done using the vector summation theory. The numerical construction of Fizeau fringes in space has been calculated considering the superposition of different number of beams. The influence of the wedge angle, number of the interfered beams and the wedge gap on the intensity distribution has been investigated. The most interesting observation in this study is the intensity distribution in different planes above the interferometer in both calculated and experimental Fizeau fringes using a He-Ne laser. Some experimental interferograms have been illustrated to confirm the validity of the proposed theory.

  16. 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. PMID:24858241

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

    PubMed

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

    2015-10-01

    Curcuminoids have received great attention in the past decades due to their health benefit properties. The aim of this study is to develop a very simple, rapid, and sensitive capillary zone electrophoresis technique coupled with a laser induced fluorescence detector (LIF) for the simultaneous determination of three major curcuminoids of turmeric, namely, curcumin, demethoxy curcumin (DMC), and bisdemethoxy curcumin (BDMC). Background electrolyte was selected as borate at pH 9.6 and (2-hydroxypropyl)-β-cyclodextrin (2-HP-β-CD) was added to prevent rapid alkali degradation of curcuminoids in buffer and to increase fluorescence intensities of molecules. With the addition of 2-HP-β-CD to the separation electrolyte, the fluorescence signal intensities of curcuminoids were enhanced considerably by 30, 40, and 54 fold for curcumin, DMC, and BDMC, respectively. The three curcuminoids of turmeric were fully separated and quantified in less than 4.5 min. The repeatability of the peak areas of curcuminoids for intra-day and inter-day experiments was in the satisfactory range of 2.26 and 2.55%, respectively. The LOD and LOQ values for the developed method were equal to or less than 0.081 and 0.270 μg/mL, respectively, for all curcuminoids. The developed method was successfully applied to find curcuminoids amount in turmeric samples and herbal supplements. PMID:26178140

  18. 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

    2016-08-01

    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. PMID:27447848

  19. 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.

  20. 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). PMID:27207680

  1. 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

  2. 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

  3. 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. PMID:25461142

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

    PubMed Central

    Vandenabeele, Frank; Grevendonk, Lotte; Verboven, Kenneth; Hansen, Dominique

    2015-01-01

    Introduction Low-to-moderate intensity exercise improves muscle contractile properties and endurance capacity in multiple sclerosis (MS). The impact of high intensity exercise remains unknown. Methods Thirty-four MS patients were randomized into a sedentary control group (SED, n = 11) and 2 exercise groups that performed 12 weeks of a high intensity interval (HITR, n = 12) or high intensity continuous cardiovascular training (HCTR, n = 11), both in combination with resistance training. M.vastus lateralis fiber cross sectional area (CSA) and proportion, knee-flexor/extensor strength, body composition, maximal endurance capacity and self-reported physical activity levels were assessed before and after 12 weeks. Results Compared to SED, 12 weeks of high intensity exercise increased mean fiber CSA (HITR: +21±7%, HCTR: +23±5%). Furthermore, fiber type I CSA increased in HCTR (+29±6%), whereas type II (+23±7%) and IIa (+23±6%,) CSA increased in HITR. Muscle strength improved in HITR and HCTR (between +13±7% and +45±20%) and body fat percentage tended to decrease (HITR: -3.9±2.0% and HCTR: -2.5±1.2%). Furthermore, endurance capacity (Wmax +21±4%, time to exhaustion +24±5%, VO2max +17±5%) and lean tissue mass (+1.4±0.5%) only increased in HITR. Finally self-reported physical activity levels increased 73±19% and 86±27% in HCTR and HITR, respectively. Conclusion High intensity cardiovascular exercise combined with resistance training was safe, well tolerated and improved muscle contractile characteristics and endurance capacity in MS. Trial Registration ClinicalTrials.gov NCT01845896 PMID:26418222

  5. 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.

  6. 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.

  7. 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. PMID:27176952

  8. 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.

  9. 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.

  10. 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

    2015-01-01

    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

  11. 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.

  12. 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.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An indirect competitive fluorescence immunoassay using DNA/dye conjugate as antibody multiple labels was developed on 96-well plates for the identification and quantification of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) in aqueous samples. A hapten, 2,4,2'-tribromodiphenyl ether-4’-aldehyde was sy...

  14. 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.

  15. 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.

  16. 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.

  17. 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. PMID:24710566

  18. A conspiracy of optimism: Sustained yield, multiple use, and intensive management on the national forests, 1945-1991

    SciTech Connect

    Hirt, P.W.

    1991-01-01

    This study focuses on two core national forest management policies; sustained yield and multiple use. Public and elected officials attempt to apply principles of sustainable development to publicly-owned forest lands to ensure that a wide variety of both market and nonmarket forest values are preserved for the benefit of present and future generations. Interest groups, the Forest Service, and policy makers have conceived of sustained yield and multiple use in different and evolving ways over the years. This study explores how these principles have been variously defined and either implemented or thwarted. After World War Two, with escalating demands on national forest resources, the US Forest Service turned to intensive management as a technological method of enhancing natural forest productivity and mitigating the environmental effects of increased use. But the agency's optimistic vision of efficient, sustained production of forest commodities through technical mastery over nature has met overwhelming fiscal, environmental, technical, and political obstacles. Changing public values since the 1960s and popularization of ecology have initiated a growing skepticism toward the premises of intensive management.

  19. Intensity based methods for brain MRI longitudinal registration. A study on multiple sclerosis patients.

    PubMed

    Diez, Yago; Oliver, Arnau; Cabezas, Mariano; Valverde, Sergi; Martí, Robert; Vilanova, Joan Carles; Ramió-Torrentà, Lluís; Rovira, Alex; Lladó, Xavier

    2014-07-01

    Registration is a key step in many automatic brain Magnetic Resonance Imaging (MRI) applications. In this work we focus on longitudinal registration of brain MRI for Multiple Sclerosis (MS) patients. First of all, we analyze the effect that MS lesions have on registration by synthetically eliminating some of the lesions. Our results show how a widely used method for longitudinal registration such as rigid registration is practically unconcerned by the presence of MS lesions while several non-rigid registration methods produce outputs that are significantly different. We then focus on assessing which is the best registration method for longitudinal MRI images of MS patients. In order to analyze the results obtained for all studied criteria, we use both descriptive statistics and statistical inference: one way ANOVA, pairwise t-tests and permutation tests. PMID:24338728

  20. 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.

  1. Direct spectroscopic observation of multiple-charged-ion acceleration by an intense femtosecond-pulse laser.

    PubMed

    Zhidkov, A G; Sasaki, A; Tajima, T; Auguste, T; D'Olivera, P; Hulin, S; Monot, P; Faenov, A Y; Pikuz, T A; Skobelev, I Y

    1999-09-01

    We have observed evidence of the emission of energetic He-and H-like ions of fluorine more than 1 MeV produced via the optical field ionization (OFI) from a solid target irradiated by an intense I=(2-4)x10(18) W/cm(2) (60 fs, lambda=800 nm), obliquely incident p-polarized pulse laser. The measured blue wing of He(alpha), He(beta), and Ly(alpha) lines of fluorine shows a feature of the Doppler-shifted spectrum due to the self-similar ion expansion dominated by superthermal electrons with the temperature T(h) approximately 100 keV. Using a collisional particle-in-cell simulation, which incorporates the nonlocal-thermodynamic-equilibrium ionization including OFI, we have obtained the plasma temperature, line shape, and maximal energy of accelerated ions, which agree well with those determined from the experimental spectra. The red wing of ion spectra gives the temperature of bulk plasma electrons. PMID:11970139

  2. Automatic Registration of Multiple Laser Scans Using Panoramic RGB and Intensity Images

    NASA Astrophysics Data System (ADS)

    Alba, M.; Barazzetti, L.; Scaioni, M.; Remondino, F.

    2011-09-01

    This paper presents an automated methodology able to register laser scanning point clouds using their panoramic images derived from intensity values or RGB data, the latter obtained from a co-registered camera. Starting from the panorama of each laser scan, a Feature-Based Matching (FBM) algorithm is pairwise applied to extract corresponding key-points. Robust estimators are then used to remove outliers through a generalized rejection procedure encompassing several geometric models. After tracking the twofold key- points across different scan pairs in order to increase the local redundancies, a global Least Squares block adjustment is computed for all scans. Ground control points can also be included at this stage for datum definition and control of block's stability. The proposed method was tested on real case studies and the experiments showed that the procedure is able to deliver the registration of all scans in a fully automatic way. On the other hand, if a higher accuracy is required this solution needs a further ICP refinement.

  3. Heterodimerization, Altered Subcellular Localization, and Function of Multiple Zinc Transporters in Viable Cells Using Bimolecular Fluorescence Complementation

    PubMed Central

    Golan, Yarden; Berman, Bluma; Assaraf, Yehuda G.

    2015-01-01

    Zinc plays a crucial role in numerous key physiological functions. Zinc transporters (ZnTs) mediate zinc efflux and compartmentalization in intracellular organelles; thus, ZnTs play a central role in zinc homeostasis. We have recently shown the in situ dimerization and function of multiple normal and mutant ZnTs using bimolecular fluorescence complementation (BiFC). Prompted by these findings, we here uncovered the heterodimerization, altered subcellular localization, and function of multiple ZnTs in live cells using this sensitive BiFC technique. We show that ZnT1, -2, -3, and -4 form stable heterodimers at distinct intracellular compartments, some of which are completely different from their homodimer localization. Specifically, unlike the plasma membrane (PM) localization of ZnT1 homodimers, ZnT1-ZnT3 heterodimers localized at intracellular vesicles. Furthermore, upon heterodimerization with ZnT1, the zinc transporters ZnT2 and ZnT4 surprisingly localized at the PM, as opposed to their vesicular homodimer localization. We further demonstrate the deleterious effect that the G87R-ZnT2 mutation, associated with transient neonatal zinc deficiency, has on ZnT1, ZnT3, and ZnT4 upon heterodimerization. The functionality of the various ZnTs was assessed by the dual BiFC-Zinquin assay. We also undertook a novel transfection competition assay with ZnT cDNAs to confirm that the driving force for heterodimer formation is the core structure of ZnTs and not the BiFC tags. These findings uncover a novel network of homo- and heterodimers of ZnTs with distinct subcellular localizations and function, hence highlighting their possible role in zinc homeostasis under physiological and pathological conditions. PMID:25657003

  4. 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.

  5. Psychosocial factors associated with pain intensity, pain-related interference, and psychological functioning in persons with multiple sclerosis and pain.

    PubMed

    Osborne, Travis L; Jensen, Mark P; Ehde, Dawn M; Hanley, Marisol A; Kraft, George

    2007-01-01

    Biopsychosocial models of chronic pain that recognize psychological and environmental factors as important aspects of adjustment to pain have been proposed for understanding chronic pain and related suffering in persons with multiple sclerosis (MS), but such models have not been empirically tested. The objective of this study was to test such a model by evaluating the associations of several psychosocial variables (i.e., pain-related catastrophizing, perceived social support, pain beliefs, and pain coping) with pain intensity, pain interference with functioning, and psychological functioning in persons with chronic pain and MS, after controlling for demographic and disease-related factors. Participants were 125 community-dwelling persons with MS and pain who completed a mailed questionnaire that included measures of pain intensity and interference, psychological functioning, catastrophizing, social support, and pain beliefs and coping. The psychosocial variables accounted for an additional 25% of the variance in average pain intensity after controlling for demographic and disease-related variables (p<.001). These variables explained an additional 22% of the variance in pain-related interference (p<.001) and 43% of the variance in psychological functioning (p<.001), after adjusting for demographic and MS-related variables and average pain intensity. Catastrophizing was consistently and independently associated with all criterion measures, whereas social support, pain beliefs, and pain coping were associated with some criterion measures but not others. The results provide empirical support for a biopsychosocial understanding of chronic pain in MS and suggest that specific psychosocial factors (e.g., catastrophizing) may be important regarding adjustment to pain in persons with MS. PMID:16950570

  6. 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

  7. Fast-electron transport and heating of solid targets in high-intensity laser interactions measured by Kα fluorescence

    NASA Astrophysics Data System (ADS)

    Martinolli, E.; Koenig, M.; Baton, S. D.; Santos, J. J.; Amiranoff, F.; Batani, D.; Perelli-Cippo, E.; Scianitti, F.; Gremillet, L.; Mélizzi, R.; Decoster, A.; Rousseaux, C.; Hall, T. A.; Key, M. H.; Snavely, R.; MacKinnon, A. J.; Freeman, R. R.; King, J. A.; Stephens, R.; Neely, D.; Clarke, R. J.

    2006-04-01

    We present experimental results on fast-electron energy deposition into solid targets in ultrahigh intensity laser-matter interaction. X-ray Kα emission spectroscopy with absolute photon counting served to diagnose fast-electron propagation in multilayered targets. Target heating was measured from ionization-shifted Kα emission. Data show a 200μm fast-electron range in solid Al. The relative intensities of spectrally shifted AlKα lines imply a mean temperature of a few tens of eV up to a 100μm depth. Experimental results suggest refluxing of the electron beam at target rear side. They were compared with the predictions of both a collisional Monte Carlo and a collisional-electromagnetic, particle-fluid transport code. The validity of the code modeling of heating in such highly transient conditions is discussed.

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

    NASA Astrophysics Data System (ADS)

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

    2011-02-01

    In this report the optical properties and energy-transfer frequency upconversion luminescence of Er3+/Yb3+-codoped laponite-derived powders under 975 nm infrared excitation is investigated. The 75%(laponite):25%(PbF2) samples doped with erbium and ytterbium ions, generated high intensity red emission around 660 nm and lower intensity green emission around 525, and 545 nm. The observed emission signals were examined as a function of the excitation power and annealing temperature. The results indicate that energy-transfer, and excited-state absorption are the major upconversion excitation mechanism for the erbium excited-state red emitting level. The precursor glass samples were also heat treated at annealing temperatures of 300 °C, 400 °C, 500 °C, and 600 °C, for a 2h period. The dependence of the visible upconversion luminescence emission upon the annealing temperature indicated the existence of an optimum temperature which leads to the generation of the most intense and spectrally pure red emission signal.

  9. 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.

  10. Theoretical and Experimental Study on Boron β-Diketonate Complexes with Intense Two-Photon-Induced Fluorescence in Solution and in the Solid State.

    PubMed

    Lanoë, Pierre-Henri; Mettra, Bastien; Liao, Yuan Yuan; Calin, Nathalie; D'Aléo, Anthony; Namikawa, Tomotaka; Kamada, Kenji; Fages, Fréderic; Monnereau, Cyrille; Andraud, Chantal

    2016-07-18

    Three boron diketonate chromophores with extended π-conjugated backbone were prepared and their spectroscopic features were investigated through a combined theoretical/experimental study. It was shown that these complexes, which undergo very large electronic reorganization upon photoexcitation, combine large two-photon absorption cross section with an emission energy and quantum efficiency in solution that is strongly dependent on solvent polarity. The strong positive influence of boron complexation on the magnitude of the two-photon absorption was clearly established, and it was shown that the two-photon absorption properties were dominated by the quadrupolar term. For one of the synthesized compounds, intense one- and two-photon-induced solid-state emission (fluorescence quantum yield of 0.65 with maximum wavelength of 610 nm) was obtained as a result of antiparallel J-aggregate crystal packing. PMID:26990918

  11. Fast electron heating in ultra-intense laser-solid interaction by shifted Kα line fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Martinolli, E.; Koenig, M.; Santos, J. J.; Amiranoff, F.; Baton, S. D.; Batani, D.; Perelli, E.; Scianitti, F.; Gremillet, L.; Rabec, M.; Rousseaux, C.; Hall, T. A.; Key, M. H.; MacKinnon, A. J.; Koch, J. A.; Freeman, R. R.; Snavely, R. A.; King, J. A.; Andersen, C.; Hill, J. M.; Stephens, R. B.; Cowan, T. E.; Ng, A.; Ao, T.

    2002-11-01

    In the context of the fast ignition studies[1], the heating of the dense fuel by fast electrons appears to be one of the most relevant aspects currently investigated [2]. In order to estimate the energy deposition and the efficiency of the fast electron transport in solid targets, we have performed experiments on LULI and RAL high power lasers, at irradiances up to a few 10^19 W/cm^2. Shifted Kα lines from an aluminum fluorescer layer buried at different depths in multilayered targets were detected using a Bragg conical-crystal spectrograph. The results were used to infer the ionization stage of the Al layer. Monte Carlo and hybrid transport codes[3] were used to study fast electron energy release by collisions and ohmic effect. The energy coupling to the target is described within an ionization model for dense matter[4] and compared to the experimental data. Despite some uncertainties of the modeling, the results give an indication of a deep heating of the target up to 30 eV after propagation in 100 μm Al. [1] M Tabak et al., Phys. of Plasmas 1, 1626 (1994) [2] E Martinolli et al., submitted to PRL, may 2002 [3] L Gremillet et al. Phys. of Plasmas 9, 941, (2002) [4] G Chiu and A Ng, PRE 59, 1024, (1999)

  12. 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.

  13. 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.

  14. 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.

  15. 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. PMID:26790523

  16. 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

  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. 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.

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

    PubMed

    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

  20. 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

  1. 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

    2016-04-01

    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. PMID:26865384

  2. Single-Isocenter Frameless Intensity-Modulated Stereotactic Radiosurgery for Simultaneous Treatment of Multiple Brain Metastases: Clinical Experience

    SciTech Connect

    Nath, Sameer K.; Lawson, Joshua D.; Simpson, Daniel R.

    2010-09-01

    Purpose: To describe our clinical experience using a unique single-isocenter technique for frameless intensity-modulated stereotactic radiosurgery (IM-SRS) to treat multiple brain metastases. Methods and Materials: 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. Results: 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 had a better 6-month LC than those >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 {>=}3 toxicity occurred in 2 patients (8%). Conclusion: 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.

  3. 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. PMID:24668857

  4. 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

  5. Detection of 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, M. A.; Bonev, B. P.; Magee-Sauer, K.; Dello Russo, N.; Mumma, M. J.; Reuter, D. C.; Villanueva, G. L.

    2006-10-01

    Formaldehyde (H2CO) was observed in comet C/2002 T7 (LINEAR) with spectral resolving power λ/Δλ~2.5×104 using the Cryogenic Echelle Spectrometer (CSHELL) at the NASA Infrared Telescope Facility, on UT 2004 May 5, 7, and 9. The observations, which sampled emission in the ν1 and ν5 rovibrational bands between 3.53 and 3.62 μm, represent the first spectrally resolved detection, at infrared wavelengths, of monomeric H2CO spanning a range of rotational energies. A comparison of measured line intensities with an existing fluorescence model permitted extraction of rotational temperatures and production rates. Two complementary approaches were used: (1) a correlation analysis that provided a direct global comparison of the observed cometary emissions with the model and (2) an excitation analysis that provided a robust line-by-line comparison. Our results validate the fluorescence model. The overall correlation coefficient was near or above 0.9 in our two principal grating settings. The excitation analysis provided accurate measures of rotational excitation (rotational temperature) on all three dates, with retrieved values of Trot clustering near 100 K. Through simultaneous measurement of OH prompt emission, which we use as a proxy for H2O, we obtained native production rates and mixing ratios for H2CO. The native production of H2CO varied from day to day, but its abundance relative to H2O, Xnative, remained approximately constant within the errors, which may suggest an overall homogeneous composition of the nucleus. We measured a mean mixing ratio Xnative= (0.79+/-0.09) × 10-2 for the three dates.

  6. 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

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

    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. PMID:22182472

  8. 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.

  9. 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. PMID:19203602

  10. UvrD limits the number and intensities of RecA-green fluorescent protein structures in Escherichia coli K-12.

    PubMed

    Centore, Richard C; Sandler, Steven J

    2007-04-01

    RecA is important for recombination, DNA repair, and SOS induction. In Escherichia coli, RecBCD, RecFOR, and RecJQ prepare DNA substrates onto which RecA binds. UvrD is a 3'-to-5' helicase that participates in methyl-directed mismatch repair and nucleotide excision repair. uvrD deletion mutants are sensitive to UV irradiation, hypermutable, and hyper-rec. In vitro, UvrD can dissociate RecA from single-stranded DNA. Other experiments suggest that UvrD removes RecA from DNA where it promotes unproductive reactions. To test if UvrD limits the number and/or the size of RecA-DNA structures in vivo, an uvrD mutation was combined with recA-gfp. This recA allele allows the number of RecA structures and the amount of RecA at these structures to be assayed in living cells. uvrD mutants show a threefold increase in the number of RecA-GFP foci, and these foci are, on average, nearly twofold higher in relative intensity. The increased number of RecA-green fluorescent protein foci in the uvrD mutant is dependent on recF, recO, recR, recJ, and recQ. The increase in average relative intensity is dependent on recO and recQ. These data support an in vivo role for UvrD in removing RecA from the DNA. PMID:17259317

  11. 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

  12. 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.

  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. 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.

  15. Leaf gas exchange, chlorophyll fluorescence and pigment indexes of Eugenia uniflora L. in response to changes in light intensity and soil flooding.

    PubMed

    Mielke, Marcelo S; Schaffer, Bruce

    2010-01-01

    The interactive effects of changing light intensity and soil flooding on the photosynthetic performance of Eugenia uniflora L. (Myrtaceae) seedlings in containers were examined. Two hypotheses were tested: (i) the photosynthetic apparatus of shade-adapted leaves can be rapidly acclimated to high light after transfer from shade to full sun, and (ii) photosynthetic acclimation to changing light intensity may be influenced by soil flooding. Seedlings cultivated in a shade house (40% of full sun, approximately 12 mol m(-)(2) day(-)(1)) for 6 months were transferred to full sun (20-40 mol m(-2) day(-1)) or shade (30% of full sun, approximately 8 mol m(-2) day(-1)) and subjected to soil flooding for 23 days or not flooded. Chlorophyll content index (CCI), chlorophyll fluorescence, leaf weight per area (LWA), photosynthetic light-response curves and leaf reflectance indexes were measured during soil flooding and after plants were unflooded. The CCI values increased throughout the experiment in leaves of shaded plants and decreased in leaves of plants transferred to full sun. There were no significant interactions between light intensity and flooding treatments for most of the variables analyzed, with the exception of Fv/Fm 22 days after plants were flooded and 5 days after flooded plants were unflooded. The light environment significantly affected LWA, and light environment and soil flooding significantly affected the light-saturated gross CO(2) assimilation rate expressed on area and dry weight bases (A(max-area) and A(max-wt), respectively), stomatal conductance of water vapor (g(ssat)) and intrinsic water use efficiency (A/g(s)). Five days after flooded plants were unflooded, the normalized difference vegetation index (NDVI) and the scaled photochemical reflectance index (sPRI) were significantly higher in shade than in sun leaves. Thirty days after transferring plants from the shade house to the light treatment, LWA was 30% higher in sun than in shade leaves, and A

  16. 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

  17. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2013-04-01

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

  19. 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 ).

  20. 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 ).

  1. 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.

  2. 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

  3. Decay of multispin multiple-quantum coherent states in the NMR of a solid and the stabilization of their intensity profile with time

    SciTech Connect

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

    2011-12-15

    Variations, experimentally observed in [14], in the intensity profiles of multiple-quantum (MQ) coherences in the presence of two special types of perturbations are explained on the basis of the theory, earlier developed by the authors, of the growth of the effective size of correlated clusters (the number of correlated spins) and the relaxation of MQ coherent states [23]. The intensity and the character of perturbation were controlled by the experimenters. It is shown that the observed stabilization of profiles with time is not associated with the stabilization of the cluster size. Quite the contrary, a cluster of correlated spins monotonically grows, while the observed variations in the intensity profile and its stabilization with time are attributed to the dependence of the decay rate of an MQ coherence on its order (its position in the MQ spectrum). The results of the theory are in good agreement with the experimental data.

  4. Effect of polarization and geometric factors on quantitative laser-induced fluorescence- to-Raman intensity ratios of water samples and a new calibration technique

    NASA Astrophysics Data System (ADS)

    Sivaprakasam, Vasanthi; Killinger, Dennis K.

    2003-09-01

    A 266-nm laser-induced fluorescence system was used to study the effect of polarization of the excitation source and geometry of the collection optics on the ratio of the signal from a fluorescence standard, quinine sulfate, and the Raman scatter from water. Although the ratio is sometimes considered to be a constant and is used for intersystem comparisons, our studies showed that the Raman signal and, thus, the ratio can vary by a factor of up to 3.6. These experimental values agree with previous studies by others involving gas and flame Raman spectroscopy and suggest a new calibration method for intersystem comparison of different fluorescence systems.

  5. 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...

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

    PubMed

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

    2015-07-27

    A new highly efficient and versatile poly(benzyl ether) dendritic organogelator HPB-G1 with 2-(2'-hydroxyphenyl)benzoxazole (HPB) at the focal point has been designed and synthesized. HPB-G1 can form stable organogels toward various apolar and polar organic solvents. Further studies revealed that intermolecular multiple π-π stacking interactions are the main driving forces for the formation of the organogels. Notably, dendron HPB-G1 exhibited a significantly enhanced emission in the gel state in contrast to weak emission in solution. Most interestingly, these dendritic organogels exhibited multiple stimuli-responsive behaviors upon exposure to environmental stimuli, including temperature, sonication, shear stress, and the presence of anions, metal cations, acids/bases, thus leading to reversible sol-gel phase transitions. PMID:26095320

  7. Simultaneous multicolor imaging of wide-field epi-fluorescence microscopy with four-bucket detection

    PubMed Central

    Park, Kwan Seob; Kim, Dong Uk; Lee, Jooran; Kim, Geon Hee; Chang, Ki Soo

    2016-01-01

    We demonstrate simultaneous imaging of multiple fluorophores using wide-field epi-fluorescence microscopy with a monochrome camera. The intensities of the three lasers are modulated by a sinusoidal waveform in order to excite each fluorophore with the same modulation frequency and a different time-delay. Then, the modulated fluorescence emissions are simultaneously detected by a camera operating at four times the excitation frequency. We show that two different fluorescence beads having crosstalk can be clearly separated using digital processing based on the phase information. In addition, multiple organelles within multi-stained single cells are shown with the phase mapping method, demonstrating an improved dynamic range and contrast compared to the conventional fluorescence image. These findings suggest that wide-field epi-fluorescence microscopy with four-bucket detection could be utilized for high-contrast multicolor imaging applications such as drug delivery and fluorescence in situ hybridization. PMID:27375944

  8. Simultaneous multicolor imaging of wide-field epi-fluorescence microscopy with four-bucket detection.

    PubMed

    Park, Kwan Seob; Kim, Dong Uk; Lee, Jooran; Kim, Geon Hee; Chang, Ki Soo

    2016-06-01

    We demonstrate simultaneous imaging of multiple fluorophores using wide-field epi-fluorescence microscopy with a monochrome camera. The intensities of the three lasers are modulated by a sinusoidal waveform in order to excite each fluorophore with the same modulation frequency and a different time-delay. Then, the modulated fluorescence emissions are simultaneously detected by a camera operating at four times the excitation frequency. We show that two different fluorescence beads having crosstalk can be clearly separated using digital processing based on the phase information. In addition, multiple organelles within multi-stained single cells are shown with the phase mapping method, demonstrating an improved dynamic range and contrast compared to the conventional fluorescence image. These findings suggest that wide-field epi-fluorescence microscopy with four-bucket detection could be utilized for high-contrast multicolor imaging applications such as drug delivery and fluorescence in situ hybridization. PMID:27375944

  9. 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. PMID:21104484

  10. 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.