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Sample records for dual fluorescent single

  1. A dual-wavelength single particle aerosol fluorescence monitor

    Science.gov (United States)

    Kaye, Paul H.; Stanley, Warren R.; Foot, Virginia; Baxter, Karen; Barrington, Stephen J.

    2005-10-01

    Laser diodes and light-emitting diodes capable of continuous sub-300 nm radiation emission will ultimately represent optimal excitation sources for compact and fieldable bio-aerosol monitors. However, until such devices are routinely available and whilst solid-state UV lasers remain relatively expensive, other low-cost sources of UV can offer advantages. This paper describes one such prototype that employs compact xenon discharge UV sources to excite intrinsic fluorescence from individual particles within an ambient aerosol sample. The prototype monitor samples ambient air via a laminar sheathed-flow arrangement such that particles within the sample flow column are rendered in single file as they intersect the beam from a continuous-wave 660nm diode laser. Each individual particle produces a scattered light signal from which an estimate of particle size (down to ~1 um) may be derived. This same signal also initiates the sequential firing (~10 us apart) of two xenon sources which irradiate the particle with UV pulses centred upon ~280 nm and ~370 nm wavelength, optimal for excitation of bio-fluorophores tryptophan and NADH respectively. For each excitation wavelength, fluorescence is detected across two bands embracing the peak emissions of the same bio-fluorophores. Thus, for each particle, a 2-dimensional fluorescence excitation-emission matrix is recorded together with an estimate of particle size. Current measurement rates are up to ~125 particles/s (limited by the xenon recharge time), corresponding to all particles for concentrations up to ~2 x 104 particles/l. Developments to increase this to ~500 particles/s are in hand. Analysis of results from aerosols of E.coli, BG spores, and a variety of non-biological materials are given.

  2. Solid-phase single molecule biosensing using dual-color colocalization of fluorescent quantum dot nanoprobes

    Science.gov (United States)

    Liu, Jianbo; Yang, Xiaohai; Wang, Kemin; Wang, Qing; Liu, Wei; Wang, Dong

    2013-10-01

    The development of solid-phase surface-based single molecule imaging technology has attracted significant interest during the past decades. Here we demonstrate a sandwich hybridization method for highly sensitive detection of a single thrombin protein at a solid-phase surface based on the use of dual-color colocalization of fluorescent quantum dot (QD) nanoprobes. Green QD560-modified thrombin binding aptamer I (QD560-TBA I) were deposited on a positive poly(l-lysine) assembled layer, followed by bovine serum albumin blocking. It allowed the thrombin protein to mediate the binding of the easily detectable red QD650-modified thrombin binding aptamer II (QD650-TBA II) to the QD560-TBA I substrate. Thus, the presence of the target thrombin can be determined based on fluorescent colocalization measurements of the nanoassemblies, without target amplification or probe separation. The detection limit of this assay reached 0.8 pM. This fluorescent colocalization assay has enabled single molecule recognition in a separation-free detection format, and can serve as a sensitive biosensing platform that greatly suppresses the nonspecific adsorption false-positive signal. This method can be extended to other areas such as multiplexed immunoassay, single cell analysis, and real time biomolecule interaction studies.The development of solid-phase surface-based single molecule imaging technology has attracted significant interest during the past decades. Here we demonstrate a sandwich hybridization method for highly sensitive detection of a single thrombin protein at a solid-phase surface based on the use of dual-color colocalization of fluorescent quantum dot (QD) nanoprobes. Green QD560-modified thrombin binding aptamer I (QD560-TBA I) were deposited on a positive poly(l-lysine) assembled layer, followed by bovine serum albumin blocking. It allowed the thrombin protein to mediate the binding of the easily detectable red QD650-modified thrombin binding aptamer II (QD650-TBA II) to

  3. Bimane: A Visible Light Induced Fluorescent Photoremovable Protecting Group for the Single and Dual Release of Carboxylic and Amino Acids.

    Science.gov (United States)

    Chaudhuri, Amrita; Venkatesh, Yarra; Behara, Krishna Kalyani; Singh, N D Pradeep

    2017-03-10

    A series of ester conjugates of carboxylic and amino acids were synthesized based on bimane fluorescent photoremovable protecting group (FPRPG). The photorelease of single and dual (same as well as different) carboxylic and amino acids is demonstrated from a single bimane molecule on irradiation with visible light (λ ≥ 410 nm). The detailed mechanistic study of photorelease revealed that the release of two caged acids is simultaneous but in a stepwise pathway.

  4. High-Resolution "Fleezers": Dual-Trap Optical Tweezers Combined with Single-Molecule Fluorescence Detection.

    Science.gov (United States)

    Whitley, Kevin D; Comstock, Matthew J; Chemla, Yann R

    2017-01-01

    Recent advances in optical tweezers have greatly expanded their measurement capabilities. A new generation of hybrid instrument that combines nanomechanical manipulation with fluorescence detection-fluorescence optical tweezers, or "fleezers"-is providing a powerful approach to study complex macromolecular dynamics. Here, we describe a combined high-resolution optical trap/confocal fluorescence microscope that can simultaneously detect sub-nanometer displacements, sub-piconewton forces, and single-molecule fluorescence signals. The primary technical challenge to these hybrid instruments is how to combine both measurement modalities without sacrificing the sensitivity of either one. We present general design principles to overcome this challenge and provide detailed, step-by-step instructions to implement them in the construction and alignment of the instrument. Lastly, we present a set of protocols to perform a simple, proof-of-principle experiment that highlights the instrument capabilities.

  5. Combined system for high-time-resolution dual-excitation fluorescence photometry and fluorescence imaging of calcium transients in single normal and diseased skeletal muscle fibers

    Science.gov (United States)

    Uttenweiler, Dietmar; Wojciechowski, Reinhold; Makabe, Makoto; Veigel, Claudia; Fink, Rainer H.

    1994-12-01

    Fast photometric measurements and video-imaging of fluorescent indicators both are powerful tools in measuring the intracellular free calcium concentration of muscle and many other cells. as photometric systems yield a high temporal resolution, calcium imaging systems have high spatial but significantly reduced temporal resolution. Therefore we have developed an integrated system combining both methods and based mostly on standard components. As a common, sensitive Ca2+- indicator we used the fluorescent probe Fura-2, which is alternatingly excited for ratio measurements at 340/380 nm. We used a commercially available dual excitation photometric system (OSP-3; Olympus) for attaching a CCD-camera and a frame grabber board. To achieve the synchronization we had to design circuitries for external triggering, synchronization and accurate control of the filter changer, which we added to the system. Additionally, the software for a triggered image acquisition was developed. With this integrated setup one can easily switch between the fast photometric mode (ratio frequency 100 Hz) and the imaging mode (ratio frequency 4.17 Hz). The calcium images are correlated with the 25 times faster spot measurements and are analyzed by means of image processing. With this combined system we study release and uptake of calcium ions of normal and diseased skeletal muscle from mdx mice. Such a system will also be important for other cellular studies in which fluorescence indicators are used to monitor similar time dependent alterations as well as changes in cellular distributions of calcium.

  6. Dual-wavelength-excitation single-particle fluorescence spectrometer/particle sorter for real-time measurement of organic carbon and biological aerosols

    Science.gov (United States)

    Pan, Yong-Le; Pinnick, Ron G.; Hill, Steven C.; Huang, Hermes; Chang, Richard K.

    2008-10-01

    We report the development of a Single-Particle Fluorescence Spectrometer (SPFS) system capable of measuring two UV-laser excited fluorescence spectra from a single particle on-the-fly. The two dispersed fluorescence spectra are obtained from excitation by two lasers at different wavelengths (263 nm and 351 nm). The SPFS samples single particles with sizes primarily in the 1-10 μm range. The fluorescence spectra are recorded from 280 nm to 600 nm (in 20 channels) for 263 nm excitation and from 370 nm to 700 nm (in 22 channels) for 351 nm excitation. The elastic scattering (channel 4 and 9) is also recorded for sizing each particle. A time stamp for single particles is marked with a variable time interval resolution from 10 ms to 10 minutes. The SPFS employs a virtual-impactor concentrator to concentrate respirable-sized particles with a resulting (size-dependent) effective flow rate of around 100 liters/min. The SPFS can measure single-particle spectra at a maximum rate of 90,000/sec, although the highest rates we have experienced for the ambient are only several hundred/sec. When the SPFS is combined with an aerodynamic deflector (puffer) to sort particles according to their fluorescence spectral characteristics, the SPFS/puffer system can selectively deflect and collect an enriched sample of targeted particles (at rates limited by the puffer) of 1200 particles/sec, for further examination. In laboratory tests, aerosol particles with similar UV-LIF spectra (e.g. B. subtilis and E.coli) are puffed into the reservoir of a micro-fluidic cell, where fluorescent-labeled antibodies bind to them and were classified by their labeled fluorescence. Measurements of the background ambient aerosol with the SPFS system made at sites with different regional climate (Connecticut, Maryland, and New Mexico) were clustered (unstructured hierarchical analysis) into 8-10 groups, with over 90% of all the fluorescent particles contained within these clusters (threshold dot product=0

  7. Solid-State Carbon Dots with Red Fluorescence and Efficient Construction of Dual-Fluorescence Morphologies.

    Science.gov (United States)

    He, Jiangling; He, Youling; Chen, Yonghao; Lei, Bingfu; Zhuang, Jianle; Xiao, Yong; Liang, Yeru; Zheng, Mingtao; Zhang, Haoran; Liu, Yingliang

    2017-07-01

    Stable solid-state red fluorescence from organosilane-functionalized carbon dots (CDs) with sizes around 3 nm is reported for the first time. Meanwhile, a novel method is also first reported for the efficient construction of dual-fluorescence morphologies. The quantum yield of these solid-state CDs and their aqueous solution is 9.60 and 50.7%, respectively. The fluorescence lifetime is 4.82 ns for solid-state CDs, and 15.57 ns for their aqueous solution. These CDs are detailedly studied how they can exhibit obvious photoluminescence overcoming the self-quenching in solid state. Luminescent materials are constructed with dual fluorescence based on as-prepared single emissive CDs (red emission) and nonfluorescence media (starch, Al2 O3 , and RnOCH3 COONa), with the characteristic peaks located at nearly 440 and 600 nm. Tunable photoluminescence can be successfully achieved by tuning the mass ratio of CDs to solid matrix (such as starch). These constructed dual-fluorescence CDs/starch composites can also be applied in white light-emitting diodes with UV chips (395 nm), and oxygen sensing. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Dual Lifetimes for Complexes between Glutathione-S-transferase (hGSTA1-1) and Product-like Ligands Detected by Single-Molecule Fluorescence Imaging.

    Science.gov (United States)

    Pettersson, John R; Lanni, Frederick; Rule, Gordon S

    2017-08-08

    Single-molecule fluorescence techniques were used to characterize the binding of products and inhibitors to human glutathione S-transferase A1-1 (hGSTA1-1). The identification of at least two different bound states for the wild-type enzyme suggests that there are at least two conformations of the protein, consistent with the model that ligand binding promotes closure of the carboxy-terminal helix over the active site. Ligand induced changes in ensemble fluorescence energy transfer support this proposed structural change. The more predominant state in the ensemble of single molecules shows a significantly faster off-rate, suggesting that the carboxy-terminal helix is delocalized in this state, permitting faster exit of the bound ligand. A point mutation (I219A), which is known to interfere with the association of the carboxy-terminal helix with the enzyme, shows increased rates of interconversion between the open and closed state. Kinematic traces of fluorescence from single molecules show that a single molecule readily samples a number of different conformations, each with a characteristic off-rate.

  9. 2000-fold parallelized dual-color STED fluorescence nanoscopy.

    Science.gov (United States)

    Bergermann, Fabian; Alber, Lucas; Sahl, Steffen J; Engelhardt, Johann; Hell, Stefan W

    2015-01-12

    Stimulated Emission Depletion (STED) nanoscopy enables multi-color fluorescence imaging at the nanometer scale. Its typical single-point scanning implementation can lead to long acquisition times. In order to unleash the full spatiotemporal resolution potential of STED nanoscopy, parallelized scanning is mandatory. Here we present a dual-color STED nanoscope utilizing two orthogonally crossed standing light waves as a fluorescence switch-off pattern, and providing a resolving power down to 30 nm. We demonstrate the imaging capabilities in a biological context for immunostained vimentin fibers in a circular field of view of 20 µm diameter at 2000-fold parallelization (i.e. 2000 "intensity minima"). The technical feasibility of massively parallelizing STED without significant compromises in resolution heralds video-rate STED nanoscopy of large fields of view, pending the availability of suitable high-speed detectors.

  10. Single-primer fluorescent sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Ruth, J.L.; Morgan, C.A.; Middendorf, L.R.; Grone, D.L.; Brumbaugh, J.A.

    1987-05-01

    Modified linker arm oligonucleotides complementary to standard M13 priming sites were synthesized, labelled with either one, two, or three fluoresceins, and purified by reverse-phase HPLC. When used as primers in standard dideoxy M13 sequencing with /sup 32/P-dNTPs, normal autoradiographic patterns were obtained. To eliminate the radioactivity, direct on-line fluorescence detection was achieved by the use of a scanning 10 mW Argon laser emitting 488 nm light. Fluorescent bands were detected directly in standard 0.2 or 0.35 mm thick polyacrylamide gels at a distance of 24 cm from the loading wells by a photomultiplier tube filtered at 520 nm. Horizontal and temporal location of each band was displayed by computer as a band in real time, providing visual appearance similar to normal 4-lane autoradiograms. Using a single primer labelled with two fluoresceins, sequences of between 500 and 600 bases have been read in a single loading with better than 98% accuracy; up to 400 bases can be read reproducibly with no errors. More than 50 sequences have been determined by this method. This approach requires only 1-2 ug of cloned template, and produces continuous sequence data at about one band per minute.

  11. Silica nanocapsules of fluorescent conjugated polymers and superparamagnetic nanocrystals for dual-mode cellular imaging.

    Science.gov (United States)

    Tan, Happy; Wang, Miao; Yang, Chang-Tong; Pant, Shilpa; Bhakoo, Kishore Kumar; Wong, Siew Yee; Chen, Zhi-Kuan; Li, Xu; Wang, John

    2011-06-01

    We describe here a facile and benign synthetic strategy to integrate the fluorescent behavior of conjugated polymers and superparamagnetic properties of iron oxide nanocrystals into silica nanocapsules, forming a new type of bifunctional magnetic fluorescent silica nanocapsule (BMFSN). The resultant BMFSNs are uniform, colloidally stable in aqueous medium, and exhibit the desired dual functionality of fluorescence and superparamagnetism in a single entity. Four conjugated polymers with different emissions were used to demonstrate the versatility of employing this class of fluorescent materials for the preparation of BMFSNs. The applicability of BMFSNs in cellular imaging was studied by incubating them with human liver cancer cells, the result of which demonstrated that the cells could be visualized by dual-mode fluorescence and magnetic resonance imaging. Furthermore, the superparamagnetic behavior of the BMFSNs was exploited for in vitro magnetic-guided delivery of the nanocapsules into the cancer cells, thereby highlighting their potential for targeting biomedical applications.

  12. Fluorescence calibration method for single-particle aerosol fluorescence instruments

    Science.gov (United States)

    Shipley Robinson, Ellis; Gao, Ru-Shan; Schwarz, Joshua P.; Fahey, David W.; Perring, Anne E.

    2017-05-01

    Real-time, single-particle fluorescence instruments used to detect atmospheric bioaerosol particles are increasingly common, yet no standard fluorescence calibration method exists for this technique. This gap limits the utility of these instruments as quantitative tools and complicates comparisons between different measurement campaigns. To address this need, we have developed a method to produce size-selected particles with a known mass of fluorophore, which we use to calibrate the fluorescence detection of a Wideband Integrated Bioaerosol Sensor (WIBS-4A). We use mixed tryptophan-ammonium sulfate particles to calibrate one detector (FL1; excitation = 280 nm, emission = 310-400 nm) and pure quinine particles to calibrate the other (FL2; excitation = 280 nm, emission = 420-650 nm). The relationship between fluorescence and mass for the mixed tryptophan-ammonium sulfate particles is linear, while that for the pure quinine particles is nonlinear, likely indicating that not all of the quinine mass contributes to the observed fluorescence. Nonetheless, both materials produce a repeatable response between observed fluorescence and particle mass. This procedure allows users to set the detector gains to achieve a known absolute response, calculate the limits of detection for a given instrument, improve the repeatability of the instrumental setup, and facilitate intercomparisons between different instruments. We recommend calibration of single-particle fluorescence instruments using these methods.

  13. In Vivo Dual Fluorescence Imaging to Detect Joint Destruction.

    Science.gov (United States)

    Cho, Hongsik; Bhatti, Fazal-Ur-Rehman; Lee, Sangmin; Brand, David D; Yi, Ae-Kyung; Hasty, Karen A

    2016-10-01

    Diagnosis of cartilage damage in early stages of arthritis is vital to impede the progression of disease. In this regard, considerable progress has been made in near-infrared fluorescence (NIRF) optical imaging technique. Arthritis can develop due to various mechanisms but one of the main contributors is the production of matrix metalloproteinases (MMPs), enzymes that can degrade components of the extracellular matrix. Especially, MMP-1 and MMP-13 have main roles in rheumatoid arthritis and osteoarthritis because they enhance collagen degradation in the process of arthritis. We present here a novel NIRF imaging strategy that can be used to determine the activity of MMPs and cartilage damage simultaneously by detection of exposed type II collagen in cartilage tissue. In this study, retro-orbital injection of mixed fluorescent dyes, MMPSense 750 FAST (MMP750) dye and Alexa Fluor 680 conjugated monoclonal mouse antibody immune-reactive to type II collagen, was administered in the arthritic mice. Both dyes were detected with different intensity according to degree of joint destruction in the animal. Thus, our dual fluorescence imaging method can be used to detect cartilage damage as well as MMP activity simultaneously in early stage arthritis.

  14. Enhanced genome editing in mammalian cells with a modified dual-fluorescent surrogate system.

    Science.gov (United States)

    Zhou, Yan; Liu, Yong; Hussmann, Dianna; Brøgger, Peter; Al-Saaidi, Rasha Abdelkadhem; Tan, Shuang; Lin, Lin; Petersen, Trine Skov; Zhou, Guang Qian; Bross, Peter; Aagaard, Lars; Klein, Tino; Rønn, Sif Groth; Pedersen, Henrik Duelund; Bolund, Lars; Nielsen, Anders Lade; Sørensen, Charlotte Brandt; Luo, Yonglun

    2016-07-01

    Programmable DNA nucleases such as TALENs and CRISPR/Cas9 are emerging as powerful tools for genome editing. Dual-fluorescent surrogate systems have been demonstrated by several studies to recapitulate DNA nuclease activity and enrich for genetically edited cells. In this study, we created a single-strand annealing-directed, dual-fluorescent surrogate reporter system, referred to as C-Check. We opted for the Golden Gate Cloning strategy to simplify C-Check construction. To demonstrate the utility of the C-Check system, we used the C-Check in combination with TALENs or CRISPR/Cas9 in different scenarios of gene editing experiments. First, we disrupted the endogenous pIAPP gene (3.0 % efficiency) by C-Check-validated TALENs in primary porcine fibroblasts (PPFs). Next, we achieved gene-editing efficiencies of 9.0-20.3 and 4.9 % when performing single- and double-gene targeting (MAPT and SORL1), respectively, in PPFs using C-Check-validated CRISPR/Cas9 vectors. Third, fluorescent tagging of endogenous genes (MYH6 and COL2A1, up to 10.0 % frequency) was achieved in human fibroblasts with C-Check-validated CRISPR/Cas9 vectors. We further demonstrated that the C-Check system could be applied to enrich for IGF1R null HEK293T cells and CBX5 null MCF-7 cells with frequencies of nearly 100.0 and 86.9 %, respectively. Most importantly, we further showed that the C-Check system is compatible with multiplexing and for studying CRISPR/Cas9 sgRNA specificity. The C-Check system may serve as an alternative dual-fluorescent surrogate tool for measuring DNA nuclease activity and enrichment of gene-edited cells, and may thereby aid in streamlining programmable DNA nuclease-mediated genome editing and biological research.

  15. Bioaerosol detection and classification using dual excitation wavelength laser-induced fluorescence

    Science.gov (United States)

    Jonsson, Per; Wästerby, Pär.; Gradmark, Per-Åke; Hedborg, Julia; Larsson, Anders; Landström, Lars

    2015-05-01

    We present results obtained by a detection system designed to measure laser-induced fluorescence from individual aerosol particles using dual excitation wavelengths. The aerosol is sampled from ambient air and via a 1 mm diameter nozzle, surrounded by a sheath air flow, confined into a particle beam. A continuous wave blue laser at 404 nm is focused on the aerosol beam and two photomultiplier tubes monitor the presence of individual particles by simultaneous measuring the scattered light and any induced fluorescence. When a particle is present in the detection volume, a laser pulse is triggered from an ultraviolet laser at 263 nm and the corresponding fluorescence spectrum is acquired with a spectrometer based on a diffraction grating and a 32 channel photomultiplier tube array with single-photon sensitivity. The spectrometer measures the fluorescence spectra in the wavelength region from 250 to 800 nm. In the present report, data were measured on different monodisperse reference aerosols, simulants of biological warfare agents, and different interference aerosol particles, e.g. pollen. In the analysis of the experimental data, i.e., the time-resolved scattered and fluorescence signals from 404 nm c.w. light excitation and the fluorescence spectra obtained by a pulsed 263 nm laser source, we use multivariate data analysis methods to classify each individual aerosol particle.

  16. Dual-wavelength excitation to reduce background fluorescence for fluorescence spectroscopic quantitation of erythrocyte zinc protoporphyrin-IX and protoporphyrin-IX from whole blood and oral mucosa

    Science.gov (United States)

    Hennig, Georg; Vogeser, Michael; Holdt, Lesca M.; Homann, Christian; Großmann, Michael; Stepp, Herbert; Gruber, Christian; Erdogan, Ilknur; Hasmüller, Stephan; Hasbargen, Uwe; Brittenham, Gary M.

    2014-02-01

    Erythrocyte zinc protoporphyrin-IX (ZnPP) and protoporphyrin-IX (PPIX) accumulate in a variety of disorders that restrict or disrupt the biosynthesis of heme, including iron deficiency and various porphyrias. We describe a reagent-free spectroscopic method based on dual-wavelength excitation that can measure simultaneously both ZnPP and PPIX fluorescence from unwashed whole blood while virtually eliminating background fluorescence. We further aim to quantify ZnPP and PPIX non-invasively from the intact oral mucosa using dual-wavelength excitation to reduce the strong tissue background fluorescence while retaining the faint porphyrin fluorescence signal originating from erythrocytes. Fluorescence spectroscopic measurements were made on 35 diluted EDTA blood samples using a custom front-face fluorometer. The difference spectrum between fluorescence at 425 nm and 407 nm excitation effectively eliminated background autofluorescence while retaining the characteristic porphyrin peaks. These peaks were evaluated quantitatively and the results compared to a reference HPLC-kit method. A modified instrument using a single 1000 μm fiber for light delivery and detection was used to record fluorescence spectra from oral mucosa. For blood measurements, the ZnPP and PPIX fluorescence intensities from the difference spectra correlated well with the reference method (ZnPP: Spearman's rho rs = 0.943, p erythrocyte ZnPP and PPIX.

  17. A dual-stimuli-responsive fluorescent switch ultrathin film

    Science.gov (United States)

    Li, Zhixiong; Liang, Ruizheng; Liu, Wendi; Yan, Dongpeng; Wei, Min

    2015-10-01

    Stimuli-responsive fluorescent switches have shown broad applications in optical devices, biological materials and intelligent responses. Herein, we describe the design and fabrication of a dual-stimuli-responsive fluorescent switch ultrathin film (UTF) via a three-step layer-by-layer (LBL) technique: (i) encapsulation of spiropyran (SP) within an amphiphilic block copolymer (PTBEM) to give the (SP@PTBEM) micelle; (ii) the mixture of riboflavin (Rf) and poly(styrene 4-sulfonate) (PSS) to enhance the adhesion ability of small molecules; (iii) assembly of negatively charged SP@PTBEM and Rf-PSS with cationic layered double hydroxide (LDH) nanoplatelets to obtain the (Rf-PSS/LDH/SP@PTBEM)n UTFs (n: bilayer number). The assembly process of the UTFs and their luminescence properties, as monitored by fluorescence spectroscopy and scanning electron microscopy (SEM), present a uniform and ordered layered structure with stepwise growth. The resulting Rf-PSS/LDH/SP@PTBEM UTF serves as a three-state switchable multicolor (green, yellow, and red) luminescent system based on stimulation from UV/Vis light and pH, with an acceptable reversibility. Therefore, this work provides a facile way to fabricate stimuli-responsive solid-state film switches with tunable-color luminescence, which have potential applications in the areas of displays, sensors, and rewritable optical memory and fluorescent logic devices.Stimuli-responsive fluorescent switches have shown broad applications in optical devices, biological materials and intelligent responses. Herein, we describe the design and fabrication of a dual-stimuli-responsive fluorescent switch ultrathin film (UTF) via a three-step layer-by-layer (LBL) technique: (i) encapsulation of spiropyran (SP) within an amphiphilic block copolymer (PTBEM) to give the (SP@PTBEM) micelle; (ii) the mixture of riboflavin (Rf) and poly(styrene 4-sulfonate) (PSS) to enhance the adhesion ability of small molecules; (iii) assembly of negatively charged SP

  18. Simultaneous live cell imaging using dual FRET sensors with a single excitation light.

    Directory of Open Access Journals (Sweden)

    Yusuke Niino

    Full Text Available Fluorescence resonance energy transfer (FRET between fluorescent proteins is a powerful tool for visualization of signal transduction in living cells, and recently, some strategies for imaging of dual FRET pairs in a single cell have been reported. However, these necessitate alteration of excitation light between two different wavelengths to avoid the spectral overlap, resulting in sequential detection with a lag time. Thus, to follow fast signal dynamics or signal changes in highly motile cells, a single-excitation dual-FRET method should be required. Here we reported this by using four-color imaging with a single excitation light and subsequent linear unmixing to distinguish fluorescent proteins. We constructed new FRET sensors with Sapphire/RFP to combine with CFP/YFP, and accomplished simultaneous imaging of cAMP and cGMP in single cells. We confirmed that signal amplitude of our dual FRET measurement is comparable to of conventional single FRET measurement. Finally, we demonstrated to monitor both intracellular Ca(2+ and cAMP in highly motile cardiac myocytes. To cancel out artifacts caused by the movement of the cell, this method expands the applicability of the combined use of dual FRET sensors for cell samples with high motility.

  19. Photoconversion of purified fluorescent proteins and dual-probe optical highlighting in live cells.

    Science.gov (United States)

    Kremers, Gert-Jan; Piston, David

    2010-06-26

    Photoconvertible fluorescent proteins (pc-FPs) are a class of fluorescent proteins with "optical highlighter" capability, meaning that the color of fluorescence can be changed by exposure to light of a specific wavelength. Optical highlighting allows noninvasive marking of a subpopulation of fluorescent molecules, and is therefore ideal for tracking single cells or organelles. Critical parameters for efficient photoconversion are the intensity and the exposure time of the photoconversion light. If the intensity is too low, photoconversion will be slow or not occur at all. On the other hand, too much intensity or too long exposure can photobleach the protein and thereby reduce the efficiency of photoconversion. This protocol describes a general approach how to set up a confocal laser scanning microscope for pc-FP photoconversion applications. First, we describe a procedure for preparing purified protein droplet samples. This sample format is very convenient for studying the photophysical behavior of fluorescent proteins under the microscope. Second, we will use the protein droplet sample to show how to configure the microscope for photoconversion. And finally, we will show how to perform optical highlighting in live cells, including dual-probe optical highlighting with mOrange2 and Dronpa.

  20. Dual Fluorescence in GFP Chromophore Analogues: Chemical Modulation of Charge Transfer and Proton Transfer Bands.

    Science.gov (United States)

    Chatterjee, Tanmay; Mandal, Mrinal; Das, Ananya; Bhattacharyya, Kalishankar; Datta, Ayan; Mandal, Prasun K

    2016-04-14

    Dual fluorescence of GFP chromophore analogues has been observed for the first time. OHIM (o-hydroxy imidazolidinone) shows only a charge transfer (CT) band, CHBDI (p-cyclicamino o-hydroxy benzimidazolidinone) shows a comparable intensity CT and PT (proton transfer) band, and MHBDI (p-methoxy o-hydroxy benzimidazolidinone) shows a higher intensity PT band. It could be shown that the differential optical behavior is not due to conformational variation in the solid or solution phase. Rather, control of the excited state electronic energy level and excited state acidity constant by functional group modification could be shown to be responsible for the differential optical behavior. Chemical modification-induced electronic control over the relative intensity of the charge transfer and proton transfer bands could thus be evidenced. Support from single-crystal X-ray structure, NMR, femtosecond to nanosecond fluorescence decay analysis, and TDDFT-based calculation provided important information and thus helped us understand the photophysics better.

  1. Single Molecule Spectroscopy of Fluorescent Proteins

    NARCIS (Netherlands)

    Blum, Christian; Subramaniam, Vinod

    2009-01-01

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

  2. A Single Switch Dual Output Non-Isolated Boost Converter

    DEFF Research Database (Denmark)

    Klimczak, Pawel; Munk-Nielsen, Stig

    2008-01-01

    very simple dual output non-isolated boost converter is presented. Single active switch is used to control both, positive and negative output voltages. The converter is desired to boost unregulated low input voltage 25-50 Vdc to regulated high voltage ±400 Vdc in dual dc-link. In this paper proposed...

  3. Dual-Colored DNA Comb Polymers for Single Molecule Rheology

    Science.gov (United States)

    Mai, Danielle; Marciel, Amanda; Schroeder, Charles

    2014-03-01

    We report the synthesis and characterization of branched biopolymers for single molecule rheology. In our work, we utilize a hybrid enzymatic-synthetic approach to graft ``short'' DNA branches to ``long'' DNA backbones, thereby producing macromolecular DNA comb polymers. The branches and backbones are synthesized via polymerase chain reaction with chemically modified deoxyribonucleotides (dNTPs): ``short'' branches consist of Cy5-labeled dNTPs and a terminal azide group, and ``long'' backbones contain dibenzylcyclooctyne-modified (DBCO) dNTPs. In this way, we utilize strain-promoted, copper-free cycloaddition ``click'' reactions for facile grafting of azide-terminated branches at DBCO sites along backbones. Copper-free click reactions are bio-orthogonal and nearly quantitative when carried out under mild conditions. Moreover, comb polymers can be labeled with an intercalating dye (e.g., YOYO) for dual-color fluorescence imaging. We characterized these materials using gel electrophoresis, HPLC, and optical microscopy, with atomic force microscopy in progress. Overall, DNA combs are suitable for single molecule dynamics, and in this way, our work holds the potential to improve our understanding of topologically complex polymer melts and solutions.

  4. R26R-GR: a Cre-activable dual fluorescent protein reporter mouse.

    Directory of Open Access Journals (Sweden)

    You-Tzung Chen

    Full Text Available Green fluorescent protein (GFP and its derivatives are the most widely used molecular reporters for live cell imagining. The development of organelle-specific fusion fluorescent proteins improves the labeling resolution to a higher level. Here we generate a R26 dual fluorescent protein reporter mouse, activated by Cre-mediated DNA recombination, labeling target cells with a chromatin-specific enhanced green fluorescence protein (EGFP and a plasma membrane-anchored monomeric cherry fluorescent protein (mCherry. This dual labeling allows the visualization of mitotic events, cell shapes and intracellular vesicle behaviors. We expect this reporter mouse to have a wide application in developmental biology studies, transplantation experiments as well as cancer/stem cell lineage tracing.

  5. In vivo tomographic imaging with fluorescence and MRI using tumor-targeted dual-labeled nanoparticles

    Directory of Open Access Journals (Sweden)

    Zhang Y

    2013-12-01

    Full Text Available Yue Zhang,1 Bin Zhang,1 Fei Liu,1,2 Jianwen Luo,1,3 Jing Bai1 1Department of Biomedical Engineering, School of Medicine, 2Tsinghua-Peking Center for Life Sciences, 3Center for Biomedical Imaging Research, Tsinghua University, Beijing, People's Republic of China Abstract: Dual-modality imaging combines the complementary advantages of different modalities, and offers the prospect of improved preclinical research. The combination of fluorescence imaging and magnetic resonance imaging (MRI provides cross-validated information and direct comparison between these modalities. Here, we report on the application of a novel tumor-targeted, dual-labeled nanoparticle (NP, utilizing iron oxide as the MRI contrast agent and near infrared (NIR dye Cy5.5 as the fluorescent agent. Results of in vitro experiments verified the specificity of the NP to tumor cells. In vivo tumor targeting and uptake of the NPs in a mouse model were visualized by fluorescence and MR imaging collected at different time points. Quantitative analysis was carried out to evaluate the efficacy of MRI contrast enhancement. Furthermore, tomographic images were also acquired using both imaging modalities and cross-validated information of tumor location and size between these two modalities was revealed. The results demonstrate that the use of dual-labeled NPs can facilitate the dual-modal detection of tumors, information cross-validation, and direct comparison by combing fluorescence molecular tomography (FMT and MRI. Keywords: dual-modality, fluorescence molecular tomography (FMT, magnetic resonance imaging (MRI, nanoparticle

  6. A new approach to dual-color two-photon microscopy with fluorescent proteins

    Directory of Open Access Journals (Sweden)

    Rebane Aleks

    2010-02-01

    Full Text Available Abstract Background Two-photon dual-color imaging of tissues and cells labeled with fluorescent proteins (FPs is challenging because most two-photon microscopes only provide one laser excitation wavelength at a time. At present, methods for two-photon dual-color imaging are limited due to the requirement of large differences in Stokes shifts between the FPs used and their low two-photon absorption (2PA efficiency. Results Here we present a new method of dual-color two-photon microscopy that uses the simultaneous excitation of the lowest-energy electronic transition of a blue fluorescent protein and a higher-energy electronic transition of a red fluorescent protein. Conclusion Our method does not require large differences in Stokes shifts and can be extended to a variety of FP pairs with larger 2PA efficiency and more optimal imaging properties.

  7. Precise diagnosis in different scenarios using photoacoustic and fluorescence imaging with dual-modality nanoparticles

    Science.gov (United States)

    Peng, Dong; Du, Yang; Shi, Yiwen; Mao, Duo; Jia, Xiaohua; Li, Hui; Zhu, Yukun; Wang, Kun; Tian, Jie

    2016-07-01

    Photoacoustic imaging and fluorescence molecular imaging are emerging as important research tools for biomedical studies. Photoacoustic imaging offers both strong optical absorption contrast and high ultrasonic resolution, and fluorescence molecular imaging provides excellent superficial resolution, high sensitivity, high throughput, and the ability for real-time imaging. Therefore, combining the imaging information of both modalities can provide comprehensive in vivo physiological and pathological information. However, currently there are limited probes available that can realize both fluorescence and photoacoustic imaging, and advanced biomedical applications for applying this dual-modality imaging approach remain underexplored. In this study, we developed a dual-modality photoacoustic-fluorescence imaging nanoprobe, ICG-loaded Au@SiO2, which was uniquely designed, consisting of gold nanorod cores and indocyanine green with silica shell spacer layers to overcome fluorophore quenching. This nanoprobe was examined by both PAI and FMI for in vivo imaging on tumor and ischemia mouse models. Our results demonstrated that the nanoparticles can specifically accumulate at the tumor and ischemic areas and be detected by both imaging modalities. Moreover, this dual-modality imaging strategy exhibited superior advantages for a precise diagnosis in different scenarios. The new nanoprobe with the dual-modality imaging approach holds great potential for diagnosis and stage classification of tumor and ischemia related diseases.Photoacoustic imaging and fluorescence molecular imaging are emerging as important research tools for biomedical studies. Photoacoustic imaging offers both strong optical absorption contrast and high ultrasonic resolution, and fluorescence molecular imaging provides excellent superficial resolution, high sensitivity, high throughput, and the ability for real-time imaging. Therefore, combining the imaging information of both modalities can provide

  8. Gd(iii)-doped carbon dots as a dual fluorescent-MRI probe

    KAUST Repository

    Bourlinos, Athanasios B.

    2012-01-01

    We describe the synthesis of Gd(iii)-doped carbon dots as dual fluorescence-MRI probes for biomedical applications. The derived Gd(iii)-doped carbon dots show uniform particle size (3-4 nm) and gadolinium distribution and form stable dispersions in water. More importantly, they exhibit bright fluorescence, strong T1-weighted MRI contrast and low cytotoxicity. © The Royal Society of Chemistry 2012.

  9. Direct Characterization of Amyloidogenic Oligomers by Single-Molecule Fluorescence

    National Research Council Canada - National Science Library

    Angel Orte; Neil R. Birkett; Richard W. Clarke; Glyn L. Devlin; Christopher M. Dobson; David Klenerman

    2008-01-01

    .... We describe here the application of a two-color single-molecule fluorescence technique to examine the assembly of oligomeric species formed during the aggregation of the SH3 domain of PI3 kinase...

  10. Lanthanide based dual-emission fluorescent probe for detection of mercury (II) in milk.

    Science.gov (United States)

    Tan, Hongliang; Li, Qian; Ma, Chanjiao; Song, Yonghai; Xu, Fugang; Chen, Shouhui; Wang, Li

    2015-01-15

    It is highly desirable to develop a simple and sensitive method for Hg(2+) detection because of the dangerous nature of Hg(2+). In this work, we prepared a dual-emission fluorescent probe for Hg(2+) detection by combining two lanthanide chelates with different emission wavelengths. Green-emitting terbium (Tb(3+)) chelates as reference signals were embedded into SiO2 nanoparticles and red-emitting europium (Eu(3+)) chelates as response units were covalently linked to the surface of silica shell. Upon the addition of Hg(2+), the fluorescence of Eu(3+) chelates can be selectively quenched, while the fluorescence of Tb(3+) chelates remained unchanged. As a kind of Hg(2+) nanosensor, the dual-emission fluorescent probe exhibited excellent selectivity to Hg(2+) and high sensitivity up to 7.07 nM detection limit. The Hg(2+) levels in drinking water and milk samples were determined by using the dual-emission fluorescent probe with satisfied recovery. Additionally, our probe has a long enough fluorescence lifetime, which can avoid the interference from autofluorescence of the biological samples. We envision that the proposed probe could find great potential applications for ultrasensitive time-resolved fluorometric assays and biomedical imaging in the future.

  11. Specific survivin dual fluorescence resonance energy transfer molecular beacons for detection of human bladder cancer cells

    Institute of Scientific and Technical Information of China (English)

    Zhi-qiang WANG; Jun ZHAO; Jin ZENG; Kai-jie WU; Yu-le CHEN; Xin-ya ng WANG; Luke S CHANG; Da-lin HE

    2011-01-01

    Survivin molecular beacons can be used to detectbladder cancer cells in urine samples non-invasively.The aim of this study is to improve the specificity of detection of bladder cancer cells using survivin dual fluorescence resonance energy transfer molecular beacons (FRET MBs) that have fluorophores forming one donor-acceptor pair.Methods:Survivin-targeting dual fluorescence resonance energy transfer molecular beacons with unique target sequences were designed,which had no overlap with the other genes in the apoptosis inhibitor protein family.Human bladder cancer cell lines 5637,253J and T24,as well as the exfoliated cells in the urine of healthy adults and patients with bladder cancer were examined.Images of cells were taken using a laser scanning confocal fluorescence microscope.For assays using dual FRET MBs,the excitation wavelength was 488 nm,and the emission detection wavelengths were 520+20 nm and 560+20 nm,respectively.Results:The human bladder cancer cell lines and exfoliated cells in the urine of patients with bladder cancer incubated with the survivin dual FRET MBs exhibited strong fluorescence signals.In contrast,no fluorescence was detected in the survivin-negative human dermal fibroblasts-adult (HDF-a) cells or exfoliated cells in the urine of healthy adults incubated with the survivin dual FRET MBs.Conclusion:The results suggest that the survivin dual FRET MBs may be used as a specific and non-invasive method for early detection and follow-up of patients with bladder cancer.

  12. Fluorescence imaging of single Kinesin motors on immobilized microtubules.

    Science.gov (United States)

    Korten, Till; Nitzsche, Bert; Gell, Chris; Ruhnow, Felix; Leduc, Cécile; Diez, Stefan

    2011-01-01

    Recent developments in optical microscopy and nanometer tracking have greatly improved our understanding of cytoskeletal motor proteins. Using fluorescence microscopy, dynamic interactions are now routinely observed in vitro on the level of single molecules mainly using a geometry, where fluorescently labeled motors move on surface-immobilized filaments. In this chapter, we review recent methods related to single-molecule kinesin motility assays. In particular, we aim to provide practical advice on: how to set up the assays, how to acquire high-precision data from fluorescently labeled kinesin motors and attached quantum dots, and how to analyze data by nanometer tracking.

  13. Dual fluorescent labelling of the human malaria parasite Plasmodium falciparum for the analysis of the ABC type transporter pfmdr2.

    Science.gov (United States)

    Rosental, Benyamin; Hadad, Uzi; Sinay, Rosa; Braiman, Alex; Porgador, Angel; Pollack, Yaakov

    2012-11-08

    The study of the Plasmodium falciparum heavy metal transporter gene pfmdr2 employed radioactive labelled heavy metal. As the use of radioactive isotopes shrank considerably during the last few years, resulting in the cessation of the production of some isotopes, amongst them Cadmium109 which was used for that purpose, a different approach had to be developed. Herein, a dual fluorescent labelling of heavy metals accumulation in the P. falciparum parasite is proposed as an alternative to the use of radioactive labelled heavy metals. Plasmodium falciparum Cd resistant and sensitive strains at the trophozoite stage were used in this study. The cells were cultured at different CdCl2 concentrations and for different time periods followed by staining of the infected red blood cells with Fluo-3/AM for Cd detection and Hoechst 33342 for parasite DNA labelling. The fluorescent analysis was done by flow cytometry and confocal microscopy. The results show that the sensitive strain has a higher Fluo-3/AM fluorescence in a Cd concentration and time dependent manner, whereas in the resistant strain Fluo-3/AM fluorescence levels were negligible and increased only at high concentrations of Cd and at long incubation periods, but to a much lesser extent than the sensitive strain. No Cd uptake is observed in uninfected red blood cells populations originating from cultures infected with either sensitive or resistant strain. In addition, confocal microscopy overlay of Fluo-3/AM and Hoechst staining shows that the Cd metal accumulates in the parasite itself. The dual fluorescent labelling is a valid method for detecting heavy metal accumulation in P. falciparum. Furthermore, in contrast to the use of radioactive labelled heavy metal, the fluorescent labelling enables us to differentiate between the different populations existing in a P. falciparum infected red blood cells cultures and thus actually study a phenomenon at the level of a single cell.

  14. Dual fluorescent labelling of the human malaria parasite Plasmodium falciparum for the analysis of the ABC type transporter pfmdr2

    Directory of Open Access Journals (Sweden)

    Rosental Benyamin

    2012-11-01

    Full Text Available Abstract Background The study of the Plasmodium falciparum heavy metal transporter gene pfmdr2 employed radioactive labelled heavy metal. As the use of radioactive isotopes shrank considerably during the last few years, resulting in the cessation of the production of some isotopes, amongst them Cadmium109 which was used for that purpose, a different approach had to be developed. Herein, a dual fluorescent labelling of heavy metals accumulation in the P. falciparum parasite is proposed as an alternative to the use of radioactive labelled heavy metals. Methods Plasmodium falciparum Cd resistant and sensitive strains at the trophozoite stage were used in this study. The cells were cultured at different CdCl2 concentrations and for different time periods followed by staining of the infected red blood cells with Fluo-3/AM for Cd detection and Hoechst 33342 for parasite DNA labelling. The fluorescent analysis was done by flow cytometry and confocal microscopy. Results The results show that the sensitive strain has a higher Fluo-3/AM fluorescence in a Cd concentration and time dependent manner, whereas in the resistant strain Fluo-3/AM fluorescence levels were negligible and increased only at high concentrations of Cd and at long incubation periods, but to a much lesser extent than the sensitive strain. No Cd uptake is observed in uninfected red blood cells populations originating from cultures infected with either sensitive or resistant strain. In addition, confocal microscopy overlay of Fluo-3/AM and Hoechst staining shows that the Cd metal accumulates in the parasite itself. Conclusions The dual fluorescent labelling is a valid method for detecting heavy metal accumulation in P. falciparum. Furthermore, in contrast to the use of radioactive labelled heavy metal, the fluorescent labelling enables us to differentiate between the different populations existing in a P. falciparum infected red blood cells cultures and thus actually study a phenomenon at

  15. Precise diagnosis in different scenarios using photoacoustic and fluorescence imaging with dual-modality nanoparticles.

    Science.gov (United States)

    Peng, Dong; Du, Yang; Shi, Yiwen; Mao, Duo; Jia, Xiaohua; Li, Hui; Zhu, Yukun; Wang, Kun; Tian, Jie

    2016-08-14

    Photoacoustic imaging and fluorescence molecular imaging are emerging as important research tools for biomedical studies. Photoacoustic imaging offers both strong optical absorption contrast and high ultrasonic resolution, and fluorescence molecular imaging provides excellent superficial resolution, high sensitivity, high throughput, and the ability for real-time imaging. Therefore, combining the imaging information of both modalities can provide comprehensive in vivo physiological and pathological information. However, currently there are limited probes available that can realize both fluorescence and photoacoustic imaging, and advanced biomedical applications for applying this dual-modality imaging approach remain underexplored. In this study, we developed a dual-modality photoacoustic-fluorescence imaging nanoprobe, ICG-loaded Au@SiO2, which was uniquely designed, consisting of gold nanorod cores and indocyanine green with silica shell spacer layers to overcome fluorophore quenching. This nanoprobe was examined by both PAI and FMI for in vivo imaging on tumor and ischemia mouse models. Our results demonstrated that the nanoparticles can specifically accumulate at the tumor and ischemic areas and be detected by both imaging modalities. Moreover, this dual-modality imaging strategy exhibited superior advantages for a precise diagnosis in different scenarios. The new nanoprobe with the dual-modality imaging approach holds great potential for diagnosis and stage classification of tumor and ischemia related diseases.

  16. Dual-modality, fluorescent, PLGA encapsulated bismuth nanoparticles for molecular and cellular fluorescence imaging and computed tomography

    Science.gov (United States)

    Swy, Eric R.; Schwartz-Duval, Aaron S.; Shuboni, Dorela D.; Latourette, Matthew T.; Mallet, Christiane L.; Parys, Maciej; Cormode, David P.; Shapiro, Erik M.

    2014-10-01

    Reports of molecular and cellular imaging using computed tomography (CT) are rapidly increasing. Many of these reports use gold nanoparticles. Bismuth has similar CT contrast properties to gold while being approximately 1000-fold less expensive. Herein we report the design, fabrication, characterization, and CT and fluorescence imaging properties of a novel, dual modality, fluorescent, polymer encapsulated bismuth nanoparticle construct for computed tomography and fluorescence imaging. We also report on cellular internalization and preliminary in vitro and in vivo toxicity effects of these constructs. 40 nm bismuth(0) nanocrystals were synthesized and encapsulated within 120 nm Poly(dl-lactic-co-glycolic acid) (PLGA) nanoparticles by oil-in-water emulsion methodologies. Coumarin-6 was co-encapsulated to impart fluorescence. High encapsulation efficiency was achieved ~70% bismuth w/w. Particles were shown to internalize within cells following incubation in culture. Bismuth nanocrystals and PLGA encapsulated bismuth nanoparticles exhibited >90% and >70% degradation, respectively, within 24 hours in acidic, lysosomal environment mimicking media and both remained nearly 100% stable in cytosolic/extracellular fluid mimicking media. μCT and clinical CT imaging was performed at multiple X-ray tube voltages to measure concentration dependent attenuation rates as well as to establish the ability to detect the nanoparticles in an ex vivo biological sample. Dual fluorescence and CT imaging is demonstrated as well. In vivo toxicity studies in rats revealed neither clinically apparent side effects nor major alterations in serum chemistry and hematology parameters. Calculations on minimal detection requirements for in vivo targeted imaging using these nanoparticles are presented. Indeed, our results indicate that these nanoparticles may serve as a platform for sensitive and specific targeted molecular CT and fluorescence imaging.Reports of molecular and cellular imaging using

  17. A new azine derivative colorimetric and fluorescent dual-channel probe for cyanide detection

    Science.gov (United States)

    Yu, Bin; Li, Chun-Yu; Sun, Yin-Xia; Jia, Hao-Ran; Guo, Jian-Qiang; Li, Jing

    2017-09-01

    A novel azine derivative colorimetric and fluorescent dual-channel probe salicylaldehyde hydrazine-3,5-dibromosalicylaldehyde (1) has been designed, synthesized and characterized. The probe 1 is confirmed to have especial selectivity and good sensitivity on detecting CN- via UV-vis absorption and fluorescence spectrum in aqueous solution (H2O/DMSO, 1:4, v/v). This colorimetric and fluorescent dual-channel probe response to CN- owed to the deprotonation process and established the mechanism by using 1H NMR spectroscopy. Further researches showed that the detection limit of the probe 1 to CN- anions is 8.01 × 10- 9 M, significantly lower than the maximum level 1.9 × 10- 6 M in potable water from WHO guidelines.

  18. A dual oxygenation and fluorescence imaging platform for reconstructive surgery

    Science.gov (United States)

    Ashitate, Yoshitomo; Nguyen, John N.; Venugopal, Vivek; Stockdale, Alan; Neacsu, Florin; Kettenring, Frank; Lee, Bernard T.; Frangioni, John V.; Gioux, Sylvain

    2013-03-01

    There is a pressing clinical need to provide image guidance during surgery. Currently, assessment of tissue that needs to be resected or avoided is performed subjectively, leading to a large number of failures, patient morbidity, and increased healthcare costs. Because near-infrared (NIR) optical imaging is safe, noncontact, inexpensive, and can provide relatively deep information (several mm), it offers unparalleled capabilities for providing image guidance during surgery. These capabilities are well illustrated through the clinical translation of fluorescence imaging during oncologic surgery. In this work, we introduce a novel imaging platform that combines two complementary NIR optical modalities: oxygenation imaging and fluorescence imaging. We validated this platform during facial reconstructive surgery on large animals approaching the size of humans. We demonstrate that NIR fluorescence imaging provides identification of perforator arteries, assesses arterial perfusion, and can detect thrombosis, while oxygenation imaging permits the passive monitoring of tissue vital status, as well as the detection and origin of vascular compromise simultaneously. Together, the two methods provide a comprehensive approach to identifying problems and intervening in real time during surgery before irreparable damage occurs. Taken together, this novel platform provides fully integrated and clinically friendly endogenous and exogenous NIR optical imaging for improved image-guided intervention during surgery.

  19. Fluobodies : green fluorescent single-chain Fv fusion proteins

    NARCIS (Netherlands)

    Griep, R.A.; Twisk, van C.; Wolf, van der J.M.; Schots, A.

    1999-01-01

    An expression system (pSKGFP), which permits the expression of single-chain variable fragments as fusion proteins with modified green fluorescent proteins, was designed. This expression system is comparable to frequently used phage display vectors and allows single-step characterization of the selec

  20. Correlated spontaneous emission of fluorescent emitters mediated by single plasmons

    CERN Document Server

    Bouchet, Dorian; Ithurria, Sandrine; Gulinatti, Angelo; Rech, Ivan; Carminati, Rémi; De Wilde, Yannick; Krachmalnicoff, Valentina

    2016-01-01

    Manipulating the spontaneous emission of a fluorescent emitter can be achieved by placing the emitter in a nanostructured environment. A privileged spot is occupied by plasmonic structures that provide a strong confinement of the electromagnetic field, which results in an enhancement of the emitter-environment interaction. While plasmonic nanostructures have been widely exploited to control the emission properties of single photon emitters, performing the coupling between quantum emitters with plasmons poses a huge challenge. In this Letter we report on a first crucial step towards this goal by the observation of correlated emission between a single CdSe/CdS/ZnS quantum dot exhibiting single photon statistics and a fluorescent nanobead located micrometers apart. This is accomplished by coupling both emitters to a silver nanowire. Single-plasmons are created on the latter from the quantum dot, and transfer energy to excite in turn the fluorescent nanobead.

  1. Facile synthesis of CdTe@GdS fluorescent-magnetic nanoparticles for tumor-targeted dual-modal imaging.

    Science.gov (United States)

    Zhang, Fei; Kong, Xiu-Qi; Li, Qiong; Sun, Ting-Ting; Chai, Chao; Shen, Wen; Hong, Zhang-Yong; He, Xi-Wen; Li, Wen-You; Zhang, Yu-Kui

    2016-01-01

    Multimodal imaging has made great contribution for diagnosis and therapy of disease since it can provide more effective and complementary information in comparison to any single imaging modality. The design and fabrication of fluorescent-magnetic nanoparticles for multimodal imaging has rapidly developed over the years. Herein, we demonstrate the facile synthesis of GdS coated CdTe nanoparticles (CdTe@GdS NPs) as multimodal agents for fluorescence (FL) and T1-weighted magnetic resonance (MR) imaging. These nanoparticles obtain both prominent fluorescent and paramagnetic properties by coating the GdS shell on the surface of CdTe core via a simple room-temperature route in aqueous solution directly. It is shown that the as-prepared CdTe@GdS NPs have high quantum yield (QY) value of 12% and outstanding longitudinal relaxation rate (r1) of 11.25 mM s(-1), which allow them to be employed as FL/MR dual-modal imaging contrast agents. They also exhibit small particle size of 5 nm, excellent colloidal stability and low cellular toxicity for concentrations up to 750 μg mL(-1). In addition, with the conjugation of folic acid, the nanoparticles were successfully used for tumor-targeted FL/MR dual-modal imaging in vitro and in vivo.

  2. Pyrene-based dual-mode fluorescence switches and logic gates that function in solution and film.

    Science.gov (United States)

    Zhou, Weidong; Li, Yongjun; Li, Yuliang; Liu, Huibiao; Wang, Shu; Li, Cuihong; Yuan, Mingjian; Liu, Xiaofeng; Zhu, Daoben

    2006-07-17

    A dual-mode fluorescence switch controlled by external inputs such as protons and metal ions is described, and each state corresponds to a specific fluorescent emission peak. Based on the reversible changes of the fluorescence emission of the switch responding to different external stimuli, the corresponding integrated logic gates and communication networks have been constructed in solid film or in solution.

  3. Fluorescence preselection of bioaerosol for single-particle mass spectrometry

    Science.gov (United States)

    Stowers, M. A.; van Wuijckhuijse, A. L.; Marijnissen, J. C. M.; Kientz, Ch. E.; Ciach, T.

    2006-11-01

    We have designed, constructed, and tested a system that preselects the biological fraction of airborne particles from the overall aerosol. The preselection is based on fluorescence emission excited by a continuous 266 nm laser beam. This beam is one of two cw beams used to measure the aerodynamic particle size of sampled particles. The intention in our system is that single particles, based on size and fluorescence emission, can be selected and further examined for chemical composition by mass spectrometry.

  4. Fluorescence preselection of bioaerosol for single-particle mass spectrometry

    OpenAIRE

    Stowers, M.A.; van Wuijckhuijse, A.L.; Marijnissen, J.C.M.; Kientz, C.E.; Ciach, T.

    2006-01-01

    We have designed, constructed, and tested a system that preselects the biological fraction of airborne particles from the overall aerosol. The preselection is based on fluorescence emission excited by a continuous 266 nm laser beam. This beam is one of two cw beams used to measure the aerodynamic particle size of sampled particles. The intention in our system is that single particles, based on size and fluorescence emission, can be selected and further examined for chemical composition by mas...

  5. Common fluorescent proteins for single-molecule localization microscopy

    Science.gov (United States)

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

    2015-07-01

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

  6. Dielectrophoretic manipulation of fluorescing single-walled carbon nanotubes.

    Science.gov (United States)

    Mureau, Natacha; Mendoza, Ernest; Silva, S Ravi P

    2007-05-01

    We investigate the behavior of fluorescing single-walled carbon nanotubes (SWCNTs) under dielectrophoretic conditions and demonstrate their collection with fluorescence microscopy. SWCNTs are dispersed in water with the aid of a nonionic surfactant, Triton X-100, and labeled through noncovalent binding with the dye 3,3'-dihexyloxacarbocyanine iodide (diOC(6)). The chromophore's affinity to the SWCNTs is due to pi-stacking interactions. Carbon nanotube (CNT) localization is clearly identified on the fluorescence images, showing that the nanotubes concentrate between the electrodes and align along the electric field lines.

  7. Single cell genomic quantification by non-fluorescence nonlinear microscopy

    Science.gov (United States)

    Kota, Divya; Liu, Jing

    2017-02-01

    Human epidermal growth receptor 2 (Her2) is a gene which plays a major role in breast cancer development. The quantification of Her2 expression in single cells is limited by several drawbacks in existing fluorescence-based single molecule techniques, such as low signal-to-noise ratio (SNR), strong autofluorescence and background signals from biological components. For rigorous genomic quantification, a robust method of orthogonal detection is highly desirable and we demonstrated it by two non-fluorescent imaging techniques -transient absorption microscopy (TAM) and second harmonic generation (SHG). In TAM, gold nanoparticles (AuNPs) are chosen as an orthogonal probes for detection of single molecules which gives background-free quantifications of single mRNA transcript. In SHG, emission from barium titanium oxide (BTO) nanoprobes was demonstrated which allows stable signal beyond the autofluorescence window. Her2 mRNA was specifically labeled with nanoprobes which are conjugated with antibodies or oligonucleotides and quantified at single copy sensitivity in the cancer cells and tissues. Furthermore, a non-fluorescent super-resolution concept, named as second harmonic super-resolution microscopy (SHaSM), was proposed to quantify individual Her2 transcripts in cancer cells beyond the diffraction limit. These non-fluorescent imaging modalities will provide new dimensions in biomarker quantification at single molecule sensitivity in turbid biological samples, offering a strong cross-platform strategy for clinical monitoring at single cell resolution.

  8. Synthesis and fluorescence studies of nine 1,5-benzodiazepine-2,4-dione derivatives: Dual emission and excimer fluorescence

    Science.gov (United States)

    Qomi, Hamid Reza; Habibi, Azizollah; Shahcheragh, Seyyed Mohammad

    2017-03-01

    The photophysical properties of nine 1,5-benzodiazepine-2,4-dione (BZD) derivatives were investigated using absorption and fluorescence spectral techniques in dimethyl sulfoxide. The trend of red shifts caused by the substitutions had full compliance with the trend of decreasing the calculated band gap (ΔELUMO-HOMO) by semi-empirical AM1 and DFT/B3LYP/6-311 + G* computational methods. The positive solvatochromism of BZD a demonstrated the π-π* nature of the singlet excited state. Dual fluorescence was observed in the emission spectra of BZD f and g, while their spectrum in different concentration showed only one peak short wavelength (SW) in dilute solutions. The main peak in SW around 370 nm was attributed to the monomer of BZD (f* or g*) and the broader emission shifted to the visible region around 400 nm in middle wavelength (MW) to the intermolecular excimer emission of BZD ([f/f]*or [g/g]*). The observed phenomena, such as solvatochromism, dual fluorescence, some red shifts caused by substitution, and larger Stokes shift indicated the existence of intramolecular charge transfer (ICT) in the BZDs series. The phosphorescence emission of the BZDs demonstrated their intersystem crossing (ISC) process.

  9. A reusable biosensor chip for SERS-fluorescence dual mode immunoassay

    Science.gov (United States)

    Wu, Lei; Wang, Zhuyuan; Fan, Kequan; Zong, Shenfei; Cui, Yiping

    2015-05-01

    Research continues in an effort to develop a versatile platform for clinical diagnosis with easy operation and low cost. In the present study, a biosensor chip has been designed and fabricated for surface enhanced Raman scattering (SERS)- fluorescence dual mode immunoassay. Here, a dual channel microfluidic chip was employed for simultaneous SERS and fluorescence detection. Unlike previously reported microfluidic immunoassays using fluorescence or SERS method independently, the proposed dual mode biosensor combined the advantages of these two optical detection techniques. The fluorescence mode can be used for fast screening of biomolecules while the SERS mode can be employed for accurate and sensitive quantitative analysis. In addition, the chip-based microfluidic platform greatly reduced the reagents cost and complicated operation. The whole detection process from sample preparation to optical detection can be finished in 90 min. Moreover, the reversibly bonded biosensor chip could be reused after cleaning, which further reduced the cost for each assay. All these merits make it a potential powerful tool for practical clinical diagnosis.

  10. Single-screen single-emulsion versus dual-screen dual-emulsion system in mammography; Comparison of imaging properties and detection of microcalcifications

    Energy Technology Data Exchange (ETDEWEB)

    Katsuda, Noboru; Hiai, Yasuhiro (Kumamoto Univ. (Japan). Hospital); Higashida, Yoshiharu (and others)

    1993-05-01

    A dual-screen/dual-emulsion film combination (Kodak MinR Fast screen/TMG film) that allows for a decrease in patient exposure was evaluated with widely used single-screen/single-emulsion systems (Kodak MinR/OM, Kodak MinR/MinR, and Toshiba MM6/MINC film) in contrast to mammography. Clustered microcalcifications randomly superimposed on a breast specimen were detected, and the locations were determined by eight observers. The dual screen-film system provided about a 64% reduction in patient exposure compared with the single-screen/single-emulsion system (MM6/MINC), but it failed to equal the single-system in detection of simulated microcalcifications. Our results suggested that the conventional single-screen/single-emulsion systems showed better detectability of microcalcifications than the dual-screen/dual-emulsion film combinations. (author).

  11. Single-fluorophore membrane transport activity sensors with dual-emission read-out.

    Science.gov (United States)

    Ast, Cindy; De Michele, Roberto; Kumke, Michael U; Frommer, Wolf B

    2015-06-19

    We recently described a series of genetically encoded, single-fluorophore-based sensors, termed AmTrac and MepTrac, which monitor membrane transporter activity in vivo (De Michele et al., 2013). However, being intensiometric, AmTrac and Meptrac are limited in their use for quantitative studies. Here, we characterized the photophysical properties (steady-state and time-resolved fluorescence spectroscopy as well as anisotropy decay analysis) of different AmTrac sensors with diverging fluorescence properties in order to generate improved, ratiometric sensors. By replacing key amino acid residues in AmTrac we constructed a set of dual-emission AmTrac sensors named deAmTracs. deAmTracs show opposing changes of blue and green emission with almost doubled emission ratio upon ammonium addition. The response ratio of the deAmTracs correlated with transport activity in mutants with altered capacity. Our results suggest that partial disruption of distance-dependent excited-state proton transfer is important for the successful generation of single-fluorophore-based dual-emission sensors.

  12. Genetically-encoded yellow fluorescent cAMP indicator with an expanded dynamic range for dual-color imaging.

    Directory of Open Access Journals (Sweden)

    Haruki Odaka

    Full Text Available Cyclic AMP is a ubiquitous second messenger, which mediates many cellular responses mainly initiated by activation of cell surface receptors. Various Förster resonance energy transfer-based ratiometric cAMP indicators have been created for monitoring the spatial and temporal dynamics of cAMP at the single-cell level. However, single fluorescent protein-based cAMP indicators have been poorly developed, with improvement required for dynamic range and brightness. Based on our previous yellow fluorescent protein-based cAMP indicator, Flamindo, we developed an improved yellow fluorescent cAMP indicator named Flamindo2. Flamindo2 has a 2-fold expanded dynamic range and 8-fold increased brightness compared with Flamindo by optimization of linker peptides in the vicinity of the chromophore. We found that fluorescence intensity of Flamindo2 was decreased to 25% in response to cAMP. Live-cell cAMP imaging of the cytosol and nucleus in COS7 cells using Flamindo2 and nlsFlamindo2, respectively, showed that forskolin elevated cAMP levels in each compartment with different kinetics. Furthermore, dual-color imaging of cAMP and Ca2+ with Flamindo2 and a red fluorescent Ca2+ indicator, R-GECO, showed that cAMP and Ca2+ elevation were induced by noradrenaline in single HeLa cells. Our study shows that Flamindo2, which is feasible for multi-color imaging with other intracellular signaling molecules, is useful and is an alternative tool for live-cell imaging of intracellular cAMP dynamics.

  13. Genetically-Encoded Yellow Fluorescent cAMP Indicator with an Expanded Dynamic Range for Dual-Color Imaging

    Science.gov (United States)

    Odaka, Haruki; Arai, Satoshi; Inoue, Takafumi; Kitaguchi, Tetsuya

    2014-01-01

    Cyclic AMP is a ubiquitous second messenger, which mediates many cellular responses mainly initiated by activation of cell surface receptors. Various Förster resonance energy transfer-based ratiometric cAMP indicators have been created for monitoring the spatial and temporal dynamics of cAMP at the single-cell level. However, single fluorescent protein-based cAMP indicators have been poorly developed, with improvement required for dynamic range and brightness. Based on our previous yellow fluorescent protein-based cAMP indicator, Flamindo, we developed an improved yellow fluorescent cAMP indicator named Flamindo2. Flamindo2 has a 2-fold expanded dynamic range and 8-fold increased brightness compared with Flamindo by optimization of linker peptides in the vicinity of the chromophore. We found that fluorescence intensity of Flamindo2 was decreased to 25% in response to cAMP. Live-cell cAMP imaging of the cytosol and nucleus in COS7 cells using Flamindo2 and nlsFlamindo2, respectively, showed that forskolin elevated cAMP levels in each compartment with different kinetics. Furthermore, dual-color imaging of cAMP and Ca2+ with Flamindo2 and a red fluorescent Ca2+ indicator, R-GECO, showed that cAMP and Ca2+ elevation were induced by noradrenaline in single HeLa cells. Our study shows that Flamindo2, which is feasible for multi-color imaging with other intracellular signaling molecules, is useful and is an alternative tool for live-cell imaging of intracellular cAMP dynamics. PMID:24959857

  14. Fluorescence-Raman Dual Modal Endoscopic System for Multiplexed Molecular Diagnostics

    Science.gov (United States)

    Jeong, Sinyoung; Kim, Yong-Il; Kang, Homan; Kim, Gunsung; Cha, Myeong Geun; Chang, Hyejin; Jung, Kyung Oh; Kim, Young-Hwa; Jun, Bong-Hyun; Hwang, Do Won; Lee, Yun-Sang; Youn, Hyewon; Lee, Yoon-Sik; Kang, Keon Wook; Lee, Dong Soo; Jeong, Dae Hong

    2015-03-01

    Optical endoscopic imaging, which was recently equipped with bioluminescence, fluorescence, and Raman scattering, allows minimally invasive real-time detection of pathologies on the surface of hollow organs. To characterize pathologic lesions in a multiplexed way, we developed a dual modal fluorescence-Raman endomicroscopic system (FRES), which used fluorescence and surface-enhanced Raman scattering nanoprobes (F-SERS dots). Real-time, in vivo, and multiple target detection of a specific cancer was successful, based on the fast imaging capability of fluorescence signals and the multiplex capability of simultaneously detected SERS signals using an optical fiber bundle for intraoperative endoscopic system. Human epidermal growth factor receptor 2 (HER2) and epidermal growth factor receptor (EGFR) on the breast cancer xenografts in a mouse orthotopic model were successfully detected in a multiplexed way, illustrating the potential of FRES as a molecular diagnostic instrument that enables real-time tumor characterization of receptors during routine endoscopic procedures.

  15. An Optic/Proton Dual-Controlled Fluorescence Switch based on Novel Photochromic Bithienylethene Derivatives

    Institute of Scientific and Technical Information of China (English)

    张佳琦; 靳家玉; 张隽佶; 邹雷

    2012-01-01

    A simple method for the synthesis of new bithienylethenes bearing a functional group on the cyclopentene moi- ety is developed. Three new photochromic compounds (4a, 4b, 4c) have been successfully synthesized through this simple method, and exhibit good photochromic properties with alternate irradiation of ultraviolet and visible light. Furthermore, the fluorescence of compound 4a, which bears a quinoline unit on the cyclopentene, can be modulated via optic and proton dual inputs. Upon excitation by 320 nm light, 4a emits a strong fluorescence at 404 nm. After irradiation with 254 nm light, the emission intensity is reduced due to the fluorescence resonance energy transfers (FRET) from quinoline unit to bithienylethene unit. Moreover, on addition of H~, the fluorescence is quenched completely due to the protonation of the quinoline. In addition, both the FRET and protonation process are reversi- ble, which indicates a potential application in molecular switches and logic gates.

  16. Fluorescence spectroscopy of single molecules at room temperature and its applications

    Energy Technology Data Exchange (ETDEWEB)

    Ha, Taekjip [Univ. of California, Berkeley, CA (United States)

    1996-12-01

    We performed fluorescence spectroscopy of single and pairs of dye molecules on a surface at room temperature. Near field scanning optical microscope (NSOM) and far field scanning optical microscope with multi-color excitation/detection capability were built. The instrument is capable of optical imaging with 100nm resolution and has the sensitivity necessary for single molecule detection. A variety of dynamic events which cannot be observed from an ensemble of molecules is revealed when the molecules are probed one at a time. They include (1) spectral jumps correlated with dark states, (2) individually resolved quantum jumps to and from the meta-stable triplet state, (3) rotational jumps due to desorption/readsorption events of single molecules on the surface. For these studies, a computer controlled optical system which automatically and rapidly locates and performs spectroscopic measurements on single molecules was developed. We also studied the interaction between closely spaced pairs of molecules. In particular, fluorescence resonance energy transfer between a single resonant pair of donor and acceptor molecules was measured. Photodestruction dynamics of the donor or acceptor were used to determine the presence and efficiency of energy transfer Dual molecule spectroscopy was extended to a non-resonant pair of molecules to obtain high resolution differential distance information. By combining NSOM and dual color scheme, we studied the co-localization of parasite proteins and host proteins on a human red blood cell membrane infected with malaria. These dual-molecule techniques can be used to measure distances, relative orientations, and changes in distances/orientations of biological macromolecules with very good spatial, angular and temporal resolutions, hence opening new capabilities in the study of such systems.

  17. Modulation of the excited state intramolecular electron transfer reaction and dual fluorescence of crystal violet lactone in room temperature ionic liquids.

    Science.gov (United States)

    Santhosh, Kotni; Samanta, Anunay

    2010-07-22

    The influence of polarity, viscosity, and hydrogen bond donating ability of the medium on the fluorescence behavior of crystal violet lactone (CVL), which undergoes excited state electron transfer reaction and exhibits dual fluorescence from two different electronic states, termed as CT(A) and CT(B), has been studied in six different room temperature ionic liquids (ILs) using steady state and time-resolved emission techniques. It is shown that the excited state CT(A) --> CT(B) transformation and dual fluorescence of CVL can be controlled by appropriate choice of the ILs. While dual fluorescence of CVL is clearly observed in pyrrolidinium IL, the molecule exhibits a single fluorescence band in ammonium IL. While the second emission from the CT(B) state can barely be seen in 1,3-dialkylimidazolium ILs, dual fluorescence is quite prominent in 1-butyl-2,3-dimethylimidazolium IL, [bmMim][Tf(2)N]. These contrasting results have been explained taking into account the hydrogen bonding interactions of the 1,3-dialkylimidazolium ions (mediated through the C(2)-hydrogen) with CVL and the viscosity of the ILs. The excited state CT(A) --> CT(B) reaction kinetics has been studied in IL by monitoring the time-evolution of the CT(B) emission in [bmMim][Tf(2)N]. The solvation dynamics in this IL has been studied by following the dynamic fluorescence Stokes shift of C153, which is used as a probe molecule. A comparison of the excited state reaction time and solvation time suggests that the rate of the CT(A) --> CT(B) reaction in moderately viscous ILs is primarily dictated by the rate of solvation. Very little or negligible excitation wavelength dependence of the emission behavior of CVL can be observed in these ILs.

  18. Dual-Color Fluorescence Imaging of Magnetic Nanoparticles in Live Cancer Cells Using Conjugated Polymer Probes.

    Science.gov (United States)

    Sun, Minjie; Sun, Bin; Liu, Yun; Shen, Qun-Dong; Jiang, Shaojun

    2016-03-02

    Rapid growth in biological applications of nanomaterials brings about pressing needs for exploring nanomaterial-cell interactions. Cationic blue-emissive and anionic green-emissive conjugated polymers are applied as dual-color fluorescence probes to the surface of negatively charged magnetic nanoparticles through sequentially electrostatic adsorption. These conjugated polymers have large extinction coefficients and high fluorescence quantum yield (82% for PFN and 62% for ThPFS). Thereby, one can visualize trace amount (2.7 μg/mL) of fluorescence-labeled nanoparticles within cancer cells by confocal laser scanning microscopy. Fluorescence labeling by the conjugated polymers is also validated for quantitative determination of the internalized nanoparticles in each individual cell by flow cytometry analysis. Extensive overlap of blue and green fluorescence signals in the cytoplasm indicates that both conjugated polymer probes tightly bind to the surface of the nanoparticles during cellular internalization. The highly charged and fluorescence-labeled nanoparticles non-specifically bind to the cell membranes, followed by cellular uptake through endocytosis. The nanoparticles form aggregates inside endosomes, which yields a punctuated staining pattern. Cellular internalization of the nanoparticles is dependent on the dosage and time. Uptake efficiency can be enhanced three-fold by application of an external magnetic field. The nanoparticles are low cytotoxicity and suitable for simultaneously noninvasive fluorescence and magnetic resonance imaging application.

  19. Fluorescence microscopy of single autofluorescent proteins for cellular biology

    CERN Document Server

    Cognet, Laurent; Choquet, Daniel; Lounis, Brahim

    2002-01-01

    In this paper we review the applicability of autofluorescent proteins for single-molecule imaging in biology. The photophysical characteristics of several mutants of the Green Fluorescent Protein (GFP) and those of DsRed are compared and critically discussed for their use in cellular biology. The alternative use of two-photon excitation at the single-molecule level or Fluorescence Correlation Spectroscopy is envisaged for the study of individual autofluorescent proteins. Single-molecule experiments performed in live cells using eGFP and preferably eYFP fusion proteins are reviewed. Finally, the first use at the single-molecule level of citrine, a more photostable variant of the eYFP is reported when fused to a receptor for neurotransmitter in live cells.

  20. Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

    NARCIS (Netherlands)

    Tanner, Nathan A.; Loparo, Joseph J.; Oijen, Antoine M. van

    2009-01-01

    We describe a simple fluorescence microscopy-based real-time method for observing DNA replication at the single-molecule level. A circular, forked DNA template is attached to a functionalized glass coverslip and replicated extensively after introduction of replication proteins and nucleotides. The

  1. Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

    NARCIS (Netherlands)

    Tanner, Nathan A.; Loparo, Joseph J.; Oijen, Antoine M. van

    2009-01-01

    We describe a simple fluorescence microscopy-based real-time method for observing DNA replication at the single-molecule level. A circular, forked DNA template is attached to a functionalized glass coverslip and replicated extensively after introduction of replication proteins and nucleotides. The g

  2. Single-Band and Dual-Band Infrared Detectors

    Science.gov (United States)

    Ting, David Z. (Inventor); Gunapala, Sarath D. (Inventor); Soibel, Alexander (Inventor); Nguyen, Jean (Inventor); Khoshakhlagh, Arezou (Inventor)

    2017-01-01

    Bias-switchable dual-band infrared detectors and methods of manufacturing such detectors are provided. The infrared detectors are based on a back-to-back heterojunction diode design, where the detector structure consists of, sequentially, a top contact layer, a unipolar hole barrier layer, an absorber layer, a unipolar electron barrier, a second absorber, a second unipolar hole barrier, and a bottom contact layer. In addition, by substantially reducing the width of one of the absorber layers, a single-band infrared detector can also be formed.

  3. Strong Fluorescent Smart Organogel as a Dual Sensing Material for Volatile Acid and Organic Amine Vapors.

    Science.gov (United States)

    Xue, Pengchong; Yao, Boqi; Wang, Panpan; Gong, Peng; Zhang, Zhenqi; Lu, Ran

    2015-11-23

    An L-phenylalanine derivative (C12PhBPCP) consisting of a strong emission fluorophore with benzoxazole and cyano groups is designed and synthesized to realize dual responses to volatile acid and organic amine vapors. The photophysical properties and self-assembly of the said derivative in the gel phase are also studied. C12PhBPCP can gelate organic solvents and self-assemble into 1 D nanofibers in the gels. UV/Vis absorption spectral results show H-aggregate formation during gelation, which indicates strong exciton coupling between fluorophores. Both wet gel and xerogel emit strong green fluorescence because the cyano group suppresses fluorescence quenching in the self-assemblies. Moreover, the xerogel film with strong green fluorescence can be used as a dual chemosensor for quantitative detection of volatile acid and organic amine vapors with fast response times and low detection limits owing to its large surface area and amplified fluorescence quenching. The detection limits are 796 ppt and 25 ppb for gaseous aniline and trifluoroacetic acid (TFA), respectively.

  4. High-Crystallinity Covalent Organic Framework with Dual Fluorescence Emissions and Its Ratiometric Sensing Application.

    Science.gov (United States)

    Qian, Hai-Long; Dai, Cong; Yang, Cheng-Xiong; Yan, Xiu-Ping

    2017-07-26

    High crystallinity of covalent organic frameworks (COFs) with dual fluorescence emissions has not been reported so far. Here, we show the rational design and preparation of high-crystallinity COF TzDa via the synergetic interaction of docking sites and hydrogen bonds: 4,4',4″-(1,3,5-Triazine-2,4,6-triyl)trianiline (Tz) with the docking site and 2,5-dihydroxyterephthalaldehyde (Da) with the OH group are employed to synthesize the imine-linked two-dimensional high-crystallinity layered structure TzDa. The prepared mesoporous TzDa (ca. 36 Å) exhibits high thermal and chemical stability. The intramolecular charge transfer (ICT) and excited-state intramolecular proton transfer (ESIPT) effects bring TzDa two main fluorescence emissions around 500 and 590 nm. Water molecules can interfere with the ICT and ESIPT effects, allowing the development of a ratiometric fluorescent sensor for water in organic solvents. The proposed sensor shows high sensitivity to trace water in conventional organic solvents. The high stability of TzDa allows its recyclable uses for trace water detection. This work not only offers a platform for the construction of high-crystallinity COFs, but also provides a rational design of COFs with dual fluorescence emissions for ratiometric sensing applications.

  5. In vivo magnetic resonance and fluorescence dual imaging of tumor sites by using dye-doped silica-coated iron oxide nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Jang, Haeyun; Lee, Chaedong [Seoul National University, Program in Nano Science and Technology, Graduate School of Convergence Science and Technology (Korea, Republic of); Nam, Gi-Eun [University of Ulsan College of Medicine, Department of Radiology, Asan Medical Center (Korea, Republic of); Quan, Bo [Seoul National University, Program in Nano Science and Technology, Graduate School of Convergence Science and Technology (Korea, Republic of); Choi, Hyuck Jae [University of Ulsan College of Medicine, Department of Radiology, Asan Medical Center (Korea, Republic of); Yoo, Jung Sun [Seoul National University, Department of Transdisciplinary Studies, Graduate School of Convergence Science and Technology, Smart Humanity Convergence Center (Korea, Republic of); Piao, Yuanzhe, E-mail: parkat9@snu.ac.kr [Seoul National University, Program in Nano Science and Technology, Graduate School of Convergence Science and Technology (Korea, Republic of)

    2016-02-15

    The difficulty in delineating tumor is a major obstacle for better outcomes in cancer treatment of patients. The use of single-imaging modality is often limited by inadequate sensitivity and resolution. Here, we present the synthesis and the use of monodisperse iron oxide nanoparticles coated with fluorescent silica nano-shells for fluorescence and magnetic resonance dual imaging of tumor. The as-synthesized core–shell nanoparticles were designed to improve the accuracy of diagnosis via simultaneous tumor imaging with dual imaging modalities by a single injection of contrast agent. The iron oxide nanocrystals (∼11 nm) were coated with Rhodamine B isothiocyanate-doped silica shells via reverse microemulsion method. Then, the core–shell nanoparticles (∼54 nm) were analyzed to confirm their size distribution by transmission electron microscopy and dynamic laser scattering. Photoluminescence spectroscopy was used to characterize the fluorescent property of the dye-doped silica shell-coated nanoparticles. The cellular compatibility of the as-prepared nanoparticles was confirmed by a trypan blue dye exclusion assay and the potential as a dual-imaging contrast agent was verified by in vivo fluorescence and magnetic resonance imaging. The experimental results show that the uniform-sized core–shell nanoparticles are highly water dispersible and the cellular toxicity of the nanoparticles is negligible. In vivo fluorescence imaging demonstrates the capability of the developed nanoparticles to selectively target tumors by the enhanced permeability and retention effects and ex vivo tissue analysis was corroborated this. Through in vitro phantom test, the core/shell nanoparticles showed a T2 relaxation time comparable to Feridex{sup ®} with smaller size, indicating that the as-made nanoparticles are suitable for imaging tumor. This new dual-modality-nanoparticle approach has promised for enabling more accurate tumor imaging.

  6. In vivo magnetic resonance and fluorescence dual imaging of tumor sites by using dye-doped silica-coated iron oxide nanoparticles

    Science.gov (United States)

    Jang, Haeyun; Lee, Chaedong; Nam, Gi-Eun; Quan, Bo; Choi, Hyuck Jae; Yoo, Jung Sun; Piao, Yuanzhe

    2016-02-01

    The difficulty in delineating tumor is a major obstacle for better outcomes in cancer treatment of patients. The use of single-imaging modality is often limited by inadequate sensitivity and resolution. Here, we present the synthesis and the use of monodisperse iron oxide nanoparticles coated with fluorescent silica nano-shells for fluorescence and magnetic resonance dual imaging of tumor. The as-synthesized core-shell nanoparticles were designed to improve the accuracy of diagnosis via simultaneous tumor imaging with dual imaging modalities by a single injection of contrast agent. The iron oxide nanocrystals ( 11 nm) were coated with Rhodamine B isothiocyanate-doped silica shells via reverse microemulsion method. Then, the core-shell nanoparticles ( 54 nm) were analyzed to confirm their size distribution by transmission electron microscopy and dynamic laser scattering. Photoluminescence spectroscopy was used to characterize the fluorescent property of the dye-doped silica shell-coated nanoparticles. The cellular compatibility of the as-prepared nanoparticles was confirmed by a trypan blue dye exclusion assay and the potential as a dual-imaging contrast agent was verified by in vivo fluorescence and magnetic resonance imaging. The experimental results show that the uniform-sized core-shell nanoparticles are highly water dispersible and the cellular toxicity of the nanoparticles is negligible. In vivo fluorescence imaging demonstrates the capability of the developed nanoparticles to selectively target tumors by the enhanced permeability and retention effects and ex vivo tissue analysis was corroborated this. Through in vitro phantom test, the core/shell nanoparticles showed a T2 relaxation time comparable to Feridex® with smaller size, indicating that the as-made nanoparticles are suitable for imaging tumor. This new dual-modality-nanoparticle approach has promised for enabling more accurate tumor imaging.

  7. Detection of Embryo Sex Chromosome by Dual Color Fluorescent In-Situ Hybridization

    Institute of Scientific and Technical Information of China (English)

    刘群; 朱桂金

    2003-01-01

    Summary: In order to evaluate the effects of sex chromosomal mosaicism on the accuracy of single-cell gender diagnosis, sex chromosomes of 21 normal fertilized embryos were detected by dual colorfluorescent in-situ hybridization (FISH). The results showed that 4 embryos had sex chromosomalmosaicism (19%) and the remaining 17 showed uniformly XX or XY signals in all blastomeres. Inconclusion, identification of sex by dual color FISH analysis of a single cell was accurate and efficient,and sex chromosomal mosaicism would not affect preimplantation gender diagnosis.

  8. High sensitivity fluorescent single particle and single molecule detection apparatus and method

    Energy Technology Data Exchange (ETDEWEB)

    Mathies, Richard A. (Contra Costa County, CA); Peck, Konan (Contra Costa County, CA); Stryer, Lubert (Santa Clara County, CA)

    1990-01-01

    Apparatus is described for ultrasensitive detection of single fluorescent particles down to the single fluorescent molecule limit in a fluid or on a substrate comprising means for illuminating a predetermined volume of the fluid or area of the substrate whereby to emit light including background light from the fluid and burst of photons from particles residing in the area. The photon burst is detected in real time to generate output representative signal. The signal is received and the burst of energy from the fluorescent particles is distinguished from the background energy to provide an indication of the number, location or concentration of the particles or molecules.

  9. Cyclic up-regulation fluorescence of pyrene excimer for studying polynucleotide kinase activity based on dual amplification.

    Science.gov (United States)

    Xu, Jing; Gao, Yanfang; Li, Baoxin; Jin, Yan

    2016-06-15

    Due to its important biological and clinical roles of polynucleotide kinase (PNK), accurate monitoring of PNK activity and inhibition is highly desirable. Herein, a homogeneous and sensitive fluorescence assay has been proposed for the detection of PNK activity by integrating target recycling signal amplification of DNA toehold strand displacement reaction (TSDR) with gamma-cyclodextrin (γ-CD) enhancement of pyrene excimer. A label-free hairpin DNA1 (H1) and two singly pyrene-labelled DNA, H2 and H3, are designed. Accompanying the occurrence of the efficient enzyme reactions, namely phosphorylation-actuated λ exonuclease reaction, a single-stranded DNA as a trigger DNA (tDNA) of TSDR can be released from H1. Then, tDNA drives circulatory interactions between H2 and H3 to continuously form H2/H3 duplex, resulting in formation of pyrene excimer and a "turn on" fluorescence signal of pyrene excimer. Furthermore, the fluorescence of pyrene excimer is further amplified by introducing gamma-cyclodextrin (γ-CD), which can regulate the space proximity of two pyrene molecules. Thus, TSDR-induced cyclic formation of pyrene excimer and γ-CD enhancement can specifically up-regulate the fluorescence of pyrene excimer for detection of PNK activity, the detection limit is 9.3 × 10(-5)UmL(-1), which is superior to those of most existing approaches. Moreover, the proposed strategy can also be successfully utilized to study inhibition efficiency of different PNK inhibitors as well. Therefore, a dual amplification approach is provided for nucleic acid phosphorylation related researches.

  10. A fluorescence ratiometric sensor for hypochlorite based on a novel dual-fluorophore response approach.

    Science.gov (United States)

    Long, Lingliang; Zhang, Dongdong; Li, Xiufen; Zhang, Jinfang; Zhang, Chi; Zhou, Liping

    2013-05-02

    A fluorescence ratiometric sensor for OCl(-) has been developed based on a novel dual fluorophore response approach. The sensor molecule contains a coumarin fluorophore and a rhodamine fluorophore, and the two fluorophores are directly linked to an OCl(-) recognition group. The structure of the sensor was characterized by ESI-MS, NMR, and X-ray crystallographic analysis. Upon treatment with OCl(-), both fluorophores in the sensor responded simultaneously at two separate optical windows, with large enhancement of the fluorescence ratio (I578/I501) from 0.01 to 39.55. The fluorescence ratios for the sensor showed a good linearity with the concentration of OCl(-) in the range of 0.2-40 μM and the detection limits is 0.024 μM (SN(-1)=3). Investigation of reaction products indicated that the sensor reaction with OCl(-) produced two new fluorescent molecules, which were responsible for the fluorescence changes in two optical windows. In addition, the sensor showed high selectivity to OCl(-) over other reactive oxygen species, reactive nitrogen species, cations, and anions. The sensor has also been successfully applied to detection of OCl(-) in natural water samples with satisfactory recovery.

  11. Single electron counting using a dual MCP assembly

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Yuzhen [School of Physics, Nanjing University, Nanjing 210093 (China); Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); State Key Laboratory of Particle Detection and Electronics, Beijing 100049 (China); Liu, Shulin, E-mail: liusl@ihep.ac.cn [Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); State Key Laboratory of Particle Detection and Electronics, Beijing 100049 (China); Zhao, Tianchi, E-mail: tianchi@u.washington.edu [Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); State Key Laboratory of Particle Detection and Electronics, Beijing 100049 (China); Department of Physics, University of Washington, Seattle, WA 98195 (United States); Yan, Baojun; Wang, Peiliang [Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); State Key Laboratory of Particle Detection and Electronics, Beijing 100049 (China); Yu, Yang [Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); State Key Laboratory of Particle Detection and Electronics, Beijing 100049 (China); School of Science, Xi’an University of Technology, Xi’an 710048 (China); Lei, Xiangcui [Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); State Key Laboratory of Particle Detection and Electronics, Beijing 100049 (China); Yang, Luping [Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); State Key Laboratory of Particle Detection and Electronics, Beijing 100049 (China); School of Physics and Electronics, Henan University, Kaifeng 475004 (China); Wen, Kaile [Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); State Key Laboratory of Particle Detection and Electronics, Beijing 100049 (China); Qi, Ming [School of Physics, Nanjing University, Nanjing 210093 (China); and others

    2016-09-11

    The gain, pulse height resolution and peak-to-valley ratio of single electrons detected by using a Chevron configured Microchannel Plate (MCP) assembly are studied. The two MCPs are separated by a 280 µm gap and are biased by four electrodes. The purpose of the study is to determine the optimum bias voltage arrangements for single electron counting. By comparing the results of various bias voltage combinations, we conclude that good performance for the electron counting can be achieved by operating the MCP assembly in saturation mode. In addition, by applying a small reverse bias voltage across the gap while adjusting the bias voltages of the MCPs, optimum performance of electron counting can be obtained. - Highlights: • Dual MCPs assembly with four electrodes using different voltage combinations has been investigated for single electron counting. • Both the MCP voltages and the gap voltage can affect the gain, pulse height resolution and P/V ratio. • A high gain of the first stage MCP, a saturation mode of the second stage MCP and an appropriately reverse gap voltage can improve the resolution greatly. • The optimum voltage arrangements is significant for the design of MCP detectors in single electron counting applications.

  12. Dual wavelength fluorescent ratiometric pH measurement by scanning near-field optical microscopy

    Science.gov (United States)

    Li, Yongbo; Shinohara, Ryosuke; Iwami, Kentaro; Ohta, Yoshihiro; Umeda, Norihiro

    2010-08-01

    A novel method to observe pH distribution by dual wavelength fluorescent ratiometric pH measurement by scanning near-field optical microscopy (SNOM) is developed. In this method, in order to investigate not only the pH of mitochondrial membrane but also its distribution in the vicinity, a pH sensitive fluorescent reagent covers mitochondria instead of injecting it to mitochondria. This method utilizes a dual-emission pH sensitive dye and SNOM with a themally-pulled and metal-coated optical fiber to improve the spatial resolution. Time-dependence of Fluorescent intensity ratio (FIR) under acid addition is investigated. As the distances between the dropped point and the SNOM probe becomes closer, the time when FIR changes becomes earlier. The response of mitochondria under supplement of nutrition is studied by using this method. While the probe is near to mitochondria, the ratio quickly becomes to increase. In conclusion, it was confirmed that the temporal variation of pH can be detected by this method, and pH distribution in the vicinity of mitochondria is able to be measured by this method.

  13. Resolution enhancement of digital laser scanning fluorescence microscopy with a dual-lens optical pickup head

    Science.gov (United States)

    Tsai, Rung-Ywan; Chen, Jung-Po; Lee, Yuan-Chin; Chiang, Hung-Chih; Huang, Tai-Ting; Huang, Chun-Chieh; Cheng, Chih-Ming; Cheng, Chung-Ta; Lo, Feng-Hsiang; Tiao, Golden

    2016-10-01

    The resolution of the cell fluorescence image captured by a digital laser scanning microscopy with a modified dual-lens BD-ROM optical pickup head is enhanced by image registration and double sample frequency. A dual objective lens of red (655 nm) and blue (405 or 488 nm) laser sources with numerical apertures of 0.6 and 0.85 is used for sample focusing and position tracking and cell fluorescence image capturing, respectively. The image registration and capturing frequency are based on the address-coded patterns of a sample slide. The address-coded patterns are designed as a string of binary code, which comprises a plurality of base-straight lands and grooves and data-straight grooves. The widths of the base-straight lands, base-straight grooves, and data-straight grooves are 0.38, 0.38, and 0.76 μm, respectively. The numbers of sample signals in the x-direction are measured at every intersection point by intersecting the base intensity of the push-pull signal of the address-coded patterns, which has a minimum spacing of 0.38 μm. After taking a double sample frequency, the resolution of the measured cell fluorescence image is enhanced from 0.38 μm to the diffraction limit of the objective lens.

  14. Dual modality of non-contact photoacoustic tomography and fluorescence imaging using double cladding fiber

    Science.gov (United States)

    Eom, Jonghyun; Park, Seong Jun; Kim, Ju Wan; Park, Soongho; Lee, Byeong Ha

    2015-03-01

    We present a fiber-based dual-modal imaging system that combines non-contact photoacoustic tomography (NCPAT) and fluorescence imaging by using double cladding fiber (DCF). The NCPAT system utilizing an all-fiber heterodyne interferometer as an ultrasound detector measures the photoacoustic signal at the sample surface without physical contact. Fluorescence imaging system is composed of fiber-optics to deliver the excitation light and the emission light. For combined system the probe consists of a specially fabricated DCF coupler and a lensed fiber so that we can simultaneously acquire the signals of two systems with the same probe. The DCF has a core and two claddings, inner and outer, which allows two concentric light-guiding channels via the core and the inner cladding. The lensed fiber of the DCF probe is compactly fabricated to focus the interferometer light and the excitation light, and to efficiently collect the fluorescence signal. To demonstrate the feasibility of the proposed dual-modal system, we have conducted phantom experiments using tissue mimicking phantoms which contained a couple of tubes filled with fluorescein solution and black ink, respectively. The proposed imaging system is implanted with fiber-optic configurations so that it has the potential for minimally invasive and improved diagnosis and guided treatment of diseases.

  15. Measuring diffusion with polarization-modulation dual-focus fluorescence correlation spectroscopy.

    Science.gov (United States)

    Korlann, You; Dertinger, Thomas; Michalet, Xavier; Weiss, Shimon; Enderlein, Jörg

    2008-09-15

    We present a new technique, polarization-modulation dual-focus fluorescence correlation spectroscopy (pmFCS), based on the recently intro-duced dual-focus fluorescence correlation spectroscopy (2fFCS) to measure the absolute value of diffusion coefficients of fluorescent molecules at pico- to nanomolar concentrations. Analogous to 2fFCS, the new technique is robust against optical saturation in yielding correct values of the diffusion coefficient. This is in stark contrast to conventional FCS where optical saturation leads to an apparent decrease in the determined diffusion coefficient with increasing excitation power. However, compared to 2fFCS, the new technique is simpler to implement into a conventional confocal microscope setup and is compatible with cw-excitation, only needing as add-ons an electro-optical modulator and a differential interference contrast prism. With pmFCS, the measured diffusion coefficient (D) for Atto655 maleimide in water at 25?C is determined to be equal to (4.09 +/- 0.07) x 10(-6)cm(2)/s, in good agreement with the value of 4.04 x 10-6cm2/s as measured by 2fFCS.

  16. Studying DNA translocation in nanocapillaries using single molecule fluorescence

    CERN Document Server

    Thacker, Vivek V; Hernández-Ainsa, Silvia; Bell, Nicholas A W; Keyser, Ulrich F; 10.1063/1.4768929

    2013-01-01

    We demonstrate simultaneous measurements of DNA translocation into glass nanopores using ionic current detection and fluorescent imaging. We verify the correspondence between the passage of a single DNA molecule through the nanopore and the accompanying characteristic ionic current blockage. By tracking the motion of individual DNA molecules in the nanocapillary perpendicular to the optical axis and using a model, we can extract an effective mobility constant for DNA in our geometry under high electric fields.

  17. Inorganic single crystalline fibers for dual-readout calorimetry

    CERN Document Server

    Pauwels, Kristof; Lecoq, Paul

    This thesis focuses on the improvement of the energy resolution of hadron calorimeters. The approach is based on dual-readout, which consists in the simultaneous detection of both scintillation and Cherenkov light. The comparison of these two signals allows a compensation of the energy fluctuations, which are inherent to the detection of hadronic showers. Lutetium aluminium garnets (LuAG), which are efficient scintillators when activated with rare-earth dopants (i.e. Cerium), can also act as Cherenkov radiators when undoped. Both undoped and doped crystals can then be assembled to build an efficient dual-readout calorimeter. With the objective to investigate the feasibility of this concept, the effects of the doping concentration and the use of various co-dopant on the light output and the timing properties of LuAG were studied. The growth method was demonstrated to induce significant differences in the nature and concentration of structural defects. The optimum geometry, which is based on single-crystals sha...

  18. Single/Dual-Polarized Infrared Rectenna for Solar Energy Harvesting

    Directory of Open Access Journals (Sweden)

    S. H. Zainud-Deen

    2016-05-01

    Full Text Available Single and dual linearly-polarized receiving mode nanoantennas are designed for solar energy harvesting at 28.3 THz. The infrared rectennas are used to harvest the solar energy and converting it to electrical energy.  The proposed infrared rectenna is a thin dipole made of gold and printed on a silicon dioxide substrate. Different shapes of the dipole arms have been investigated for maximum collected energy. The two poles of the dipole have been determined in a rectangular, circular and rhombus shapes. The rectenna dipole is used to concentrate the electromagnetic energy into a small localized area at the inner tips of the gap between the dipole arms. The dimensions of the different dipole shapes are optimized for maximum near electric field intensity at a frequency of 28.3 THz. A Metal Insulator Metal (MIM diode is incorporated with the nanoantenna dipole to rectify the received energy. The receiving efficiency of the solar energy collector with integrated MIM diode has been investigated. A dual-polarized, four arms, rhombus shaped nanoantenna dipole for solar energy harvesting has been designed and optimized for 28.3 THz applications.

  19. Single Self-Dual Chiral Boson: Augmented Superfield Approach

    CERN Document Server

    Shukla, D; Malik, R P

    2013-01-01

    We exploit the standard tools and techniques of the augmented version of Bonora-Tonin (BT) superfield formalism to derive the off-shell nilpotent and absolutely anticommuting (anti-)BRST and (anti-)co-BRST symmetry transformations for the Becchi-Rouet-Stora-Tyutin (BRST) invariant Lagrangian density of a single self-dual bosonic system. In the derivation of the full set of the above symmetry transformations, we invoke the (dual-)horizontality conditions, (anti-)BRST and (anti-)co-BRST invariant restrictions on the superfields that are defined on the (2, 2)-dimensional supermanifold. The latter is parameterized by the bosonic variable x^\\mu\\,(\\mu = 0,\\, 1) and a pair of Grassmanian variables \\theta and \\bar\\theta (with \\theta^2 = \\bar\\theta^2 = 0 and \\theta\\bar\\theta + \\bar\\theta\\theta = 0). The dynamics of this system is such that, instead of the full (2, 2) dimensional superspace coordinates (x^\\mu, \\theta, \\bar\\theta), we require {\\it only} the specific (1, 2)-dimensional super-subspace variables (t, \\theta...

  20. Single Walled Carbon Nanotubes Exhibit Dual-Phase Regulation to Exposed Arabidopsis Mesophyll Cells

    Science.gov (United States)

    Yuan, Hengguang; Hu, Shanglian; Huang, Peng; Song, Hua; Wang, Kan; Ruan, Jing; He, Rong; Cui, Daxiang

    2011-12-01

    Herein we are the first to report that single-walled carbon nanotubes (SWCNTs) exhibit dual-phase regulation to Arabidopsis mesophyll cells exposed to different concentration of SWCNTs. The mesophyll protoplasts were prepared by enzyme digestion, and incubated with 15, 25, 50, 100 μg/ml SWCNTs for 48 h, and then were observed by optical microscopy and transmission electron microscopy, the reactive oxygen species (ROS) generation was measured. Partial protoplasts were stained with propidium iodide and 4'-6- diamidino-2-phenylindole, partial protoplasts were incubated with fluorescein isothiocyanate-labeled SWCNTs, and observed by fluorescence microscopy. Results showed that SWCNTs could traverse both the plant cell wall and cell membrane, with less than or equal to 50 μg/ml in the culture medium, SWCNTs stimulated plant cells to grow out trichome clusters on their surface, with more than 50 μg/ml SWCNTs in the culture medium, SWCNTs exhibited obvious toxic effects to the protoplasts such as increasing generation of ROS, inducing changes of protoplast morphology, changing green leaves into yellow, and inducing protoplast cells' necrosis and apoptosis. In conclusion, single walled carbon nanotubes can get through Arabidopsis mesophyll cell wall and membrane, and exhibit dose-dependent dual-phase regulation to Arabidopsis mesophyll protoplasts such as low dose stimulating cell growth, and high dose inducing cells' ROS generation, necrosis or apoptosis.

  1. Single Walled Carbon Nanotubes Exhibit Dual-Phase Regulation to Exposed Arabidopsis Mesophyll Cells

    Directory of Open Access Journals (Sweden)

    Huang Peng

    2011-01-01

    Full Text Available Abstract Herein we are the first to report that single-walled carbon nanotubes (SWCNTs exhibit dual-phase regulation to Arabidopsis mesophyll cells exposed to different concentration of SWCNTs. The mesophyll protoplasts were prepared by enzyme digestion, and incubated with 15, 25, 50, 100 μg/ml SWCNTs for 48 h, and then were observed by optical microscopy and transmission electron microscopy, the reactive oxygen species (ROS generation was measured. Partial protoplasts were stained with propidium iodide and 4'-6- diamidino-2-phenylindole, partial protoplasts were incubated with fluorescein isothiocyanate-labeled SWCNTs, and observed by fluorescence microscopy. Results showed that SWCNTs could traverse both the plant cell wall and cell membrane, with less than or equal to 50 μg/ml in the culture medium, SWCNTs stimulated plant cells to grow out trichome clusters on their surface, with more than 50 μg/ml SWCNTs in the culture medium, SWCNTs exhibited obvious toxic effects to the protoplasts such as increasing generation of ROS, inducing changes of protoplast morphology, changing green leaves into yellow, and inducing protoplast cells' necrosis and apoptosis. In conclusion, single walled carbon nanotubes can get through Arabidopsis mesophyll cell wall and membrane, and exhibit dose-dependent dual-phase regulation to Arabidopsis mesophyll protoplasts such as low dose stimulating cell growth, and high dose inducing cells' ROS generation, necrosis or apoptosis.

  2. Nanoscale fluorescence lifetime imaging with a single diamond NV center

    CERN Document Server

    Beams, Ryan; Johnson, Timothy W; Oh, Sang-Hyun; Novotny, Lukas; Vamivakas, Nick

    2013-01-01

    Solid-state quantum emitters, such as artificially engineered quantum dots or naturally occurring defects in solids, are being investigated for applications ranging from quantum information science and optoelectronics to biomedical imaging. Recently, these same systems have also been studied from the perspective of nanoscale metrology. In this letter we study the near-field optical properties of a diamond nanocrystal hosting a single nitrogen vacancy center. We find that the nitrogen vacancy center is a sensitive probe of the surrounding electromagnetic mode structure. We exploit this sensitivity to demonstrate nanoscale fluorescence lifetime imaging microscopy (FLIM) with a single nitrogen vacancy center by imaging the local density of states of an optical antenna.

  3. Single-, Dual- and Triple-band Frequency Reconfigurable Antenna

    Directory of Open Access Journals (Sweden)

    I. H. Idris

    2014-09-01

    Full Text Available The paper presents a frequency reconfigurable slot dipole antenna. The antenna is capable of being switched between single-band, dual-band or triple-band operation. The antenna incorporates three pairs of pin-diodes which are located within the dipole arms. The antenna was designed to operate at 2.4 GHz, 3.5 GHz and 5.2 GHz using the aid of CST Microwave Studio. The average measured gains are 1.54, 2.92 and 1.89 dBi for low, mid and high band respectively. A prototype was then constructed in order to verify the performance of the device. A good level of agreement was observed between simulation and measurement.

  4. Plasmon-enhanced emission from single fluorescent proteins

    Science.gov (United States)

    Donehue, Jessica E.; Haas, Beth L.; Wertz, Esther; Talicska, Courtney N.; Biteen, Julie S.

    2013-02-01

    In this work, we use evaporated gold nanoparticle films (GNPFs) as substrates for plasmon-enhanced imaging of two fluorescent proteins (FPs): mCherry and YFP. Through single-molecule epifluorescence microscopy, we show enhancement of single FP emission in the presence of GNPFs. The gold-coupled FPs demonstrate emission up to four times brighter and seven times longer lived, yielding order-of-magnitude enhancements in total photons detected. Ultimately, this results in increased localization accuracies for single-molecule imaging. Furthermore, we introduce preliminary results for enhancement of mCherry-labeled TcpP membrane proteins inside live Vibrio cholerae cells coupled to GNPFs. Our work indicates that plasmonic substrates are uniquely advantageous for super-resolution imaging and that plasmon-enhanced imaging is a promising technique for improving live cell single-molecule microscopy.

  5. In Vivo Photoacoustic and Fluorescence Cystography Using Clinically Relevant Dual Modal Indocyanine Green

    Directory of Open Access Journals (Sweden)

    Sungjo Park

    2014-10-01

    Full Text Available Conventional X-ray-based cystography uses radio-opaque materials, but this method uses harmful ionizing radiation and is not sensitive. In this study, we demonstrate nonionizing and noninvasive photoacoustic (PA and fluorescence (FL cystography using clinically relevant indocyanine green (ICG in vivo. After transurethral injection of ICG into rats through a catheter, their bladders were photoacoustically and fluorescently visualized. A deeply positioned bladder below the skin surface (i.e., ~1.5–5 mm was clearly visible in the PA and FL image using a laser pulse energy of less than 2 mJ/cm2 (1/15 of the safety limit. Then, the in vivo imaging results were validated through in situ studies. Our results suggest that dual modal cystography can provide a nonionizing and noninvasive imaging tool for bladder mapping.

  6. Probing and controlling fluorescence blinking of single semiconductor nanoparticles

    Directory of Open Access Journals (Sweden)

    Hsien-Chen Ko

    2011-02-01

    Full Text Available In this review we present an overview of the experimental and theoretical development on fluorescence intermittency (blinking and the roles of electron transfer in semiconductor crystalline nanoparticles. Blinking is a very interesting phenomenon commonly observed in single molecule/particle experiments. Under continuous laser illumination, the fluorescence time trace of these single nanoparticles exhibit random light and dark periods. Since its first observation in the mid-1990s, this intriguing phenomenon has attracted wide attention among researchers from many disciplines. We will first present the historical background of the discovery and the observation of unusual inverse power-law dependence for the waiting time distributions of light and dark periods. Then, we will describe our theoretical modeling efforts to elucidate the causes for the power-law behavior, to probe the roles of electron transfer in blinking, and eventually to control blinking and to achieve complete suppression of the blinking, which is an annoying feature in many applications of quantum dots as light sources and fluorescence labels for biomedical imaging.

  7. Upconversion ratiometric fluorescence and colorimetric dual-readout assay for uric acid.

    Science.gov (United States)

    Fang, Aijin; Wu, Qiongqiong; Lu, Qiujun; Chen, Hongyu; Li, Haitao; Liu, Meiling; Zhang, Youyu; Yao, Shouzhuo

    2016-12-15

    A new upconversion colorimetric and ratiometric fluorescence detection method for uric acid (UA) has been designed. Yb(3+), Er(3+) and Tm(3+) co-doped NaYF4 nanoparticles (UCNPs) was synthesized. The co-doped NaYF4 nanoparticles, emit upconversion fluorescence with four typical emission peaks centered at 490nm, 557nm, 670nm and 705nm under the 980nm near-infrared (NIR) irradiation. The ZnFe2O4 magnetic nanoparticles (MNPs) possessing excellent peroxidase-like activity was prepared and used to catalyze oxidation the coupling of N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium salt (TOPS) and 4-amino-antipyrine (4-AAP) in the presence of H2O2 to form purple products (compound 1) which has a characteristic absorption peak located at 550nm. The upconversion fluorescence at 557nm was quenched by the compound 1 while the upconversion emission at 705nm was essentially unchanged, the fluorescence ratio ((I557/I705)0/(I557/I705)) is positively proportional to UA concentration in existence of uricase. More importantly, colorimetric signal can be easily observed and applied to directly distinguish the concentration of UA by the naked eye. Under the optimized conditions, the linear range of colorimetric and ratiometric fluorescence sensing towards UA was 0.01-1mM, the detection limits were as low as 5.79μM and 2.86μM (S/N=3), respectively. The proposed method has been successfully applied to the analysis of UA in human serum. These results indicate that the colorimetric and ratiometric fluorescence dual-readout assay method has great potential for applications in physiological and pathological diagnosis.

  8. Single-exposure dual-energy computed radiography.

    Science.gov (United States)

    Stewart, B K; Huang, H K

    1990-01-01

    This paper focuses on analysis and development of a single-exposure dual-energy digital radiographic method using computed radiography (Fuji FCR-101 storage phosphor system). A detector sandwich consisting of storage phosphor imaging plates and an interdetector filter is used. The goal of this process is to provide a simple dual-energy method using typical plane-projection radiographic equipment and techniques. This approach exploits the transparency of the storage phosphor plates, using radiographic information that would be otherwise lost, to provide energy selective information essentially as a by-product of the radiographic examination. In order to effectively make use of the large dynamic range of the storage phosphor imaging plates (10,000:1), a computed radiography image reading mode of fixed analog-to-digital converter gain and variable photomultiplier sensitivity provides image data which can be related to relative incident exposure for export to the decomposition algorithm. Scatter rejection requirements necessitated crossed 12:1 grids for a field size of 36 x 36 cm. Optimal technique parameters obtained from computer simulation through minimization of the aluminum and Plexiglas equivalent image uncertainty under conditions of constant absorbed does resulted as: 100 kVp using a 0.15-mm-thick tin (Sn) interdetector filter for the lung field. This yields a surface exposure of 23 mR and a surface absorbed dose of 0.26 mGy for a 23-cm-thick chest. Clinical application in evaluation of the solitary pulmonary nodule is discussed, along with an image set demonstrating this application.

  9. Fluorescence and Magnetic Resonance Dual-Modality Imaging-Guided Photothermal and Photodynamic Dual-Therapy with Magnetic Porphyrin-Metal Organic Framework Nanocomposites

    Science.gov (United States)

    Zhang, Hui; Li, Yu-Hao; Chen, Yang; Wang, Man-Man; Wang, Xue-Sheng; Yin, Xue-Bo

    2017-01-01

    Phototherapy shows some unique advantages in clinical application, such as remote controllability, improved selectivity, and low bio-toxicity, than chemotherapy. In order to improve the safety and therapeutic efficacy, imaging-guided therapy seems particularly important because it integrates visible information to speculate the distribution and metabolism of the probe. Here we prepare biocompatible core-shell nanocomposites for dual-modality imaging-guided photothermal and photodynamic dual-therapy by the in situ growth of porphyrin-metal organic framework (PMOF) on Fe3O4@C core. Fe3O4@C core was used as T2-weighted magnetic resonance (MR) imaging and photothermal therapy (PTT) agent. The optical properties of porphyrin were well remained in PMOF, and PMOF was therefore selected for photodynamic therapy (PDT) and fluorescence imaging. Fluorescence and MR dual-modality imaging-guided PTT and PDT dual-therapy was confirmed with tumour-bearing mice as model. The high tumour accumulation of Fe3O4@C@PMOF and controllable light excitation at the tumour site achieved efficient cancer therapy, but low toxicity was observed to the normal tissues. The results demonstrated that Fe3O4@C@PMOF was a promising dual-imaging guided PTT and PDT dual-therapy platform for tumour diagnosis and treatment with low cytotoxicity and negligible in vivo toxicity. PMID:28272454

  10. Combined analysis of intracellular calcium with dual excitation fluorescence photometry and imaging

    Science.gov (United States)

    Uttenweiler, Dietmar; Wojciechowski, Reinhold; Makabe, Makoto; Veigel, Claudia; Fink, Rainer H.

    1995-10-01

    We have developed an integrated microscopy system combining fast dual-excitation fluorescence photometry and digital image analysis with high spatial resolution, based mainly on standard components. With the combination of these well-established techniques in one setup it is possible to monitor intracellular calcium with both sufficiently high temporal and high spatial resolution on the same preparation for many biological applications. Our system consists of a commercially available dual-excitation photometric system, an attached ICCD camera, and a frame grabber board. With this integrated setup one can easily switch between the fast photometric mode and the imaging mode. We used the system to record Fura-2 calcium images (340/380 nm ratios), which were correlated with the faster spot measurements and were analyzed by means of image processing. As an example for its application we reconstructed caffeine-induced calcium transient released from the sarcoplasmic reticulum of isolated and permeabilized skeletal muscle fiber preparations. Such a combined technique will also be important for cellular studies using other fluorescence indicators. Additionally, the described system has an external trigger facility that enables combination with other cell physiological methods, e.g., electrophysiological techniques.

  11. Single vs. dual mode stimulation in spinal cord stimulation - what is the difference?

    NARCIS (Netherlands)

    Manola, L.; Holsheimer, J.

    2006-01-01

    Objectives. Stimulation with dual percutaneous leads is often used in SCS. Either a single generator gives pulses to both leads simultaneously (single mode), or two generators give pulses to each lead alternately (dual mode) [1]. The aim was to compare theoretically the performance of these stimul

  12. Project Title: Radiochemical Analysis by High Sensitivity Dual-Optic Micro X-ray Fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Havrilla, George J.; Gao, Ning

    2003-06-01

    A novel dual-optic micro X-ray fluorescence instrument will be developed to do radiochemical analysis of high-level radioactive wastes at DOE sites such as Savannah River Site and Hanford. This concept incorporates new X-ray optical elements such as monolithic polycapillaries, which focus X-rays. The polycapillary optic can be used to focus X-rays emitted by the X-ray tube thereby increasing the X-ray flux on the sample over 1000 times. The polycapillary optic will also be used to collect the X-rays from the excitation site. This will effectively screen the radiation background from the radioactive species in the specimen. This dual-optic approach significantly reduces the background and increases the analyte signal thereby increasing the sensitivity of the analysis. This dual-capillary design is essentially a confocal (having the same foci) design, i.e. the detected X-rays are only emitted from the overlap of the two focal spots. This increases spatial resolution and reduce s background. The integration of the X-ray optics increases the signal-to-noise and thereby increases the sensitivity of the analysis for low-level analytes. This work will address a key need for radiochemical analysis of high-level waste using a non-destructive, multi-element, and rapid method in a radiation environment. There is significant potential that this instrumentation could be capable of on-line analysis for process waste stream characterization at DOE sites.

  13. Radiochemical Analysis by High Sensitivity Dual-Optic Micro X-ray Fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Havrilla, George J.; Gao, Ning

    2004-06-01

    A novel dual-optic micro X-ray fluorescence instrument will be developed to do radiochemical analysis of high-level radioactive wastes at DOE sites such as Savannah River Site and Hanford. This concept incorporates new X-ray optical elements such as monolithic polycapillaries, which focus X-rays. The polycapillary optic can be used to focus X-rays emitted by the X-ray tube thereby increasing the X-ray flux on the sample over 1000 times. The polycapillary optic will also be used to collect the X-rays from the excitation site. This will effectively screen the radiation background from the radioactive species in the specimen. This dual-optic approach significantly reduces the background and increases the analyte signal thereby increasing the sensitivity of the analysis. This dual-capillary design is essentially a confocal (having the same foci) design, i.e. the detected X-rays are only emitted from the overlap of the two focal spots. This increases spatial resolution and reduces background. The integration of the X-ray optics increases the signal-to-noise and thereby increases the sensitivity of the analysis for low-level analytes. This work will address a key need for radiochemical analysis of high-level waste using a non-destructive, multi-element, and rapid method in a radiation environment. There is significant potential that this instrumentation could be capable of on-line analysis for process waste stream characterization at DOE sites.

  14. The application of image cytometry to viability assessment in dual fluorescence-stained fish spermatozoa.

    Science.gov (United States)

    Flajshans, Martin; Cosson, Jacky; Rodina, Marek; Linhart, Otomar

    2004-01-01

    The viability of spermatozoa has been assessed using SYBR 14 staining for DNA of living cells and propidium iodide staining for DNA of degenerate cells. This dual staining was performed on four fish species (Siberian sturgeon, Acipenser baerii; common carp, Cyprinus carpio; tench, Tinca tinca and wels, Silurus glanis) and the proportions of live and dead spermatozoa were assessed by epifluorescence microscopy and image cytometry. Ten phase contrast and epifluorescent images were recorded per sample, corresponding images were overlaid, and the blended images were evaluated for live and dead spermatozoa, represented by green and red fluorescence signals. Live/dead proportions were assessed, after dual thresholding, by imaging software that counted absolute numbers of objects and computed their frequencies. All sperm heads were found to be labelled, emitting either green or red light. Mean numbers of spermatozoa per image were in the ranges 32-113, 61-105, 48-104 and 29-91 for Siberian sturgeon, common carp, tench and wels, respectively. The corresponding proportions of live spermatozoa were in the ranges 83.56-94.59%, 93.92-97.02%, 76.14-97.76% and 79.45-83.76%. Standard deviations did not exceed 5% of the means. The image cytometric system using dual staining with SYBR 14 and propidium iodide was clearly suitable for assessing the viability of freshwater fish spermatozoa.

  15. Dual energy with dual source CT and kVp switching with single source CT: a comparison of dual energy performance

    Science.gov (United States)

    Grasruck, M.; Kappler, S.; Reinwand, M.; Stierstorfer, K.

    2009-02-01

    Stimulated by the introduction of clinical dual source CT, the interest in dual energy methods has been increasing in the past years. Whereas the potential of material decomposition by dual energy methods is known since the early 1980ies, the realization of dual energy methods is a wide field of today's research. Energy separation can be achieved with energy selective detectors or by varying X-ray source spectra. This paper focuses on dual energy techniques with varying X-ray spectra. These can be provided by dual source CT devices, operated with different kVp settings on each tube. Excellent spectral separation is the key property for use in clinical routine. The drawback of higher cost for two tubes and two detectors leads to an alternative realization, where a single source CT yields different spectra by fast kVp switching from reading to reading. This provides access to dual-energy methods in single source CT. However, this technique comes with some intrinsic limitations. The maximum X-ray flux is reduced in comparison to the dual source system. The kVp rise and fall time between each reading reduces the spectral separation. In comparison to dual source CT, for a constant number of projections per energy spectrum the temporal resolution is reduced; a reasonable trade of between reduced numbers of projection and limited temporal resolution has to be found. The overall dual energy performance is the guiding line for our investigations. We present simulations and measurements which benchmark both solutions in terms of spectral behavior, especially of spectral separation.

  16. Single chip SPR and fluorescent ELISA assay of prostate specific antigen.

    Science.gov (United States)

    Breault-Turcot, J; Poirier-Richard, H-P; Couture, M; Pelechacz, D; Masson, J-F

    2015-12-01

    A multi-channel system combining fluidics and micropatterned plasmonic materials with wavelength interrogation surface plasmon resonance (SPR) and fluorescence detection was integrated from the combination of a small and motorized fluorescence microscope mounted on a portable 4-channel SPR instrument. The SPR and fluorescent measurements were performed based on the same detection area in a multi-channel fluidic, with a sensing scheme for prostate-specific antigen (PSA) consisting of a sandwich assay with a capture anti-PSA immobilized onto the SPR sensor and a detection anti-PSA modified with horseradish peroxidase (HRP). In this dual-detection instrument, fluorescence was measured from the solution side of the micropatterned gold film, while the interface between the glass prism and the gold film served to interrogate the SPR response. The SPR sensors were comprised of microhole arrays fabricated by photolithography to enhance the instrumental response for PSA detection by approximately a factor of 2 to 3 and they were coated with a self-assembled monolayer of a peptide (3-MPA-HHHDD-OH) to minimize nonspecific adsorption. PSA was successfully detected at clinical concentrations from 10 pM to 50 nM with this integrated system in a single assay lasting 12 minutes, almost centering on the desired range for PSA diagnostic tests (>4 ng mL(-1) or >150 pM). The combination of two robust techniques in a single chip and instrument has led to a simple and effective assay that can be carried out on a small and portable instrument providing rapid biodetection of an important cancer biomarker with a dynamic range of nearly 4 orders of magnitude in the clinical range.

  17. Melanin-originated carbonaceous dots for triple negative breast cancer diagnosis by fluorescence and photoacoustic dual-mode imaging.

    Science.gov (United States)

    Xiao, Wei; Li, Yuan; Hu, Chuan; Huang, Yuan; He, Qin; Gao, Huile

    2017-07-01

    Carbonaceous dots exhibit increasing applications in diagnosis and drug delivery due to excellent photostability and biocompatibility properties. However, relative short excitation and emission of melanin carbonaceous dots (MCDs) limit the applicability in fluorescence bioimaging. Furthermore, the generally poor spatial resolution of fluorescence imaging limits potential in vivo applications. Due to a variety of beneficial properties, in this study, MCDs were prepared exhibiting great potential in fluorescence and photoacoustic dual-mode bioimaging. The MCDs exhibited a long excitation peak at 615nm and emission peak at 650nm, further highlighting the applicability in fluorescence imaging, while the absorbance peak at 633nm renders MCDs suitable for photoacoustic imaging. In vivo, the photoacoustic signal of MCDs was linearly correlated with the concentration of MCDs. Moreover, the MCDs were shown to be taken up into triple negative breast cancer cell line 4T1 in both a time- and concentration-dependent manner. In vivo fluorescence and photoacoustic imaging of subcutaneous 4T1 tumor demonstrated that MCDs could passively target triple negative breast cancer tissue by enhanced permeability and retention effects and may therefore be used for tumor dual-mode imaging. Furthermore, fluorescence distribution in tissue slices suggested that MCDs may distribute in 4T1 tumor with high efficacy. In conclusion, the MCDs studied offer potential application in fluorescence and photoacoustic dual-mode imaging.

  18. Structural and Luminescent Properties of [N-benzyl-N-(2-pyridyl) methylamine]dichlorozinc(II): Dual Fluorescence of N-benzyl- N-(2-pyridyl)methylamine

    Energy Technology Data Exchange (ETDEWEB)

    Song, Young-Kwang; Kim, Young-Inn [Department of Chemistry Education and BK21 Plus Team for Advanced Chemical Materials, Seoul (Korea, Republic of); Kang, Sung Kwon [Pusan National University, Busan (Korea, Republic of)

    2015-01-15

    According to the TICT model, a molecule with an electron donor and acceptor group connected by a single bond may exist as a twisted structure and exhibit an ICT state upon excitation. Such molecules transform from the exited state to the twisted structure via a radiationless process, in which the donor group is perpendicular to the rest of the molecular frame generating a complete decoupling state. Such molecules generally exhibit dual emission, which is observed as two well-separated emission bands. Furthermore, the ratio of the intensities of these two bands is not dependent on its concentration. We designed and synthesized a novel dual fluorescent bpma molecule and its Zn(II) chloride complex Zn(bpma)Cl{sub 2}. The dual fluorescence exhibited by bpma was attributed to the formation of a TICT state in a polar medium. Furthermore, Zn(bpma)Cl{sub 2} was found to quench the emission from the TICT state, resulting in a single emission attributed to the LE state in the blue region. These findings indicate that Zn(bpma)Cl{sub 2} is a possible candidate for blue emissive photoactive material and Zn(II) recognition sensor.

  19. Influence of different environments on the excited-state proton transfer and dual fluorescence of fisetin

    Science.gov (United States)

    Guharay, Jayanti; Dennison, S. Moses; Sengupta, Pradeep K.

    1999-05-01

    The influence of different protic and aprotic solvent environments on the excited-state intramolecular proton transfer (ESIPT) leading to a dual fluorescence behaviour of a biologically important, naturally occurring, polyhydroxyflavone, fisetin (3,3',4',7-tetrahydroxyflavone), has been investigated. The normal fluorescence band, in particular, is extremely sensitive to solvent polarity with νmax shifting from 24 510 cm -1 in dioxane ( ET(30)=36.0) to 20 790 cm -1 in methanol ( ET(30)=55.5). This is rationalized in terms of solvent dipolar relaxation process, which also accounts for the red edge excitation shifts (REES) observed in viscous environments such as glycerol at low temperatures. Significant solvent dependence of the tautomer fluorescence properties ( νmax, yield and decay kinetics) reveals the influence of external hydrogen bonding perturbation on the internal hydrogen bond of the molecule. These excited-state relaxation phenomena and their relevant parameters have been used to probe the microenvironment of fisetin in a membrane mimetic system, namely AOT reverse micelles in n-heptane at different water/surfactant molar ratio ( w0).

  20. Characterization of a natural triple-tandem c-di-GMP riboswitch and application of the riboswitch-based dual-fluorescence reporter.

    Science.gov (United States)

    Zhou, Hang; Zheng, Cao; Su, Jianmei; Chen, Bo; Fu, Yang; Xie, Yuqun; Tang, Qing; Chou, Shan-Ho; He, Jin

    2016-02-19

    c-di-GMP riboswitches are structured RNAs located in the 5'-untranslated regions (5'-UTRs) of mRNAs that regulate expression of downstream genes in response to changing concentrations of the second messenger c-di-GMP. We discovered three complete c-di-GMP riboswitches (Bc3, Bc4 and Bc5 RNA) with similar structures, which are arranged in tandem to constitute a triple-tandem (Bc3-5 RNA) riboswitch in the 5'-UTR of the cspABCDE mRNA in Bacillus thuringiensis subsp. chinensis CT-43. Our results showed that this natural triple-tandem riboswitch controlled the expression of the reporter gene more stringently and digitally than the double-tandem or single riboswitch. A sandwich-like dual-fluorescence reporter was further constructed by fusing the Bc3-5 RNA gene between the two fluorescence protein genes amcyan and turborfp. This reporter strain was found to exhibit detectable fluorescence color changes under bright field in response to intracellular c-di-GMP level altered by induced expression of diguanylate cyclase (DGC) PleD. Using this system, two putative membrane-bound DGCs from B. thuringiensis and Xanthomonas oryzae were verified to be functional by replacing pleD with the corresponding DGC genes. This report represented the first native triple-tandem riboswitch that was applied to serve as a riboswitch-based dual-fluorescence reporter for the efficient and convenient verification of putative DGC activity in vivo.

  1. Exploring single-molecule dynamics with fluorescence nanoscopy

    Energy Technology Data Exchange (ETDEWEB)

    Ringemann, Christian; Harke, Ben; Von Middendorff, Claas; Medda, Rebecca; Leutenegger, Marcel; Schoenle, Andreas; W Hell, Stefan; Eggeling, Christian [Department of Nanobiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Goettingen (Germany); Honigmann, Alf; Wagner, Richard [Biophysik, University Osnabrueck, FB Biologie/Chemie, Osnabrueck (Germany)], E-mail: ceggeli@gwdg.de

    2009-10-15

    The study of molecular dynamics at the single-molecule level with fluorescence correlation spectroscopy (FCS) and far-field optics has contributed greatly to the functional understanding of complex systems. Unfortunately, such studies are restricted to length scales of >200 nm because diffraction does not allow further reduction of the measurement volume. This sets an upper limit on the applicable concentration of fluorescently labeled molecules and even more importantly, averages out details of nanoscale dynamics. By combining FCS and fluorescence intensity distribution analysis (FIDA) with sub-diffraction-resolution stimulated emission depletion (STED) nanoscopy, we remove this restriction and obtain open measurement volumes of nanoscale dimensions which are tunable in size. As a consequence, single-molecule studies can now be extended to nanoscale dynamics and may be applied to much larger, often endogenous concentrations. In solution, low-brightness signal from axial out-of-focus volume shells was taken into account by using both FCS and FIDA in conjunction to analyze the data. In two-dimensional systems, such as lipid membranes, the background is greatly reduced and measurements feature excellent signal-to-noise ratios. Measurement foci of down to 30 nm in diameter directly reveal anomalous diffusion of lipids in the plasma membrane of living cells and allow for the determination of on/off rates of the binding of lipids to other membrane constituents. Such important insight into the prominent biological question of lipid membrane organization or 'lipid rafts' shows that combining fluctuation analysis with STED-engineered ultra-small measurement volumes is a viable and powerful new approach to probing molecular dynamics on the nanoscale.

  2. Data analysis considerations in probing single quantum dot fluorescence intermittency

    Science.gov (United States)

    Krogmeier, Jeffrey R.; Hwang, Jeeseong

    2005-04-01

    The fluorescence intermittency of single, bare, CdSe/ZnS quantum dots was probed using single molecule confocal microscopy and found to demonstrate power law kinetics. Various threshold values and line fitting parameters are employed in the data analysis and their effects on the extracted power law exponents, moff and mon, are presented. The threshold is found to be critical in determining moff while having no significant effect on mon. The mean plus 2σ threshold, calculated from the background noise in the measurement, results in a more negative moff slope in comparison to the mean plus 3σ or mean plus 4σ thresholds. This is likely due to the mean plus 2σ threshold lying within the background noise outliers which mimic short on events. In contrast, the mean plus 4σ threshold is above 99.99% of the background noise while adequately below the fluorescence signal. Additionally, it is found that fitting only the ten most probable data points rather than all the data points in the log-log probability density graphs results in no significant change in moff and mon.

  3. Comparing the da Vinci si single console and dual console in teaching novice surgeons suturing techniques.

    Science.gov (United States)

    Crusco, Salvatore; Jackson, Tiffany; Advincula, Arnold

    2014-01-01

    Robot-assisted laparoscopic surgery is often taught with the surgical mentor at the surgeon console and the trainee at the patient's bedside. The da Vinci dual console (Intuitive Surgical, Sunnyvale, California) allows a surgical mentor to teach with both the mentor and the trainee working at a surgeon console simultaneously. The purpose of this study is to evaluate the effectiveness of the dual console versus the single console for teaching medical students robotic tasks. Forty novice medical students were randomized to either the da Vinci single-console or dual-console group and were taught 4 knot-tying techniques by a surgical mentor. The students were timed while performing the tasks. No statistically significant differences in mean task times were observed between the single- and dual-console groups: interrupted stitch with a 2-handed knot (300 seconds for single vs 294 seconds for dual, P=.59), interrupted stitch with a 1-handed knot (198 seconds for single vs 212 seconds for dual, P=.88), figure-of-8 stitch with a 2-handed knot (261 seconds for single vs 219 seconds for dual, P=.20), and figure-of-8 stitch with a 1-handed knot (200 seconds for single vs 199 seconds for dual, P=.53). No significant difference was observed in performance time when teaching knot-tying techniques to medical students using the da Vinci dual console compared with the single console. More research needs to be performed on the utility of the da Vinci dual console in surgical training.

  4. Multiplex fluorescence melting curve analysis for mutation detection with dual-labeled, self-quenched probes.

    Directory of Open Access Journals (Sweden)

    Qiuying Huang

    Full Text Available Probe-based fluorescence melting curve analysis (FMCA is a powerful tool for mutation detection based on melting temperature generated by thermal denaturation of the probe-target hybrid. Nevertheless, the color multiplexing, probe design, and cross-platform compatibility remain to be limited by using existing probe chemistries. We hereby explored two dual-labeled, self-quenched probes, TaqMan and shared-stem molecular beacons, in their ability to conduct FMCA. Both probes could be directly used for FMCA and readily integrated with closed-tube amplicon hybridization under asymmetric PCR conditions. Improved flexibility of FMCA by using these probes was illustrated in three representative applications of FMCA: mutation scanning, mutation identification and mutation genotyping, all of which achieved improved color-multiplexing with easy probe design and versatile probe combination and all were validated with a large number of real clinical samples. The universal cross-platform compatibility of these probes-based FMCA was also demonstrated by a 4-color mutation genotyping assay performed on five different real-time PCR instruments. The dual-labeled, self-quenched probes offered unprecedented combined advantage of enhanced multiplexing, improved flexibility in probe design, and expanded cross-platform compatibility, which would substantially improve FMCA in mutation detection of various applications.

  5. Ultimate statistical physics: fluorescence of a single atom

    Science.gov (United States)

    Pomeau, Yves; Le Berre, Martine; Ginibre, Jean

    2016-10-01

    We discuss the statistics of emission of photons by a single atom or ion illuminated by a laser beam at the frequency of quasi-resonance between two energy levels, a situation that corresponds to real experiments. We extend this to the case of two laser beams resonant with the energy differences between two excited levels and the ground state (three level atom in V-configuration). We use a novel approach to this type of problem by considering a Kolmogorov equation for the probability distribution of the atomic state, which takes into account first the deterministic evolution of this state under the effect of the incoming laser beam and second the random emission of photons during the spontaneous decay of the excited state(s) to the ground state. This approach yields solvable equations in the two level atom case. For the three level atom case we set the problem and clearly define its frame. The results obtained are valid in both opposite limits of rare and frequent spontaneous decay, compared to the period of the optical Rabi oscillations due to the interaction between resonant excitation and atomic levels. Our analysis gives access to various statistical properties of the fluorescence light, including one showing that its fluctuations in time are not invariants under time reversal. This result makes evident the fundamentally irreversible character of quantum measurements, represented here by the emission of photons of fluorescence.

  6. Fluorescence cross-correlation spectroscopy using single wavelength laser

    Institute of Scientific and Technical Information of China (English)

    Chao XIE; Chaoqing DONG; Jicun REN

    2009-01-01

    In this paper, we first introduced the basic principle of fluorescence cross-correlation spectroscopy (FCCS) and then established an FCCS setup using a single wavelength laser. We systematically optimized the setup, and the detection volume reached about 0.7 fL. The home-built setup was successfully applied for the study of the binding reaction of human immunoglobulin G with goat antihuman immunoglobulin G. Using quantum dots (745 nm emission wavelength) and Rhodamine B (580 nm emission wavelength) as labeling probes and 532 nm laser beam as an excitation source, the cross-talk effect was almost completely suppressed. The molecule numbers in a highly focused volume, the concentration, and the diffusion time and hydrodynamic radii of the reaction products can be determined by FCCS system.

  7. Fluorescence-excitation and Emission Spectroscopy on Single FMO Complexes.

    Science.gov (United States)

    Löhner, Alexander; Ashraf, Khuram; Cogdell, Richard J; Köhler, Jürgen

    2016-08-22

    In green-sulfur bacteria sunlight is absorbed by antenna structures termed chlorosomes, and transferred to the RC via the Fenna-Matthews-Olson (FMO) complex. FMO consists of three monomers arranged in C3 symmetry where each monomer accommodates eight Bacteriochlorophyll a (BChl a) molecules. It was the first pigment-protein complex for which the structure has been determined with high resolution and since then this complex has been the subject of numerous studies both experimentally and theoretically. Here we report about fluorescence-excitation spectroscopy as well as emission spectroscopy from individual FMO complexes at low temperatures. The individual FMO complexes are subjected to very fast spectral fluctuations smearing out any possible different information from the ensemble data that were recorded under the same experimental conditions. In other words, on the time scales that are experimentally accessible by single-molecule techniques, the FMO complex exhibits ergodic behaviour.

  8. Single-gene dual-color reporter cell line to analyze RNA synthesis in vivo.

    Science.gov (United States)

    Palangat, Murali; Larson, Daniel R

    2016-07-01

    RNA synthesis occurs through the multi-step process of transcription which consists of initiation, elongation, termination, and cleavage of the nascent RNA. In recent years, post-initiation events have attracted considerable attention as regulatory steps in gene expression. In particular, changes in elongation rate have been proposed to alter RNA fate either through changes in RNA secondary structure or recruitment of trans-acting factors, but systematic approaches for perturbing and measuring elongation rate are currently lacking. Here, we describe a system for precisely measuring elongation dynamics for single nascent transcripts at a single gene locus in human cell lines. The system is based on observing the production of fluorescently labeled RNA stem loops which flank a region of interest. The region of interest can be altered using flp recombinases, thus allowing one to study the effects of cis-acting sequences on transcription rate. The dual-color RNAs which are made during this process are exported and translated, thus enabling visualization of each step in gene expression.

  9. Ocean wave parameters and spectrum estimated from single and dual high-frequency radar systems

    Science.gov (United States)

    Hisaki, Yukiharu

    2016-09-01

    The high-frequency (HF) radar inversion algorithm for spectrum estimation (HIAS) can estimate ocean wave directional spectra from both dual and single radar. Wave data from a dual radar and two single radars are compared with in situ observations. The agreement of the wave parameters estimated from the dual radar with those from in situ observations is the best of the three. In contrast, the agreement of the wave parameters estimated from the single radar in which no Doppler spectra are observed in the cell closest to the in situ observation point is the worst among the three. Wave data from the dual radar and the two single radars are compared. The comparison of the wave heights estimated from the single and dual radars shows that the area sampled by the Doppler spectra for the single radar is more critical than the number of Doppler spectra in terms of agreement with the dual-radar-estimated wave heights. In contrast, the comparison of the wave periods demonstrates that the number of Doppler spectra observed by the single radar is more critical for agreement of the wave periods than the area of the Doppler spectra. There is a bias directed to the radar position in the single radar estimated wave direction.

  10. Laser Induced Dual Fluorescence Ratiometric Technique for Mixing Characterization in Microfluidic Systems

    Science.gov (United States)

    Bedding, David; Hidrovo, Carlso

    2016-11-01

    Increasing the rate of mixing within microfluidic systems is vitally important in understanding biological and chemical reaction kinetics and mechanisms. The small length scales characteristic of these systems which translate into highly viscous, Stokes flows result in mixing that is primarily dominated by diffusion. In order to counteract this, an approach that utilizes inertial droplet collisions to promote chaotic advection between two mixing species has been developed. A Laser-Induced Dual Fluorescence (LIDF) system in conjunction with a high-speed camera and appropriate optics are used to capture two intensity fields providing information about the mixing process as well as the excitation intensity field over the volume of interest. The rate of mixing for the coalescing droplets was quantified by taking the standard deviation of the first intensity field over time, while the second intensity field provides information about the intensity field. A ratiometric imaging approach allows removal of mixing fluorescence signal noise in the form of variation in excitation intensity, primarily from the lasing patterns and lensing effects within the interrogation volume. NSF CAREER Award Grant CBET - 1151091.

  11. Developing DNA nanotechnology using single-molecule fluorescence.

    Science.gov (United States)

    Tsukanov, Roman; Tomov, Toma E; Liber, Miran; Berger, Yaron; Nir, Eyal

    2014-06-17

    CONSPECTUS: An important effort in the DNA nanotechnology field is focused on the rational design and manufacture of molecular structures and dynamic devices made of DNA. As is the case for other technologies that deal with manipulation of matter, rational development requires high quality and informative feedback on the building blocks and final products. For DNA nanotechnology such feedback is typically provided by gel electrophoresis, atomic force microscopy (AFM), and transmission electron microscopy (TEM). These analytical tools provide excellent structural information; however, usually they do not provide high-resolution dynamic information. For the development of DNA-made dynamic devices such as machines, motors, robots, and computers this constitutes a major problem. Bulk-fluorescence techniques are capable of providing dynamic information, but because only ensemble averaged information is obtained, the technique may not adequately describe the dynamics in the context of complex DNA devices. The single-molecule fluorescence (SMF) technique offers a unique combination of capabilities that make it an excellent tool for guiding the development of DNA-made devices. The technique has been increasingly used in DNA nanotechnology, especially for the analysis of structure, dynamics, integrity, and operation of DNA-made devices; however, its capabilities are not yet sufficiently familiar to the community. The purpose of this Account is to demonstrate how different SMF tools can be utilized for the development of DNA devices and for structural dynamic investigation of biomolecules in general and DNA molecules in particular. Single-molecule diffusion-based Förster resonance energy transfer and alternating laser excitation (sm-FRET/ALEX) and immobilization-based total internal reflection fluorescence (TIRF) techniques are briefly described and demonstrated. To illustrate the many applications of SMF to DNA nanotechnology, examples of SMF studies of DNA hairpins and

  12. Dual-emission fluorescent silica nanoparticle-based probe for ultrasensitive detection of Cu2+.

    Science.gov (United States)

    Zong, Chenghua; Ai, Kelong; Zhang, Guo; Li, Hongwei; Lu, Lehui

    2011-04-15

    An effective dual-emission fluorescent silica nanoparticle-based probe has been constructed for rapid and ultrasensitive detection of Cu(2+). In this nanoprobe, a dye-doped silica core served as a reference signal, thus providing a built-in correction for environmental effects. A response dye was covalently grafted on the surface of the silica nanoparticles through a chelating reagent for Cu(2+). The fluorescence of the response dye could be selectively quenched in the presence of Cu(2+), accompanied by a visual orange-to-green color switch of the nanoprobe. The nanoprobe provided an effective platform for reliable detection of Cu(2+) with a detection limit as low as 10 nM, which is nearly 2 × 10(3) times lower than the maximum level (∼20 μM) of Cu(2+) in drinking water permitted by the U.S. Environmental Protection Agency (EPA). The high sensitivity was attributed to the strong chelation of Cu(2+) with polyethyleneimine (PEI) and a signal amplification effect. The nanoprobe constructed by this method was very stable, enabling the rapid detection of Cu(2+) in real water samples. Good linear correlations were obtained over the concentration range from 1 × 10(-7) to 8 × 10(-7) (R(2) = 0.99) with recoveries of 103.8-99.14% and 95.5-95.14% for industrial wastewater and lake water, respectively. Additionally, the long-wavelength emission of the response dye can avoid the interference of the autofluorescence of the biosystems, which facilitated their applications in monitoring Cu(2+) in cells. Furthermore, the nanoprobe showed a good reversibility; the fluorescence can be switched "off" and "on" by an addition of Cu(2+) and EDTA, respectively. © 2011 American Chemical Society

  13. A dual-modality optical coherence tomography and fluorescence lifetime imaging microscopy system for simultaneous morphological and biochemical tissue characterization.

    Science.gov (United States)

    Park, Jesung; Jo, Javier A; Shrestha, Sebina; Pande, Paritosh; Wan, Qiujie; Applegate, Brian E

    2010-07-16

    Most pathological conditions elicit changes in the tissue optical response that may be interrogated by one or more optical imaging modalities. Any single modality typically only furnishes an incomplete picture of the tissue optical response, hence an approach that integrates complementary optical imaging modalities is needed for a more comprehensive non-destructive and minimally-invasive tissue characterization. We have developed a dual-modality system, incorporating optical coherence tomography (OCT) and fluorescence lifetime imaging microscopy (FLIM), that is capable of simultaneously characterizing the 3-D tissue morphology and its biochemical composition. The Fourier domain OCT subsystem, at an 830 nm center wavelength, provided high-resolution morphological volumetric tissue images with an axial and lateral resolution of 7.3 and 13.4 µm, respectively. The multispectral FLIM subsystem, based on a direct pulse-recording approach (upon 355 nm laser excitation), provided two-dimensional superficial maps of the tissue autofluorescence intensity and lifetime at three customizable emission bands with 100 µm lateral resolution. Both subsystems share the same excitation/illumination optical path and are simultaneously raster scanned on the sample to generate coregistered OCT volumes and FLIM images. The developed OCT/FLIM system was capable of a maximum A-line rate of 59 KHz for OCT and a pixel rate of up to 30 KHz for FLIM. The dual-modality system was validated with standard fluorophore solutions and subsequently applied to the characterization of two biological tissue types: postmortem human coronary atherosclerotic plaques, and in vivo normal and cancerous hamster cheek pouch epithelial tissue.

  14. Application of dual-fuel propulsion to a single stage AMLS vehicle

    Science.gov (United States)

    Lepsch, Roger A., Jr.; Stanley, Douglas O.; Unal, Resit

    1993-01-01

    As part of NASA's Advanced Manned Launch System (AMLS) study to determine a follow-on, or complement, to the Space Shuttle, a reusable single-stage-to-orbit concept utilizing dual-fuel rocket propulsion has been examined. Several dual-fuel propulsion concepts were investigated. These include: a separate engine concept combining Russian RD-170 kerosene-fueled engines with SSME-derivative engines; the kerosene and hydrogen-fueled Russian RD-701 engine concept; and a dual-fuel, dual-expander engine concept. Analysis to determine vehicle weight and size characteristics was performed using conceptual level design techniques. A response surface methodology for multidisciplinary design was utilized to optimize the dual-fuel vehicle concepts with respect to several important propulsion system and vehicle design parameters in order to achieve minimum empty weight. Comparisons were then made with a hydrogen-fueled reference, single-stage vehicle. The tools and methods employed in the analysis process are also summarized.

  15. Comparison of Orbicularis Oculi Muscle Activity during Computer Work with Single and Dual Monitors

    OpenAIRE

    Yoo, Won-gyu

    2014-01-01

    [Purpose] This study compared the orbicularis oculi muscle activity during computer work with single and dual monitors. [Subjects] Ten computer workers 22–27 years of age were included in this study. [Methods] Subjects performed computer work with single or dual monitors, and the activity of the right orbicularis oculi muscle was measured with a MP150 system. [Results] The muscle activity of the orbicularis oculi under condition 1 was significantly decreased compared with that under condition...

  16. A simple optical fiber device for quantitative fluorescence microscopy of single living cells

    NARCIS (Netherlands)

    Graft, van Marja; Oosterhuis, Bernard; Werf, van der Kees O.; Grooth, de Bart G.; Greve, Jan

    1993-01-01

    simple and relatively inexpensive system is described for obtaining quantitative fluorescence measurements on single living cells loaded with a fluorescent probe to study cell physiological processes. The light emitted from the fluorescent cells is captured by and transported through an optical fibe

  17. Indocyanine Green Loaded Reduced Graphene Oxide for In Vivo Photoacoustic/Fluorescence Dual-Modality Tumor Imaging.

    Science.gov (United States)

    Chen, Jingqin; Liu, Chengbo; Zeng, Guang; You, Yujia; Wang, Huina; Gong, Xiaojing; Zheng, Rongqin; Kim, Jeesu; Kim, Chulhong; Song, Liang

    2016-12-01

    Multimodality imaging based on multifunctional nanocomposites holds great promise to fundamentally augment the capability of biomedical imaging. Specifically, photoacoustic and fluorescence dual-modality imaging is gaining much interest because of their non-invasiveness and the complementary nature of the two modalities in terms of imaging resolution, depth, sensitivity, and speed. Herein, using a green and facile method, we synthesize indocyanine green (ICG) loaded, polyethylene glycol (PEG)ylated, reduced nano-graphene oxide nanocomposite (rNGO-PEG/ICG) as a new type of fluorescence and photoacoustic dual-modality imaging contrast. The nanocomposite is shown to have minimal toxicity and excellent photoacoustic/fluorescence signals both in vitro and in vivo. Compared with free ICG, the nanocomposite is demonstrated to possess greater stability, longer blood circulation time, and superior passive tumor targeting capability. In vivo study shows that the circulation time of rNGO-PEG/ICG in the mouse body can sustain up to 6 h upon intravenous injection; while after 1 day, no obvious accumulation of rNGO-PEG/ICG is found in any major organs except the tumor regions. The demonstrated high fluorescence/photoacoustic dual contrasts, together with its low toxicity and excellent circulation life time, suggest that the synthesized rNGO-PEG/ICG can be a promising candidate for further translational studies on both the early diagnosis and image-guided therapy/surgery of cancer.

  18. Indocyanine Green Loaded Reduced Graphene Oxide for In Vivo Photoacoustic/Fluorescence Dual-Modality Tumor Imaging

    Science.gov (United States)

    Chen, Jingqin; Liu, Chengbo; Zeng, Guang; You, Yujia; Wang, Huina; Gong, Xiaojing; Zheng, Rongqin; Kim, Jeesu; Kim, Chulhong; Song, Liang

    2016-02-01

    Multimodality imaging based on multifunctional nanocomposites holds great promise to fundamentally augment the capability of biomedical imaging. Specifically, photoacoustic and fluorescence dual-modality imaging is gaining much interest because of their non-invasiveness and the complementary nature of the two modalities in terms of imaging resolution, depth, sensitivity, and speed. Herein, using a green and facile method, we synthesize indocyanine green (ICG) loaded, polyethylene glycol (PEG)ylated, reduced nano-graphene oxide nanocomposite (rNGO-PEG/ICG) as a new type of fluorescence and photoacoustic dual-modality imaging contrast. The nanocomposite is shown to have minimal toxicity and excellent photoacoustic/fluorescence signals both in vitro and in vivo. Compared with free ICG, the nanocomposite is demonstrated to possess greater stability, longer blood circulation time, and superior passive tumor targeting capability. In vivo study shows that the circulation time of rNGO-PEG/ICG in the mouse body can sustain up to 6 h upon intravenous injection; while after 1 day, no obvious accumulation of rNGO-PEG/ICG is found in any major organs except the tumor regions. The demonstrated high fluorescence/photoacoustic dual contrasts, together with its low toxicity and excellent circulation life time, suggest that the synthesized rNGO-PEG/ICG can be a promising candidate for further translational studies on both the early diagnosis and image-guided therapy/surgery of cancer.

  19. Theoretical investigation on dual fluorescence and intramolecular charge transfer of 5-phenyl-5H-phenanthridin-6-one

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Quantum-chemical calculations with the time-dependent density function theory (TDDFT) have been carried out for 5-phenyl-5H-phenanthridin-6-one (PP). For this molecule, dual fluorescence and in- tramolecular charge transfer (ICT) were experimentally observed. The B3LYP functional with 6-311+G (2d, p) basis set has been used for the theoretical calculations. The solvent effects have been described within the polarizable continuum model (PCM). Ground-state geometry optimization reveals that the phenyl/phenanthridinone dihedral angle equals 90.0°, a nearly perpendicular structure. Vertical ab- sorption energy calculations characterize the lower singlet excited states both in gas phase and in solvents. It can be found that the lower excited states have locally excitation (LE) feature. Through constructing the potential energy curves of both isolated and solvated systems describing the LE→ICT reaction and fluorescence emission, we obtain the enthalpy difference ΔH between the LE and ICT states, energy barrier Ea, and energy difference δEFC, indicating the structural changes taking place during the ICT reaction. Potential curve and calculated emission energies for both isolated and sol- vated systems show a dual fluorescence phenomenon, consisting of a LE emission band and a red-shifted ICT band. Our calculations including the solvent effects indicate that the dual fluorescence is brought about by the change in molecular structure connected with the planarization of the twisted N-phenylphenanthridinone during the ICT reaction.

  20. Project Title: Radiochemical Analysis by High Sensitivity Dual-Optic Micro X-ray Fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Havrilla, George J.; Gao, Ning

    2002-06-01

    A novel dual-optic micro X-ray fluorescence instrument will be developed to do radiochemical analysis of high-level radioactive wastes at DOE sites such as Savannah River Site and Hanford. This concept incorporates new X-ray optical elements such as monolithic polycapillaries and double bent crystals, which focus X-rays. The polycapillary optic can be used to focus X-rays emitted by the X-ray tube thereby increasing the X-ray flux on the sample over 1000 times. Polycapillaries will also be used to collect the X-rays from the excitation site and screen the radiation background from the radioactive species in the specimen. This dual-optic approach significantly reduces the background and increases the analyte signal thereby increasing the sensitivity of the analysis. A doubly bent crystal used as the focusing optic produces focused monochromatic X-ray excitation, which eliminates the bremsstrahlung background from the X-ray source. The coupling of the doubly bent crystal for monochromatic excitation with a polycapillary for signal collection can effectively eliminate the noise background and radiation background from the specimen. The integration of these X-ray optics increases the signal-to-noise and thereby increases the sensitivity of the analysis for low-level analytes. This work will address a key need for radiochemical analysis of high-level waste using a non-destructive, multi-element, and rapid method in a radiation environment. There is significant potential that this instrumentation could be capable of on-line analysis for process waste stream characterization at DOE sites.

  1. A single dual-emissive nanofluorophore test paper for highly sensitive colorimetry-based quantification of blood glucose.

    Science.gov (United States)

    Huang, Xiaoyan; Zhou, Yujie; Liu, Cui; Zhang, Ruilong; Zhang, Liying; Du, Shuhu; Liu, Bianhua; Han, Ming-Yong; Zhang, Zhongping

    2016-12-15

    Fluorescent test papers are promising for the wide applications in the assays of diagnosis, environments and foods, but unlike classical dye-absorption-based pH test paper, they are usually limited in the qualitative yes/no type of detection by fluorescent brightness, and the colorimetry-based quantification remains a challenging task. Here, we report a single dual-emissive nanofluorophore probe to achieve the consecutive color variations from blue to red for the quantification of blood glucose on its as-prepared test papers. Red quantum dots were embedded into silica nanoparticles as a stable internal standard emission, and blue carbon dots (CDs) were further covalently linked onto the surface of silica, in which the ratiometric fluorescence intensity of blue to red is controlled at 5:1. While the oxidation of glucose induced the formation of Fe(3+) ions, the blue emission of CDs was thus quenched by the electron transfer from CDs to Fe(3+), displaying a serial of consecutive color variations from blue to red with the dosage of glucose. The high-quality test papers printed by the probe ink exhibited a dosage-sensitive allochromatic capability with the clear differentiations of ~5, 7, 9, 11mM glucose in human serum (normal: 3-8mM). The blood glucose determined by the test paper was almost in accordance with that measured by a standard glucometer. The method reported here opens a window to the wide applications of fluorescent test paper in biological assays.

  2. Plasmonic-enhanced two-photon fluorescence with single gold nanoshell

    Science.gov (United States)

    Zhang, TianYue; Lu, GuoWei; Shen, HongMing; Perriat, P.; Martini, M.; Tillement, O.; Gong, QiHuang

    2014-06-01

    Single gold nanoshell with mutilpolar plasmon resonances is proposed to enhance two-photon fluorescence efficiently. The single emitter single nanoshell configuration is studied systematically by employing the finite-difference time-domain method. The emitter located inside or outside the nanoshell at various positions leads to a significantly different enhancement effect. The fluorescent emitter placed outside the nanoshell can achieve large fluorescence intensity given that both the position and orientation of the emission dipole are optimally controlled. In contrast, for the case of the emitter placed inside the nanoshell, it can experience substantial two-photon fluorescence enhancement without strict requirements upon the position and dipole orientations. Metallic nanoshell encapsulating many fluorescent emitters should be a promising nanocomposite configuration for bright two-photon fluorescence label. The results provide a comprehensive understanding about the plasmonic-enhanced two-photon fluorescence behaviors, and the nanocomposite configuration has great potential for optical detecting, imaging and sensing in biological applications.

  3. [Cloning goat producing human lactoferrin with genetically modified donor cells selected by single or dual markers].

    Science.gov (United States)

    An, Liyou; Yuan, Yuguo; Yu, Baoli; Yang, Tingjia; Cheng, Yong

    2012-12-01

    We compared the efficiency of cloning goat using human lactoferrin (hLF) with genetically modified donor cells marked by single (Neo(r)) or double (Neo(r)/GFP) markers. Single marker expression vector (pBLC14) or dual markers expression vector (pAPLM) was delivered to goat fetal fibroblasts (GFF), and then the transgenic GFF was used as donor cells to produce transgenic goats. Respectively, 58.8% (20/34) and 86.7% (26/30) resistant cell lines confirmed the transgenic integration by PCR. Moreover, pAPLM cells lines were subcultured with several passages, only 20% (6/30) cell lines was observed fluorescence from each cell during the cell passage. Somatic cell nuclear transfer using the donor cells harbouring pBLC14 or pAPLM construct, resulting in a total of 806 reconstructed embryos, a pregnancy rate at 35 d (53.8%, 39.1%) and 60 d (26.9%, 21.7%), and an offspring birth rate (1.9%, 1.4%) with 5 and 7 newborn cloned goats, respectively. Transgene was confirmed by PCR and southern-blot in all cloned offspring. There were no significant differences at the reconstructed embryo fusion rates, pregnancy rates and the birth rate (P > 0.05) between single and double markers groups. The Neo(r)/GFP double markers could improve the reliability for accurately and efficiently selecting the genetically modified donor cells. No adverse effect was observed on the efficiency of transgenic goat production by SCNT using somatic cells transfected with double (Neo(r)/GFP) markers vector.

  4. Single molecule localization microscopy of the distribution of chromatin using Hoechst and DAPI fluorescent probes

    OpenAIRE

    Szczurek, Aleksander T; PRAKASH, KIRTI; Lee, Hyun-Keun; Żurek-Biesiada, Dominika J; Best, Gerrit; Hagmann, Martin; Dobrucki, Jurek W; Cremer, Christoph; Birk, Udo

    2014-01-01

    Several approaches have been described to fluorescently label and image DNA and chromatin in situ on the single-molecule level. These superresolution microscopy techniques are based on detecting optically isolated, fluorescently tagged anti-histone antibodies, fluorescently labeled DNA precursor analogs, or fluorescent dyes bound to DNA. Presently they suffer from various drawbacks such as low labeling efficiency or interference with DNA structure. In this report, we demonstrate that DNA mino...

  5. Testing a dual-fluorescence assay to monitor the viability of filamentous cyanobacteria.

    Science.gov (United States)

    Johnson, Tylor J; Hildreth, Michael B; Gu, Liping; Zhou, Ruanbao; Gibbons, William R

    2015-06-01

    Filamentous cyanobacteria are currently being engineered to produce long-chain organic compounds, including 3rd generation biofuels. Because of their filamentous morphology, standard methods to quantify viability (e.g., plate counts) are not possible. This study investigated a dual-fluorescence assay based upon the LIVE/DEAD® BacLight™ Bacterial Viability Kit to quantify the percent viability of filamentous cyanobacteria using a microplate reader in a high throughput 96-well plate format. The manufacturer's protocol calls for an optical density normalization step to equalize the numbers of viable and non-viable cells used to generate calibration curves. Unfortunately, the isopropanol treatment used to generate non-viable cells released a blue pigment that altered absorbance readings of the non-viable cell solution, resulting in an inaccurate calibration curve. Thus we omitted this optical density normalization step, and carefully divided cell cultures into two equal fractions before the isopropanol treatment. While the resulting calibration curves had relatively high correlation coefficients, their use in various experiments resulted in viability estimates ranging from below 0% to far above 100%. We traced this to the apparent inaccuracy of the propidium iodide (PI) dye that was to stain only non-viable cells. Through further analysis via microplate reader, as well as confocal and wide-field epi-fluorescence microscopy, we observed non-specific binding of PI in viable filamentous cyanobacteria. While PI will not work for filamentous cyanobacteria, it is possible that other fluorochrome dyes could be used to selectively stain non-viable cells. This will be essential in future studies for screening mutants and optimizing photobioreactor system performance for filamentous cyanobacteria.

  6. Efficacy and safety of single versus dual antiplatelet therapy for coiling of unruptured aneurysms.

    Science.gov (United States)

    Nishikawa, Yusuke; Satow, Tetsu; Takagi, Toshinori; Murao, Kenichi; Miyamoto, Susumu; Iihara, Koji

    2013-07-01

    Although the efficacy of antiplatelet therapy for coiling of unruptured cerebral aneurysms has been reported, regimens for this therapy are not yet well established. The aim of this retrospective study was to analyze correlations among the modes of antiplatelet use, aneurysmal configuration, coiling methods, and complications to elucidate the optimal antiplatelet therapy for coiling. The study population comprised 154 patients with unruptured aneurysms who underwent coiling with antiplatelet therapy at our institution between 2001 and 2009. The patients were categorized by mode of antiplatelet therapy (single [n = 64] or dual [n = 90]), neck size (wide [n = 80] or narrow [n = 74]), and technique used (simple [n = 42] or adjunctive [n = 112]). The incidences of hemorrhagic/ischemic complications and abnormalities on postprocedural diffusion-weighted magnetic resonance imaging (DWI) in each group were statistically assessed. Hemorrhagic complications occurred in 1 case (1.5%) with single antiplatelet therapy and in 2 cases (2.2%) with dual antiplatelet therapy. Symptomatic ischemic complications occurred in 5 cases (7.8%) with single therapy and in 4 cases (4.4%) with dual therapy. Abnormalities were detected by DWI in 27 cases (42%) with single therapy and in 31 cases (34%) with dual therapy. No significant difference was found between modes of antiplatelet therapy even when the technique used was taken into account. In cases of wide neck, however, there were significant differences in the rate of symptomatic ischemic complications (single, 21.7%; dual, 3.5%; P = .014) and DWI abnormalities (single, 37.8%; dual, 20.9%; P = .048). Our data suggest that dual antiplatelet therapy may better prevent ischemic complications from coiling for wide-necked aneurysms compared with single antiplatelet therapy. Copyright © 2013 National Stroke Association. Published by Elsevier Inc. All rights reserved.

  7. Three-photon fluorescence imaging of melanin with a dual-wedge confocal scanning system

    Science.gov (United States)

    Mega, Yair; Kerimo, Joseph; Robinson, Joseph; Vakili, Ali; Johnson, Nicolette; DiMarzio, Charles

    2012-03-01

    Confocal microscopy can be used as a practical tool in non-invasive applications in medical diagnostics and evaluation. In particular, it is being used for the early detection of skin cancer to identify pathological cellular components and, potentially, replace conventional biopsies. The detection of melanin and its spatial location and distribution plays a crucial role in the detection and evaluation of skin cancer. Our previous work has shown that the visible emission from melanin is strong and can be easily observed with a near-infrared CW laser using low power. This is due to a unique step-wise, (SW) three-photon excitation of melanin. This paper shows that the same SW, 3-photon fluorescence can also be achieved with an inexpensive, continuous-wave laser using a dual-prism scanning system. This demonstrates that the technology could be integrated into a portable confocal microscope for clinical applications. The results presented here are in agreement with images obtained with the larger and more expensive femtosecond laser system used earlier.

  8. Plasmonic antennas and zero mode waveguides to enhance single molecule fluorescence detection and fluorescence correlation spectroscopy towards physiological concentrations

    CERN Document Server

    Punj, Deep; Moparthi, Satish Babu; de Torres, Juan; Grigoriev, Victor; Rigneault, Hervé; Wenger, Jérôme

    2014-01-01

    Single-molecule approaches to biology offer a powerful new vision to elucidate the mechanisms that underpin the functioning of living cells. However, conventional optical single molecule spectroscopy techniques such as F\\"orster fluorescence resonance energy transfer (FRET) or fluorescence correlation spectroscopy (FCS) are limited by diffraction to the nanomolar concentration range, far below the physiological micromolar concentration range where most biological reaction occur. To breach the diffraction limit, zero mode waveguides and plasmonic antennas exploit the surface plasmon resonances to confine and enhance light down to the nanometre scale. The ability of plasmonics to achieve extreme light concentration unlocks an enormous potential to enhance fluorescence detection, FRET and FCS. Single molecule spectroscopy techniques greatly benefit from zero mode waveguides and plasmonic antennas to enter a new dimension of molecular concentration reaching physiological conditions. The application of nano-optics...

  9. Impact of acid and oxidative modifications, single or dual, of sorghum starch on biodegradable films.

    Science.gov (United States)

    Biduski, Bárbara; Silva, Francine Tavares da; Silva, Wyller Max da; Halal, Shanise Lisie de Mello El; Pinto, Vania Zanella; Dias, Alvaro Renato Guerra; Zavareze, Elessandra da Rosa

    2017-01-01

    The objective of this study was to evaluate the effects of acid and oxidation modifications on sorghum starch, as well as the effect of dual modification of starch on the physical, morphological, mechanical, and barrier properties of biodegradable films. The acid modification was performed with 3% lactic acid and the oxidation was performed with 1.5% active chlorine. For dual modification, the acid modification was performed first, followed by oxidation under the same conditions as above. Both films of the oxidized starches, single and dual, had increased stiffness, providing a higher tensile strength and lower elongation when compared to films based on native and single acid modified starches. However, the dual modification increased the water vapor permeability of the films without changing their solubility. The increase in sorghum starch concentration in the filmogenic solution increased the thickness, water vapor permeability, and elongation of the films.

  10. Dual-Fuel Propulsion in Single-Stage Advanced Manned Launch System Vehicle

    Science.gov (United States)

    Lepsch, Roger A., Jr.; Stanley, Douglas O.; Unal, Resit

    1995-01-01

    As part of the United States Advanced Manned Launch System study to determine a follow-on, or complement, to the Space Shuttle, a reusable single-stage-to-orbit concept utilizing dual-fuel rocket propulsion has been examined. Several dual-fuel propulsion concepts were investigated. These include: a separate-engine concept combining Russian RD-170 kerosene-fueled engines with space shuttle main engine-derivative engines: the kerosene- and hydrogen-fueled Russian RD-701 engine; and a dual-fuel, dual-expander engine. Analysis to determine vehicle weight and size characteristics was performed using conceptual-level design techniques. A response-surface methodology for multidisciplinary design was utilized to optimize the dual-fuel vehicles with respect to several important propulsion-system and vehicle design parameters, in order to achieve minimum empty weight. The tools and methods employed in the analysis process are also summarized. In comparison with a reference hydrogen- fueled single-stage vehicle, results showed that the dual-fuel vehicles were from 10 to 30% lower in empty weight for the same payload capability, with the dual-expander engine types showing the greatest potential.

  11. A Starting Point for Fluorescence-Based Single-Molecule Measurements in Biomolecular Research

    Directory of Open Access Journals (Sweden)

    Alexander Gust

    2014-09-01

    Full Text Available Single-molecule fluorescence techniques are ideally suited to provide information about the structure-function-dynamics relationship of a biomolecule as static and dynamic heterogeneity can be easily detected. However, what type of single-molecule fluorescence technique is suited for which kind of biological question and what are the obstacles on the way to a successful single-molecule microscopy experiment? In this review, we provide practical insights into fluorescence-based single-molecule experiments aiming for scientists who wish to take their experiments to the single-molecule level. We especially focus on fluorescence resonance energy transfer (FRET experiments as these are a widely employed tool for the investigation of biomolecular mechanisms. We will guide the reader through the most critical steps that determine the success and quality of diffusion-based confocal and immobilization-based total internal reflection fluorescence microscopy. We discuss the specific chemical and photophysical requirements that make fluorescent dyes suitable for single-molecule fluorescence experiments. Most importantly, we review recently emerged photoprotection systems as well as passivation and immobilization strategies that enable the observation of fluorescently labeled molecules under biocompatible conditions. Moreover, we discuss how the optical single-molecule toolkit has been extended in recent years to capture the physiological complexity of a cell making it even more relevant for biological research.

  12. A starting point for fluorescence-based single-molecule measurements in biomolecular research.

    Science.gov (United States)

    Gust, Alexander; Zander, Adrian; Gietl, Andreas; Holzmeister, Phil; Schulz, Sarah; Lalkens, Birka; Tinnefeld, Philip; Grohmann, Dina

    2014-09-30

    Single-molecule fluorescence techniques are ideally suited to provide information about the structure-function-dynamics relationship of a biomolecule as static and dynamic heterogeneity can be easily detected. However, what type of single-molecule fluorescence technique is suited for which kind of biological question and what are the obstacles on the way to a successful single-molecule microscopy experiment? In this review, we provide practical insights into fluorescence-based single-molecule experiments aiming for scientists who wish to take their experiments to the single-molecule level. We especially focus on fluorescence resonance energy transfer (FRET) experiments as these are a widely employed tool for the investigation of biomolecular mechanisms. We will guide the reader through the most critical steps that determine the success and quality of diffusion-based confocal and immobilization-based total internal reflection fluorescence microscopy. We discuss the specific chemical and photophysical requirements that make fluorescent dyes suitable for single-molecule fluorescence experiments. Most importantly, we review recently emerged photoprotection systems as well as passivation and immobilization strategies that enable the observation of fluorescently labeled molecules under biocompatible conditions. Moreover, we discuss how the optical single-molecule toolkit has been extended in recent years to capture the physiological complexity of a cell making it even more relevant for biological research.

  13. Single aflatoxin contaminated corn kernel analysis with fluorescence hyperspectral image

    Science.gov (United States)

    Yao, Haibo; Hruska, Zuzana; Kincaid, Russell; Ononye, Ambrose; Brown, Robert L.; Cleveland, Thomas E.

    2010-04-01

    Aflatoxins are toxic secondary metabolites of the fungi Aspergillus flavus and Aspergillus parasiticus, among others. Aflatoxin contaminated corn is toxic to domestic animals when ingested in feed and is a known carcinogen associated with liver and lung cancer in humans. Consequently, aflatoxin levels in food and feed are regulated by the Food and Drug Administration (FDA) in the US, allowing 20 ppb (parts per billion) limits in food and 100 ppb in feed for interstate commerce. Currently, aflatoxin detection and quantification methods are based on analytical tests including thin-layer chromatography (TCL) and high performance liquid chromatography (HPLC). These analytical tests require the destruction of samples, and are costly and time consuming. Thus, the ability to detect aflatoxin in a rapid, nondestructive way is crucial to the grain industry, particularly to corn industry. Hyperspectral imaging technology offers a non-invasive approach toward screening for food safety inspection and quality control based on its spectral signature. The focus of this paper is to classify aflatoxin contaminated single corn kernels using fluorescence hyperspectral imagery. Field inoculated corn kernels were used in the study. Contaminated and control kernels under long wavelength ultraviolet excitation were imaged using a visible near-infrared (VNIR) hyperspectral camera. The imaged kernels were chemically analyzed to provide reference information for image analysis. This paper describes a procedure to process corn kernels located in different images for statistical training and classification. Two classification algorithms, Maximum Likelihood and Binary Encoding, were used to classify each corn kernel into "control" or "contaminated" through pixel classification. The Binary Encoding approach had a slightly better performance with accuracy equals to 87% or 88% when 20 ppb or 100 ppb was used as classification threshold, respectively.

  14. Composite nanoparticle of Au and quantum dots for X-ray computed tomography and fluorescence dual-mode imaging in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Song, Ji-Tao; Yang, Xiao-Quan; Zhang, Xiao-Shuai; Yan, Dong-Mei; Yao, Ming-Hao; Qin, Meng-Yao; Zhao, Yuan-Di, E-mail: zydi@mail.hust.edu.cn [Huazhong University of Science and Technology, Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics - Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology (China)

    2015-12-15

    In this study, composite nanoparticles comprising Au nanoparticle and quantum dots were built and used for contrast-enhanced computed tomography imaging (CT) and fluorescence dual-mode imaging in vivo. The nanoparticle exhibited good monodispersity and good biocompatibility, and had excellent CT contrast-enhancement effect and fluorescence imaging capability. They were appropriate for being used as dual-mode imaging probe in vivo.

  15. Multiplexing PKA and ERK1&2 kinases FRET biosensors in living cells using single excitation wavelength dual colour FLIM

    Science.gov (United States)

    Demeautis, Claire; Sipieter, François; Roul, Julien; Chapuis, Catherine; Padilla-Parra, Sergi; Riquet, Franck B.; Tramier, Marc

    2017-01-01

    Monitoring of different signalling enzymes in a single assay using multiplex biosensing provides a multidimensional workspace to elucidate biological processes, signalling pathway crosstalk, and determine precise sequence of events at the single living cell level. In this study, we interrogate the complexity in cAMP/PKA-MAPK/ERK1&2 crosstalk by using multi-parameter biosensing experiments to correlate biochemical activities simultaneously in time and space. Using a single excitation wavelength dual colour FLIM method we are able to detect fluorescence lifetime images of two donors to simultaneously measure PKA and ERK1&2 kinase activities in the same cellular localization by using FRET biosensors. To this end, we excite two FRET donors mTFP1 and LSSmOrange with a 440 nm wavelength and we alleviate spectral bleed-through associated limitations with the very dim-fluorescent acceptor ShadowG for mTFP1 and the red-shifted mKate2 for LSSmOrange. The simultaneous recording of PKA and ERK1&2 kinase activities reveals concomitant EGF-mediated activations of both kinases in HeLa cells. Under these conditions the subsequent Forskolin-induced cAMP release reverses the transient increase of EGF-mediated ERK1&2 kinase activity while reinforcing PKA activation. Here we propose a validated methodology for multiparametric kinase biosensing in living cells using FRET-FLIM. PMID:28106114

  16. Iodinated silica/porphyrin hybrid nanoparticles for X-ray computed tomography/fluorescence dual-modal imaging of tumors

    Directory of Open Access Journals (Sweden)

    Koichiro Hayashi

    2014-12-01

    Full Text Available Silica nanoparticles containing covalently linked iodine and a near-infrared (NIR fluorescence dye, namely porphyrin, have been synthesized through a one-pot sol–gel reaction. These particles are called iodinated silica/porphyrin hybrid nanoparticles (ISP HNPs. The ISP HNPs have both high X-ray absorption coefficient and NIR fluorescence. The ISP HNPs modified with folic acid (FA and polyethylene glycol (PEG, denoted as FA-PEG-ISP HNPs, enabled the successful visualization of tumors in mice by both X-ray computed tomography (CT and fluorescence imaging (FI. Thus, the FA-PEG-ISP HNPs are useful as contrast agents or probes for CT/FI dual-modal imaging.

  17. Polypyrrole-based nanotheranostics for activatable fluorescence imaging and chemo/photothermal dual therapy of triple-negative breast cancer

    Science.gov (United States)

    Park, Dongjin; Ahn, Kyung-Ohk; Jeong, Kyung-Chae; Choi, Yongdoo

    2016-05-01

    Here, we fabricated polypyrrole nanoparticles (PPys) (termed HA10-PPy, HA20-PPy, and HA40-PPy) doped with different average molecular weight hyaluronic acids (HAs) (10, 20, and 40 kDa, respectively), and evaluated the effect of molecular weight of doped HA on photothermal induction, fluorescence quenching, and drug loading efficiencies. Doxorubicin-loaded HA-doped PPys (DOX@HA-PPys) could be used for imaging and therapy of triple-negative breast cancer (TNBC). Fluorescence turn-on, stimuli-responsive drug release, and photo-induced heating of DOX@HA-PPys enabled not only activatable fluorescence imaging but also subsequent chemo/photothermal dual therapy for TNBC. In particular, we illustrated the potential usefulness of the photothermal effect of the nanoparticles for overcoming chemoresistance in TNBC.

  18. Potential of protoporphyrin IX and metal derivatives for single molecule fluorescence studies

    Energy Technology Data Exchange (ETDEWEB)

    Hu Yi; Geissinger, Peter [Department of Chemistry and Biochemistry, Laboratory for Surface Studies, University of Wisconsin-Milwaukee, 3210 N. Cramer Street, Milwaukee, WI 53211 (United States); Woehl, Joerg C., E-mail: woehl@uwm.ed [Department of Chemistry and Biochemistry, Laboratory for Surface Studies, University of Wisconsin-Milwaukee, 3210 N. Cramer Street, Milwaukee, WI 53211 (United States)

    2011-03-15

    Metalloporphyrins are cofactors of a variety of proteins, and are often used as spectroscopic probes of the active site. Many high resolution techniques, such as single molecule spectroscopy, are based on fluorescence contrast and require the replacement of the native metalloporphyrin by a fluorescent analog. We have investigated the potential of several fluorescent analogs of heme, namely free-base protoporphyrin IX and its metal derivatives containing Zn, Sn, and Mg, for single molecule fluorescence studies by determining their room-temperature molecular absorption cross sections and fluorescence quantum yields. According to these data, free-base protoporphyrin IX and its Zn derivative, which have the highest fluorescence quantum yields, are the most suitable heme analogs for single molecule fluorescence studies. - Research highlights: Protoporphyrin IX and fluorescent metal derivatives for single molecule detection. Measurement of room temperature absorption cross sections for Q bands. Measurement of room temperature fluorescence quantum yields for Q bands. PPIX and Zn derivative have highest quantum yields for lowest-energy transition.

  19. Dual-band Fabry-Perot lasing from single ZnO microbelt

    Science.gov (United States)

    Zhu, Qiuxiang; Qin, Feifei; Lu, Junfeng; Zhu, Zhu; Shi, Zengliang; Xu, Chunxiang

    2016-10-01

    Dual-band semiconductor microbelt lasing are promising for multifunctional applications ranging from optical communication to spectroscopy analysis. Here, we demonstrated a dual-band Fabry-Perot (F-P) lasing from both length and width directions in a single ZnO microbelt. The lasing performance, spectral variation and mode structure significantly depended on the cavity size, which corresponded to the length and width of the ZnO microbelts. The resonant process and mechanism were investigated systematically through the experimental analysis and numerically FDTD simulation. The results of the dual-band F-P lasing modes and wide lasing wavelength are helpful to design the dual-wavelength electronic and optoelectronic devices.

  20. Single-layer dual-band terahertz filter with weak coupling between two neighboring cross slots

    Institute of Scientific and Technical Information of China (English)

    亓丽梅; 李超; 方广有; 李士超

    2015-01-01

    A dual-band terahertz (THz) filter consisting of two different cross slots is designed and fabricated in a single molyb-denum layer. Experimental verification by THz time-domain spectroscopy indicates good agreement with the simulation results. Owing to the weak coupling between the two neighboring cross slots in the unit cell, good selectivity performance can be easily achieved, both in the lower and higher bands, by tuning the dimensions of the two crosses. The physical mechanisms of the dual-band resonant are clarified by using three differently configured filters and electric field distribu-tion diagrams. Owing to the rotational symmetry of the cross-shaped filter, the radiation at normal incidence is insensitive to polarization. Compared with the THz dual-band filters that were reported earlier, these filters also have the advantages of easy fabrication and low cost, which would find applications in dual-band sensors, THz communication systems, and emerging THz technologies.

  1. Dual-Mode Encoded Magnetic Composite Microsphere Based on Fluorescence Reporters and Raman Probes as Covert Tag for Anticounterfeiting Applications.

    Science.gov (United States)

    Li, Ruimin; Zhang, Yuting; Tan, Jing; Wan, Jiaxun; Guo, Jia; Wang, Changchun

    2016-04-13

    Utilizing fluorescence reporters and SERS probes as the security labels, a series of dual-mode encoded magnetic composite microspheres with micrometer size was designed and prepared for anticounterfeiting applications. At first, the micro-meter-sized melamine formaldehyde microspheres with different fluorescence molecules (FMF) were prepared by precipitation polymerization, and then the magnetite composite microspheres (FMF/MNPs) were fabricated by direct immobilization of magnetic nanoparticles (MNPs) onto the surface of FMF microspheres. After deposition of Ag nanoparticles (Ag-NPs) onto FMF/MNPs microspheres, the SERS probes were absorbed onto the surface of Ag-NPs, and then a protection layer of silica was coated on the composite microspheres by Stöber method. The combination of different fluorescence reporters and SERS probes greatly increased the encoding complexity and volume for high-level anticounterfeiting. The structure of the dual-encoded FMF/MNPs/Ag-NPs/SiO2 composite microspheres was characterized by FESEM, TEM, FLS(fluorescence spectrometer), XRD, VSM, UV-vis and EDS. The embedded magnetic nanoparticles enable the composite microspheres to be quickly isolated from the marked latex paint by magnet at the concentration of as low as 1 ppm, and the covert tag information can be read out even from one composite microsphere. In addition, the covert security information in the marked coating film can be also read out in situ and the existence of the composite microspheres does not influence the visible appearance of the coating film. All the above outstanding properties will make these dual-mode encoded composite microspheres as advanced security tags for next-generation anticounterfeiting applications.

  2. Photonic mode density effects on single-molecule fluorescence blinking

    CERN Document Server

    Stefani, F D; Bocchio, N; Gaul, F; Pomozzi, A; Kreiter, M

    2006-01-01

    We investigated the influence of the photonic mode density (PMD) on the triplet dynamics of individual chromophores on a dielectric interface by comparing their response in the presence and absence of a nearby gold film. Lifetimes of the excited singlet state were evaluated in ordet to measure directly the PMD at the molecules position. Triplet state lifetimes were simultaneously determined by statistical analysis of the detection time of the fluorescence photons. The observed singlet decay rates are in agreement with the predicted PMD for molecules with different orientations. The triplet decay rate is modified in a fashion correlated to the singlet decay rate. These results show that PMD engineering can lead to an important suppression of the fluorescence, introducing a novel aspect of the physical mechanism to enhance fluorescence intensity in PMD-enhancing systems such as plasmonic devices.

  3. Single-Walled Carbon Nanotubes as Fluorescence Biosensors for Pathogen Recognition in Water Systems

    Directory of Open Access Journals (Sweden)

    Venkata K. K. Upadhyayula

    2008-01-01

    Full Text Available The possibility of using single-walled carbon nanotubes (SWCNTs aggregates as fluorescence sensors for pathogen recognition in drinking water treatment applications has been studied. Batch adsorption study is conducted to adsorb large concentrations of Staphylococcus aureus aureus SH 1000 and Escherichia coli pKV-11 on single-walled carbon nanotubes. Subsequently the immobilized bacteria are detected with confocal microscopy by coating the nanotubes with fluorescence emitting antibodies. The Freundlich adsorption equilibrium constant (k for S.aureus and E.coli determined from batch adsorption study was found to be 9×108 and 2×108 ml/g, respectively. The visualization of bacterial cells adsorbed on fluorescently modified carbon nanotubes is also clearly seen. The results indicate that hydrophobic single-walled carbon nanotubes have excellent bacterial adsorption capacity and fluorescent detection capability. This is an important advancement in designing fluorescence biosensors for pathogen recognition in water systems.

  4. Single molecule fluorescence fluctuations of the cyanine dyes linked covalently to DNA

    Institute of Scientific and Technical Information of China (English)

    AUMILER; Damir

    2009-01-01

    The intersystem crossing and isomerization dynamics of free-Cy3,Cy3-ssDNA,free-Cy5 and Cy5-ssDNA are obtained through simple analysis of rapid on/off blinking from single molecule fluorescence intensity time-traces and the fluorescence correlation spectroscopy(FCS).The on-and off-times observed in fluorescence time traces of single cyanine dyes are due to the formation of the triplet state and isomerization,where both the interaction with DNA and long central polymethine chain of cyanine dyes increase the barriers of isomerization,leading to long off-time.The results indicate that the single molecule fluorescence fluctuation together with the resulting second autocorrelation analysis are powerful methods for determining the triplet state and isomerization dynamics,which could be the simple techniques and complementary to other spectroscopic techniques,such as fluorescence decay measurement and laser flash photolysis to study the photophysical processes of complex molecules.

  5. Dual in vivo Photoacoustic and Fluorescence Imaging of HER2 Expression in Breast Tumors for Diagnosis, Margin Assessment, and Surgical Guidance

    Directory of Open Access Journals (Sweden)

    Azusa Maeda

    2015-01-01

    Full Text Available Biomarker-specific imaging probes offer ways to improve molecular diagnosis, intraoperative margin assessment, and tumor resection. Fluorescence and photoacoustic imaging probes are of particular interest for clinical applications because the combination enables deeper tissue penetration for tumor detection while maintaining imaging sensitivity compared to a single optical imaging modality. Here we describe the development of a human epidermal growth factor receptor 2 (HER2-targeting imaging probe to visualize differential levels of HER2 expression in a breast cancer model. Specifically, we labeled trastuzumab with Black Hole Quencher 3 (BHQ3 and fluorescein for photoacoustic and fluorescence imaging of HER2 overexpression, respectively. The dual-labeled trastuzumab was tested for its ability to detect HER2 overexpression in vitro and in vivo. We demonstrated an over twofold increase in the signal intensity for HER2-overexpressing tumors in vivo, compared to low–HER2-expressing tumors, using photoacoustic imaging. Furthermore, we demonstrated the feasibility of detecting tumors and positive surgical margins by fluorescence imaging. These results suggest that multimodal HER2-specific imaging of breast cancer using the BHQ3-fluorescein trastuzumab enables molecular-level detection and surgical margin assessment of breast tumors in vivo. This technique may have future clinical impact for primary lesion detection, as well as intraoperative molecular-level surgical guidance in breast cancer.

  6. Probing single processive molecular motors with high-speed optical tweezers and fluorescence microscopy

    Science.gov (United States)

    Gardini, L.; Pavone, F. S.; Capitanio, M.

    2017-02-01

    Here we present development of optical techniques for the study of single processive myosin motors based on the combination of high-speed optical tweezers force spectroscopy and single molecule fluorescence imaging. Ultrafast force-clamp spectroscopy1 is applied to study the dependence of single chemo-mechanical steps of processive myosin motors on the applied load. On the other hand, single molecule localization through FIONA (Fluorescence Imaging with One Nanometer Accuracy)2, 3 is applied to in vitro motility assay to measure parameters such as the runlength, velocity and step size of single myosin V motors, labeled with Quantum Dots, under unloaded conditions.

  7. Fluorescence preselection of bioaerosol for single-particle mass spectrometry

    NARCIS (Netherlands)

    Stowers, M.A.; Van Wuijckhuijse, A.L.; Marijnissen, J.C.M.; Kientz, C.E.; Ciach, T.

    2006-01-01

    We have designed, constructed, and tested a system that preselects the biological fraction of airborne particles from the overall aerosol. The preselection is based on fluorescence emission excited by a continuous 266 nm laser beam. This beam is one of two cw beams used to measure the aerodynamic pa

  8. Dual collection mode optical microscope with single-pixel detection

    Science.gov (United States)

    Rodríguez, A. D.; Clemente, P.; Fernández-Alonso, Mercedes; Tajahuerce, E.; Lancis, J.

    2015-07-01

    In this work we have developed a single-pixel optical microscope that provides both re ection and transmission images of the sample under test by attaching a diamond pixel layout DMD to a commercial inverted microscope. Our system performs simultaneous measurements of re ection and transmission modes. Besides, in contrast with a conventional system, in our single-element detection system both images belong, unequivocally, to the same plane of the sample. Furthermore, we have designed an algorithm to modify the shape of the projected patterns that improves the resolution and prevents the artifacts produced by the diamond pixel architecture.

  9. General Calibration Algorithm for Single-transmitting-dual-receiving Polarimetric SAR System

    Directory of Open Access Journals (Sweden)

    Chen Lin

    2012-09-01

    Full Text Available The single-transmitting-dual-receiving polarimetric SAR system has only two receving channels, which induces lack of prior information for calibration. Due to the polarization diversity of this kind of system (which operates on different dual and compact polarimetric modes, there has not a general algorithm currently. In this paper, a new general calibration algorithm is proposed for single-transmitting-dual-receiving polarimetric SAR system, which can be widely applied to diverse polarization modes. The transmitting and receiving distortion can be estimated using trihedral, 0° dihedral and 45° dihedral as ideal point targets without any assumption on the scene and system. The basic principle of this new algorithm is induced theoretically. The effect of calibrator error on the distortion estimation is analyzed by simulation. Point targets’calibration results and polarization signatures verify this algorithm.

  10. A dual-responsive fluorescence method for the detection of clenbuterol based on BSA-protected gold nanoclusters.

    Science.gov (United States)

    Cao, Xueling; Li, Hongwei; Lian, Lili; Xu, Na; Lou, Dawei; Wu, Yuqing

    2015-04-29

    The illegal feeding of clenbuterol (CLB) to domestic animals and the potential harm of it to human health lead an urgent requirement for the efficient detection of CLB, especially in the edible meat. In this paper we reported a new fluorescence method for the detection of trace amount of CLB by using the BSA-protected gold nanoclusters (AuNCs@BSA). Under the excitation of either 280 or 500 nm the emission of AuNCs@BSA was quenched obviously by diazotized CLB, supplying a dual-responsive fluorescence method to detect CLB in aqueous solution. In addition, the linear response of the fluorescence intensity of AuNCs@BSA to diazotized CLB allowed the quantitative detection of CLB in a range of 4.0 nM-300 μM upon excitation at two wavelength, and the limit of detection for CLB was 3.0 nM upon 280 nm excitation and 1.6 nM upon 500 nm excitation, respectively. In addition, the dual-responsive mechanism of AuNCs@BSA to CLB was investigated in detail by using several CLB analogues and reference compounds. Particularly, the proposed method was successfully applied to detect CLB in pork mince and the results were validated well by HPLC, illustrating it could be used as a reliable, rapid, and cost-effective technique for the determination of CLB residues in real samples.

  11. In situ, dual-mode monitoring of organ-on-a-chip with smartphone-based fluorescence microscope.

    Science.gov (United States)

    Cho, Soohee; Islas-Robles, Argel; Nicolini, Ariana M; Monks, Terrence J; Yoon, Jeong-Yeol

    2016-12-15

    The use of organ-on-a-chip (OOC) platforms enables improved simulation of the human kidney's response to nephrotoxic drugs. The standard method of analyzing nephrotoxicity from existing OOC has majorly consisted of invasively collecting samples (cells, lysates, media, etc.) from an OOC. Such disruptive analyses potentiate contamination, disrupt the replicated in vivo environment, and require expertize to execute. Moreover, traditional analyses, including immunofluorescence microscopy, immunoblot, and microplate immunoassay are essentially not in situ and require substantial time, resources, and costs. In the present work, the incorporation of fluorescence nanoparticle immunocapture/immunoagglutination assay into an OOC enabled dual-mode monitoring of drug-induced nephrotoxicity in situ. A smartphone-based fluorescence microscope was fabricated as a handheld in situ monitoring device attached to an OOC. Both the presence of γ-glutamyl transpeptidase (GGT) on the apical brush-border membrane of 786-O proximal tubule cells within the OOC surface, and the release of GGT to the outflow of the OOC were evaluated with the fluorescence scatter detection of captured and immunoagglutinated anti-GGT conjugated nanoparticles. This dual-mode assay method provides a novel groundbreaking tool to enable the internal and external in situ monitoring of the OOC, which may be integrated into any existing OOCs to facilitate their subsequent analyses. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. A dual mode targeting probe for distinguishing HER2-positive breast cancer cells using silica-coated fluorescent magnetic nanoparticles

    Science.gov (United States)

    Li, Jia; An, Yan-Li; Zang, Feng-Chao; Zong, Shen-Fei; Cui, Yi-Ping; Teng, Gao-Jun

    2013-10-01

    We report a composite nanoprobe based on silica-coated magnetic nanoparticles (NPs) for distinguishing breast cancers at different HER2 statuses. The nanoprobe has a core-shell structure, with Fe3O4 NPs as the magnetic core and dye-embedded silica as the fluorescent shell, whose average size is about 150 nm. Besides, the outmost surfaces of the probes were modified with specific antibodies to endow the probe with a targeting ability. With such a structure, the nanoprobe can accomplish dual mode targeting of human breast cancer cells based on fluorescence and magnetic resonance imaging (MRI). In the experiments, three human breast cancer cell lines were used to test the targeting ability of the nanoprobe. Specifically, SKBR3 cells with a high HER2 expression level were used as the model target cells, while MCF7 cells with a lower HER2 expression levels and HER2-negative MDA-MB-231 cells were used as the controls. Both the fluorescence and MRI imaging results confirmed that the nanoprobe can distinguish three cancer cell lines with different HER2 expression levels. With the dual mode imaging and specific targeting properties, we anticipate that the presented nanoprobe may have a great potential in the diagnosis and treatment of cancerous diseases.

  13. Single versus Dual Paycheck: Married Parents' Attitudes about Maternal Employment.

    Science.gov (United States)

    Ryckebusch, Jenna-Lyn; Miller, Heather; Fulmer, Kimberly; Fontanez, Mary; Ellis, Trisha; DiBlasi, Francis Paul; Carey, Brandi; Chambliss, Catherine

    This study examines attitudes about maternal employment by comparing the responses of married parents from single versus two-paycheck families. Participants in this study were 138 mothers and 120 fathers given the Beliefs About the Consequences of Maternal Employment for Children Scale (BACMEC), which assesses views about maternal employment.…

  14. Synthesis of single walled carbon nanotubes by dual laser vaporization

    CSIR Research Space (South Africa)

    Moodley, MK et al.

    2006-02-27

    Full Text Available Single-walled carbon nanotubes were synthesised by the laser vaporisation of graphite composite targets in a tube furnace. Two pulsed Nd:YAG lasers operating at fundamental (1 064 nm) and 2nd harmonic (532 nm) were combined, focused and evaporated...

  15. Homogeneous vs heterogeneous polymerization catalysis revealed by single-particle fluorescence microscopy.

    Science.gov (United States)

    Esfandiari, N Melody; Blum, Suzanne A

    2011-11-16

    A high-sensitivity and high-resolution single-particle fluorescence microscopy technique differentiated between homogeneous and heterogeneous metathesis polymerization catalysis by imaging the location of the early stages of polymerization. By imaging single polymers and single crystals of Grubbs II, polymerization catalysis was revealed to be solely homogeneous rather than heterogeneous or both.

  16. Single-frequency, dual-GNSS versus dual-frequency, single-GNSS: a low-cost and high-grade receivers GPS-BDS RTK analysis

    Science.gov (United States)

    Odolinski, Robert; Teunissen, Peter J. G.

    2016-11-01

    The concept of single-frequency, dual-system (SF-DS) real-time kinematic (RTK) positioning has become feasible since, for instance, the Chinese BeiDou Navigation Satellite System (BDS) has become operational in the Asia-Pacific region. The goal of the present contribution is to investigate the single-epoch RTK performance of such a dual-system and compare it to a dual-frequency, single-system (DF-SS). As the SF-DS we investigate the L1 GPS + B1 BDS model, and for DF-SS we take L1, L2 GPS and B1, B2 BDS, respectively. Two different locations in the Asia-Pacific region are analysed with varying visibility of the BDS constellation, namely Perth in Australia and Dunedin in New Zealand. To emphasize the benefits of such a model we also look into using low-cost ublox single-frequency receivers and compare such SF-DS RTK performance to that of a DF-SS, based on much more expensive survey-grade receivers. In this contribution a formal and empirical analysis is given. It will be shown that with the SF-DS higher elevation cut-off angles than the conventional 10° or 15° can be used. The experiment with low-cost receivers for the SF-DS reveals (for the first time) that it has the potential to achieve comparable ambiguity resolution performance to that of a DF-SS (L1, L2 GPS), based on the survey-grade receivers.

  17. Contrasting single and multi-component working-memory systems in dual tasking

    NARCIS (Netherlands)

    Nijboer, Menno; Borst, Jelmer; van Rijn, Hedderik; Taatgen, Niels

    2016-01-01

    Working memory can be a major source of interference in dual tasking. However, there is no consensus on whether this interference is the result of a single working memory bottleneck, or of interactions between different working memory components that together form a complete working-memory system.

  18. Shock efficacy of single and dual coil electrodes-new insights from the NORDIC ICD Trial

    DEFF Research Database (Denmark)

    Bänsch, Dietmar; Bonnemeier, Hendrik; Brandt, Johan

    2017-01-01

    Aims: Dual coil (DC) electrodes are preferred to single coil (SC) electrodes because of an assumed higher shock efficacy. However, DC-electrodes may be associated with an increased difficulty and risk of lead extraction. We aimed to compare SC- and DC-electrodes with respect to the first shock ef...

  19. 14 CFR 105.43 - Use of single-harness, dual-parachute systems.

    Science.gov (United States)

    2010-01-01

    ... making the next jump with that parachute, or a non-certificated person under the direct supervision of a... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Use of single-harness, dual-parachute... TRANSPORTATION (CONTINUED) AIR TRAFFIC AND GENERAL OPERATING RULES PARACHUTE OPERATIONS Parachute Equipment...

  20. Ultrasound-guided single-penetration dual-injection block for leg and foot surgery

    DEFF Research Database (Denmark)

    Børglum, Jens; Johansen, Karina; Christensen, Karen Margrethe;

    2014-01-01

    We describe a new approach to blocking the sciatic and saphenous nerves in the proximal thigh (level of the lesser trochanter or immediately below) using a single-penetration dual-injection (SPEDI) technique. The popliteal-sciatic approach necessitates repositioning of the leg exposing...

  1. Change detection in quad and dual pol, single- and bi-frequency SAR data

    DEFF Research Database (Denmark)

    Nielsen, Allan Aasbjerg; Conradsen, Knut; Skriver, Henning

    2015-01-01

    -value are given. In a case study airborne EMISAR C- and L-band SAR images covering agricultural fields and wooded areas near Foulum, Denmark, are used in single- and bi-frequency, bi-temporal change detection with full and dual polarimetry data. © (2015) COPYRIGHT Society of Photo-Optical Instrumentation...

  2. Contrasting single and multi-component working-memory systems in dual tasking

    NARCIS (Netherlands)

    Nijboer, Menno; Borst, Jelmer; van Rijn, Hedderik; Taatgen, Niels

    2016-01-01

    Working memory can be a major source of interference in dual tasking. However, there is no consensus on whether this interference is the result of a single working memory bottleneck, or of interactions between different working memory components that together form a complete working-memory system. W

  3. protoDUNE-Single Phase and protDUNE-DualPhase

    CERN Multimedia

    Brice, Maximilien

    2016-01-01

    At the EHN1 two big 8m x 8m x8m detector prototypes (protoDUNE-Single Phase and protDUNE-DualPhase) are being constructed. The aim is to test technologies and detector performances for DUNE, a new generation of LBN neutr

  4. BRONCHIAL LAVAGE AND BRONCHOALVEOLAR LAVAGE IN ALLERGEN-INDUCED SINGLE EARLY AND DUAL ASTHMATIC RESPONDERS

    NARCIS (Netherlands)

    AALBERS, R; KAUFFMAN, HF; VRUGT, B; SMITH, M; KOETER, GH; TIMENS, W; DEMONCHY, JGR

    1993-01-01

    The phenotypic cellular profile of bronchial lavage (BL) and bronchoalveolar lavage (BAL) was studied in 7 single early (SR) and 10 dual asthmatic responders (DR). Lavage was performed, after previously having determined bronchial hyperresponsiveness to histamine and the response to house dust mite

  5. BRONCHIAL LAVAGE AND BRONCHOALVEOLAR LAVAGE IN ALLERGEN-INDUCED SINGLE EARLY AND DUAL ASTHMATIC RESPONDERS

    NARCIS (Netherlands)

    AALBERS, R; KAUFFMAN, HF; VRUGT, B; SMITH, M; KOETER, GH; TIMENS, W; DEMONCHY, JGR

    The phenotypic cellular profile of bronchial lavage (BL) and bronchoalveolar lavage (BAL) was studied in 7 single early (SR) and 10 dual asthmatic responders (DR). Lavage was performed, after previously having determined bronchial hyperresponsiveness to histamine and the response to house dust mite

  6. Room-temperature single-photon sources based on nanocrystal fluorescence in photonic/plasmonic nanostructures

    Science.gov (United States)

    Lukishova, S. G.; Winkler, J. M.; Bissell, L. J.; Mihaylova, D.; Liapis, Andreas C.; Shi, Z.; Goldberg, D.; Menon, V. M.; Boyd, R. W.; Chen, G.; Prasad, P.

    2014-10-01

    Results are presented here towards robust room-temperature SPSs based on fluorescence in nanocrystals: colloidal quantum dots, color-center diamonds and doped with trivalent rare-earth ions (TR3+). We used cholesteric chiral photonic bandgap and Bragg-reflector microcavities for single emitter fluorescence enhancement. We also developed plasmonic bowtie nanoantennas and 2D-Si-photonic bandgap microcavities. The paper also provides short outlines of other technologies for room-temperature single-photon sources.

  7. Detecting pad-wafer contact in CMP using dual emission laser induced fluorescence

    Science.gov (United States)

    Gray, Caprice

    Chemical Mechanical Polishing (CMP) is a technique used for planarizing a wide range of surfaces, including metal and dielectric materials, during the manufacturing of integrated circuits (ICs). Material removal in CMP occurs with the combind chemical and mechanical action of the slurry and polishing pad. For inter-layer dielectric (ILD) polishing, particle-wafer contact is believed responsible for planarization. The particles sit atop the polishing pad asperities, meaning removal rate should correlate with pad-wafer contact area. In this thesis, we presents in-situ measurements of slurry layer thickness and pad-wafer contact using Dual Emission Laser Induced Fluorescence (DELIF). The DELIF technique for measuring the thickness of thin fluid films has been adapted to make instantaneous measurements of slurry thickness in CMP at high spatial resolution (2.9 mum per pixel). Modeling and calibration experiments confirm that there is a linear relationship between CMP DELIF image intensity and fluid layer thickness over a range of 0 to 133 mum. The slurry thickness in the contacting portions of a Cabot Microelectronics D100 polishing pad is 0-60 mum for polishing speeds ≤ 0.62 m/s and applied wafer load ≤ 1.7 psi. As the polishing pad becomes fully conditioned, the slurry layer thickness approaches a saturation point. The time to saturation is approximately 1 hour. DELIF images of the slurry thickness can be analyzed for contact by plotting the image intensity histogram. This histogram is representative of the pad asperity height distribution. When the pad is in contact with the wafer, the asperity peaks flatten and give rise to a secondary peak in the low intensity extreme of the distribution representative of contact. Data simulations show that the secondary peak is sharp at low noise:signal ratio (<10% noise), and is gradually smoothed as more noise is added increasing measurement errors. Even though DELIF is a high noise measurement technique (≥ 15

  8. Single- and two-photon fluorescence recovery after photobleaching.

    Science.gov (United States)

    Sullivan, Kelley D; Majewska, Ania K; Brown, Edward B

    2015-01-05

    Fluorescence recovery after photobleaching (FRAP) is a microscopy technique for measuring the kinetics of fluorescently labeled molecules and can be applied both in vitro and in vivo for two- and three-dimensional systems. This introduction discusses the three basic FRAP methods: traditional FRAP, multiphoton FRAP (MPFRAP), and FRAP with spatial Fourier analysis (SFA-FRAP). Each discussion is accompanied by a description of the mathematical analysis appropriate for situations in which the recovery kinetics is dictated by free diffusion. In some experiments, the recovery kinetics is dictated by the boundary conditions of the system, and FRAP is then used to quantify the connectivity of various compartments. Because the appropriate mathematical analysis is independent of the bleaching method, the analysis of compartmental connectivity is discussed last, in a separate section.

  9. Platinum plasmonic nanostructure arrays for massively parallel single-molecule detection based on enhanced fluorescence measurements.

    Science.gov (United States)

    Saito, Toshiro; Takahashi, Satoshi; Obara, Takayuki; Itabashi, Naoshi; Imai, Kazumichi

    2011-11-04

    We fabricated platinum bowtie nanostructure arrays producing fluorescence enhancement and evaluated their performance using two-photon photoluminescence and single-molecule fluorescence measurements. A comprehensive selection of suitable materials was explored by electromagnetic simulation and Pt was chosen as the plasmonic material for visible light excitation near 500 nm, which is preferable for multicolor dye-labeling applications like DNA sequencing. The observation of bright photoluminescence (λ = 500-600 nm) from each Pt nanostructure, induced by irradiation at 800 nm with a femtosecond laser pulse, clearly indicates that a highly enhanced local field is created near the Pt nanostructure. The attachment of a single dye molecule was attempted between the Pt triangles of each nanostructure by using selective immobilization chemistry. The fluorescence intensities of the single dye molecule localized on the nanostructures were measured. A highly enhanced fluorescence, which was increased by a factor of 30, was observed. The two-photon photoluminescence intensity and fluorescence intensity showed qualitatively consistent gap size dependence. However, the average fluorescence enhancement factor was rather repressed even in the nanostructure with the smallest gap size compared to the large growth of photoluminescence. The variation of the position of the dye molecule attached to the nanostructure may influence the wide distribution of the fluorescence enhancement factor and cause the rather small average value of the fluorescence enhancement factor.

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

    Science.gov (United States)

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

    2007-03-01

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

  11. A fluorescence-Raman dual-imaging platform based on complexes of conjugated polymers and carbon nanotubes.

    Science.gov (United States)

    Liu, Yun; Huang, Jun; Sun, Min-Jie; Yu, Ji-Cheng; Chen, Yu-Lei; Zhang, Yu-Qi; Jiang, Shao-Jun; Shen, Qun-Dong

    2014-01-01

    The present study describes a flexible nanoplatform based on electrostatic assembly of conjugated polyelectrolytes (CPEs) and carboxylated multi-walled carbon nanotubes (cMWNTs). It is demonstrated that the obtained nanocomposites inherit intrinsic optical properties of CPEs and characteristic Raman vibration modes of MWNTs, providing a fluorescence-Raman dual-imaging method for intracellular tracking and locating of MWNTs. We suggest that the cellular internalization of the CPE-cMWNT nanocomposites is a surface charge-dependent process. The strengths of this nanoplatform include satisfying biocompatibility, enhanced protein-repellent property, and ease of implementation, making it available for both in vitro and in vivo applications.

  12. Perspectives about Family Meals from Single-Headed and Dual-Headed Households: A Qualitative Analysis

    Science.gov (United States)

    Berge, Jerica M.; Hoppmann, Caroline; Hanson, Carrie; Neumark-Sztainer, Dianne

    2013-01-01

    Cross-sectional and longitudinal research has shown that family meals are protective for adolescent healthful eating behaviors. However, little is known about what parents think of these findings and whether parents from single- versus dual-headed households have differing perspectives about the findings. Additionally, parents’ perspectives regarding barriers to applying the findings on family meals in their own homes and suggestions for more wide-spread adoption of the findings are unknown. The current study aimed to identify single- and dual-headed household parents’ perspectives regarding the research findings on family meals, barriers to applying the findings in their own homes and suggestions for helping families have more family meals. The current qualitative study included 59 parents who participated in sub-study of two linked multi-level studies—EAT 2010 (Eating and Activity in Teens) and Families and Eating and Activity in Teens (F-EAT). Parents (91.5% female) were racially/ethnically and socio-economically diverse. Data were analyzed using a grounded theory approach. Results from the current study suggest that parents from both single- and dual-headed households have similar perspectives regarding why family meals are protective for healthful eating habits for adolescents (e.g., provides structure/routine, opportunities for communication, connection), but provide similar and different reasons for barriers to family meals (e.g., single-headed=cost vs. dual-headed=lack of creativity) and ideas and suggestions for how to increase the frequency of family meals (e.g., single-headed=give fewer options vs. dual-headed=include children in the meal preparation). Findings may help inform public health intervention researchers and providers who work with adolescents and their families to understand how to approach discussions regarding reasons for having family meals, barriers to carrying out family meals and ways to increase family meals depending on family

  13. Perspectives about family meals from single-headed and dual-headed households: a qualitative analysis.

    Science.gov (United States)

    Berge, Jerica M; Hoppmann, Caroline; Hanson, Carrie; Neumark-Sztainer, Dianne

    2013-12-01

    Cross-sectional and longitudinal research has shown that family meals are protective for adolescent healthful eating behaviors. However, little is known about what parents think of these findings and whether parents from single- vs dual-headed households have differing perspectives about the findings. In addition, parents' perspectives regarding barriers to applying the findings on family meals in their own homes and suggestions for more widespread adoption of the findings are unknown. The current study aimed to identify single- and dual-headed household parents' perspectives regarding the research findings on family meals, barriers to applying the findings in their own homes, and suggestions for helping families have more family meals. The current qualitative study included 59 parents who participated in substudy of two linked multilevel studies-EAT 2010 (Eating and Activity in Teens) and Families and Eating and Activity in Teens (F-EAT). Parents (91.5% female) were racially/ethnically and socioeconomically diverse. Data were analyzed using a grounded theory approach. Results from the current study suggest that parents from both single- and dual-headed households have similar perspectives regarding why family meals are protective for healthful eating habits for adolescents (eg, provides structure/routine, opportunities for communication, connection), but provide similar and different reasons for barriers to family meals (eg, single-headed=cost vs dual-headed=lack of creativity) and ideas and suggestions for how to increase the frequency of family meals (eg, single-headed=give fewer options vs dual-headed=include children in the meal preparation). Findings can help inform public health intervention researchers and providers who work with adolescents and their families to understand how to approach discussions regarding reasons for having family meals, barriers to carrying out family meals, and ways to increase family meals depending on family structure.

  14. Systematic Review and Meta-analysis of Dual Versus Single Antiplatelet Therapy in Carotid Interventions.

    Science.gov (United States)

    Barkat, M; Hajibandeh, S; Hajibandeh, S; Torella, F; Antoniou, G A

    2017-01-01

    The importance of antiplatelet therapy for the management and prevention of ischaemic stroke cannot be overstated. Despite the established guidelines, there is no clear consensus on how to manage antiplatelet therapy during and after carotid interventions. The objective was to undertake a systematic literature review and perform a meta-analysis to assess the effects of dual antiplatelet therapy in carotid endarterectomy (CEA) and stenting (CAS). Electronic information sources (MEDLINE, EMBASE, CINAHL, CENTRAL) and bibliographic reference lists were searched to identify randomised controlled trials (RCTs) and observational studies reporting comparative outcomes of dual versus single antiplatelet therapy in CEA and CAS. Primary outcomes were mortality and stroke within 30 days of intervention. Secondary outcomes were transient ischaemic attack (TIA), major bleeding, groin or neck haematoma, and myocardial infarction (MI). Dichotomous outcome measures were reported using the risk difference (RD) and 95% confidence interval (CI). Combined overall treatment effects were calculated using fixed-effect or random-effects models. Three RCTs and seven observational studies were identified reporting a total of 36,881 CEAs and 150 CAS procedures. In CEA, there were no differences in stroke/TIA/death between single and dual antiplatelet therapy, but there was a significant risk of major bleeding (RD, 0.00; 95% CI, 0.00-0.01; p = .0003) and neck haematoma with dual therapy (RD, 0.04; 95% CI, 0.01-0.06; p = .001). In addition, the rate of MI was higher in the dual therapy group than the single therapy group (RD, 0.00; 95% CI, 0.00-0.01; p = .003). In CAS, there was no difference in major bleeding or haematoma formation, but a significant difference in TIA in favour of dual therapy was identified (RD -0.13, 95% CI, -0.22 to -0.05; p = .003). Dual antiplatelet therapy demonstrates advantages over single therapy only in CAS, as indicated by a reduced risk of TIA. Dual

  15. Performance Analysis Of Single-Pumped And Dual-Pumped Parametric Optical Amplifier

    Directory of Open Access Journals (Sweden)

    Sandar Myint

    2015-06-01

    Full Text Available Abstract In this study we present a performance analysis of single-pumped and dual- pumped parametric optical amplifier and present the analysis of gain flatness in dual- pumped Fiber Optical Parametric Amplifier FOPA based on four-wave mixing FWM. Result shows that changing the signal power and pump power give the various gains in FOPA. It is also found out that the parametric gain increase with increase in pump power and decrease in signal power. .Moreover in this paper the phase matching condition in FWM plays a vital role in predicting the gain profile of the FOPAbecause the parametric gain is maximum when the total phase mismatch is zero.In this paper single-pumped parametric amplification over a 50nm gain bandwidth is demonstrated using 500 nm highly nonlinear fiber HNLF and signal achieves about 31dB gain. For dual-pumped parametric amplification signal achieves 26.5dB gains over a 50nm gain bandwidth. Therefore dual-pumped parametric amplifier can provide relatively flat gain over a much wider bandwidth than the single-pumped FOPA.

  16. Fluorescence-Magnetism Functional EuS Nanocrystals with Controllable Morphologies for Dual Bioimaging.

    Science.gov (United States)

    Sun, Yuanqing; Wang, Dandan; Zhao, Tianxin; Jiang, Yingnan; Zhao, Yueqi; Wang, Chuanxi; Sun, Hongchen; Yang, Bai; Lin, Quan

    2016-12-14

    Multiple functions incorporated in one single component material offer important applications in biosystems. Here we prepared a divalent state of rare earth EuS nanocrystals (NCs), which provides luminescent and magnetic properties, using both 1-Dodecanethiol (DT) and oleylamine (OLA) as reducing agents. The resultant EuS NCs exhibit controllable shapes, uniform size, and bright luminescence with a quantum yield as high as 3.5%. OLA as a surface ligand plays an important role in tunable morphologies, such as nanowires, nanorods, nanospheres et al. Another attractive nature of the EuS NCs is their paramagnetism at room temperature. In order to expand the biological applications, the resultant EuS NCs were modified with amphiphilic block copolymer F127 and transferred from oil to water phase. The excellent biocompatibility of EuS NCs is demonstrated as well as preservation of their luminescence and paramagnetic properties. The EuS NCs offer multifunction and great advantages of bright luminescence, paramagnetic, controllable morphologies, and good biocompatibility promising applications in the field of simultaneous magnetic resonance and fluorescence bioimaging.

  17. Construction of a Dual-Fluorescence Reporter System to Monitor the Dynamic Progression of Pluripotent Cell Differentiation.

    Science.gov (United States)

    Sun, Wu-Sheng; Chun, Ju-Lan; Do, Jeong-Tae; Kim, Dong-Hwan; Ahn, Jin-Seop; Kim, Min-Kyu; Hwang, In-Sul; Kwon, Dae-Jin; Hwang, Seong-Soo; Lee, Jeong-Woong

    2016-01-01

    Oct4 is a crucial germ line-specific transcription factor expressed in different pluripotent cells and downregulated in the process of differentiation. There are two conserved enhancers, called the distal enhancer (DE) and proximal enhancer (PE), in the 5' upstream regulatory sequences (URSs) of the mouse Oct4 gene, which were demonstrated to control Oct4 expression independently in embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs). We analyzed the URSs of the pig Oct4 and identified two similar enhancers that were highly consistent with the mouse DE and PE. A dual-fluorescence reporter was later constructed by combining a DE-free-Oct4-promoter-driven EGFP reporter cassette with a PE-free-Oct4-promoter-driven mCherry reporter cassette. Then, it was tested in a mouse ESC-like cell line (F9) and a mouse EpiSC-like cell line (P19) before it is formally used for pig. As a result, a higher red fluorescence was observed in F9 cells, while green fluorescence was primarily detected in P19 cells. This fluorescence expression pattern in the two cell lines was consistent with that in the early naïve pluripotent state and late primed pluripotent state during differentiation of mouse ESCs. Hence, this reporter system will be a convenient tool for screening out ESC-like naïve pluripotent stem cells from other metastable state cells in a heterogenous population.

  18. Construction of a Dual-Fluorescence Reporter System to Monitor the Dynamic Progression of Pluripotent Cell Differentiation

    Directory of Open Access Journals (Sweden)

    Wu-Sheng Sun

    2016-01-01

    Full Text Available Oct4 is a crucial germ line-specific transcription factor expressed in different pluripotent cells and downregulated in the process of differentiation. There are two conserved enhancers, called the distal enhancer (DE and proximal enhancer (PE, in the 5′ upstream regulatory sequences (URSs of the mouse Oct4 gene, which were demonstrated to control Oct4 expression independently in embryonic stem cells (ESCs and epiblast stem cells (EpiSCs. We analyzed the URSs of the pig Oct4 and identified two similar enhancers that were highly consistent with the mouse DE and PE. A dual-fluorescence reporter was later constructed by combining a DE-free-Oct4-promoter-driven EGFP reporter cassette with a PE-free-Oct4-promoter-driven mCherry reporter cassette. Then, it was tested in a mouse ESC-like cell line (F9 and a mouse EpiSC-like cell line (P19 before it is formally used for pig. As a result, a higher red fluorescence was observed in F9 cells, while green fluorescence was primarily detected in P19 cells. This fluorescence expression pattern in the two cell lines was consistent with that in the early naïve pluripotent state and late primed pluripotent state during differentiation of mouse ESCs. Hence, this reporter system will be a convenient tool for screening out ESC-like naïve pluripotent stem cells from other metastable state cells in a heterogenous population.

  19. Magnetically engineered Cd-free quantum dots as dual-modality probes for fluorescence/magnetic resonance imaging of tumors.

    Science.gov (United States)

    Ding, Ke; Jing, Lihong; Liu, Chunyan; Hou, Yi; Gao, Mingyuan

    2014-02-01

    Magnetically engineered Cd-free CuInS2@ZnS:Mn quantum dots (QDs) were designed, synthesized, and evaluated as potential dual-modality probes for fluorescence and magnetic resonance imaging (MRI) of tumors in vivo. The synthesis of Mn-doped core-shell structured CuInS2@ZnS mainly comprised three steps, i.e., the preparation of fluorescent CuInS2 seeds, the particle surface coating of ZnS, and the Mn-doping of the ZnS shells. Systematic spectroscopy studies were carried out to illustrate the impacts of ZnS coating and the following Mn-doping on the optical properties of the QDs. In combination with conventional fluorescence, fluorescence excitation, and time-resolved fluorescence measurements, the structure of CuInS2@ZnS:Mn QDs prepared under optimized conditions presented a Zn gradient CuInS2 core and a ZnS outer shell, while Mn ions were mainly located in the ZnS shell, which well balanced the optical and magnetic properties of the resultant QDs. For the following in vivo imaging experiments, the hydrophobic CuInS2@ZnS:Mn QDs were transferred into water upon ligand exchange reactions by replacing the 1-dodecanethiol ligand with dihydrolipoic acid-poly(ethylene glycol) (DHLA-PEG) ligand. The MTT assays based on HeLa cells were carried out to evaluate the cytotoxicity of the current Cd-free CuInS2@ZnS:Mn QDs for comparing with that of water soluble CdTe QDs. Further in vivo fluorescence and MR imaging experiments suggested that the PEGylated CuInS2@ZnS:Mn QDs could well target both subcutaneous and intraperitoneal tumors in vivo.

  20. A dual-channel fluorescent chemosensor for discriminative detection of glutathione based on functionalized carbon quantum dots.

    Science.gov (United States)

    Huang, Yuanyuan; Zhou, Jin; Feng, Hui; Zheng, Jieyu; Ma, Hui-Min; Liu, Weidong; Tang, Cong; Ao, Hang; Zhao, Meizhi; Qian, Zhaosheng

    2016-12-15

    A convenient, fluorescent dual-channel chemosensor on the basis of bis(3-pyridylmethyl)amine-functionalized carbon quantum dots (BPMA-CQDs) nanoprobe was constructed, and it can discriminatively detect glutathione from its analogues cysteine and homocysteine based on two distinctive strategies. Two distinct fluorescence responses of BPMA-CQDs probe to Cu(II) and Ag(I) were identified and further employed to achieve selective detection of Cu(II) and Ag(I) respectively. Based on the BPMA-CQDs/Cu(II) conjugate, discriminative detection of GSH was achieved in terms of correlation between the amounts of GSH and fluorescence recovery. The addition of GSH into BPMA-CQDs/Cu(II) system induces the reduction of Cu(II) to Cu(I), which could efficiently block PET process resulting in the following fluorescence recovery. Based on the BPMA-CQDs/Ag(I) conjugate, GSH assay could also be established on the basis of fluorescence response to GSH. The introduction of GSH into the preceding system triggers the competitive coordination to Ag(I) between BPMA and GSH, and silver ions are finally taken away by GSH from the probe, where the fluorescence is restored to its original weak state. Both of the detection strategies can achieve discriminative detection of GSH from Cys and Hcy. The assays showed good stability and repeatability, and covered a broad linear range of up to 13.3μM with a lowest detection limit of 42.0nM. Moreover, both of them were utilized to monitor GSH level in live cells.

  1. Cyanine 5.5 conjugated nanobubbles as a tumor selective contrast agent for dual ultrasound-fluorescence imaging in a mouse model.

    Directory of Open Access Journals (Sweden)

    Liyi Mai

    Full Text Available Nanobubbles and microbubbles are non-invasive ultrasound imaging contrast agents that may potentially enhance diagnosis of tumors. However, to date, both nanobubbles and microbubbles display poor in vivo tumor-selectivity over non-targeted organs such as liver. We report here cyanine 5.5 conjugated nanobubbles (cy5.5-nanobubbles of a biocompatible chitosan-vitamin C lipid system as a dual ultrasound-fluorescence contrast agent that achieved tumor-selective imaging in a mouse tumor model. Cy5.5-nanobubble suspension contained single bubble spheres and clusters of bubble spheres with the size ranging between 400-800 nm. In the in vivo mouse study, enhancement of ultrasound signals at tumor site was found to persist over 2 h while tumor-selective fluorescence emission was persistently observed over 24 h with intravenous injection of cy5.5-nanobubbles. In vitro cell study indicated that cy5.5-flurescence dye was able to accumulate in cancer cells due to the unique conjugated nanobubble structure. Further in vivo fluorescence study suggested that cy5.5-nanobubbles were mainly located at tumor site and in the bladder of mice. Subsequent analysis confirmed that accumulation of high fluorescence was present at the intact subcutaneous tumor site and in isolated tumor tissue but not in liver tissue post intravenous injection of cy5.5-nanobubbles. All these results led to the conclusion that cy5.5-nanobubbles with unique crosslinked chitosan-vitamin C lipid system have achieved tumor-selective imaging in vivo.

  2. A nomogram for deconvolution of single exponential fluorescence decays.

    OpenAIRE

    Rockley, M.G.

    1980-01-01

    An extremely rapid technique for deconvolving single exponential luminescence decay data is described that involves essentially no mathematical manipulation of the experimental data. The method permits "real time" measurement of deconvolved luminescence lifetimes with conventional pulsed, lifetime-fluorometers and phosphorimeters. The method assumes that the true luminescence decay of the chromophore is accurately represented by a single exponential decay function.

  3. Comparison of migration behavior between single and dual lag screw implants for intertrochanteric fracture fixation

    Directory of Open Access Journals (Sweden)

    Katonis Pavlos G

    2009-05-01

    Full Text Available Abstract Background Lag screw cut-out failure following fixation of unstable intertrochanteric fractures in osteoporotic bone remains an unsolved challenge. This study tested if resistance to cut-out failure can be improved by using a dual lag screw implant in place of a single lag screw implant. Migration behavior and cut-out resistance of a single and a dual lag screw implant were comparatively evaluated in surrogate specimens using an established laboratory model of hip screw cut-out failure. Methods Five dual lag screw implants (Endovis, Citieffe and five single lag screw implants (DHS, Synthes were tested in the Hip Implant Performance Simulator (HIPS of the Legacy Biomechanics Laboratory. This model simulated osteoporotic bone, an unstable fracture, and biaxial rocking motion representative of hip loading during normal gait. All constructs were loaded up to 20,000 cycles of 1.45 kN peak magnitude under biaxial rocking motion. The migration kinematics was continuously monitored with 6-degrees of freedom motion tracking system and the number of cycles to implant cut-out was recorded. Results The dual lag screw implant exhibited significantly less migration and sustained more loading cycles in comparison to the DHS single lag screw. All DHS constructs failed before 20,000 cycles, on average at 6,638 ± 2,837 cycles either by cut-out or permanent screw bending. At failure, DHS constructs exhibited 10.8 ± 2.3° varus collapse and 15.5 ± 9.5° rotation around the lag screw axis. Four out of five dual screws constructs sustained 20,000 loading cycles. One dual screw specimens sustained cut-out by medial migration of the distal screw after 10,054 cycles. At test end, varus collapse and neck rotation in dual screws implants advanced to 3.7 ± 1.7° and 1.6 ± 1.0°, respectively. Conclusion The single and double lag screw implants demonstrated a significantly different migration resistance in surrogate specimens under gait loading simulation with

  4. Electrophysiological spread of excitation and pitch perception for dual and single electrodes using the Nucleus Freedom cochlear implant.

    Science.gov (United States)

    Busby, Peter A; Battmer, Rolf D; Pesch, Joerg

    2008-12-01

    The first objective of the study was to determine whether there were any consistent differences in the electrophysiological spread of excitation (SOE) function, as measured using the electrically evoked compound action potential (ECAP), between dual and single electrode stimulation with the Nucleus Freedom cochlear implant system. Dual electrode stimulation is produced by electrically coupling two adjacent single electrodes. The second objective was to determine whether there were any relationships between the SOE functions and psychophysically measured pitch ranking of dual and single electrodes. Nine adult cochlear implant subjects participated in the study. ECAPs for dual and single electrode stimulation were measured using the forward masking paradigm, as also used in the Neural Response Telemetry (NRT) software with the Nucleus implant. Research software was used to generate the dual and single electrode stimuli and record the ECAPs. Spread of excitations (SOEs) were measured on a dual electrode and the two adjacent single electrodes, at three positions on the array: apical, mid, and basal. Compared were the ECAP amplitudes at the peak of the SOE functions, the widths of the scaled SOE functions at the 75% point, and the electrode positions at the peak of the SOE function and at the 75%, 50%, and 25% points on apical and basal sides of the scaled functions. Pitch ranking was measured for the same sets of dual and single electrodes. A two-alternative forced choice procedure was used, with the electrodes in each set paired with each other as AB and BA pairs. The subject indicated which of the two stimuli had the higher pitch. Dual electrode SOEs could be successfully obtained using the same methods as used to measure single electrode SOEs. The shapes of the dual and single electrodes SOEs were similar. There was a trend of a higher ECAP amplitude for the dual electrode at the peak of the SOE function, but this was only significant for two comparisons at the

  5. Dual fluorescent molecular substrates selectively report the activation, sustainability and reversibility of cellular PKB/Akt activity

    Science.gov (United States)

    Shen, Duanwen; Bai, Mingfeng; Tang, Rui; Xu, Baogang; Ju, Xiaoming; Pestell, Richard G.; Achilefu, Samuel

    2013-04-01

    Using a newly developed near-infrared (NIR) dye that fluoresces at two different wavelengths (dichromic fluorescence, DCF), we discovered a new fluorescent substrate for Akt, also known as protein kinase B, and a method to quantitatively report this enzyme's activity in real time. Upon insulin activation of cellular Akt, the enzyme multi-phosphorylated a single serine residue of a diserine DCF substrate in a time-dependent manner, culminating in monophospho- to triphospho-serine products. The NIR DCF probe was highly selective for the Akt1 isoform, which was demonstrated using Akt1 knockout cells derived from MMTV-ErbB2 transgenic mice. The DCF mechanism provides unparalleled potential to assess the stimulation, sustainability, and reversibility of Akt activation longitudinally. Importantly, NIR fluorescence provides a pathway to translate findings from cells to living organisms, a condition that could eventually facilitate the use of these probes in humans.

  6. Single-Photon Source for Quantum Information Based on Single Dye Molecule Fluorescence in Liquid Crystal Host

    Energy Technology Data Exchange (ETDEWEB)

    Lukishova, S.G.; Knox, R.P.; Freivald, P.; McNamara, A.; Boyd, R.W.; Stroud, Jr., C.R.; Schmid, A.W.; Marshall, K.L.

    2006-08-18

    This paper describes a new application for liquid crystals: quantum information technology. A deterministically polarized single-photon source that efficiently produces photons exhibiting antibunching is a pivotal hardware element in absolutely secure quantum communication. Planar-aligned nematic liquid crystal hosts deterministically align the single dye molecules which produce deterministically polarized single (antibunched) photons. In addition, 1-D photonic bandgap cholesteric liquid crystals will increase single-photon source efficiency. The experiments and challenges in the observation of deterministically polarized fluorescence from single dye molecules in planar-aligned glassy nematic-liquid-crystal oligomer as well as photon antibunching in glassy cholesteric oligomer are described for the first time.

  7. Multicolor bioluminescence boosts malaria research: quantitative dual-color assay and single-cell imaging in Plasmodium falciparum parasites.

    Science.gov (United States)

    Cevenini, Luca; Camarda, Grazia; Michelini, Elisa; Siciliano, Giulia; Calabretta, Maria Maddalena; Bona, Roberta; Kumar, T R Santha; Cara, Andrea; Branchini, Bruce R; Fidock, David A; Roda, Aldo; Alano, Pietro

    2014-09-02

    New reliable and cost-effective antimalarial drug screening assays are urgently needed to identify drugs acting on different stages of the parasite Plasmodium falciparum, and particularly those responsible for human-to-mosquito transmission, that is, the P. falciparum gametocytes. Low Z' factors, narrow dynamic ranges, and/or extended assay times are commonly reported in current gametocyte assays measuring gametocyte-expressed fluorescent or luciferase reporters, endogenous ATP levels, activity of gametocyte enzymes, or redox-dependent dye fluorescence. We hereby report on a dual-luciferase gametocyte assay with immature and mature P. falciparum gametocyte stages expressing red and green-emitting luciferases from Pyrophorus plagiophthalamus under the control of the parasite sexual stage-specific pfs16 gene promoter. The assay was validated with reference antimalarial drugs and allowed to quantitatively and simultaneously measure stage-specific drug effects on parasites at different developmental stages. The optimized assay, requiring only 48 h incubation with drugs and using a cost-effective luminogenic substrate, significantly reduces assay cost and time in comparison to state-of-the-art analogous assays. The assay had a Z' factor of 0.71 ± 0.03, and it is suitable for implementation in 96- and 384-well microplate formats. Moreover, the use of a nonlysing D-luciferin substrate significantly improved the reliability of the assay and allowed one to perform, for the first time, P. falciparum bioluminescence imaging at single-cell level.

  8. Optimization of Single- and Dual-Color Immunofluorescence Protocols for Formalin-Fixed, Paraffin-Embedded Archival Tissues.

    Science.gov (United States)

    Kajimura, Junko; Ito, Reiko; Manley, Nancy R; Hale, Laura P

    2016-02-01

    Performance of immunofluorescence staining on archival formalin-fixed paraffin-embedded human tissues is generally not considered to be feasible, primarily due to problems with tissue quality and autofluorescence. We report the development and application of procedures that allowed for the study of a unique archive of thymus tissues derived from autopsies of individuals exposed to atomic bomb radiation in Hiroshima, Japan in 1945. Multiple independent treatments were used to minimize autofluorescence and maximize fluorescent antibody signals. Treatments with NH3/EtOH and Sudan Black B were particularly useful in decreasing autofluorescent moieties present in the tissue. Deconvolution microscopy was used to further enhance the signal-to-noise ratios. Together, these techniques provide high-quality single- and dual-color fluorescent images with low background and high contrast from paraffin blocks of thymus tissue that were prepared up to 60 years ago. The resulting high-quality images allow the application of a variety of image analyses to thymus tissues that previously were not accessible. Whereas the procedures presented remain to be tested for other tissue types and archival conditions, the approach described may facilitate greater utilization of older paraffin block archives for modern immunofluorescence studies.

  9. Single particle multichannel bio-aerosol fluorescence sensor.

    Science.gov (United States)

    Kaye, P; Stanley, W R; Hirst, E; Foot, E V; Baxter, K L; Barrington, S J

    2005-05-16

    We describe a prototype low-cost multi-channel aerosol fluorescence sensor designed for unattended deployment in medium to large area bio-aerosol detection networks. Individual airborne particles down to ~1mum in size are detected and sized by measurement of light scattered from a continuous-wave diode laser (660nm). This scatter signal is then used to trigger the sequential firing of two xenon sources which irradiate the particle with UV pulses at ~280 nm and ~370 nm, optimal for excitation of bio-fluorophores tryptophan and NADH (nicotinamide adenine dinucleotide) respectively. For each excitation wavelength, fluorescence is detected across two bands embracing the peak emissions of the same bio-fluorophores. Current measurement rates are up to ~125 particles/s, corresponding to all particles for concentrations up to 1.3 x 104 particles/l. Developments to increase this to ~500 particles/s are in hand. Device sensitivity is illustrated in preliminary data recorded from aerosols of E.coli, BG spores, and a variety of non-biological materials.

  10. Single particle multichannel bio-aerosol fluorescence sensor

    Science.gov (United States)

    Kaye, P. H.; Stanley, W. R.; Hirst, E.; Foot, E. V.; Baxter, K. L.; Barrington, S. J.

    2005-05-01

    We describe a prototype low-cost multi-channel aerosol fluorescence sensor designed for unattended deployment in medium to large area bio-aerosol detection networks. Individual airborne particles down to ~1μm in size are detected and sized by measurement of light scattered from a continuous-wave diode laser (660nm). This scatter signal is then used to trigger the sequential firing of two xenon sources which irradiate the particle with UV pulses at ~280 nm and ~370 nm, optimal for excitation of bio-fluorophores tryptophan and NADH (nicotinamide adenine dinucleotide) respectively. For each excitation wavelength, fluorescence is detected across two bands embracing the peak emissions of the same bio-fluorophores. Current measurement rates are up to ~125 particles/s, corresponding to all particles for concentrations up to 1.3 x 104 particles/l. Developments to increase this to ~500 particles/s are in hand. Device sensitivity is illustrated in preliminary data recorded from aerosols of E.coli, BG spores, and a variety of non-biological materials.

  11. Single-transducer dual-frequency ultrasound generation to enhance acoustic cavitation.

    Science.gov (United States)

    Liu, Hao-Li; Hsieh, Chao-Ming

    2009-03-01

    Dual- or multiple-frequency ultrasound stimulation is capable of effectively enhancing the acoustic cavitation effect over single-frequency ultrasound. Potential application of this sonoreactor design has been widely proposed such as on sonoluminescence, sonochemistry enhancement, and transdermal drug release enhancement. All currently available sonoreactor designs employed multiple piezoelectric transducers for generating single-frequency ultrasonic waves separately and then these waves were mixed and interfered in solutions. The purpose of this research is to propose a novel design of generating dual-frequency ultrasonic waves with single piezoelectric elements, thereby enhancing acoustic cavitation. Macroscopic bubbles were detected optically, and they were quantified at either a single-frequency or for different frequency combinations for determining their efficiency for enhancing acoustic cavitation. Visible bubbles were optically detected and hydrogen peroxide was measured to quantify acoustic cavitation. Test water samples with different gas concentrations and different power levels were used to determine the efficacy of enhancing acoustic cavitation of this design. The spectrum obtained from the backscattered signals was also recorded and examined to confirm the occurrence of stable cavitation. The results confirmed that single-element dual-frequency ultrasound stimulation can enhance acoustic cavitation. Under certain testing conditions, the generation of bubbles can be enhanced up to a level of five times higher than the generation of bubbles in single-frequency stimulation, and can increase the hydrogen peroxide production up to an increase of one fold. This design may serve as a useful alternative for future sonoreactor design owing to its simplicity to produce dual- or multiple-frequency ultrasound.

  12. Characterization of cytoplasmic Gag-gag interactions by dual-color z-scan fluorescence fluctuation spectroscopy.

    Science.gov (United States)

    Fogarty, Keir H; Chen, Yan; Grigsby, Iwen F; Macdonald, Patrick J; Smith, Elizabeth M; Johnson, Jolene L; Rawson, Jonathan M; Mansky, Louis M; Mueller, Joachim D

    2011-03-16

    Fluorescence fluctuation spectroscopy (FFS) quantifies the interactions of fluorescently-labeled proteins inside living cells by brightness analysis. However, the study of cytoplasmic proteins that interact with the plasma membrane is challenging with FFS. If the cytoplasmic section is thinner than the axial size of the observation volume, cytoplasmic and membrane-bound proteins are coexcited, which leads to brightness artifacts. This brightness bias, if not recognized, leads to erroneous interpretation of the data. We have overcome this challenge by introducing dual-color z-scan FFS and the addition of a distinctly colored reference protein. Here, we apply this technique to study the cytoplasmic interactions of the Gag proteins from human immunodeficiency virus type 1 (HIV-1) and human T-lymphotropic virus type 1 (HTLV-1). The Gag protein plays a crucial role in the assembly of retroviruses and is found in both membrane and cytoplasm. Dual-color z-scans demonstrate that brightness artifacts are caused by a dim nonpunctate membrane-bound fraction of Gag. We perform an unbiased brightness characterization of cytoplasmic Gag by avoiding the membrane-bound fraction and reveal previously unknown differences in the behavior of the two retroviral Gag species. HIV-1 Gag exhibits concentration-dependent oligomerization in the cytoplasm, whereas HTLV-1 Gag lacks significant cytoplasmic Gag-Gag interactions. Copyright © 2011 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  13. A Dual-Modality System for Both Multi-Color Ultrasound-Switchable Fluorescence and Ultrasound Imaging

    Science.gov (United States)

    Kandukuri, Jayanth; Yu, Shuai; Cheng, Bingbing; Bandi, Venugopal; D’Souza, Francis; Nguyen, Kytai T.; Hong, Yi; Yuan, Baohong

    2017-01-01

    Simultaneous imaging of multiple targets (SIMT) in opaque biological tissues is an important goal for molecular imaging in the future. Multi-color fluorescence imaging in deep tissues is a promising technology to reach this goal. In this work, we developed a dual-modality imaging system by combining our recently developed ultrasound-switchable fluorescence (USF) imaging technology with the conventional ultrasound (US) B-mode imaging. This dual-modality system can simultaneously image tissue acoustic structure information and multi-color fluorophores in centimeter-deep tissue with comparable spatial resolutions. To conduct USF imaging on the same plane (i.e., x-z plane) as US imaging, we adopted two 90°-crossed ultrasound transducers with an overlapped focal region, while the US transducer (the third one) was positioned at the center of these two USF transducers. Thus, the axial resolution of USF is close to the lateral resolution, which allows a point-by-point USF scanning on the same plane as the US imaging. Both multi-color USF and ultrasound imaging of a tissue phantom were demonstrated. PMID:28165390

  14. Smartphone-Based Dual-Modality Imaging System for Quantitative Detection of Color or Fluorescent Lateral Flow Immunochromatographic Strips

    Science.gov (United States)

    Hou, Yafei; Wang, Kan; Xiao, Kun; Qin, Weijian; Lu, Wenting; Tao, Wei; Cui, Daxiang

    2017-04-01

    Nowadays, lateral flow immunochromatographic assays are increasingly popular as a diagnostic tool for point-of-care (POC) test based on their simplicity, specificity, and sensitivity. Hence, quantitative detection and pluralistic popular application are urgently needed in medical examination. In this study, a smartphone-based dual-modality imaging system was developed for quantitative detection of color or fluorescent lateral flow test strips, which can be operated anywhere at any time. In this system, the white and ultra-violet (UV) light of optical device was designed, which was tunable with different strips, and the Sobel operator algorithm was used in the software, which could enhance the identification ability to recognize the test area from the background boundary information. Moreover, this technology based on extraction of the components from RGB format (red, green, and blue) of color strips or only red format of the fluorescent strips can obviously improve the high-signal intensity and sensitivity. Fifty samples were used to evaluate the accuracy of this system, and the ideal detection limit was calculated separately from detection of human chorionic gonadotropin (HCG) and carcinoembryonic antigen (CEA). The results indicated that smartphone-controlled dual-modality imaging system could provide various POC diagnoses, which becomes a potential technology for developing the next-generation of portable system in the near future.

  15. Biomimetic Preparation and Dual-Color Bioimaging of Fluorescent Silicon Nanoparticles.

    Science.gov (United States)

    Wu, Sicong; Zhong, Yiling; Zhou, Yanfeng; Song, Bin; Chu, Binbin; Ji, Xiaoyuan; Wu, Yanyan; Su, Yuanyuan; He, Yao

    2015-11-25

    Fluorescent silicon nanoparticles (SiNPs), as the most important zero-dimensional silicon nanostructures, hold high promise for long-awaited silicon-based optic applications. There currently remain major challenges for the green, inexpensive, and mass production of fluorescent SiNPs, resulting in difficulties in sufficiently exploiting the properties of these remarkable materials. Here, we show that fluorescent small-sized (∼3.8 nm) SiNPs can be produced through biomimetic synthesis in rapid (10 min), low-cost, and environmentally benign manners. The as-prepared SiNPs simultaneously feature bright fluorescence (quantum yield (QY), ∼15-20%), narrow emission spectral width (full width at half-maximum (fwhm), ∼30 nm), and nontoxicity, making them as high-quality fluorescent probes for biological imaging in vitro and in vivo.

  16. Semiconductor Quantum Rods as Single Molecule FluorescentBiological Labels

    Energy Technology Data Exchange (ETDEWEB)

    Fu, Aihua; Gu, Weiwei; Boussert, Benjamine; Koski, Kristie; Gerion, Daniele; Manna, Liberato; Le Gros, Mark; Larabell, Carolyn; Alivisatos, A. Paul

    2006-05-29

    In recent years, semiconductor quantum dots have beenapplied with great advantage in a wide range of biological imagingapplications. The continuing developments in the synthesis of nanoscalematerials and specifically in the area of colloidal semiconductornanocrystals have created an opportunity to generate a next generation ofbiological labels with complementary or in some cases enhanced propertiescompared to colloidal quantum dots. In this paper, we report thedevelopment of rod shaped semiconductor nanocrystals (quantum rods) asnew fluorescent biological labels. We have engineered biocompatiblequantum rods by surface silanization and have applied them fornon-specific cell tracking as well as specific cellular targeting. Theproperties of quantum rods as demonstrated here are enhanced sensitivityand greater resistance for degradation as compared to quantum dots.Quantum rods have many potential applications as biological labels insituations where their properties offer advantages over quantumdots.

  17. Method for rapid multidiameter single-fiber reflectance and fluorescence spectroscopy through a fiber bundle

    NARCIS (Netherlands)

    Amelink, A.; Hoy, C.L.; Gamm, U.A.; Sterenborg, H.J.C.M.; Robinson, D.J.

    2014-01-01

    We have recently demonstrated a means for quantifying the absorption and scattering properties of biological tissue through multidiameter single-fiber reflectance (MDSFR) spectroscopy. These measurements can be used to correct single-fiber fluorescence (SFF) spectra for the influence of optical prop

  18. Method for rapid multidiameter single-fiber reflectance and fluorescence spectroscopy through a fiber bundle

    NARCIS (Netherlands)

    Amelink, A.; Hoy, C.L.; Gamm, U.A.; Sterenborg, H.J.C.M.; Robinson, D.J.

    2014-01-01

    We have recently demonstrated a means for quantifying the absorption and scattering properties of biological tissue through multidiameter single-fiber reflectance (MDSFR) spectroscopy. These measurements can be used to correct single-fiber fluorescence (SFF) spectra for the influence of optical

  19. Fluorescent electrochemistry : towards controlled redox-switching of a single metalloprotein

    NARCIS (Netherlands)

    Akkılıç, Namık

    2013-01-01

    The availability of single-molecule fluorescence detection techniques has brought about a breakthrough in the optical studies of biomolecular properties and functions by enabling the study of molecules one at a time. In this thesis, single-molecule detection is used to obtain detailed information of

  20. Recent Advances in Biological Single-Molecule Applications of Optical Tweezers and Fluorescence Microscopy

    NARCIS (Netherlands)

    Hashemi Shabestari, M; Meijering, A E C; Roos, W H; Wuite, G J L; Peterman, E J G

    2017-01-01

    Over the past two decades, single-molecule techniques have evolved into robust tools to study many fundamental biological processes. The combination of optical tweezers with fluorescence microscopy and microfluidics provides a powerful single-molecule manipulation and visualization technique that

  1. Abnormal Ventral and Dorsal Attention Network Activity During Single and Dual Target Detection in Schizophrenia

    Directory of Open Access Journals (Sweden)

    Amy M. Jimenez

    2016-03-01

    Full Text Available Early visual perception and attention are impaired in schizophrenia, and these deficits can be observed on target detection tasks. These tasks activate distinct ventral and dorsal brain networks which support stimulus-driven and goal-directed attention, respectively. We used single and dual target rapid serial visual presentation (RSVP tasks during fMRI with an ROI approach to examine regions within these networks associated with target detection and the attentional blink (AB in 21 schizophrenia outpatients and 25 healthy controls. In both tasks, letters were targets and numbers were distractors. For the dual target task, the second target (T2 was presented at 3 different lags after the first target (T1 (lag1=100ms, lag3=300ms, lag7=700ms. For both single and dual target tasks, patients identified fewer targets than controls. For the dual target task, both groups showed the expected AB effect with poorer performance at lag 3 than at lags 1 or 7, and there was no group by lag interaction. During the single target task, patients showed abnormally increased deactivation of the temporo-parietal junction (TPJ, a key region of the ventral network. When attention demands were increased during the dual target task, patients showed overactivation of the posterior intraparietal cortex, a key dorsal network region, along with failure to deactivate TPJ. Results suggest inefficient and faulty suppression of salience-oriented processing regions, resulting in increased sensitivity to stimuli in general, and difficulty distinguishing targets from non-targets.

  2. Dual joint space arthrography in temporomandibular joint disorders: Comparison with single inferior joint space arthrography

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Hyung Sik; Chang, Duk Soo; Lee, Kyung Soo; Kim, Woo Sun; Sung, Jung Ho; Jun, Young Hwan [Capital Armed Forces General Hospital, Seoul (Korea, Republic of)

    1989-02-15

    The temporomandibular joint(TMJ) is really a complex of two synovial space separated by fibrocartilaginous disc. Single inferior joint space arthrography is commonly performed for evaluation of TMJ disorders, which is known to be superior in demonstrating joint dynamics. But it reveals only the inferior surface of the disc. Therefore, dual space arthrography is superior to demonstrate the soft tissue anatomic feature of the joint such as disc position and shape. Authors performed 83 TMJ arthrograms in TMJ problems. Initially, the inferior joint space was done and then the superior space was sequentially contrasted. The follow results were noted: 1. In all cases, dual space arthrography revealed accurate disc shape and positions. 2. Concordant findings between the two techniques: 68 cases (82%). Discordance between the two techniques: 15 cases (18%) 3. Possible causes of discordance between inferior and dual space arthrography. a) Normal varians of anterior recess: 3 cases b) Posterior disc displacement: 4 cases c) Influence of the patient's head position change :4 cases d) False perforation: 2 cases e) Reduction change: 2 cases 4. In 5 cases with anterior displacement, dual space arthrography gave additional findings such as adhesion within the superior space, which could not be evaluated by single inferior space.

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

    Science.gov (United States)

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

    2009-02-01

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

  4. A Google Glass navigation system for ultrasound and fluorescence dual-mode image-guided surgery

    Science.gov (United States)

    Zhang, Zeshu; Pei, Jing; Wang, Dong; Hu, Chuanzhen; Ye, Jian; Gan, Qi; Liu, Peng; Yue, Jian; Wang, Benzhong; Shao, Pengfei; Povoski, Stephen P.; Martin, Edward W.; Yilmaz, Alper; Tweedle, Michael F.; Xu, Ronald X.

    2016-03-01

    Surgical resection remains the primary curative intervention for cancer treatment. However, the occurrence of a residual tumor after resection is very common, leading to the recurrence of the disease and the need for re-resection. We develop a surgical Google Glass navigation system that combines near infrared fluorescent imaging and ultrasonography for intraoperative detection of sites of tumor and assessment of surgical resection boundaries, well as for guiding sentinel lymph node (SLN) mapping and biopsy. The system consists of a monochromatic CCD camera, a computer, a Google Glass wearable headset, an ultrasonic machine and an array of LED light sources. All the above components, except the Google Glass, are connected to a host computer by a USB or HDMI port. Wireless connection is established between the glass and the host computer for image acquisition and data transport tasks. A control program is written in C++ to call OpenCV functions for image calibration, processing and display. The technical feasibility of the system is tested in both tumor simulating phantoms and in a human subject. When the system is used for simulated phantom resection tasks, the tumor boundaries, invisible to the naked eye, can be clearly visualized with the surgical Google Glass navigation system. This system has also been used in an IRB approved protocol in a single patient during SLN mapping and biopsy in the First Affiliated Hospital of Anhui Medical University, demonstrating the ability to successfully localize and resect all apparent SLNs. In summary, our tumor simulating phantom and human subject studies have demonstrated the technical feasibility of successfully using the proposed goggle navigation system during cancer surgery.

  5. Fluorescent detection of single tracks of alpha particles using lithium fluoride crystals

    Science.gov (United States)

    Bilski, P.; Marczewska, B.

    2017-02-01

    Lithium fluoride single crystals were successfully used for fluorescent imaging of single tracks of alpha particles. This was realized with a standard wide-field fluorescent microscope equipped with a 100× objective. Alpha particles create F2 and F3+ color centers in LiF crystals. The subsequent illumination with the blue light (wavelength around 445 nm), excites these centers and produces fluorescence with a broad band peaked at 670 nm. The observed tracks of alpha particles have diameter of about 500 nm. Focusing of the microscope at different depths in a LiF crystal, enables imaging changes of shape and position of tracks, allowing for visualization of their paths. These encouraging results are the first step towards practical application of LiF as fluorescent nuclear track detectors.

  6. Super-resolution Localization and Defocused Fluorescence Microscopy on Resonantly Coupled Single-Molecule, Single-Nanorod Hybrids.

    Science.gov (United States)

    Su, Liang; Yuan, Haifeng; Lu, Gang; Rocha, Susana; Orrit, Michel; Hofkens, Johan; Uji-i, Hiroshi

    2016-02-23

    Optical antennas made of metallic nanostructures dramatically enhance single-molecule fluorescence to boost the detection sensitivity. Moreover, emission properties detected at the optical far field are dictated by the antenna. Here we study the emission from molecule-antenna hybrids by means of super-resolution localization and defocused imaging. Whereas gold nanorods make single-crystal violet molecules in the tip's vicinity visible in fluorescence, super-resolution localization on the enhanced molecular fluorescence reveals geometrical centers of the nanorod antenna instead. Furthermore, emission angular distributions of dyes linked to the nanorod surface resemble that of nanorods in defocused imaging. The experimental observations are consistent with numerical calculations using the finite-difference time-domain method.

  7. Single-molecule analysis of fluorescent carbon dots towards localization-based super-resolution microscopy

    Science.gov (United States)

    Verma, Navneet C.; Khan, Syamantak; Nandi, Chayan K.

    2016-12-01

    The advancement of high-resolution bioimaging has always been dependent on the discovery of bright and easily available fluorescent probes. Fluorescent carbon nanodots, an interesting class of relatively new nanomaterials, have emerged as a versatile alternative due to their superior optical properties, non-toxicity, cell penetrability and easy routes to synthesis. Although a plethora of reports is available on bioimaging using carbon dots, single-molecule-based super-resolution imaging is rare in the literature. In this study, we have systematically characterized the single-molecule fluorescence of three carbon dots and compared them with a standard fluorescent probe. Each of these carbon dots showed a long-lived dark state in the presence of an electron acceptor. The electron transfer mechanism was investigated in single-molecule as well as in ensemble experiments. The average on-off rate between the fluorescent bright and dark states, which is one of the important parameters for single-molecule localization-based super-resolution microscopy, was measured by changing the laser power. We report that the photon budget and on-off rate of these carbon dots were good enough to achieve single-molecule localization with a precision of ~35 nm.

  8. [Impulse cytofluorometry of the redox activity of single cells using fluorescent formazan].

    Science.gov (United States)

    Severin, E; Stellmach, J

    1984-01-01

    The first application of a newly developed fluorescent formazan in flow cytometry is described. The cell surface redox activity of isolated mouse hepatocytes after incubation with the tetrazolium salt forming the new fluorescent formazan and the DNA content after Hoechst staining have been measured simultaneously. 2 parametric distribution patterns have been obtained. This new and sensitive fluorometric technique can be used for automatic measurements of single cells correlating redox activity with other cell parameters.

  9. Quantitative single-molecule detection of protein based on DNA tetrahedron fluorescent nanolabels.

    Science.gov (United States)

    Ding, Yongshun; Liu, Xingti; Zhu, Jing; Wang, Lei; Jiang, Wei

    2014-07-01

    A highly sensitive method for single-molecule quantitative detection of human IgG is presented by the employment of a new fluorescent nanolabel. In this method, fluorescent nanolabels were assembled by inserting SYBR Green I into DNA tetrahedron nanostructure. The bio-nanolabels were attached to the streptavidin-antihuman antibody by a specific reaction between biotin and streptavidin. The antibody was combined with the target antigen, human IgG, which was immobilized on the silanized glass subtrate surface. Finally, epi-fluorescence microscopy (EFM) coupled with an electron multiplying charge-coupled device was employed for fluorescence imaging. The fluorescent spots corresponding to single protein molecule on images were counted and further used for the quantitative detection. It was found that the new nanolabel shows good photostability, biocompatiblity and exhibits no blinking compared to traditional labels like fluorescence dyes and quantum dot (QDs). In addition, the number of fluorescence spots on the images has a linear relationship with the concentration of human IgG in the range of 3.0×10(-14) to 1.0×10(-12)mol L(-1). What is more, this method showed an excellent specificity and a low matrix effect.

  10. Monitoring the aggregation of single casein micelles using fluorescence microscopy

    DEFF Research Database (Denmark)

    Bomholt, Julie; Moth-Poulsen, Kasper; Harboe, Marianne

    2011-01-01

    The aggregation of casein micelles (CMs) induced by milk-clotting enzymes is a process of fundamental importance in the dairy industry for cheese production; however, it is not well characterized on the nanoscale. Here we enabled the monitoring of the kinetics of aggregation between single CMs (30...

  11. Optical manipulation of aerosol droplets using a holographic dual and single beam trap.

    Science.gov (United States)

    Brzobohatý, Oto; Šiler, Martin; Ježek, Jan; Jákl, Petr; Zemánek, Pavel

    2013-11-15

    We present optical trapping and manipulation of pure water and salt water airborne droplets of various sizes ranging from sub-micrometers up to several tens of micrometers in a holographic dual and single beam trap. In the dual beam trap, successful fusion of droplets as well as precise delivery of many droplets and manipulation of multiple droplets are demonstrated. Furthermore, employing the transfer of the orbital angular momentum of light from Laguerre-Gaussian beams, we show that the water droplets orbit around the beam propagation axis and their tangential speed can be controlled by beam waist magnitude. We also demonstrate that sub-micrometer sized pure water droplets can be trapped and manipulated by a single beam trap with a relatively low numerical aperture. In this case, multiple stable trapping positions were observed, both theoretically and experimentally, which were due to the optical intensity oscillations in the focal region of the laser beam.

  12. Single- and dual-wavelength switchable linear polarized Yb(3+)-doped double-clad fiber laser.

    Science.gov (United States)

    Liu, Guanxiu; Feng, Dejun

    2015-05-10

    A single- and dual-wavelength switchable linear polarized Yb-doped double-clad fiber laser is proposed, in which the resonance cavity was composed of a fiber Bragg grating fabricated in a polarization-maintaining fiber and a dichromatic mirror with high reflectivity. The polarization hole burning is enhanced through selective polarization feedback by the polarization-maintaining fiber Bragg grating. The switchover of single and dual wavelengths is realized by tuning the rotation angle of a cubic polarization beam splitter that is inserted between the dichromatic mirror and the collimator in the cavity. The laser features wavelengths of 1070.08 and 1070.39 nm, output power of 1.0 W, signal to noise ratio of 45 dB, and slope efficiency of 34%, as well as a very narrow linewidth of 0.022 nm. The polarization characteristics are analyzed by measuring the laser power transmitted through a Glan-Thomson polarizer during rotation.

  13. Single- versus dual-energy quantitative computed tomography for spinal densitometry in patients with rheumatoid arthritis

    Energy Technology Data Exchange (ETDEWEB)

    Laan, R.F.J.M.; Erning, L.J.Th.O. van; Lemmens, J.A.M.; Putte, L.B.A. van de; Ruijs, S.H.J.; Riel, P.L.C.M. van (University Hospital, Nijmegen (Netherlands))

    1992-10-01

    Lumbar bone mineral density was measured by both single- and dual-energy quantitative computed tomography in 109 patients with rheumatoid arthritis. The results were corrected for the age-related increase in vertebral fat content by converting them to percentages of expected densities, using sex and energy-level specific regression equations obtained in a normal reference population. The percentages of expected density are approximately 10% lower in the single- than in the dual-energy mode, both in the patients with and without prednisone therapy. This difference is statistically highly significant, and is positively correlated with the duration of the disease and with the degree of radiological joint destruction. The data suggest that the vertebral fat content may be increased in patients with rheumatoid arthritis, as a consequence of disease-dependent mechanisms. (Author).

  14. Computed tomography with single-shot dual-energy sandwich detectors

    Science.gov (United States)

    Kim, Seung Ho; Youn, Hanbean; Kim, Daecheon; Kim, Dong Woon; Jeon, Hosang; Kim, Ho Kyung

    2016-03-01

    Single-shot dual-energy sandwich detector can produce sharp images because of subtraction of images from two sub-detector layers, which have different thick x-ray converters, of the sandwich detector. Inspired by this observation, the authors have developed a microtomography system with the sandwich detector in pursuit of high-resolution bone-enhanced small-animal imaging. The preliminary results show that the bone-enhanced images reconstructed with the subtracted projection data are better in visibility of bone details than the conventionally reconstructed images. In addition, the bone-enhanced images obtained from the sandwich detector are relatively immune to the artifacts caused by photon starvation. The microtomography with the single-shot dual-energy sandwich detector will be useful for the high-resolution bone imaging.

  15. Single and Dual Physical Link Failures Stability Effect on Degree Three WDM Networks

    DEFF Research Database (Denmark)

    Gutierrez Lopez, Jose Manuel; Georgakilas, Konstantinos; Riaz, M. Tahir

    2010-01-01

    This paper studies the effects of Single and Dual physical link failures to the stability of WDM networks when deployed as regular 3-degree structures. The failure impact on the source-destination pair connections is evaluated for the different topology scenarios. In this way it is possible...... to provide an overview of the failure effects and their relation to network metrics such as availability or cost. The results quantify how much the different network interconnection designs are affected by Single and Dual Physical link failures. The case study treats a realistic scenario, the interconnection...... of the NSFNET topology nodes. The main conclusions show that, when networks are designed under the same conditions, there is a linear relation between average downtime and ratio of connections affected by failures. Moreover, the most expensive optimized topologies to deploy provide higher availability....

  16. Femtosecond three-photon excitation and single-photon timing detection of α-NPO fluorescence

    Science.gov (United States)

    Volkmer, A.; Hatrick, D. A.; Bai, Y.; Birch, D. J. S.

    1997-04-01

    We demonstrate the application of three-photon excitation to fluorescence probe studies using time-correlated single-photon counting (TCSPC). By exciting with 120 fs Ti:sapphire laser pulses at 800 nm we have observed fluorescence emission from the scintillator 2-(1-napthyl)-5-phenyloxazole (α-NPO) in solutions and small unilamellar vesicles (SUVs) of L-α-dipalmitoylphosphatidylcholine (DPPC). In SUVs the time-resolved excimer emission and fluorescence anisotropy are consistent with a heterogeneous distribution of α-NPO molecules between isolated sites and ground state clusters in a similar manner to that which we reported previously for 2,5-diphenyloxazole (PPO).

  17. Dual-energy compared to single-energy CT in pediatric imaging: a phantom study for DECT clinical guidance

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Xiaowei; Servaes, Sabah; Darge, Kassa [The Children' s Hospital of Philadelphia, Department of Radiology, Philadelphia, PA (United States); University of Pennsylvania, The Perelman School of Medicine, Philadelphia, PA (United States); McCullough, William P. [University of Virginia Health System, Department of Radiology and Medical Imaging, Charlottesville, VA (United States); Mecca, Patricia [The Children' s Hospital of Philadelphia, Department of Radiology, Philadelphia, PA (United States)

    2016-11-15

    Dual-energy CT technology is available on scanners from several vendors and offers significant advantages over classic single-energy CT technology in multiple clinical applications. Many studies have detailed dual-energy CT applications in adults and several have evaluated the relative radiation dose performance of dual-energy CT in adult imaging. However, little has been published on dual-energy CT imaging in the pediatric population, and the relative dose performance of dual-energy CT imaging in the pediatric population is not well described. When evaluating dual-energy CT technology for implementation into a routine clinical pediatric imaging practice, the radiation dose implications must be considered, and when comparing relative CT dose performance, image quality must also be evaluated. Therefore the purpose of this study is to develop dual-energy CT scan protocols based on our optimized single-energy scan protocols and compare the dose. We scanned the head, chest and abdomen regions of pediatric-size anthropomorphic phantoms with contrast inserts, using our optimized single-energy clinical imaging protocols on a Siemens Flash {sup registered} CT scanner. We then scanned the phantoms in dual-energy mode using matching image-quality reference settings. The effective CT dose index volume (CTDI{sub vol}) of the scans was used as a surrogate for relative dose in comparing the single- and dual-energy scans. Additionally, we evaluated image quality using visual assessment and contrast-to-noise ratio. Dual-energy CT scans of the head and abdomen were dose-neutral for all three phantoms. Dual-energy CT scans of the chest showed a relative dose increase over the single-energy scan for 1- and 5-year-old child-based age-equivalent phantoms, ranging 11-20%. Quantitative analysis of image quality showed no statistically significant difference in image quality between the single-energy and dual-energy scans. There was no clinically significant difference in image quality by

  18. Heterogeneous fluorescence intermittency in single layer reduced graphene oxide

    Science.gov (United States)

    Si, Jixin; Volkan-Kacso, Sandor; Eltom, Ahmed; Morozov, Yurii; McDonald, Matthew P.; Ruth, Anthony; Kuno, Masaru; Janko, Boldizsar

    Fluorescence intermittency, or blinking, has been observed in a wide range of systems, including quantum dots, nanorods, and nanowires. Striking similarities have been documented in the optical response of these nanoscale emitters. However, the mechanism behind blinking still remains elusive. For the first time, blinking has been observed in a two-dimensional system in recent experiments on reduced graphene oxide (rGO). Here we reveal the power spectral density (PSD) of the blinking in rGO shares the same 1/f-like behavior of previously known blinking systems; meanwhile, the heterogeneous dynamic evolution and spatial correlation make rGO a unique blinking system. To investigate the origin of blinking, we self-consistently explain the evolution of rGO blinking using the phenomenological multiple recombination center (MRC) model that captures common features of nanoscale blinking. Furthermore, tight binding method and ab-initio method calculations of carbon nanodots are utilized to look for the microscopic structure corresponding to the RCs in the MRC model. M. K. thanks the American Chemical Society Petroleum Research Fund, the Army Research Office (W911NF-12-1-0578) for support. B.J. was supported in part by the U. S. DOE, Office of Science, Office of Basic Energy Sciences, under Contract W-31-109-Eng-38.

  19. High-sensitivity single-molecule fluorescence detection in theory and practice

    Energy Technology Data Exchange (ETDEWEB)

    Mathies, R.A.; Peck, K. (California Univ., Berkeley, CA (United States). Dept. of Chemistry); Stryer, L. (Stanford Univ., CA (United States). Dept. of Cell Biology)

    1989-01-01

    The number of emitted photons that can be obtained from a fluorophore increases with the incident light intensity and the duration of illumination. However, saturation of the absorption transition and photodestruction place natural limits on the ultimate signal-to-noise ratio that can be obtained. Equations have been derived to describe the fluorescence-to-background-noise ratio in the presence of saturating light intensities and photodestruction. The fluorescence lifetime and the photodestruction quantum yield are the key parameters that determine the optimum light intensity and exposure time. To test this theory we have performed single molecule detection of phycoerythrin (PE). The laser power was selected to give a mean time between absorptions approximately equal to the fluorescence decay rate. The transit time was selected to be nearly equal to the photodestruction time of {approximately}600 {mu}s. Under these conditions the photocount distribution function, the photocount autocorrelation function, and the concentration dependence clearly show that we are detecting bursts of fluorescence from individual fluorophores. A hard-wired version of this single-molecule detection system was used to measure the concentration of PE down to 10{sup {minus}15} M. This single-molecule counter is three orders-of-magnitude more sensitive than conventional fluorescence detection systems. The approach presented here should be useful in the optimization of fluorescence detected DNA sequencing gels. 17 refs., 4 figs.

  20. Performance of low resistivity single and dual-gap RPCs for LHCb

    CERN Document Server

    Adinolfi, M; Messi, R; Pacciani, L; Santovetti, E; Santovetti, Emanuele

    1999-01-01

    99-049 RPCs are strong candidates for the outer regions of the LHCb muon detector. We have tested single-gap and dual-gap detectors built with low-resistivity phenolic plates (ro = 9 x 10^9 microcm) and operated in avalanche mode. Measurements have been performed over a wide range of beam intensities and on the GIF at CERN. The results are presented and discussed, with special emphasis on the detection efficiency.

  1. Performance of low-resistivity single and dual-gap RPCs for LHCb

    CERN Document Server

    Adinolfi, M; Messi, R; Pacciani, L; Paoluzi, L; Santovetti, E

    2000-01-01

    Resistive plate chambers (RPC) are strong candidates for the outer regions of the LHCb muon detector. We have tested single-gap and dual-gap detectors built with low-resistivity phenolic plates ( rho =9*10/sup 9/ Omega cm) and operated in avalanche mode. Measurements have been performed over a wide range of beam intensities and on the GIF at CERN. The results are presented and discussed, with special emphasis on the detection efficiency. (6 refs).

  2. Onsite-effects of dual-hemisphere versus conventional single-hemisphere transcranial direct current stimulation

    OpenAIRE

    Kwon, Yong Hyun; Jang, Sung Ho

    2012-01-01

    We performed functional MRI examinations in six right-handed healthy subjects. During functional MRI scanning, transcranial direct current stimulation was delivered with the anode over the right primary sensorimotor cortex and the cathode over the left primary sensorimotor cortex using dual-hemispheric transcranial direct current stimulation. This was compared to a cathode over the left supraorbital area using conventional single-hemispheric transcranial direct current stimulation. Voxel coun...

  3. Comparison of Single Loop and Dual Loop PP Reactors and PP Product Development

    Institute of Scientific and Technical Information of China (English)

    Lu Yunfeng; Jiang Rong; Feng Suogui

    2003-01-01

    The present status of PP technology in China is presented. Through a review of single loop and dual loop PP reactor technology as well as development and utilization of PP products it is concluded that import of PP technology should be based on the demand of PP products and market trend with consideration of local conditions. The existing problems and future development of PP technology in China are discussed.

  4. Reliable measurement of E. coli single cell fluorescence distribution using a standard microscope set-up.

    Science.gov (United States)

    Cortesi, Marilisa; Bandiera, Lucia; Pasini, Alice; Bevilacqua, Alessandro; Gherardi, Alessandro; Furini, Simone; Giordano, Emanuele

    2017-01-01

    Quantifying gene expression at single cell level is fundamental for the complete characterization of synthetic gene circuits, due to the significant impact of noise and inter-cellular variability on the system's functionality. Commercial set-ups that allow the acquisition of fluorescent signal at single cell level (flow cytometers or quantitative microscopes) are expensive apparatuses that are hardly affordable by small laboratories. A protocol that makes a standard optical microscope able to acquire quantitative, single cell, fluorescent data from a bacterial population transformed with synthetic gene circuitry is presented. Single cell fluorescence values, acquired with a microscope set-up and processed with custom-made software, are compared with results that were obtained with a flow cytometer in a bacterial population transformed with the same gene circuitry. The high correlation between data from the two experimental set-ups, with a correlation coefficient computed over the tested dynamic range > 0.99, proves that a standard optical microscope- when coupled with appropriate software for image processing- might be used for quantitative single-cell fluorescence measurements. The calibration of the set-up, together with its validation, is described. The experimental protocol described in this paper makes quantitative measurement of single cell fluorescence accessible to laboratories equipped with standard optical microscope set-ups. Our method allows for an affordable measurement/quantification of intercellular variability, whose better understanding of this phenomenon will improve our comprehension of cellular behaviors and the design of synthetic gene circuits. All the required software is freely available to the synthetic biology community (MUSIQ Microscope flUorescence SIngle cell Quantification).

  5. Fluorescence assay for glycan expression on living cancer cells based on competitive strategy coupled with dual-functionalized nanobiocomposites.

    Science.gov (United States)

    Fu, Ying; Lu, Danqin; Lin, Bin; Sun, Qianqian; Liu, Kai; Xu, Lili; Zhang, Shengping; Hu, Chen; Wang, Chuangui; Xu, Zhiai; Zhang, Wen

    2013-11-21

    Cell surface glycans are a class of sophisticated biomolecules related to cancer development and progression, and their analysis is of great significance for early cancer diagnosis and treatment. In this paper, we proposed a fluorescence assay to evaluate glycan expression on living cancer cells based on a competitive strategy coupled with dual-functionalized nanobiocomposites. The competitive assay was conducted between living cancer cells and thiomannosyl derivatives using concanavalin A (Con A)-modified electrode as the interaction platform. To impart fluorescence signaling ability to competitive derivatives, quantum dots (QDs) were anchored on BSA-protected Au nanoparticles, and thiomannosyl derivatives were further immobilized on the nanoparticle surface through Au-S binding. Due to the spacing between QDs and Au nanoparticles by BSA, the {QDs-Au-BSA-mannose} nanobiocomposites maintained the fluorescence of QDs and showed binding ability with the Con A-modified electrode. Au nanorods (AuNRs)-modified electrode was used as an effective substrate to immobilize Con A. This assay was successfully applied to the analysis of two cancer cells lines (A549 and QGY-7701). The method is simple and shows promise for the study of glycan expression on living cancer cells.

  6. Dual coupled radiative transfer equation and diffusion approximation for the solution of the forward problem in fluorescence molecular imaging

    Science.gov (United States)

    Gorpas, Dimitris; Andersson-Engels, Stefan

    2012-03-01

    The solution of the forward problem in fluorescence molecular imaging is among the most important premises for the successful confrontation of the inverse reconstruction problem. To date, the most typical approach has been the application of the diffusion approximation as the forward model. This model is basically a first order angular approximation for the radiative transfer equation, and thus it presents certain limitations. The scope of this manuscript is to present the dual coupled radiative transfer equation and diffusion approximation model for the solution of the forward problem in fluorescence molecular imaging. The integro-differential equations of its weak formalism were solved via the finite elements method. Algorithmic blocks with cubature rules and analytical solutions of the multiple integrals have been constructed for the solution. Furthermore, specialized mapping matrices have been developed to assembly the finite elements matrix. As a radiative transfer equation based model, the integration over the angular discretization was implemented analytically, while quadrature rules were applied whenever required. Finally, this model was evaluated on numerous virtual phantoms and its relative accuracy, with respect to the radiative transfer equation, was over 95%, when the widely applied diffusion approximation presented almost 85% corresponding relative accuracy for the fluorescence emission.

  7. Analysis of single band and dual band graphene based patch antenna for terahertz region

    Science.gov (United States)

    George, Jemima Nissiyah; Madhan, M. Ganesh

    2017-10-01

    A microstrip patch antenna is designed using a very thin layer of graphene as the radiating patch, which is fed by a microstrip transmission line. The graphene based patch is designed on a silicon substrate having a dielectric constant of 11.9, to radiate at a single frequency of 2.6 THz. Further, this antenna is made to resonate at dual frequencies of 2.48 THz and 3.35 THz, by changing the substrate height, which is reported for the first time. Various antenna parameters such as return loss, VSWR, gain, efficiency and bandwidth are also determined for the single and dual band operation. For the single band operation, a bandwidth of 145.4 GHz and an efficiency of 92% was achieved. For dual band operation, a maximum bandwidth of 140.5 GHz was obtained at 3.35 THz and an efficiency of 87.3% was obtained at the first resonant frequency of 2.48 THz. The absorption cross section of the antenna is also analysed for various substrate heights and has maximum peaks at the corresponding resonating frequencies. The simulation has been carried out by using a full wave electromagnetic simulator based on FDTD method.

  8. Comparison of Single and Dual Target Visual Attention Tasks in Children with down Syndrome

    Directory of Open Access Journals (Sweden)

    Melanie J. Murphy

    2011-05-01

    Full Text Available Understanding the nature of attentional processing in children with Down Syndrome (DS is imperative for developing effective education practices. The aim of the current study was to investigate whether children with DS exhibit impairment in sustained, transient, single-, or dual-target continuous performance tasks. Target detection time and accuracy was compared in children with DS to Typically Developing (TD children of similar nonverbal mental age (as measured by the Raven's Coloured Progressive Matrices, on single and dual- target continuous performance tasks measuring sustained attention, a visual change detection task measuring transient attention, and feature and conjunctive visual search tasks measuring both sustained and transient attention. Results showed that children with DS performed similarly to TD children on sustained and transient attention tasks that only required the detection of a single unique target, but were impaired in overall accuracy on tasks that required dual-target detection. Findings suggest a possible impairment in attention and working memory in children with DS. Error analysis of task responses revealed differences in problem solving strategy between children with DS and TD children, despite similar overall performance. Findings have implications for the education of children with DS and understanding of the nature of intellectual disability per se.

  9. A multi-view time-domain non-contact diffuse optical tomography scanner with dual wavelength detection for intrinsic and fluorescence small animal imaging.

    Science.gov (United States)

    Lapointe, Eric; Pichette, Julien; Bérubé-Lauzière, Yves

    2012-06-01

    We present a non-contact diffuse optical tomography (DOT) scanner with multi-view detection (over 360°) for localizing fluorescent markers in scattering and absorbing media, in particular small animals. It relies on time-domain detection after short pulse laser excitation. Ultrafast time-correlated single photon counting and photomultiplier tubes are used for time-domain measurements. For light collection, seven free-space optics non-contact dual wavelength detection channels comprising 14 detectors overall are placed around the subject, allowing the measurement of time point-spread functions at both excitation and fluorescence wavelengths. The scanner is endowed with a stereo camera pair for measuring the outer shape of the subject in 3D. Surface and DOT measurements are acquired simultaneously with the same laser beam. The hardware and software architecture of the scanner are discussed. Phantoms are used to validate the instrument. Results on the localization of fluorescent point-like inclusions immersed in a scattering and absorbing object are presented. The localization algorithm relies on distance ranging based on the measurement of early photons arrival times at different positions around the subject. This requires exquisite timing accuracy from the scanner. Further exploiting this capability, we show results on the effect of a scattering hetereogenity on the arrival time of early photons. These results demonstrate that our scanner provides all that is necessary for reconstructing images of small animals using full tomographic reconstruction algorithms, which will be the next step. Through its free-space optics design and the short pulse laser used, our scanner shows unprecedented timing resolution compared to other multi-view time-domain scanners.

  10. New dual emission fluorescent sensor for pH and Pb(II) based on bis(napfthalimide) derivative

    Energy Technology Data Exchange (ETDEWEB)

    Pina-Luis, Georgina, E-mail: gpinaluis@yahoo.com [Centro de Graduados e Investigacion en Quimica, Instituto Tecnologico de Tijuana, AP 1166, Tijuana 22500, BC (Mexico); Martinez-Quiroz, Marisela; Ochoa-Teran, Adrian [Centro de Graduados e Investigacion en Quimica, Instituto Tecnologico de Tijuana, AP 1166, Tijuana 22500, BC (Mexico); Santacruz-Ortega, Hisila [Departamento de investigacion en Polimeros y Materiales, Universidad de Sonora, Hermosillo, Sonora 83000 (Mexico); Mendez-Valenzuela, Eduardo [Centro de Graduados e Investigacion en Quimica, Instituto Tecnologico de Tijuana, AP 1166, Tijuana 22500, BC (Mexico)

    2013-02-15

    This paper describes a novel dual emission bis-1,8-naphthalimide sensor for selective determination of pH and Pb{sup 2+} ions. The influence of the variability in the backbone that links the two fluorophores (naphthalimides) as a function of pH and metal ions was studied by UV-visible and fluorescence spectroscopy. Compounds 1(a-d) with different length alkyl linkers (CH{sub 2}){sub n} (n=1, 2, 4 and 6) showed no excimer formation in aqueous solution. Fluorescence emission of these derivatives varied in a narrow range of pH (5-8) and was only slightly influenced by the addition of metal ions in CH{sub 3}CN solutions. However, derivative 1e with amino-containing spacer (CH{sub 2}-NH-CH{sub 2}) showed excimer emission in aqueous solution, a wide response to pH (2.5-9.5) and fluorescence enhancement with selective behavior towards metal ions. The pH sensor based in derivative 1e has a sufficient selectivity for practical pH monitoring in the presence of Li{sup +}, Na{sup +}, K{sup +}, Cs{sup +}, Ca{sup 2+}, Mg{sup 2+}, Ba{sup 2+}, Cu{sup 2+}, Pb{sup 2+}, Ni{sup 2+}, Zn{sup 2+} and Cd{sup 2+}. The coordination chemistry of these complexes was studied by UV-Vis, fluorescence and {sup 1}H NMR. This chemosensor displayed high selectivity fluorescence enhancement toward Pb{sup 2+} ions in the presence of the metals ions mentioned in CH{sub 3}CN solutions. Competitive assays show that a 1-fold of metal cations in each case, compared with Pb{sup 2+} ions, results in less than {+-}5% fluorescence intensity changes. Linear calibration up to 1 Multiplication-Sign 10{sup -5} M for Pb(II) ions (R=0.9968) was obtained and detection limit resulted of 5.0 Multiplication-Sign 10{sup -8} M. - Highlights: Black-Right-Pointing-Pointer A novel dual emission bis-1,8-naphthalimide sensor for pH and Pb{sup 2+} ions is synthetized. Black-Right-Pointing-Pointer The excimer formation depends on the spacer that links the two naphthalimide groups. Black-Right-Pointing-Pointer Bis

  11. Dual-Utility of Digital Holography & Epi-Fluorescence to Localize Cellular Nuclei

    Science.gov (United States)

    Sheldrake, Eric

    Digital Holographic Microscopy (DHM) and Epi-Fluorescence have been combined for biological imaging. DHM is a non-invasive phase contrast microscopy technique that provides quantitative information such as the variations in refractive index and physical height. 3D physical height and refractive index profiles are imaged for HEK293 and CHO cells. These values and profiles are compared with known values for the sample gathered from literature and other microscopy techniques, including Scanning Electron Microscopy. The measured CHO cellular length is (15.31 +/- 2.60) microm. Localization of cellular nuclei is performed using the Epi-Fluorescence setup and the DNA specific fluorophore DAPI. The fluorescence intensity profile was imaged, where nuclei shape, size, and localization are compared using an A.1 Zeiss Fluorescence Microscope. The CHO cells are comparable with apoptotic cells, where the measured nucleus length is (10.91 +/- 2.27) microm. DHM and Epi-Fluorescence are combined to analyze the physical heights of the cell as well as localize its nucleus. The combination of these techniques are greatly advantageous to understand cellular functions and variability.

  12. State space approach to single molecule localization in fluorescence microscopy.

    Science.gov (United States)

    Vahid, Milad R; Chao, Jerry; Kim, Dongyoung; Ward, E Sally; Ober, Raimund J

    2017-03-01

    Single molecule super-resolution microscopy enables imaging at sub-diffraction-limit resolution by producing images of subsets of stochastically photoactivated fluorophores over a sequence of frames. In each frame of the sequence, the fluorophores are accurately localized, and the estimated locations are used to construct a high-resolution image of the cellular structures labeled by the fluorophores. Many methods have been developed for localizing fluorophores from the images. The majority of these methods comprise two separate steps: detection and estimation. In the detection step, fluorophores are identified. In the estimation step, the locations of the identified fluorophores are estimated through an iterative approach. Here, we propose a non-iterative state space-based localization method which combines the detection and estimation steps. We demonstrate that the estimated locations obtained from the proposed method can be used as initial conditions in an estimation routine to potentially obtain improved location estimates. The proposed method models the given image as the frequency response of a multi-order system obtained with a balanced state space realization algorithm based on the singular value decomposition of a Hankel matrix. The locations of the poles of the resulting system determine the peak locations in the frequency domain, and the locations of the most significant peaks correspond to the single molecule locations in the original image. The performance of the method is validated using both simulated and experimental data.

  13. Multi-functional stage-scanning fluorescence micro/nanoscope for single-lipid dynamics

    Science.gov (United States)

    Yang, Li-Ling; Hsieh, Chia-Fen; Chang, Yi-Ren; Shen, Jie-Pan; Chang, Yu-Chung; Chou, Chia-Fu

    2012-02-01

    We combine pulsed laser, supercontinuum radiation source and fast single-photon counting peripherals to obtain a multifunctional micro/nano-scope. This provides us with better spatial and temporal resolution to observe fast dynamics. Performing fluorescence correlation spectroscopy for fast dynamics (lipid dynamics in supported lipid bilayers and living cells is our goal. Lipid raft serves as a platform for recruiting signaling components of effective signal transduction. However, the dynamics of sub-200nm rapidly aggregated lipid rafts are still not elucidated in living cells. We here report our recent progress on the construction of this multi-functional stage-scanning fluorescence micro/nanoscope for single-lipid dynamics study.

  14. Opportunities and challenges in single-molecule and single-particle fluorescence microscopy for mechanistic studies of chemical reactions

    Science.gov (United States)

    Cordes, Thorben; Blum, Suzanne A.

    2013-12-01

    In recent years, single-molecule and single-particle fluorescence microscopy has emerged as a tool to investigate chemical systems. After an initial lag of over a decade with respect to biophysical studies, this powerful imaging technique is now revealing mechanisms of 'classical' organic reactions, spatial distribution of chemical reactivity on surfaces and the phase of active catalysts. The recent advance into commercial imaging systems obviates the need for home-built laser systems and thus opens this technique to traditionally trained synthetic chemists. We discuss the requisite photophysical and chemical properties of fluorescent reporters and highlight the main challenges in applying single-molecule techniques to chemical questions. The goal of this Perspective is to provide a snapshot of an emerging multidisciplinary field and to encourage broader use of this young experimental approach that aids the observation of chemical reactions as depicted in many textbooks: molecule by molecule.

  15. Opportunities and challenges in single-molecule and single-particle fluorescence microscopy for mechanistic studies of chemical reactions.

    Science.gov (United States)

    Cordes, Thorben; Blum, Suzanne A

    2013-12-01

    In recent years, single-molecule and single-particle fluorescence microscopy has emerged as a tool to investigate chemical systems. After an initial lag of over a decade with respect to biophysical studies, this powerful imaging technique is now revealing mechanisms of 'classical' organic reactions, spatial distribution of chemical reactivity on surfaces and the phase of active catalysts. The recent advance into commercial imaging systems obviates the need for home-built laser systems and thus opens this technique to traditionally trained synthetic chemists. We discuss the requisite photophysical and chemical properties of fluorescent reporters and highlight the main challenges in applying single-molecule techniques to chemical questions. The goal of this Perspective is to provide a snapshot of an emerging multidisciplinary field and to encourage broader use of this young experimental approach that aids the observation of chemical reactions as depicted in many textbooks: molecule by molecule.

  16. Corrosion protection performance of single and dual Plasma Electrolytic Oxidation (PEO) coating for aerospace applications

    Energy Technology Data Exchange (ETDEWEB)

    Madhan Kumar, A., E-mail: princemadhank@gmail.com; Kwon, Sun Hwan; Jung, Hwa Chul; Shin, Kwang Seon

    2015-01-15

    Plasma Electrolytic Oxidation (PEO) coatings are known to be one of the most appropriate method for corrosion protection of magnesium (Mg) alloy. The improvement of PEO coatings and the optimization of their surface aspects are of major importance. In this current work, the influence of dual PEO coating on strip-cast AZ31 Mg alloy substrate has been evaluated with the aim of improving the surface and corrosion protection aspects. For this purpose, AZ31 Mg substrates are subjected to single and dual PEO processing in silicate and phosphate electrolyte under similar condition. Scanning electron microscopy (SEM) analysis confirmed that the number of pores in PEO coating processed in silicate electrolyte is higher than others. X-ray diffraction analysis of PEO coatings showed that the surface coating is mainly comprised of Mg{sub 2}SiO{sub 4}, Mg{sub 3}(PO{sub 4}){sub 2} and MgO with different quantity based on PEO processing. Compared with the AZ31 Mg, the corrosion potential (E{sub corr}) of both type PEO coatings was positively shifted about 250–400 mV and the corrosion current density (i{sub corr}) was lowered by 3-4 orders of magnitude as result of adequate corrosion protection to the Mg alloy in 3.5% NaCl solution. All of the observation obviously showed that the dual PEO coating provides better corrosion protection performance than their respective single due to its synergistic beneficial effect. - Highlights: • Influence of dual PEO coating on AZ31 Mg alloy substrate was evaluated. • XRD confirmed formation of thin MgO inner, Mg{sub 3}(PO{sub 4}){sub 2} and Mg{sub 2}SiO{sub 4} outer layer. • SEM results showed uniform coating with no cracks and relatively less micro pores. • Micro hardness of dual PEO coatings is higher than single PEO coatings. • Dual coating provides superior corrosion performance due to its synergistic effect.

  17. Dual-Modal Colorimetric/Fluorescence Molecular Probe for Ratiometric Sensing of pH and Its Application.

    Science.gov (United States)

    Wu, Luling; Li, Xiaolin; Huang, Chusen; Jia, Nengqin

    2016-08-16

    As traditional pH meters cannot work well for minute regions (such as subcellular organelles) and in harsh media, molecular pH-sensitive devices for monitoring pH changes in diverse local heterogeneous environments are urgently needed. Here, we report a new dual-modal colorimetric/fluorescence merocyanine-based molecular probe (CPH) for ratiometric sensing of pH. Compared with previously reported pH probes, CPH bearing the benzyl group at the nitrogen position of the indolium group and the phenol, which is used as the acceptor for proton, could respond to pH changes immediately through both the ratiometric fluorescence signal readout and naked-eye colorimetric observation. The sensing process was highly stable and reversible. Most importantly, the suitable pKa value (6.44) allows CPH to presumably accumulate in lysosomes and become a lysosome-target fluorescent probe. By using CPH, the intralysosomal pH fluctuation stimulated by antimalaria drug chloroquine was successfully tracked in live cells through the ratiometric fluorescence images. Additionally, CPH could be immobilized on test papers, which exhibited a rapid and reversible colorimetric response to acid/base vapor through the naked-eye colorimetric analysis. This proof-of-concept study presents the potential application of CPH as a molecular tool for monitoring intralysosomal pH fluctuation in live cells, as well as paves the way for developing the economic, reusable, and fast-response optical pH meters for colorimetric sensing acid/base vapor with direct naked-eye observation.

  18. 5-Fluorotryptophan as a Dual NMR and Fluorescent Probe of α-Synuclein

    Science.gov (United States)

    Pfefferkorn, Candace M.; Lee, Jennifer C.

    2012-01-01

    i. Summary Analysis of conventional proton nuclear magnetic resonance (NMR) experiments on intrinsically disordered proteins (IDPs) is challenging because of the highly flexible and multiple rapidly exchanging conformations typifying this class of proteins. One method to circumvent some of these difficulties is to incorporate non-native fluorine (19F) nuclei at specific sites within the polypeptide. 19F NMR is particularly suitable for characterization of unfolded structures because 19F chemical shifts are highly sensitive to local environments and conformations. Furthermore, the incorporation of fluorine analogs of fluorescent amino acids such as 5-fluoro-tryptophan (5FW) allows for complementary studies of protein microenvironment via fluorescence spectroscopy. Herein we describe methods to produce, purify, characterize, and perform steady-state fluorescence and 1-D NMR experiments on 5FW analogues of the IDP α-synuclein. PMID:22760321

  19. Single-Task and Dual-Task Gait Among Collegiate Athletes of Different Sport Classifications: Implications for Concussion Management.

    Science.gov (United States)

    Howell, David R; Oldham, Jessie R; DiFabio, Melissa; Vallabhajosula, Srikant; Hall, Eric E; Ketcham, Caroline J; Meehan, William P; Buckley, Thomas A

    2017-02-01

    Gait impairments have been documented following sport-related concussion. Whether preexisting gait pattern differences exist among athletes who participate in different sport classifications, however, remains unclear. Dual-task gait examinations probe the simultaneous performance of everyday tasks (ie, walking and thinking), and can quantify gait performance using inertial sensors. The purpose of this study was to compare the single-task and dual-task gait performance of collision/contact and noncontact athletes. A group of collegiate athletes (n = 265) were tested before their season at 3 institutions (mean age= 19.1 ± 1.1 years). All participants stood still (single-task standing) and walked while simultaneously completing a cognitive test (dual-task gait), and completed walking trials without the cognitive test (single-task gait). Spatial-temporal gait parameters were compared between collision/contact and noncontact athletes using MANCOVAs; cognitive task performance was compared using ANCOVAs. No significant single-task or dual-task gait differences were found between collision/contact and noncontact athletes. Noncontact athletes demonstrated higher cognitive task accuracy during single-task standing (P = .001) and dual-task gait conditions (P = .02) than collision/contact athletes. These data demonstrate the utility of a dual-task gait assessment outside of a laboratory and suggest that preinjury cognitive task performance during dual-tasks may differ between athletes of different sport classifications.

  20. All-Dielectric Silicon Nanogap Antennas To Enhance the Fluorescence of Single Molecules.

    Science.gov (United States)

    Regmi, Raju; Berthelot, Johann; Winkler, Pamina M; Mivelle, Mathieu; Proust, Julien; Bedu, Frédéric; Ozerov, Igor; Begou, Thomas; Lumeau, Julien; Rigneault, Hervé; García-Parajó, María F; Bidault, Sébastien; Wenger, Jérôme; Bonod, Nicolas

    2016-08-10

    Plasmonic antennas have a profound impact on nanophotonics as they provide efficient means to manipulate light and enhance light-matter interactions at the nanoscale. However, the large absorption losses found in metals can severely limit the plasmonic applications in the visible spectral range. Here, we demonstrate the effectiveness of an alternative approach using all-dielectric nanoantennas based on silicon dimers to enhance the fluorescence detection of single molecules. The silicon antenna design is optimized to confine the near-field intensity in the 20 nm nanogap and reach a 270-fold fluorescence enhancement in a nanoscale volume of λ(3)/1800 with dielectric materials only. Our conclusions are assessed by combining polarization resolved optical spectroscopy of individual antennas, scanning electron microscopy, numerical simulations, fluorescence lifetime measurements, fluorescence burst analysis, and fluorescence correlation spectroscopy. This work demonstrates that all-silicon nanoantennas are a valid alternative to plasmonic devices for enhanced single molecule fluorescence sensing, with the additional key advantages of reduced nonradiative quenching, negligible heat generation, cost-efficiency, and complementary metal-oxide-semiconductor (CMOS) compatibility.

  1. Improved perovskite film quality and solar cell performances using dual single solution coating

    Science.gov (United States)

    Baltakesmez, Ali; Biber, Mehmet; Tüzemen, Sebahattin

    2017-08-01

    In this study, we present high quality perovskite CH3NH3PbI3-xClx thin films prepared by a combination of static and dynamic coating approaches, named dual single solution coating. Static coating, dynamic coating and the combination of these are comparatively studied. Scanning electron microscopy, X-ray diffraction, atomic force microscopy, UV-visible spectroscopy, and photoluminescence techniques are used for the determination of morphological, structural, and optical properties of thin films prepared using different coating approaches and deposition temperatures. All the coating approaches are repeated at room temperature and with hot deposition. The high quality and density CH3NH3PbI3-xClx films with full surface coverage are obtained using the dual single solution coating, particularly with hot-deposition. The perovskite solar cells prepared by the dual coating approach with hot deposition have better values for all the performance parameters in comparison to the other coating approaches, resulting in high efficiencies. The best device has a short circuit current of 22.03 mA/cm2, an open circuit voltage of 0.91 V, a fill factor of 0.73, and a power conversion efficiency of 14.68% from short-circuit to forward bias, and 22.39 mA/cm2, 0.91 V, 75% and 15.32% for the vice-versa, respectively.

  2. Can single-phase dual-energy CT reliably identify adrenal adenomas?

    Energy Technology Data Exchange (ETDEWEB)

    Helck, A.; Hummel, N.; Meinel, F.G.; Johnson, T.; Nikolaou, K.; Graser, A. [University of Munich, Institute for Clinical Radiology, Munich (Germany)

    2014-07-15

    To evaluate whether single-phase dual-energy-CT-based attenuation measurements can reliably differentiate lipid-rich adrenal adenomas from malignant adrenal lesions. We retrospectively identified 51 patients with adrenal masses who had undergone contrast-enhanced dual-energy-CT (140/100 or 140/80 kVp). Virtual non-contrast and colour-coded iodine images were generated, allowing for measurement of pre- and post-contrast density on a single-phase acquisition. Adrenal adenoma was diagnosed if density on virtual non-contrast images was ≤10 HU. Clinical follow-up, true non-contrast CT, PET/CT, in- and opposed-phase MRI, and histopathology served as the standard of reference. Based on the standard of reference, 46/57 (80.7 %) adrenal masses were characterised as adenomas or other benign lesions; 9 malignant lesions were detected. Based on a cutoff value of 10 HU, virtual non-contrast images allowed for correct identification of adrenal adenomas in 33 of 46 (71 %), whereas 13/46 (28 %) adrenal adenomas were lipid poor with a density ≥10 HU. Based on the threshold of 10 HU on the virtual non-contrast images, the sensitivity, specificity, and accuracy for detection of benign adrenal lesions was 73 %, 100 %, and 81 % respectively. Virtual non-contrast images derived from dual-energy-CT allow for accurate characterisation of lipid-rich adrenal adenomas and can help to avoid additional follow-up imaging. (orig.)

  3. Safety of dental extractions during uninterrupted single or dual antiplatelet treatment.

    Science.gov (United States)

    Lillis, Theodoros; Ziakas, Antonios; Koskinas, Konstantinos; Tsirlis, Anastasios; Giannoglou, George

    2011-10-01

    Optimal dental management in patients on long-term antiplatelet treatment is not clearly defined. Antiplatelet discontinuation increases the risk of thrombotic complications, whereas uninterrupted antiplatelet therapy, which is the currently recommended approach, is assumed to increase the bleeding hazard after dental procedures. We sought to prospectively compare the risk of immediate and late postextraction bleeding in patients receiving uninterrupted single or dual antiplatelet therapy. We recruited 643 consecutive patients referred for dental extractions. In total 111 (17.3%) were on clinically indicated antiplatelet therapy: aspirin (n = 42), clopidogrel (n = 36), and aspirin and clopidogrel (n = 33). Controls (n = 532, 82.7%) were not on antiplatelet treatment. Immediate and late bleeding complications were recorded. Compared to controls the risk of prolonged immediate bleeding was higher in patients on dual antiplatelet therapy (relative risk [RR] 177.3, 95% confidence interval [CI] 43.5 to 722, p patients on aspirin alone (RR = 6.3, 95% CI 0.6 to 68.4, p = 0.2) or clopidogrel alone (RR = 7.4, 95% CI 0.7 to 79.5, p = 0.18); however, all immediate bleeding complications in all treatment groups were successfully managed with local hemostatic measures. No patient developed any late hemorrhage. In conclusion, dental extractions may be safely performed in patients receiving single or dual antiplatelet therapy when appropriate local hemostatic measures are taken, thus averting thrombotic risk of temporary antiplatelet discontinuation.

  4. Ensemble and single-molecule studies on fluorescence quenching in transition metal bipyridine-complexes.

    Directory of Open Access Journals (Sweden)

    Dominik Brox

    Full Text Available Beyond their use in analytical chemistry fluorescent probes continuously gain importance because of recent applications of single-molecule fluorescence spectroscopy to monitor elementary reaction steps. In this context, we characterized quenching of a fluorescent probe by different metal ions with fluorescence spectroscopy in the bulk and at the single-molecule level. We apply a quantitative model to explain deviations from existing standard models for fluorescence quenching. The model is based on a reversible transition from a bright to a dim state upon binding of the metal ion. We use the model to estimate the stability constants of complexes with different metal ions and the change of the relative quantum yield of different reporter dye labels. We found ensemble data to agree widely with results from single-molecule experiments. Our data indicates a mechanism involving close molecular contact of dye and quenching moiety which we also found in molecular dynamics simulations. We close the manuscript with a discussion of possible mechanisms based on Förster distances and electrochemical potentials which renders photo-induced electron transfer to be more likely than Förster resonance energy transfer.

  5. Single particle fluorescence burst analysis of epsin induced membrane fission.

    Science.gov (United States)

    Brooks, Arielle; Shoup, Daniel; Kustigian, Lauren; Puchalla, Jason; Carr, Chavela M; Rye, Hays S

    2015-01-01

    Vital cellular processes, from cell growth to synaptic transmission, rely on membrane-bounded carriers and vesicles to transport molecular cargo to and from specific intracellular compartments throughout the cell. Compartment-specific proteins are required for the final step, membrane fission, which releases the transport carrier from the intracellular compartment. The role of fission proteins, especially at intracellular locations and in non-neuronal cells, while informed by the dynamin-1 paradigm, remains to be resolved. In this study, we introduce a highly sensitive approach for the identification and analysis of membrane fission machinery, called burst analysis spectroscopy (BAS). BAS is a single particle, free-solution approach, well suited for quantitative measurements of membrane dynamics. Here, we use BAS to analyze membrane fission induced by the potent, fission-active ENTH domain of epsin. Using this method, we obtained temperature-dependent, time-resolved measurements of liposome size and concentration changes, even at sub-micromolar concentration of the epsin ENTH domain. We also uncovered, at 37°C, fission activity for the full-length epsin protein, supporting the argument that the membrane-fission activity observed with the ENTH domain represents a native function of the full-length epsin protein.

  6. Ultimate Statistical Physics: fluorescence of a single atom

    CERN Document Server

    Pomeau, Yves; Ginibre, Jean

    2016-01-01

    We discuss the statistics of emission of photons by a single atom or ion illuminated by a laser beam at the frequency of quasi-resonance between two energy levels, a situation that corresponds to real experiments. We extend this to the case of two laser beams resonant with the energy differences between two excited levels and the ground state (three level atom in V-configuration). We use a novel approach of this type of problem by considering Kolmogorov equation for the probability distribution of the atomic state which takes into account first the deterministic evolution of this state under the effect of the incoming laser beam and the random emission of photons during the spontaneous decay of the excited state(s) to the ground state. This approach yields solvable equations in the two level atom case. For the three level atom case we set the problem and define clearly its frame. The results obtained are valid both in the opposite limits of rare and of frequent spontaneous decay, compared to the period of the...

  7. A novel non invasive measurement of hemodynamic parameters: Comparison of single-chamber ventricular and dual-chamber pacemaker

    Directory of Open Access Journals (Sweden)

    Ingrid M. Pardede

    2008-03-01

    Full Text Available We carried out a cross sectional study to analyze hemodynamic parameters of single-chamber ventricular pacemaker compared with dual-chamber pacemaker by using thoracic electrical bioimpedance monitoring method (Physio Flow™ - a novel simple non-invasive measurement. A total of 48 consecutive outpatients comprised of 27 single chamber pacemaker and 21 dual chamber were analyzed. We measured cardiac parameters: heart rate, stroke volume index, cardiac output index, estimated ejection fraction, end diastolic volume, early diastolic function ratio, thoracic fluid index, and systemic parameters: left cardiac work index and systemic vascular resistance index. Baseline characteristic and pacemaker indication were similar in both groups. Cardiac parameters assessment revealed no significant difference between single-chamber pacemaker and dual-chamber pacemaker in heart rate, stroke volume index, cardiac index, estimated ejection fraction, end-diastolic volume, thoracic fluid index. There was significantly higher early diastolic function ratio in single-chamber pacemaker compared to dual-chamber pacemaker: 92% (10.2-187.7% vs. 100.6% (48.7-403.2%; p=0.006. Systemic parameters assessment revealed significantly higher left cardiac work index in single-chamber group than dual-chamber group 4.9 kg.m/m² (2.8-7.6 kg.m/m² vs. 4.3 kg.m/m² (2.9-7.2 kg.m/m²; p=0.004. There was no significant difference on systemic vascular resistance in single-chamber compared to dual-chamber pacemaker. Single-chamber ventricular pacemaker provides similar stroke volume, cardiac output and left cardiac work, compared to dual-chamber pacemaker. A non-invasive hemodynamic measurement using thoracic electrical bioimpedance is feasible for permanent pacemaker outpatients. (Med J Indones 2008; 17: 25-32Keywords: Permanent pacemaker, single chamber, dual chamber, thoracic electrical bioimpedance, hemodynamic parameter

  8. Pre- and postfunctionalized self-assembled π-conjugated fluorescent organic nanoparticles for dual targeting.

    Science.gov (United States)

    Petkau, Katja; Kaeser, Adrien; Fischer, Irén; Brunsveld, Luc; Schenning, Albertus P H J

    2011-10-26

    There is currently a high demand for novel approaches to engineer fluorescent nanoparticles with precise surface properties suitable for various applications, including imaging and sensing. To this end, we report a facile and highly reproducible one-step method for generating functionalized fluorescent organic nanoparticles via self-assembly of prefunctionalized π-conjugated oligomers. The engineered design of the nonionic amphiphilic oligomers enables the introduction of different ligands at the extremities of inert ethylene glycol side chains without interfering with the self-assembly process. The intrinsic fluorescence of the nanoparticles permits the measurement of their surface properties and binding to dye-labeled target molecules via Förster resonance energy transfer (FRET). Co-assembly of differently functionalized oligomers is also demonstrated, which enables the tuning of ligand composition and density. Furthermore, nanoparticle prefunctionalization has been combined with subsequent postmodification of azide-bearing oligomers via click chemistry. This allows for expanding ligand diversity at two independent stages in the nanoparticle fabrication process. The practicability of the different methods entails greater control over surface functionality. Through labeling with different ligands, selective binding of proteins, bacteria, and functionalized beads to the nanoparticles has been achieved. This, in combination with the absence of unspecific adsorption, clearly demonstrates the broad potential of these nanoparticles for selective targeting and sequestration. Therefore, controlled bifunctionalization of fluorescent π-conjugated oligomer nanoparticles represents a novel approach with high applicability to multitargeted imaging and sensing in biology and medicine.

  9. Fluorescent magnetic Fe3 O4 /rare Earth colloidal nanoparticles for dual-modality imaging.

    Science.gov (United States)

    Zhu, Haie; Shang, Yalei; Wang, Wenhao; Zhou, Yingjie; Li, Penghui; Yan, Kai; Wu, Shuilin; Yeung, Kelvin W K; Xu, Zushun; Xu, Haibo; Chu, Paul K

    2013-09-09

    Fluorescent magnetic colloidal nanoparticles (FMCNPs) are produced by a two-step, seed emulsifier-free emulsion polymerization in the presence of oleic acid and sodium undecylenate-modified Fe3 O4 nanoparticles (NPs). The Fe3 O4 /poly(St-co-GMA) nanoparticles are first synthesized as the seed and Eu(AA)3 Phen is copolymerized with the remaining St and GMA to form the fluorescent polymer shell in the second step. The uniform core-shell structured FMCNPs with a mean diameter of 120 nm exhibit superparamagnetism with saturation magnetization of 1.92 emu/g. Red luminescence from the FMCNPs is confirmed by the salient fluorescence emission peaks of europium ions at 594 and 619 nm as well as 2-photon confocal scanning laser microscopy. The in vitro cytotoxicity test conducted using the MTT assay shows good cytocompatibility and the T2 relaxivity of the FMCNPs is 353.86 mM(-1) S(-1) suggesting its potential in magnetic resonance imaging (MRI). In vivo MRI studies based on a rat model show significantly enhanced T2 -weighted images of the liver after administration and prussian blue staining of the liver tissue slice reveals accumulation of FMCNPs in the organ. The cytocompatibility, superparamagnetism, and excellent fluorescent properties of FMCNPs make them suitable for biological imaging probes in MRI and optical imaging.

  10. The Acquisition of Past Tense in English/Norwegian Bilingual Children Single versus Dual Mechanisms

    Directory of Open Access Journals (Sweden)

    Maja Jensvoll

    2004-01-01

    Full Text Available In a study of three Norwegian/English bilingual siblings, their strategies for acquiring past tense of verbs in both languages were examined. Elicitation tests were performed in both languages and the children’s performance and error patterns were examined. These results were then compared to data from monolingual English and Norwegian speaking children. The results are discussed within the framework of the Single Mechanism Account Hypothesis, a Connectionist approach, and the Dual Mechanism Account Hypothesis, a Generative Approach. The current study suggests support for the Dual Mechanism Approach Hypothesis. It seems that the children in the current study did in fact create rules for past tense which they could apply by default.

  11. A fluoride-sensing receptor based on 2,2'-bis(indolyl)methane by dual-function of colorimetry and fluorescence.

    Science.gov (United States)

    Wei, Wei; Shao, Shi Jun; Guo, Yong

    2015-10-05

    A compound based on 2,2'-bis(indolyl)methane containing nitro group was studied as a new anion receptor. It could recognize selectively F(-) by an increasing fluorescence signal and a visible color change from colorless to blue. The introduction of nitro group induced the spectral dual-function related to the deprotonation of N-H protons.

  12. Polarisation Sensitive Single Molecule Fluorescence Detection with Linear Polarised Excitation Light and Modulated Polarisation Direction Applied to Multichromophoric Entities

    NARCIS (Netherlands)

    Gensch, T.; Hofkens, J.; Köhn, F.; Vosch, T.; Herrmann, A.; Müllen, K.; Schryver, F.C. De

    2001-01-01

    Recently, investigations of the fluorescence properties of a multichromophoric dendritic entity at the single molecule level have revealed multiple fluorescence levels, collective off-states, variations of the polarisation, large shifts in the spectral position and changes in the fluorescence decay

  13. Chemical responses of single yeast cells studied by fluorescence microspectroscopy under solution-flow conditions.

    Science.gov (United States)

    Kogi, Osamu; Kim, Haeng-Boo; Kitamura, Noboru

    2002-07-01

    A microspectroscopy system combined with a fluid manifold was developed to manipulate and analyze "single" living cells. A sample buffer solution containing living cells was introduced into a flow cell set on a thermostated microscope stage and a few cells were allowed to attach to the bottom wall of the flow cell. With these living cells being attached to the wall, other floating cells were pumped out by flowing a buffer solution. These procedures made it possible to keep a few cells in the flow cell and to analyze single cells by fluorescence microspectroscopy. The technique was applied to study the time course of staining processes of single living yeast (Saccharomyces cerevisiae) cells by using two types of a fluorescent probe. The present methodology was shown to be of primary importance for obtaining biochemical/physiological information on single living cells and also for studying cell-to-cell variations in several characteristics.

  14. Fluorescence lifetime imaging using a single photon avalanche diode array sensor (Conference Presentation)

    Science.gov (United States)

    Wargocki, Piotr M.; Spence, David J.; Goldys, Ewa M.; Charbon, Edoardo; Bruschini, Claudio E.; Antalović, Ivan Michel; Burri, Samuel

    2017-02-01

    Single photon detectors allows us work with the weakest fluorescence signals. Single photon arrays, combined with ps-controlled gating allow us to create image maps of fluorescence lifetimes, which can be used for in-vivo discrimination of tissue activity. Here we present fluorescence lifetime imaging using the `SwissSPAD' sensor, a 512-by-128-pixel array of gated single photon detectors, fabricated in a standard high-voltage 0.35 μm CMOS process. We present a protocol for spatially resolved lifetime measurements where the lifetime can be retrieved for each pixel. We demonstrate the system by imaging patterns of Fluorescein and Rhodamine B on test slides, as well as measuring mixed samples to retrieve both components of the decay lifetime. The single photon sensitivity of the sensor creates a valuable instrument to perform live cell or live animal (in vivo) measurements of the weak autofluorescent signals, for example distinguishing unlabelled free and bound NADH. Our ultimate goal is to create a real time fluorescence lifetime imaging system, possibly integrated into augmented reality goggles, which could allow immediate discrimination of in vivo tissues.

  15. Single-molecule fluorescence autocorrelation experiments on pentacene : The dependence of intersystem crossing on isotopic composition

    NARCIS (Netherlands)

    Brouwer, A.C.J.; Köhler, J.; Oijen, A.M. van; Groenen, E.J.J.; Schmidt, J.

    1999-01-01

    Single pentacene molecules containing 13C or 1H in a pentacene-d14 doped p-terphenyl crystal have been studied by fluorescence autocorrelation. The triplet dynamics has been analyzed and a systematic dependence of the S1→T1 intersystem crossing rate on isotopic composition was found. This variation

  16. Strong antenna-enhanced fluorescence of a single light-harvesting complex shows photon antibunching.

    Science.gov (United States)

    Wientjes, Emilie; Renger, Jan; Curto, Alberto G; Cogdell, Richard; van Hulst, Niek F

    2014-06-23

    The nature of the highly efficient energy transfer in photosynthetic light-harvesting complexes is a subject of intense research. Unfortunately, the low fluorescence efficiency and limited photostability hampers the study of individual light-harvesting complexes at ambient conditions. Here we demonstrate an over 500-fold fluorescence enhancement of light-harvesting complex 2 (LH2) at the single-molecule level by coupling to a gold nanoantenna. The resonant antenna produces an excitation enhancement of circa 100 times and a fluorescence lifetime shortening to ~20 ps. The radiative rate enhancement results in a 5.5-fold-improved fluorescence quantum efficiency. Exploiting the unique brightness, we have recorded the first photon antibunching of a single light-harvesting complex under ambient conditions, showing that the 27 bacteriochlorophylls coordinated by LH2 act as a non-classical single-photon emitter. The presented bright antenna-enhanced LH2 emission is a highly promising system to study energy transfer and the role of quantum coherence at the level of single complexes.

  17. ¿/4 Resonance of an Optical Monopole Antenna Probed by Single Molecule Fluorescence

    NARCIS (Netherlands)

    Taminiau, Tim H.; Moerland, R.J.; Segerink, Franciscus B.; Kuipers, L.; van Hulst, N.F.

    2007-01-01

    We present a resonant optical nanoantenna positioned at the end of a metal-coated glass fiber near-field probe. Antenna resonances, excitation conditions, and field localization are directly probed in the near field by single fluorescent molecules and compared to finite integration technique

  18. Simultaneous measurement of position and color of single fluorescent emitters using diffractive optics

    NARCIS (Netherlands)

    Broeken, J.; Rieger, B.; Stallinga, S.

    2014-01-01

    We propose a method for simultaneously measuring the position and emission color of single fluorescent emitters based on the use of a large pitch diffraction grating in the emission light path. The grating produces satellite spots adjacent to the main spot; the relative distance between the spots is

  19. pyFRET: A Python Library for Single Molecule Fluorescence Data Analysis

    OpenAIRE

    Murphy, Rebecca R.; Jackson, Sophie E.; Klenerman, David

    2014-01-01

    Single molecule F\\"orster resonance energy transfer (smFRET) is a powerful experimental technique for studying the properties of individual biological molecules in solution. However, as adoption of smFRET techniques becomes more widespread, the lack of available software, whether open source or commercial, for data analysis, is becoming a significant issue. Here, we present pyFRET, an open source Python package for the analysis of data from single-molecule fluorescence experiments from freely...

  20. Photoacoustic imaging of a near-infrared fluorescent marker based on dual wavelength pump-probe excitation

    Science.gov (United States)

    Märk, Julia; Theiss, Christoph; Schmitt, Franz-Josef; Laufer, Jan

    2014-03-01

    Photoacoustic imaging has been used to determine the spatial distribution of fluorophores, such as exogenous dyes and genetically expressed proteins, from images acquired in phantoms and in vivo. Most methods involve the acquisition of multiwavelength images and rely on differences in the absorption spectra of the tissue chromophores to estimate the spatial distribution and abundance of the latter using spectral decomposition techniques, such as model based inversion schemes. However, the inversion of 3-D images can be computationally expensive. Experimental approaches to localising contrast agents may therefore be useful, especially if quantification is not essential. This work aims to develop a method for determining the spatial distribution of a near-infrared fluorescent cell marker from images acquired using dual wavelength excitation. The excitation wavelengths coincided with the absorption and emission spectrum of the fluorophore. The contrast mechanism relies on reducing the excited state lifetime of the fluorophore by inducing stimulated emission. This changes the amount of energy thermalized by the fluorophore, and hence the photoacoustic signal amplitude. Since this is not observed in endogenous chromophores, the background may be removed by subtracting two images acquired with and without pulse delay between the pump and probe pulses. To characterise the fluorophore, the signal amplitude is measured in a cuvette as a function of pulse delay, concentration, and fluence. The spatial distribution of the fluorophore is determined from images acquired in realistic tissue phantoms. This method may be suitable for in vivo applications, such as imaging of exogenous or genetically expressed fluorescent cell markers.

  1. A dual amplification fluorescent strategy for sensitive detection of DNA methyltransferase activity based on strand displacement amplification and DNAzyme amplification.

    Science.gov (United States)

    Cui, Wanling; Wang, Lei; Jiang, Wei

    2016-03-15

    DNA methyltransferase (MTase) plays a critical role in many biological processes and has been regarded as a predictive cancer biomarker and a therapeutic target in cancer treatment. Sensitive detection of DNA MTase activity is essential for early cancer diagnosis and therapeutics. Here, we developed a dual amplification fluorescent strategy for sensitive detection of DNA MTase activity based on strand displacement amplification (SDA) and DNAzyme amplification. A trifunctional double-stranded DNA (dsDNA) probe was designed including a methylation site for DNA MTase recognition, a complementary sequence of 8-17 DNAzyme for synthesizing DNAzyme, and a nicking site for nicking enzyme cleavage. Firstly, the trifunctional dsDNA probe was methylated by DNA MTase to form the methylated dsDNA. Subsequently, HpaII restriction endonuclease specifically cleaved the residue of unmethylated dsDNA. Next, under the action of polymerase and nicking enzyme, the methylared dsDNA initiated SDA, releasing numbers of 8-17 DNAzymes. Finally, the released 8-17 DNAzymes triggered DNAzyme amplification reaction to induce a significant fluorescence enhancement. This strategy could detect DNA MTase activity as low as 0.0082U/mL. Additionally, the strategy was successfully applied for evaluating the inhibitions of DNA MTase using two anticancer drugs, 5-azacytidine and 5-aza-2'-deoxycytidine. The results indicate the proposed strategy has a potential application in early cancer diagnosis and therapeutics.

  2. Terbium-doped gadolinium oxide nanoparticles prepared by laser ablation in liquid for use as a fluorescence and magnetic resonance imaging dual-modal contrast agent.

    Science.gov (United States)

    Chen, Fei; Chen, Min; Yang, Chuan; Liu, Jun; Luo, Ningqi; Yang, Guowei; Chen, Dihu; Li, Li

    2015-01-14

    Dual-modal lanthanide-doped gadolinium nanoparticles (NPs), which exhibit an excellent magnetic resonance imaging (MRI) spatial resolution and high fluorescence imaging (FI) sensitivity, have attracted tremendous attention in biotechnology and nanomedicine applications. In this paper, terbium (Tb) ion doped gadolinium oxide (Gd2O3:Tb) NPs with varied Tb concentrations were synthesized by a laser ablation in liquid (LAL) method. The characterization of the structure, morphology, and composition shows that these NPs are spherical with excellent crystallinity. The effects of Tb ion concentration on the visible green fluorescence and longitudinal relaxivity were investigated, indicating that the fluorescence properties were significantly influenced by the Tb ion concentration, but all samples were still efficient T1-weighted contrast agents. Furthermore, the optimum Tb doping concentration was determined to be 1%. The cell viability, cellular fluorescence imaging and in vivo MRI of this dual-modal nano-probe were studied, with the results revealing that the Gd2O3:Tb NPs did not have a significant cytotoxic effect, making them good candidates for use as a dual-modal contrast agent for MRI and fluorescence imaging.

  3. Multi-ion detection and molecular switching behaviour of reversible dual fluorescent sensor

    Science.gov (United States)

    Basheer, Sabeel M.; Muralisankar, M.; Anjana, T. V.; Aneesrahman, K. N.; Sreekanth, Anandaram

    2017-07-01

    The selective chemosensing behaviour of imidazole bisthiocarbohydrazone (IBTC) towards F- and Cu2 + are studied via colorimetric, UV-Visible, fluorescence spectra studies, and binding constants were calculated. The 1H NMR titration study strongly support that the deprotonation of IBTC followed by the hydrogen bond formation via N1sbnd H1 and N2sbnd H2 protons with fluoride ion. The fluorescence inactive IBTC-Cu complex became fluorescence active in the presence of perchlorate (ClO4-) ion. The selective detection of perchlorate ion was also explained. The F- sensing mechanism of IBTC has been investigated by Density Functional Theory (DFT) and Time-Dependent Density Functional Theory (TDDFT) methods. The theoretical outcomes well reproduce the experimental results. And it concluded the Nsbnd H protons, nearby the imine group was first captured by the added F- ion and then deprotonation happened followed by the formation of hydrogen bond. The IBTC found good reversibility character with the alternative addition of Ca2 + and F-. The multi-ion detection of IBTC was used to construct the NOR, OR and INHIBITION molecular logic gates.

  4. Stable J-aggregation enabled dual photoacoustic and fluorescence nanoparticles for intraoperative cancer imaging.

    Science.gov (United States)

    Shakiba, Mojdeh; Ng, Kenneth K; Huynh, Elizabeth; Chan, Harley; Charron, Danielle M; Chen, Juan; Muhanna, Nidal; Foster, F Stuart; Wilson, Brian C; Zheng, Gang

    2016-07-07

    J-aggregates display nanoscale optical properties which enable their use in fluorescence and photoacoustic imaging applications. However, control over their optical properties in an in vivo setting is hampered by the conformational lability of the J-aggregate structure in complex biological environments. J-aggregating nanoparticles (JNP) formed by self-assembly of bacteriopheophorbide-lipid (Bchl-lipid) in lipid nanovesicles represents a novel strategy to stabilize J-aggregates for in vivo bioimaging applications. We find that 15 mol% Bchl-lipid embedded within a saturated phospholipid bilayer vesicle was optimal in terms of maximizing Bchl-lipid dye loading, while maintaining a spherical nanoparticle morphology and retaining spectral properties characteristic of J-aggregates. The addition of cholesterol maintains the stability of the J-aggregate absorption band for up to 6 hours in the presence of 90% FBS. In a proof-of-concept experiment, we successfully applied JNPs as a fluorescence contrast agent for real-time intraoperative detection of metastatic lymph nodes in a rabbit head-and-neck cancer model. Lymph node metastasis delineation was further verified by visualizing the JNP within the excised lymph node using photoacoustic imaging. Using JNPs, we demonstrate the possibility of using J-aggregates as fluorescence and photoacoustic contrast agents and may potentially spur the development of other nanomaterials that can stably induce J-aggregation for in vivo cancer bioimaging applications.

  5. Dual channel fluorescence engineering on the nano-patterened metamaterial (Conference Presentation)

    Science.gov (United States)

    Wu, Huijun; Tseng, Ming Lun; Tsai, Wei Yi; Chen, Ting-Yu; Chien, Yihsin; Chang, Yun-Chorng; Tsai, Din Ping

    2016-09-01

    Upconversion fluorescence from Lanthanide-doped nanocrystals has attracted widespread interests because of its greatly potential applications in various fields, such as photonic crystal lasers, material science, biological therapy, and so on. However, the relatively low quantum yield (typically chemical methods, plasmonic structures have been adopted as another strategy to improve the radiation efficiency and control the relaxation process of the upcovnersion nanocrystals. We designed the anti-symmetric split ring resonators with various periods and the fishnet structures. The surface plasmon resonance peaks of the structure shift as the periods varies. For example, in a multi-layered plamsonic metasurface with the period of 250nm, both the electric and magnetic modes could be generated simultaneously when excited by the incident light with proper polarization. This plasmonic structure provides two different channels for the enhancement of upconversion fluorescence. The resonance peak of 650nm is magnetic resonance mode, while the peak of 980nm is electric resonance mode. The resonance peak of 980nm coincides with the absorption band of the Lanthanide-dopoed nanocrystal, and the peak of 650nm matches with its emission band. We found that the upconversion fluorescence intensity could be enhanced more than 10 times when the electric resonance frequency of the metasurface matches with the absorption band of the upconversion nanocrystals, while the magnetic mode overlaps with its emission band. This is due to the local density of optical states was significantly enhanced by the plasmonic metasurface. The detailed results and mechanism will be discussed.

  6. Versatile single-molecule multi-color excitation and detection fluorescence setup for studying biomolecular dynamics

    KAUST Repository

    Sobhy, M. A.

    2011-11-07

    Single-molecule fluorescence imaging is at the forefront of tools applied to study biomolecular dynamics both in vitro and in vivo. The ability of the single-molecule fluorescence microscope to conduct simultaneous multi-color excitation and detection is a key experimental feature that is under continuous development. In this paper, we describe in detail the design and the construction of a sophisticated and versatile multi-color excitation and emission fluorescence instrument for studying biomolecular dynamics at the single-molecule level. The setup is novel, economical and compact, where two inverted microscopes share a laser combiner module with six individual laser sources that extend from 400 to 640 nm. Nonetheless, each microscope can independently and in a flexible manner select the combinations, sequences, and intensities of the excitation wavelengths. This high flexibility is achieved by the replacement of conventional mechanical shutters with acousto-optic tunable filter (AOTF). The use of AOTF provides major advancement by controlling the intensities, duration, and selection of up to eight different wavelengths with microsecond alternation time in a transparent and easy manner for the end user. To our knowledge this is the first time AOTF is applied to wide-field total internal reflection fluorescence (TIRF) microscopy even though it has been commonly used in multi-wavelength confocal microscopy. The laser outputs from the combiner module are coupled to the microscopes by two sets of four single-mode optic fibers in order to allow for the optimization of the TIRF angle for each wavelength independently. The emission is split into two or four spectral channels to allow for the simultaneous detection of up to four different fluorophores of wide selection and using many possible excitation and photoactivation schemes. We demonstrate the performance of this new setup by conducting two-color alternating excitation single-molecule fluorescence resonance energy

  7. Single molecule localization microscopy of the distribution of chromatin using Hoechst and DAPI fluorescent probes.

    Science.gov (United States)

    Szczurek, Aleksander T; Prakash, Kirti; Lee, Hyun-Keun; Zurek-Biesiada, Dominika J; Best, Gerrit; Hagmann, Martin; Dobrucki, Jurek W; Cremer, Christoph; Birk, Udo

    2014-01-01

    Several approaches have been described to fluorescently label and image DNA and chromatin in situ on the single-molecule level. These superresolution microscopy techniques are based on detecting optically isolated, fluorescently tagged anti-histone antibodies, fluorescently labeled DNA precursor analogs, or fluorescent dyes bound to DNA. Presently they suffer from various drawbacks such as low labeling efficiency or interference with DNA structure. In this report, we demonstrate that DNA minor groove binding dyes, such as Hoechst 33258, Hoechst 33342, and DAPI, can be effectively employed in single molecule localization microscopy (SMLM) with high optical and structural resolution. Upon illumination with low intensity 405 nm light, a small subpopulation of these molecules stochastically undergoes photoconversion from the original blue-emitting form to a green-emitting form. Using a 491 nm laser excitation, fluorescence of these green-emitting, optically isolated molecules was registered until "bleached". This procedure facilitated substantially the optical isolation and localization of large numbers of individual dye molecules bound to DNA in situ, in nuclei of fixed mammalian cells, or in mitotic chromosomes, and enabled the reconstruction of high-quality DNA density maps. We anticipate that this approach will provide new insights into DNA replication, DNA repair, gene transcription, and other nuclear processes.

  8. Single molecule localization microscopy of the distribution of chromatin using Hoechst and DAPI fluorescent probes

    Science.gov (United States)

    Szczurek, Aleksander T; Prakash, Kirti; Lee, Hyun-Keun; Żurek-Biesiada, Dominika J; Best, Gerrit; Hagmann, Martin; Dobrucki, Jurek W; Cremer, Christoph; Birk, Udo

    2014-01-01

    Several approaches have been described to fluorescently label and image DNA and chromatin in situ on the single-molecule level. These superresolution microscopy techniques are based on detecting optically isolated, fluorescently tagged anti-histone antibodies, fluorescently labeled DNA precursor analogs, or fluorescent dyes bound to DNA. Presently they suffer from various drawbacks such as low labeling efficiency or interference with DNA structure. In this report, we demonstrate that DNA minor groove binding dyes, such as Hoechst 33258, Hoechst 33342, and DAPI, can be effectively employed in single molecule localization microscopy (SMLM) with high optical and structural resolution. Upon illumination with low intensity 405 nm light, a small subpopulation of these molecules stochastically undergoes photoconversion from the original blue-emitting form to a green-emitting form. Using a 491 nm laser excitation, fluorescence of these green-emitting, optically isolated molecules was registered until “bleached”. This procedure facilitated substantially the optical isolation and localization of large numbers of individual dye molecules bound to DNA in situ, in nuclei of fixed mammalian cells, or in mitotic chromosomes, and enabled the reconstruction of high-quality DNA density maps. We anticipate that this approach will provide new insights into DNA replication, DNA repair, gene transcription, and other nuclear processes. PMID:25482122

  9. Dual-wavelength single-frequency laser emission in asymmetric coupled microdisks

    Science.gov (United States)

    Wang, Haotian; Liu, Sheng; Chen, Lin; Shen, Deyuan; Wu, Xiang

    2016-12-01

    The gain and loss in a microcavity laser play an important role for the modulation of laser spectrum. We show that dual-wavelength single mode lasing can be achieved in an asymmetric coupled system consisted of two size-mismatched microdisks. The amount of eigenmodes in this coupled-microdisk system is reduced relying on the Vernier effect. Then a single mode is selected to lase by controlling the gain branching in the supermodes. The supermodes are formed by the coupling between different transverse whispering-gallery modes (WGMs). When the gain/loss status between the two mirodisks is changed through selectively pumping process, the modulated gain branching for various supermodes leads to the switchable single-frequency laser emission. The results obtained in this work will provide the further understand for the spectral modulation mechanism in the coupled microcavity laser system.

  10. Single fluorescent protein-based Ca2+ sensors with increased dynamic range

    Directory of Open Access Journals (Sweden)

    Labas Yulii A

    2007-06-01

    Full Text Available Abstract Background Genetically encoded sensors developed on the basis of green fluorescent protein (GFP-like proteins are becoming more and more popular instruments for monitoring cellular analytes and enzyme activities in living cells and transgenic organisms. In particular, a number of Ca2+ sensors have been developed, either based on FRET (Fluorescence Resonance Energy Transfer changes between two GFP-mutants or on the change in fluorescence intensity of a single circularly permuted fluorescent protein (cpFP. Results Here we report significant progress on the development of the latter type of Ca2+ sensors. Derived from the knowledge of previously reported cpFP-based sensors, we generated a set of cpFP-based indicators with different spectral properties and fluorescent responses to changes in Ca2+ concentration. Two variants, named Case12 and Case16, were characterized by particular high brightness and superior dynamic range, up to 12-fold and 16.5-fold increase in green fluorescence between Ca2+-free and Ca2+-saturated forms. We demonstrated the high potential of these sensors on various examples, including monitoring of Ca2+ response to a prolonged glutamate treatment in cortical neurons. Conclusion We believe that expanded dynamic range, high brightness and relatively high pH-stability should make Case12 and Case16 popular research tools both in scientific studies and high throughput screening assays.

  11. Dual optical recordings for action potentials and calcium handling in induced pluripotent stem cell models of cardiac arrhythmias using genetically encoded fluorescent indicators.

    Science.gov (United States)

    Song, LouJin; Awari, Daniel W; Han, Elizabeth Y; Uche-Anya, Eugenia; Park, Seon-Hye E; Yabe, Yoko A; Chung, Wendy K; Yazawa, Masayuki

    2015-05-01

    Reprogramming of human somatic cells to pluripotency has been used to investigate disease mechanisms and to identify potential therapeutics. However, the methods used for reprogramming, in vitro differentiation, and phenotyping are still complicated, expensive, and time-consuming. To address the limitations, we first optimized a protocol for reprogramming of human fibroblasts and keratinocytes into pluripotency using single lipofection and the episomal vectors in a 24-well plate format. This method allowed us to generate multiple lines of integration-free and feeder-free induced pluripotent stem cells (iPSCs) from seven patients with cardiac diseases and three controls. Second, we differentiated human iPSCs derived from patients with Timothy syndrome into cardiomyocytes using a monolayer differentiation method. We found that Timothy syndrome cardiomyocytes showed slower, irregular contractions and abnormal calcium handling compared with the controls. The results are consistent with previous reports using a retroviral method for reprogramming and an embryoid body-based method for cardiac differentiation. Third, we developed an efficient approach for recording the action potentials and calcium transients simultaneously in control and patient cardiomyocytes using genetically encoded fluorescent indicators, ArcLight and R-GECO1. The dual optical recordings enabled us to observe prolonged action potentials and abnormal calcium handling in Timothy syndrome cardiomyocytes. We confirmed that roscovitine rescued the phenotypes in Timothy syndrome cardiomyocytes and that these findings were consistent with previous studies using conventional electrophysiological recordings and calcium imaging with dyes. The approaches using our optimized methods and dual optical recordings will improve iPSC applicability for disease modeling to investigate mechanisms underlying cardiac arrhythmias and to test potential therapeutics.

  12. A microfluidic dual-well device for high-throughput single-cell capture and culture.

    Science.gov (United States)

    Lin, Ching-Hui; Hsiao, Yi-Hsing; Chang, Hao-Chen; Yeh, Chuan-Feng; He, Cheng-Kun; Salm, Eric M; Chen, Chihchen; Chiu, Ing-Ming; Hsu, Chia-Hsien

    2015-07-21

    In vitro culture of single cells facilitates biological studies by deconvoluting complications from cell population heterogeneity. However, there is still a lack of simple yet high-throughput methods to perform single cell culture experiments. In this paper, we report the development and application of a microfluidic device with a dual-well (DW) design concept for high-yield single-cell loading (~77%) in large microwells (285 and 485 μm in diameter) which allowed for cell spreading, proliferation and differentiation. The increased single-cell loading yield is achieved by using sets of small microwells termed "capture-wells" and big microwells termed "culture-wells" according to their utilities for single-cell capture and culture, respectively. This novel device architecture allows the size of the "culture" microwells to be flexibly adjusted without affecting the single-cell loading efficiency making it useful for cell culture applications as demonstrated by our experiments of KT98 mouse neural stem cell differentiation, A549 and MDA-MB-435 cancer cell proliferation, and single-cell colony formation assay with A549 cells in this paper.

  13. Single lead atrial vs. dual chamber pacing in sick sinus syndrome

    DEFF Research Database (Denmark)

    Brandt, Niels H; Kirkfeldt, Rikke Esberg; Nielsen, Jens Cosedis

    2016-01-01

    AIMS: The DANPACE trial randomized patients with sick sinus syndrome (SSS) to single lead atrial (AAIR) or dual chamber (DDDR) pacemaker (PM). After 5 years follow-up, no difference in overall survival, stroke or heart failure (HF) was observed, whereas risk of atrial fibrillation (AF) and PM....... CONCLUSION: This register-based long-term follow-up study indicates that there is no difference in mortality among patients with SSS randomized to AAIR or DDDR pacing, even with very long follow-up. Nor is there any difference in risk of AF hospitalization, stroke or HF. The higher rate of pacing mode...

  14. Operational characteristics of dual gain single cavity Nd:YVO4 laser

    Indian Academy of Sciences (India)

    Pranab K Mukhopadhyay; Jogy George; S K Sharma; P K Gupta; T P S Nathan

    2002-01-01

    Operational characteristics of a dual gain single cavity Nd:YVO4 laser have been investigated. With semiconductor diode laser pump power of 2 W, 800 mW output was obtained with a slope efficiency of 49%. Further, by changing the relative orientation of the two crystals the polarization characteristics of the output could be varied. In particular by keeping the two Nd:YVO4 crystals with their -axes orthogonal to each other and adjusting the gain of the crystals so that both operate at approximately the same power level, completely unpolarized beams could be obtained.

  15. Charge-driven feedback loop in the resonance fluorescence of a single quantum dot

    Science.gov (United States)

    Merkel, B.; Kurzmann, A.; Schulze, J.-H.; Strittmatter, A.; Geller, M.; Lorke, A.

    2017-03-01

    We demonstrate a feedback loop that manifests itself in a strong hysteresis and bistability of the exciton resonance fluorescence signal. Field ionization of photogenerated quantum dot excitons leads to the formation of a charged interface layer that drags the emission line along over a frequency range of more than 30 GHz . These measurements are well described by a rate equation model. With a time-resolved resonance fluorescence measurement we determined the buildup times for the hole gas in the orders of milliseconds. This internal charge-driven feedback loop could be used to reduce the spectral wandering in the emission spectra of single self-assembled quantum dots.

  16. Salen-based [Zn2Ln3] complexes with fluorescence and single-molecule-magnet properties.

    Science.gov (United States)

    Burrow, Carolyn E; Burchell, Tara J; Lin, Po-Heng; Habib, Fatemah; Wernsdorfer, Wolfgang; Clérac, Rodolphe; Murugesu, Muralee

    2009-09-07

    A family of four isostructural complexes with a V-shaped pentanuclear [Zn(2)Ln(3)] core of general formula [Zn(2)Ln(3)(m-salen)(3)(N(3))(5)(OH)(2)] [Ln(III) = Tb(III) (1), Eu(III) (2), Ho(III) (3), Dy(III) (4); m-salen = N,N'-ethylenebis(3-methoxysalicylideneamine)] were isolated and structurally characterized. The fluorescence and magnetic measurements of the four compounds were investigated. Complex 1 exhibits strong fluorescence properties, while single-molecule-magnet behavior is seen in complex 4.

  17. Detection of Fluorescence from Single Chlorophyll a Molecules Absorbed on Glass Surface

    Institute of Scientific and Technical Information of China (English)

    JI Dong-Mei; HUANG Zheng-Xi; XIA An-Dong

    2005-01-01

    @@ We investigate the single molecule spectroscopy of chlorophyll a molecules on glass surface in N2-saturated environment. The basic photodynamic parameters of chlorophyll a molecules, such as fluorescence lifetime,survival time before photobleaching, on-time, and off-time, are reported. A four-level model is employed to describe the possible dynamics and photobleaching of chlorophyll a upon excitation. Broad distributions in fluorescence lifetimes and survival times are mainly due to the heterogeneities of both molecular conformation and local environment.

  18. Single molecule fluorescence fluctuations of the cyanine dyes linked covalently to DNA

    Institute of Scientific and Technical Information of China (English)

    LV Wei; CHEN XuDong; AUMILER Damir; XIA AnDong

    2009-01-01

    The intersystem crossing and isomerization dynamics of free-Cy3,Cy3-ssDNA,free-Cy5 and Cy5-ssDNA are obtained through simple analysis of rapid on/off blinking from single molecule fluo rescence intensity time-traces and the fluorescence correlation spectroscopy(FCS).The on-and off-times observed in fluorescence time traces of single cyanine dyes are due to the formation of the triplet state and isomerization,where both the interaction with DNA and long central polymethine chain of cyanine dyes increase the barriers of isomerization,leading to long off-time.The results indicate that the single molecule fluorescence fluctuation together with the resulting second autocorrelation analysis are powerful methods for determining the triplet state and isomerization dynamics,which could be the simple techniques and complementary to other spectroscopic techniques,such as fluorescence decay measurement and laser flash photolysis to study the photophysical processes of complex molecules.

  19. Recent Advances in Biological Single-Molecule Applications of Optical Tweezers and Fluorescence Microscopy.

    Science.gov (United States)

    Hashemi Shabestari, M; Meijering, A E C; Roos, W H; Wuite, G J L; Peterman, E J G

    2017-01-01

    Over the past two decades, single-molecule techniques have evolved into robust tools to study many fundamental biological processes. The combination of optical tweezers with fluorescence microscopy and microfluidics provides a powerful single-molecule manipulation and visualization technique that has found widespread application in biology. In this combined approach, the spatial (~nm) and temporal (~ms) resolution, as well as the force scale (~pN) accessible to optical tweezers is complemented with the power of fluorescence microscopy. Thereby, it provides information on the local presence, identity, spatial dynamics, and conformational dynamics of single biomolecules. Together, these techniques allow comprehensive studies of, among others, molecular motors, protein-protein and protein-DNA interactions, biomolecular conformational changes, and mechanotransduction pathways. In this chapter, recent applications of fluorescence microscopy in combination with optical trapping are discussed. After an introductory section, we provide a description of instrumentation together with the current capabilities and limitations of the approaches. Next we summarize recent studies that applied this combination of techniques in biological systems and highlight some representative biological assays to mark the exquisite opportunities that optical tweezers combined with fluorescence microscopy provide. © 2017 Elsevier Inc. All rights reserved.

  20. Stable J-aggregation enabled dual photoacoustic and fluorescence nanoparticles for intraoperative cancer imaging

    Science.gov (United States)

    Shakiba, Mojdeh; Ng, Kenneth K.; Huynh, Elizabeth; Chan, Harley; Charron, Danielle M.; Chen, Juan; Muhanna, Nidal; Foster, F. Stuart; Wilson, Brian C.; Zheng, Gang

    2016-06-01

    J-aggregates display nanoscale optical properties which enable their use in fluorescence and photoacoustic imaging applications. However, control over their optical properties in an in vivo setting is hampered by the conformational lability of the J-aggregate structure in complex biological environments. J-aggregating nanoparticles (JNP) formed by self-assembly of bacteriopheophorbide-lipid (Bchl-lipid) in lipid nanovesicles represents a novel strategy to stabilize J-aggregates for in vivo bioimaging applications. We find that 15 mol% Bchl-lipid embedded within a saturated phospholipid bilayer vesicle was optimal in terms of maximizing Bchl-lipid dye loading, while maintaining a spherical nanoparticle morphology and retaining spectral properties characteristic of J-aggregates. The addition of cholesterol maintains the stability of the J-aggregate absorption band for up to 6 hours in the presence of 90% FBS. In a proof-of-concept experiment, we successfully applied JNPs as a fluorescence contrast agent for real-time intraoperative detection of metastatic lymph nodes in a rabbit head-and-neck cancer model. Lymph node metastasis delineation was further verified by visualizing the JNP within the excised lymph node using photoacoustic imaging. Using JNPs, we demonstrate the possibility of using J-aggregates as fluorescence and photoacoustic contrast agents and may potentially spur the development of other nanomaterials that can stably induce J-aggregation for in vivo cancer bioimaging applications.J-aggregates display nanoscale optical properties which enable their use in fluorescence and photoacoustic imaging applications. However, control over their optical properties in an in vivo setting is hampered by the conformational lability of the J-aggregate structure in complex biological environments. J-aggregating nanoparticles (JNP) formed by self-assembly of bacteriopheophorbide-lipid (Bchl-lipid) in lipid nanovesicles represents a novel strategy to stabilize J

  1. Fast methods for analysis of neurotransmitters from single cell and monitoring their releases in central nervous system by capillary electrophoresis, fluorescence microscopy and luminescence imaging

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ziqiang [Iowa State Univ., Ames, IA (United States)

    1999-12-10

    Fast methods for separation and detection of important neurotransmitters and the releases in central nervous system (CNS) were developed. Enzyme based immunoassay combined with capillary electrophoresis was used to analyze the contents of amino acid neurotransmitters from single neuron cells. The release of amino acid neurotransmitters from neuron cultures was monitored by laser induced fluorescence imaging method. The release and signal transduction of adenosine triphosphate (ATP) in CNS was studied with sensitive luminescence imaging method. A new dual-enzyme on-column reaction method combined with capillary electrophoresis has been developed for determining the glutamate content in single cells. Detection was based on monitoring the laser-induced fluorescence of the reaction product NADH, and the measured fluorescence intensity was related to the concentration of glutamate in each cell. The detection limit of glutamate is down to 10-8 M level, which is 1 order of magnitude lower than the previously reported detection limit based on similar detection methods. The mass detection limit of a few attomoles is far superior to that of any other reports. Selectivity for glutamate is excellent over most of amino acids. The glutamate content in single human erythrocyte and baby rat brain neurons were determined with this method and results agreed well with literature values.

  2. Fast methods for analysis of neurotransmitters from single cell and monitoring their releases in central nervous system by capillary electrophoresis, fluorescence microscopy and luminescence imaging

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ziqiang

    1999-12-10

    Fast methods for separation and detection of important neurotransmitters and the releases in central nervous system (CNS) were developed. Enzyme based immunoassay combined with capillary electrophoresis was used to analyze the contents of amino acid neurotransmitters from single neuron cells. The release of amino acid neurotransmitters from neuron cultures was monitored by laser induced fluorescence imaging method. The release and signal transduction of adenosine triphosphate (ATP) in CNS was studied with sensitive luminescence imaging method. A new dual-enzyme on-column reaction method combined with capillary electrophoresis has been developed for determining the glutamate content in single cells. Detection was based on monitoring the laser-induced fluorescence of the reaction product NADH, and the measured fluorescence intensity was related to the concentration of glutamate in each cell. The detection limit of glutamate is down to 10{sup {minus}8} M level, which is 1 order of magnitude lower than the previously reported detection limit based on similar detection methods. The mass detection limit of a few attomoles is far superior to that of any other reports. Selectivity for glutamate is excellent over most of amino acids. The glutamate content in single human erythrocyte and baby rat brain neurons were determined with this method and results agreed well with literature values.

  3. Future challenges in single-molecule fluorescence and laser trap approaches to studies of molecular motors.

    Science.gov (United States)

    Elting, Mary Williard; Spudich, James A

    2012-12-11

    Single-molecule analysis is a powerful modern form of biochemistry, in which individual kinetic steps of a catalytic cycle of an enzyme can be explored in exquisite detail. Both single-molecule fluorescence and single-molecule force techniques have been widely used to characterize a number of protein systems. We focus here on molecular motors as a paradigm. We describe two areas where we expect to see exciting developments in the near future: first, characterizing the coupling of force production to chemical and mechanical changes in motors, and second, understanding how multiple motors work together in the environment of the cell.

  4. Single-atom-resolved fluorescence imaging of an atomic Mott insulator

    DEFF Research Database (Denmark)

    Sherson, Jacob; Weitenberg, Christof; Andres, Manuel

    2010-01-01

    in situ images of a quantum fluid in which each underlying quantum particle is detected. Here we report fluorescence imaging of strongly interacting bosonic Mott insulators in an optical lattice with single-atom and single-site resolution. From our images, we fully reconstruct the atom distribution......The reliable detection of single quantum particles has revolutionized the field of quantum optics and quantum information processing. For several years, researchers have aspired to extend such detection possibilities to larger-scale, strongly correlated quantum systems 1 , 2 in order to record...

  5. The durability of single, dual, and multiphase titanate ceramic waste forms for nuclear waste immobilization

    Science.gov (United States)

    Harkins, Devin J. H.

    A significant amount of the energy used in the United States comes from nuclear power, which produces a large amount of waste materials. Recycling nuclear waste is possible, but requires a way to permanently fix the unusable radionuclides remaining from the recycling process in a stable, leach resistant structure. Multiphase titanate ceramic waste forms are one promising option under consideration. However, there is insufficient work on the long term corrosion of the individual phases, as well as the multiphase systems of these ceramics. These multiphase titanate ceramic waste forms have three targeted phases: hollandite, pyrochlore, and zirconolite. Hollandite is a promising candidate for the incorporation of Cs, while pyrochlore is readily formed with lanthanides, such as Nd, the most prevalent lanthanide in the waste stream. The third targeted phase, zirconolite, is for the incorporation of zirconium and the actinides. This work looks into the formation of single phase systems of lanthanide titanates, formation of dual phase systems of Ga doped Ba hollandites and Nd titanate, durability of single phase hollandites and multiphase model systems using Vapor Hydration Testing (ASTM C 1663-09), dissolution of dual phase systems of Ga doped Ba hollandites and Nd titanate using Product Consistency Testing (ASTM C 1285-02), as well investigating how grain size affects amount of alterative phases formed using Vapor Hydration Testing. The dual phase systems of hollandites and Nd titanate show significant amounts of secondary phases forming, heavily influenced by the composition of hollandite used in the systems. The most significant phase present was BaNd2Ti5O14. This phase proves to be problematic due to the degradation to the hollandite structure. Using Vapor Hydration Testing to investigate single and multiphase systems presented many some possible alteration phases that could occur in the long term aging of these ceramics. Most notably, Cs rich phases were found in

  6. Single camera imaging system for color and near-infrared fluorescence image guided surgery.

    Science.gov (United States)

    Chen, Zhenyue; Zhu, Nan; Pacheco, Shaun; Wang, Xia; Liang, Rongguang

    2014-08-01

    Near-infrared (NIR) fluorescence imaging systems have been developed for image guided surgery in recent years. However, current systems are typically bulky and work only when surgical light in the operating room (OR) is off. We propose a single camera imaging system that is capable of capturing NIR fluorescence and color images under normal surgical lighting illumination. Using a new RGB-NIR sensor and synchronized NIR excitation illumination, we have demonstrated that the system can acquire both color information and fluorescence signal with high sensitivity under normal surgical lighting illumination. The experimental results show that ICG sample with concentration of 0.13 μM can be detected when the excitation irradiance is 3.92 mW/cm(2) at an exposure time of 10 ms.

  7. Fluorescent metal nanoshell and CK19 detection on single cell image

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Jian, E-mail: jian@cfs.biomet.umaryland.edu [Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201 (United States); Fu, Yi [Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201 (United States); Li, Ge [Division of Molecular Pathology, Department of Pathology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 (United States); Lakowicz, Joseph R. [Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201 (United States); Zhao, Richard Y., E-mail: rzhao@som.umaryland.edu [Division of Molecular Pathology, Department of Pathology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 (United States); Department of Microbiology-Immunology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 (United States); Institute of Human Virology, University of Maryland School of Medicine, 10 South Pine Street, Baltimore, MD 21201 (United States)

    2011-09-16

    Highlights: {yields} Novel metal nanoshell as fluorescence imaging agent. {yields} Fluorescent mAb-metal complex with enhanced intensity and shortened lifetime. {yields} Immuno-interactions of mAb-metal complexes with CK19 molecules on CNCAP and HeLa cell surfaces. {yields} Isolation of conjugated mAb-metal complexes from cellular autofluorescence on cell image. -- Abstract: In this article, we report the synthesis strategy and optical properties of a novel type of fluorescence metal nanoshell when it was used as imaging agent for fluorescence cell imaging. The metal nanoshells were made with 40 nm silica cores and 10 nm silver shells. Unlike typical fluorescence metal nanoshells which contain the organic dyes in the cores, novel metal nanoshells were composed of Cy5-labelled monoclonal anti-CK19 antibodies (mAbs) on the external surfaces of shells. Optical measurements to the single nanoparticles showed that in comparison with the metal free labelled mAbs, the mAb-Ag complexes displayed significantly enhanced emission intensity and dramatically shortened lifetime due to near-field interactions of fluorophores with metal. These metal nanoshells were found to be able to immunoreact with target cytokeratin 19 (CK19) molecules on the surfaces of LNCAP and HeLa cells. Fluorescence cell images were recorded on a time-resolved confocal microscope. The emissions from the metal nanoprobes could be clearly isolated from the cellular autofluorescence backgrounds on the cell images as either individuals or small clusters due to their stronger emission intensities and shorter lifetimes. These emission signals could also be precisely counted on single cell images. The count number may provide an approach for quantifying the target molecules in the cells.

  8. Photophysics of Fluorescent Probes for Single-Molecule Biophysics and Super-Resolution Imaging

    Science.gov (United States)

    Ha, Taekjip; Tinnefeld, Philip

    2012-05-01

    Single-molecule fluorescence spectroscopy and super-resolution microscopy are important elements of the ongoing technical revolution to reveal biochemical and cellular processes in unprecedented clarity and precision. Demands placed on the photophysical properties of the fluorophores are stringent and drive the choice of appropriate probes. Such fluorophores are not simple light bulbs of a certain color and brightness but instead have their own “personalities” regarding spectroscopic parameters, redox properties, size, water solubility, photostability, and several other factors. Here, we review the photophysics of fluorescent probes, both organic fluorophores and fluorescent proteins, used in applications such as particle tracking, single-molecule FRET, stoichiometry determination, and super-resolution imaging. Of particular interest is the thiol-induced blinking of Cy5, a curse for single-molecule biophysical studies that was later overcome using Trolox through a reducing/oxidizing system but a boon for super-resolution imaging owing to the controllable photoswitching. Understanding photophysics is critical in the design and interpretation of single-molecule experiments.

  9. Single and dual-Gregorian reflector antenna shaped beam far-field synthesis

    Science.gov (United States)

    Mehler, M. J.

    The direct far-field G.O. synthesis of shaped beam reflector antennas has recently been treated by Mehler, Tun and Adatia (1986). These authors use a synthesis technique which exploits complex coordinates and which is based on a method originally considered by Norris and Westcott (1976). They describe the synthesis of single reflector antennas which radiate both elliptical beams and European coverage patterns. Here this technique is extended to consider a class of dual reflector antennas which possess shaped main reflectors and conic subreflectors. An example is given of a Gregorian duel reflector antenna which radiates a cross-polar field significantly smaller than that radiated by single shaped reflector antennas. In addition, the behavior of the radiation pattern as a function of the reflector diameter is investigated.

  10. FEM analysis of an single stator dual PM rotors axial synchronous machine

    Science.gov (United States)

    Tutelea, L. N.; Deaconu, S. I.; Popa, G. N.

    2017-01-01

    The actual e - continuously variable transmission (e-CVT) solution for the parallel Hybrid Electric Vehicle (HEV) requires two electric machines, two inverters, and a planetary gear. A distinct electric generator and a propulsion electric motor, both with full power converters, are typical for a series HEV. In an effort to simplify the planetary-geared e-CVT for the parallel HEV or the series HEV we hereby propose to replace the basically two electric machines and their two power converters by a single, axial-air-gap, electric machine central stator, fed from a single PWM converter with dual frequency voltage output and two independent PM rotors. The proposed topologies, the magneto-motive force analysis and quasi 3D-FEM analysis are the core of the paper.

  11. STEEL CORROSION AT 600°C IN SINGLE AND DUAL CONDITION IN OXYFUEL ATMOSPHERE

    Directory of Open Access Journals (Sweden)

    Daniel Massari de Souza Coelho

    2014-10-01

    Full Text Available Coal-fired power plants using the Oxyfuel process are being developed to produce electricity with zero CO2 emission. Steels used in this and other processes are often exposed to different atmospheres in each side of the material, especially in heat exchangers and solid oxide fuel cells. Some studies have shown that steels exposed to different hydrogen partial pressures in each side have a different corrosion behavior from steels exposed to a single atmosphere condition. In this investigation, two experimental steels were studied at 600°C and 1 atm in dual atmospheres containing water vapor in one side and flue gas in the other and they were compared to steels oxidized in single atmospheres. The gas composition used is similar to the ones found in Oxyfuel coal power plants, where there is a great concentration of CO2, and also H2O and SO2. Analyses were made using SEM and TEM.

  12. Comparative study of shale-gas production using single- and dual-continuum approaches

    KAUST Repository

    El-Amin, Mohamed

    2017-07-06

    In this paper, we explore the possibility of specifying the ideal hypothetical positions of matrices blocks and fractures in fractured porous media as a single-continuum reservoir model in a way that mimics the dual-porosity dual-permeability (DPDP) configuration. In order to get an ideal mimic, we use the typical configuration and geometrical hypotheses of the DPDP model for the SDFM. Unlike the DPDP model which consists of two equations for the two-continuum coupled by a transfer term, the proposed single-domain fracture model (SDFM) model consists of a single equation for the single-continuum. Each one of the two models includes slippage effect, adsorption, Knudsen diffusion, geomechanics, and thermodynamics deviation factor. For the thermodynamics calculations, the cubic Peng-Robinson equation of state is employed. The diffusion model is verified by calculating the total mass flux through a nanopore by combination of slip flow and Knudsen diffusion and compared with experimental data. A semi-implicit scheme is used for the time discretization while the thermodynamics equations are updated explicitly. The spatial discretization is done using the cell-centered finite difference (CCFD) method. Finally, numerical experiments are performed under variations of the physical parameters. Several results are discussed such as pressure, production rate and cumulative production. We compare the results of the two models using the same dimensions and physical and computational parameters. We found that the DPDP and the SDFM models production rate and cumulative production behave similarly with approximately the same slope but with some differences in values. Moreover, we found that the poroelasticity effect reduces the production rate and consequently the cumulative production rate but in the SDFM model the reservoir takes more time to achieve depletion than the DPDP model. The normal fracture factor which appears in the transfer term of the DPDP model is adjusted against

  13. Fluorescent metal nanoshell and CK19 detection on single cell image.

    Science.gov (United States)

    Zhang, Jian; Fu, Yi; Li, Ge; Lakowicz, Joseph R; Zhao, Richard Y

    2011-09-16

    In this article, we report the synthesis strategy and optical properties of a novel type of fluorescence metal nanoshell when it was used as imaging agent for fluorescence cell imaging. The metal nanoshells were made with 40 nm silica cores and 10nm silver shells. Unlike typical fluorescence metal nanoshells which contain the organic dyes in the cores, novel metal nanoshells were composed of Cy5-labelled monoclonal anti-CK19 antibodies (mAbs) on the external surfaces of shells. Optical measurements to the single nanoparticles showed that in comparison with the metal free labelled mAbs, the mAb-Ag complexes displayed significantly enhanced emission intensity and dramatically shortened lifetime due to near-field interactions of fluorophores with metal. These metal nanoshells were found to be able to immunoreact with target cytokeratin 19 (CK19) molecules on the surfaces of LNCAP and HeLa cells. Fluorescence cell images were recorded on a time-resolved confocal microscope. The emissions from the metal nanoprobes could be clearly isolated from the cellular autofluorescence backgrounds on the cell images as either individuals or small clusters due to their stronger emission intensities and shorter lifetimes. These emission signals could also be precisely counted on single cell images. The count number may provide an approach for quantifying the target molecules in the cells.

  14. A comparison of single-lead atrial pacing with dual-chamber pacing in sick sinus syndrome

    DEFF Research Database (Denmark)

    Nielsen, Jens Cosedis; Thomsen, Poul Erik B; Højberg, Søren

    2011-01-01

    In patients with sick sinus syndrome, bradycardia can be treated with a single-lead pacemaker or a dual-chamber pacemaker. Previous trials have revealed that pacing modes preserving atrio-ventricular synchrony are superior to single-lead ventricular pacing, but it remains unclear if there is any ...

  15. Dual-Modal Nanoprobes for Imaging of Mesenchymal Stem Cell Transplant by MRI and Fluorescence Imaging

    Energy Technology Data Exchange (ETDEWEB)

    Sung, Chang Kyu; Hong, Kyung Ah; Lin, Shun Mei [Seoul Metropolitan Boramae Medical Center, Seoul (Korea, Republic of)] (and others)

    2009-12-15

    To determine the feasibility of labeling human mesenchymal stem cells (hMSCs) with bifunctional nanoparticles and assessing their potential as imaging probes in the monitoring of hMSC transplantation. The T1 and T2 relaxivities of the nanoparticles (MNP SiO{sub 2}[RITC]-PEG) were measured at 1.5T and 3T magnetic resonance scanner. Using hMSCs and the nanoparticles, labeling efficiency, toxicity, and proliferation were assessed. Confocal laser scanning microscopy and transmission electron microscopy were used to specify the intracellular localization of the endocytosed iron nanoparticles. We also observed in vitro and in vivo visualization of the labeled hMSCs with a 3T MR scanner and optical imaging. MNP SiO{sub 2}(RITC)-PEG showed both superparamagnetic and fluorescent properties. The r{sub 1} and r{sub 2} relaxivity values of the MNP SiO{sub 2}(RITC)-PEG were 0.33 and 398 mM{sup -1} s{sup -1} at 1.5T, respectively, and 0.29 and 453 mM{sup -1} s{sup -1} at 3T, respectively. The effective internalization of MNP SiO{sub 2}(RITC)-PEG into hMSCs was observed by confocal laser scanning fluorescence microscopy. The transmission electron microscopy images showed that MNP SiO{sub 2}(RITC)-PEG was internalized into the cells and mainly resided in the cytoplasm. The viability and proliferation of MNP SiO{sub 2}(RITC)-PEG-labeled hMSCs were not significantly different from the control cells. MNP SiO{sub 2}(RITC)-PEG-labeled hMSCs were observed in vitro and in vivo with optical and MR imaging. MNP SiO{sub 2}(RITC)-PEG can be a useful contrast agent for stem cell imaging, which is suitable for a bimodal detection by MRI and optical imaging.

  16. Fluorescence monitoring of riboswitch transcription regulation using a dual molecular beacon assay.

    Science.gov (United States)

    Chinnappan, Raja; Dubé, Audrey; Lemay, Jean-François; Lafontaine, Daniel A

    2013-05-01

    Riboswitches are mRNA elements that specifically bind cellular metabolites and control gene expression by modifying their structure. As riboswitches often control essential genes in pathogenic bacteria, riboswitches have been proposed as new targets for antibiotics. High-throughput screening provides a powerful approach to identify riboswitch ligand analogs that could act as powerful antibacterial drugs. Biochemical assays have already been used to find riboswitch-binding analogs, but those methods do take into account the transcriptional context for riboswitch regulation. As the importance of co-transcriptional ligand binding has been shown for several riboswitches, it is vital to develop an assay that screens riboswitch-binding analogs during the transcriptional process. Here, we describe the development of a dual molecular beacon system monitoring the transcriptional regulation activity of the Bacillus subtilis pbuE adenine riboswitch. This system relies on two molecular beacons that enable the monitoring of transcription efficiency, as well as the regulatory activity of the riboswitch. Different analogs were tested using our system, and a good correlation was observed between riboswitch activity and reported metabolite affinities. This method is specific, reliable and could be applied at the high-throughput level for the identification of new potential antibiotics targeting any riboswitch-regulating gene expression at the mRNA level.

  17. Removal of Out-of-Plane Fluorescence for Single Cell Visualization and Quantification in Cryo-Imaging

    OpenAIRE

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

    2009-01-01

    We developed a cryo-imaging system, which alternates between sectioning (10–40 μm) and imaging bright field and fluorescence block-face image volumes with micronscale-resolution. For applications requiring single-cell detection of fluorescently labeled cells anywhere in a mouse, we are developing software for reduction of out-of-plane fluorescence. In mouse experiments, we imaged GFP-labeled cancer and stem cells, and cell-sized fluorescent microspheres. To remove out-of-plane fluorescence, w...

  18. Deciphering the Structure and Function of Nuclear Pores Using Single-Molecule Fluorescence Approaches.

    Science.gov (United States)

    Musser, Siegfried M; Grünwald, David

    2016-05-22

    Due to its central role in macromolecular trafficking and nucleocytoplasmic information transfer, the nuclear pore complex (NPC) has been studied in great detail using a wide spectrum of methods. Consequently, many aspects of its architecture, general function, and role in the life cycle of a cell are well understood. Over the last decade, fluorescence microscopy methods have enabled the real-time visualization of single molecules interacting with and transiting through the NPC, allowing novel questions to be examined with nanometer precision. While initial single-molecule studies focused primarily on import pathways using permeabilized cells, it has recently proven feasible to investigate the export of mRNAs in living cells. Single-molecule assays can address questions that are difficult or impossible to answer by other means, yet the complexity of nucleocytoplasmic transport requires that interpretation be based on a firm genetic, biochemical, and structural foundation. Moreover, conceptually simple single-molecule experiments remain technically challenging, particularly with regard to signal intensity, signal-to-noise ratio, and the analysis of noise, stochasticity, and precision. We discuss nuclear transport issues recently addressed by single-molecule microscopy, evaluate the limits of existing assays and data, and identify open questions for future studies. We expect that single-molecule fluorescence approaches will continue to be applied to outstanding nucleocytoplasmic transport questions, and that the approaches developed for NPC studies are extendable to additional complex systems and pathways within cells.

  19. Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization (FISH)

    DEFF Research Database (Denmark)

    Ronander, Elena; Bengtsson, Dominique C; Joergensen, Louise;

    2012-01-01

    and the consequence of differential binding on the clinical outcome of P. falciparum infections. Recently, the mutually exclusive transcription paradigm has been called into doubt by transcription assays based on individual P. falciparum transcript identification in single infected erythrocytic cells using RNA...... fluorescent in situ hybridization (FISH) analysis of var gene transcription by the parasite in individual nuclei of P. falciparum IE(1). Here, we present a detailed protocol for carrying out the RNA-FISH methodology for analysis of var gene transcription in single-nuclei of P. falciparum infected human...... erythrocytes. The method is based on the use of digoxigenin- and biotin- labeled antisense RNA probes using the TSA Plus Fluorescence Palette System(2) (Perkin Elmer), microscopic analyses and freshly selected P. falciparum IE. The in situ hybridization method can be used to monitor transcription...

  20. Note: Single ion imaging and fluorescence collection with a parabolic mirror trap

    Science.gov (United States)

    Chou, Chen-Kuan; Auchter, Carolyn; Lilieholm, Jennifer; Smith, Kevin; Blinov, Boris

    2017-08-01

    Efficient fluorescence collection is the most challenging part in remote entangled ion qubit state generation. To address this issue, we developed an ion trap consisting of a reflective parabolic surface and a needle electrode. This parabolic trap design covers a solid angle of 2π steradians and allows precise ion placement at the focal point of the parabola. We measured (39 ± 3)% fluorescence collection from a single ion with this trap and analyzed the mirror optical performance. We observed single ion image spot size of 3.4 times diffraction limit, improved to 2.8 times diffraction limit with the help of an external deformable mirror. The micromotion of the ion is determined to be the limiting factor, and the result is consistent with theoretical calculation.

  1. A Single-Photon Avalanche Diode Array for Fluorescence Lifetime Imaging Microscopy.

    Science.gov (United States)

    Schwartz, David Eric; Charbon, Edoardo; Shepard, Kenneth L

    2008-11-21

    We describe the design, characterization, and demonstration of a fully integrated single-photon avalanche diode (SPAD) imager for use in time-resolved fluorescence imaging. The imager consists of a 64-by-64 array of active SPAD pixels and an on-chip time-to-digital converter (TDC) based on a delay-locked loop (DLL) and calibrated interpolators. The imager can perform both standard time-correlated single-photon counting (TCSPC) and an alternative gated-window detection useful for avoiding pulse pile-up when measuring bright signal levels. To illustrate the use of the imager, we present measurements of the decay lifetimes of fluorescent dyes of several types with a timing resolution of 350 ps.

  2. Measurement of Three-Dimensional Dipole Orientation of a Single Fluorescent Nanoemitter by Emission Polarization Analysis

    OpenAIRE

    Lethiec, Clotilde; Laverdant, Julien; Vallon, Henri; Javaux, Clémentine; Dubertret, Benoît; Frigerio, Jean-Marc; Schwob, Catherine; Coolen, Laurent; Maître, Agnès

    2014-01-01

    International audience; We demonstrate theoretically and experimentally that the three-dimensional orientation of a single fluorescent nanoemitter can be determined by polarization analysis of the emitted light (while excitation polarization analysis provides only the in-plane orientation). The determination of the emitter orientation by polarimetry requires a theoretical description, including the objective numerical aperture, the 1D or 2D nature of the emitting dipole, and the environment c...

  3. Color center fluorescence and spin manipulation in single crystal, pyramidal diamond tips

    Science.gov (United States)

    Nelz, Richard; Fuchs, Philipp; Opaluch, Oliver; Sonusen, Selda; Savenko, Natalia; Podgursky, Vitali; Neu, Elke

    2016-11-01

    We investigate bright fluorescence of nitrogen (NV)- and silicon-vacancy color centers in pyramidal, single crystal diamond tips, which are commercially available as atomic force microscope probes. We coherently manipulate NV electronic spin ensembles with T2 = 7.7(3) μs. Color center lifetimes in different tip heights indicate effective refractive index effects and quenching. Using numerical simulations, we verify enhanced photon rates from emitters close to the pyramid apex rendering them promising as scanning probe sensors.

  4. Color center fluorescence and spin manipulation in single crystal, pyramidal diamond tips

    CERN Document Server

    Nelz, Richard; Opaluch, Oliver; Sonusen, Selda; Savenko, Natalia; Podgursky, Vitali; Neu, Elke

    2016-01-01

    We investigate bright fluorescence of nitrogen (NV)- and silicon-vacancy color centers in pyramidal, single crystal diamond tips which are commercially available as atomic force microscope probes. We coherently manipulate NV electronic spin ensembles with $T_2 = 7.7(3)\\,\\mu$s. Color center lifetimes in different tip heights indicate effective refractive index effects and quenching. Using numerical simulations, we verify enhanced photon rates from emitters close to the pyramid apex; a situation promising for scanning probe sensing.

  5. Single-molecule fluorescence study of the inhibition of the oncogenic functionality of STAT3

    Science.gov (United States)

    Liu, Baoxu; Badali, Daniel; Fletcher, Steven; Avadisian, Miriam; Gunning, Patrick; Gradinaru, Claudiu

    2009-06-01

    Signal-Transducer-and-Activator-of-Transcription 3 (STAT3) protein plays an important role in the onset of cancers such as leukemia and lymphoma. In this study, we aim to test the effectiveness of a novel peptide drug designed to tether STAT3 to the phospholipid bilayer of the cell membrane and thus inhibit unwanted transcription. As a first step, STAT3 proteins were successfully labelled with tetramethylrhodamine (TMR), a fluorescent dye with suitable photostability for single molecule studies. The effectiveness of labelling was determined using fluorescence correlation spectroscopy in a custom built confocal microscope, from which diffusion times and hydrodynamic radii of individual proteins were determined. A newly developed fluorescein derivative label (F-NAc) has been designed to be incorporated into the structure of the peptide drug so that peptide-STAT3 interactions can be examined. This dye is spectrally characterized and is found to be well suited for its application to this project, as well as other single-molecule studies. The membrane localization via high-affinity cholesterol-bound small-molecule binding agents can be demonstrated by encapsulating TMR-labeled STAT3 and inhibitors within a vesicle model cell system. To this end, unilaminar lipid vesicles were examined for size and encapsulation ability. Preliminary results of the efficiency and stability of the STAT3 anchoring in lipid membranes obtained via quantitative confocal imaging and single-molecule spectroscopy using a custom-built multiparameter fluorescence microscope are reported here.

  6. Dual-stimuli responsive and reversibly activatable theranostic nanoprobe for precision tumor-targeting and fluorescence-guided photothermal therapy

    Science.gov (United States)

    Zhao, Xu; Yang, Cheng-Xiong; Chen, Li-Gong; Yan, Xiu-Ping

    2017-05-01

    The integrated functions of diagnostics and therapeutics make theranostics great potential for personalized medicine. Stimulus-responsive therapy allows spatial control of therapeutic effect only in the site of interest, and offers promising opportunities for imaging-guided precision therapy. However, the imaging strategies in previous stimulus-responsive therapies are `always on' or irreversible `turn on' modality, resulting in poor signal-to-noise ratios or even `false positive' results. Here we show the design of dual-stimuli-responsive and reversibly activatable nanoprobe for precision tumour-targeting and fluorescence-guided photothermal therapy. We fabricate the nanoprobe from asymmetric cyanine and glycosyl-functionalized gold nanorods (AuNRs) with matrix metalloproteinases (MMPs)-specific peptide as a linker to achieve MMPs/pH synergistic and pH reversible activation. The unique activation and glycosyl targetibility makes the nanoprobe bright only in tumour sites with negligible background, while AuNRs and asymmetric cyanine give synergistic photothermal effect. This work paves the way to designing efficient nanoprobes for precision theranostics.

  7. Nanoscale fluorescence lifetime imaging of an optical antenna with a single diamond NV center.

    Science.gov (United States)

    Beams, Ryan; Smith, Dallas; Johnson, Timothy W; Oh, Sang-Hyun; Novotny, Lukas; Vamivakas, A Nick

    2013-08-14

    Solid-state quantum emitters, such as artificially engineered quantum dots or naturally occurring defects in solids, are being investigated for applications ranging from quantum information science and optoelectronics to biomedical imaging. Recently, these same systems have also been studied from the perspective of nanoscale metrology. In this letter, we study the near-field optical properties of a diamond nanocrystal hosting a single nitrogen vacancy center. We find that the nitrogen vacancy center is a sensitive probe of the surrounding electromagnetic mode structure. We exploit this sensitivity to demonstrate nanoscale fluorescence lifetime imaging microscopy (FLIM) with a single nitrogen vacancy center by imaging the local density of states of an optical antenna.

  8. Economic and simple system to combine single-spot photolysis and whole-field fluorescence imaging

    Science.gov (United States)

    Jaafari, Nadia; Henson, Mark; Graham, Jeremy; Canepari, Marco

    2013-06-01

    In recent years, the use of light emitting diodes (LEDs) has become commonplace in fluorescence microscopy. LEDs are economical and easy to couple to commercial microscopes, and they provide powerful and stable light that can be triggered by transistor-transistor logic pulses in the range of tens of microseconds or shorter. LEDs are usually installed on the epifluorescence port of the microscope to obtain whole-field illumination, which is ideal for fluorescence imaging. In contrast, photolysis or channelrhodopsin stimulation often requires localized illumination, typically achieved using lasers. Here we show that insertion of a long-pass (>411 nm) filter with an appropriately sized pinhole in the epifluorescence pathway, combined with dual UV/visible illumination, can produce efficient whole-field visible illumination and spot UV illumination of 15 to 20 μm. We tested our system by performing calcium imaging experiments combined with L-glutamate or N-methyl-D-aspartic acid (NMDA) photorelease in hippocampal neurons from brain slices or dissociated cultures, demonstrating the ability to obtain local activation of NMDA receptors exclusively in the illuminated spot. The very inexpensive and simple system that we report here will allow many laboratories with limited budgets to run similar experiments in a variety of physiological applications.

  9. Multi-Pivot Quicksort: an Experiment with Single, Dual, Triple, Quad, and Penta-Pivot Quicksort Algorithms in Python

    Science.gov (United States)

    Budiman, M. A.; Zamzami, E. M.; Rachmawati, D.

    2017-03-01

    Dual-pivot quicksort, which was proposed by Yaroslavsky, has been experimentally proven to be more efficient than the classical single-pivot quicksort under the Java Virtual Machine [6]. Moreover, Kushagara, López-Ortiz, and Munro [4] has shown that triple-pivot quicksort runs 7-8% faster than dual-pivot quicksort in C, mutatis mutandis. In this research, we implement and experiment with single, dual, triple, quad, and penta-pivot quicksort algorithms in Python. Our experimental results are as follows. Firstly, the quicksort with single pivot is the slowest among the five variants. Secondly, at least until five (penta) pivots are being used, it is proven that the more pivots are used in a quicksort algorithm, the faster its performance becomes. Thirdly, the increase of speed resulted by adding more pivots tends to decrease gradually.

  10. Onsite-effects of dual-hemisphere versus conventional single-hemisphere transcranial direct current stimulation: A functional MRI study.

    Science.gov (United States)

    Kwon, Yong Hyun; Jang, Sung Ho

    2012-08-25

    We performed functional MRI examinations in six right-handed healthy subjects. During functional MRI scanning, transcranial direct current stimulation was delivered with the anode over the right primary sensorimotor cortex and the cathode over the left primary sensorimotor cortex using dual-hemispheric transcranial direct current stimulation. This was compared to a cathode over the left supraorbital area using conventional single-hemispheric transcranial direct current stimulation. Voxel counts and blood oxygenation level-dependent signal intensities in the right primary sensorimotor cortex regions were estimated and compared between the two transcranial direct current stimulation conditions. Our results showed that dual-hemispheric transcranial direct current stimulation induced greater cortical activities than single-hemispheric transcranial direct current stimulation. These findings suggest that dual-hemispheric transcranial direct current stimulation may provide more effective cortical stimulation than single-hemispheric transcranial direct current stimulation.

  11. Magnetic and fluorescent Gd2O3:Yb3+/Ln3+ nanoparticles for simultaneous upconversion luminescence/MR dual modal imaging and NIR-induced photodynamic therapy

    OpenAIRE

    LIU Jun; Huang, Long; Tian, Xiumei; Chen, Xiaoming; Shao, Yuanzhi; Xie, Fukang; Chen, Dihu; Li, Li

    2016-01-01

    The development of upconversion nanoparticles (UCNs) for theranostics application is a new strategy toward the accurate diagnosis and efficient treatment of cancer. Here, magnetic and fluorescent lanthanide-doped gadolinium oxide (Gd2O3) UCNs with bright upconversion luminescence (UCL) and high longitudinal relaxivity (r 1) are used for simultaneous magnetic resonance imaging (MRI)/UCL dual-modal imaging and photodynamic therapy (PDT). In vitro and in vivo MRI studies show that these products...

  12. Dental extraction without stopping single or dual antiplatelet therapy: results of a retrospective cohort study.

    Science.gov (United States)

    Lu, S-Y; Tsai, C-Y; Lin, L-H; Lu, S-N

    2016-10-01

    The aim of this study was to investigate the incidence of bleeding after dental extraction without stopping antiplatelet therapy. Postoperative bleeding was assessed in a total of 1271 patients who were divided into two groups: a study group comprising 183 patients on antiplatelet therapy (aspirin 125 patients/185 occasions; clopidogrel 42 patients/65 occasions; dual therapy 16 patients/24 occasions) who underwent 548 dental extractions on 274 occasions, and a control group comprising 1088 patients who were not receiving any antiplatelet or anticoagulant therapy and underwent 2487 dental extractions on 1472 occasions. The incidence of postoperative bleeding was higher in the study group (5/274, 1.8%) than in the control group (10/1472, 0.7%), and also in the dual antiplatelet subgroup (1/24, 4.2%) than in the single antiplatelet subgroups (clopidogrel: 2/65, 3.1%; aspirin: 2/185, 1.1%); however, these differences were not significant. Postoperative bleeding was managed successfully by repacking with Gelfoam impregnated with tranexamic acid powder in 12 patients and by resuturing in three of the control patients undergoing extraction of impacted teeth with flap elevation. These findings indicate that there is no need to interrupt antiplatelet drugs before dental extraction. Copyright © 2016 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  13. Single file and normal dual mode diffusion in highly confined hard sphere mixtures under flow.

    Science.gov (United States)

    Wanasundara, Surajith N; Spiteri, Raymond J; Bowles, Richard K

    2012-09-14

    We use Monte Carlo simulations to study the dual-mode diffusion regime of binary and tertiary mixtures of hard spheres confined in narrow cylindrical pores under the influence of an imposed flow. The flow is introduced to the dynamics by adding a small bias directed along the long axis of the pore to the random displacement of each Monte Carlo move. As a result, the motion of the particles in all the components is dominated by a drift velocity that causes the mean squared displacements to increase quadratically in the long time limit. However, an analysis of the mean squared displacements at intermediate time scales shows that components of the mixture above and below their passing thresholds still exhibit behaviors consistent with normal and single-file diffusion, respectively. The difference between the mean squared displacements of the various components is shown to go though a maximum, suggesting there may be an optimal pore diameter for the separation of mixtures exhibiting dual-mode diffusion.

  14. SAR reduction using a single SRR superstrate for a dual-band antenna.

    Science.gov (United States)

    Rosaline, Imaculate; Singaravelu, Raghavan

    2017-01-01

    A dual-band microstrip antenna operating at GSM 900 and GSM 1800 MHz is designed initially. Then a single split ring resonator (SRR) structure is used as a superstrate for this dual-band antenna. A circular current is induced in the SRR due to the perpendicular plane wave excitation, which in turn leads to an electric excitation coupled to the magnetic resonance. It also exhibits higher order excitations at 0.9 and 1.8 GHz which ultimately resulted in specific absorption rate (SAR) reduction of human head at both the designed frequencies of the antenna. The antenna and the SRR superstrate are printed on a 1.6 mm thick FR-4 substrate of dimension 59.6 × 49.6 mm(2). Analysis of the SRR using the classic waveguide theory approach is discussed. Radiation pattern of the antenna in the presence of SRR superstrate and human head is also discussed. Prototype of the antenna along with the SRR superstrate is fabricated and measured for return loss and radiation pattern. Measurement results fairly agree with the simulated results. A human head phantom is utilized in the calculation of SAR.

  15. Photon efficiency optimization in time-correlated single photon counting technique for fluorescence lifetime imaging systems.

    Science.gov (United States)

    Turgeman, Lior; Fixler, Dror

    2013-06-01

    In time-correlated single photon counting (TCSPC) systems, the maximum signal throughput is limited by the occurrence of pile-up and other effects. In many biological applications that exhibit high levels of fluorescence intensity (FI), pile-up-related distortions yield serious distortions in the fluorescence lifetime (FLT) calculation as well as significant decrease in the signal-to-noise ratio (SNR). Recent developments that allow the use of high-repetition-rate light sources (in the range of 50-100 MHz) in fluorescence lifetime imaging (FLIM) experiments enable minimization of pile-up-related distortions. However, modern TCSPC configurations that use high-repetition-rate excitation sources for FLIM suffer from dead-time-related distortions that cause unpredictable distortions of the FI signal. In this study, the loss of SNR is described by F- value as it is typically done in FLIM systems. This F-value describes the relation of the relative standard deviation in the estimated FLT to the relative standard deviation in FI measurements. Optimization of the F-value allows minimization of signal distortion, as well as shortening of the acquisition time for certain samples. We applied this method for Fluorescein, Rhodamine B, and Erythrosine fluorescent solutions that have different FLT values (4 ns, 1.67 ns, and 140 ps, respectively).

  16. A Single-Longitudinal-Mode Dual-Wavelength cw Tm,Ho: GdVO4 Microchip Laser

    Institute of Scientific and Technical Information of China (English)

    JU You-Lun; WANG Zhen-Guo; LI Yu-Feng; WANG Yue-Zhu

    2008-01-01

    A liquid nitrogen cooled dual-wavelength Tm,Ho:GdVO4 microchip laser is reported. The output dual wavelengths are at 2038.9nm and 2050.1nm. At each wavelength, the laser has a single longitudinal mode. The threshold power is nearly 20mW and the slope efficiency is 18.7%. The single longitudinal mode output power reaches 98mW, and the ratio of power is about 60% (2038.9nm) and 40% (2050.1 nm).

  17. Reconstructing comptonization parameters using simulations of single frequency, dual-beam observations of galaxy clusters

    CERN Document Server

    Lew, Bartosz

    2016-01-01

    Systematical effects in dual-beam, differential, radio observations of extended objects are discussed in the context of the One Centimeter Receiver Array (OCRA). We use simulated samples of Sunyaev--Zel'dovich (SZ) galaxy clusters at low ($z<0.4$) and intermediate ($0.4single frequency (30 GHz) on the accuracy of extracting SZ flux densities and of reconstructing comptonization parameters with OCRA, analysing dependences on cluster mass, redshift, observation strategy, and telescope pointing accuracy. Using $Planck$ data to make primary cosmic microwave background (CMB) templates, we test the feasibility of mitigating CMB confusion effects in observations of SZ profiles at angular scales larger than the separation of the receiver beams.

  18. Performance Analysis of Optimal Single Stream Beamforming in MIMO Dual-Hop AF Systems

    CERN Document Server

    Zhong, Caijun; Jin, Shi; Wong, Kai Kit

    2012-01-01

    This paper investigates the performance of optimal single stream beamforming schemes in multiple-input multiple-output (MIMO) dual-hop amplify-and-forward (AF) systems. Assuming channel state information is not available at the source and relay, the optimal transmit and receive beamforming vectors are computed at the destination, and the transmit beamforming vector is sent to the transmitter via a dedicated feedback link. Then, a set of new closed-form expressions for the statistical properties of the maximum eigenvalue of the resultant channel is derived, i.e., the cumulative density function (cdf), probability density function (pdf) and general moments, as well as the first order asymptotic expansion and asymptotic large dimension approximations. These analytical expressions are then applied to study three important performance metrics of the system, i.e., outage probability, average symbol error rate and ergodic capacity. In addition, more detailed treatments are provided for some important special cases, ...

  19. A single channel input virtual dual-phase lock-in amplifier

    Science.gov (United States)

    Gao, Zhongjian; Zheng, Hua; Li, Lianhuang; Chen, Fang; Guo, Fuyuan

    2011-11-01

    In this article, it presents a suit of single channel input virtual Dual-Phase Lock-in Amplifier (DPLIA) that is constructed by a personal computer sound card and the LabVIEW software. The virtual DPLIA is low cost and convenient to implement. The implemented DPLIA could enhance the noise tolerance capability and lower the LOD of the optical signal detection system. A primary benefit of this instrument is it only needs one input channel; two internal reference signals of the digital PLL are generated in the LabVIEW software. It is easy to control and operate, the data processing results can be saved directly to disks. This instrument will be applied in other weak signal detection systems.

  20. Restorability on 3-connected WDM Networks Under Single and Dual Physical Link Failures

    DEFF Research Database (Denmark)

    Gutierrez Lopez, Jose Manuel; Jensen, Michael; Riaz, Tahir

    2013-01-01

    This work studies the influence the network interconnection has over restoration techniques. The way physical links are distributed to interconnect network nodes has a great impact on parameters such as path distances when failures occur and restoration is applied. The work focuses on single...... and dual physical link failures restorability on WDM transport networks. This failure scenarios are tested over several 3-connected topologies, and studied in graph theory and network planning terms. In connection with the graphs, the resulting hop path distances and lengths are evaluated. In relation...... to network planning, the trade-off network length vs. performance of the different topological options is studied. The results show how 3-connected graphs could provide a reasonable trade-off between costs, link failure rates, and restored path parameters....

  1. A dual voltage control strategy for single-phase PWM converters with power decoupling function

    DEFF Research Database (Denmark)

    Tang, Yi; Qin, Zian; Blaabjerg, Frede

    2015-01-01

    converter topology based on a symmetrical half bridge circuit is proposed to decouple the ripple power so that balanced instantaneous power flow is assured between source and load, and the required dc-link capacitance can be dramatically reduced. For proper closed-loop regulation, the small signal modeling......The inherent double line ripple power in single-phase systems is adverse to the performance of power electronics converters, e.g. limited lifetime due to the requirement of large electrolytic capacitors and low voltage control bandwidth due to harmonic disturbance. In this paper, an active...... of the proposed system is presented, and a dual voltage control strategy is then proposed, which comprises one voltage loop implemented in the synchronous reference frame for active power balancing, and another one implemented in the stationary reference frame for ripple power compensation. Special attention...

  2. Evaluation of pulmonary function using single-breath-hold dual-energy computed tomography with xenon

    Science.gov (United States)

    Kyoyama, Hiroyuki; Hirata, Yusuke; Kikuchi, Satoshi; Sakai, Kosuke; Saito, Yuriko; Mikami, Shintaro; Moriyama, Gaku; Yanagita, Hisami; Watanabe, Wataru; Otani, Katharina; Honda, Norinari; Uematsu, Kazutsugu

    2017-01-01

    Abstract Xenon-enhanced dual-energy computed tomography (xenon-enhanced CT) can provide lung ventilation maps that may be useful for assessing structural and functional abnormalities of the lung. Xenon-enhanced CT has been performed using a multiple-breath-hold technique during xenon washout. We recently developed xenon-enhanced CT using a single-breath-hold technique to assess ventilation. We sought to evaluate whether xenon-enhanced CT using a single-breath-hold technique correlates with pulmonary function testing (PFT) results. Twenty-six patients, including 11 chronic obstructive pulmonary disease (COPD) patients, underwent xenon-enhanced CT and PFT. Three of the COPD patients underwent xenon-enhanced CT before and after bronchodilator treatment. Images from xenon-CT were obtained by dual-source CT during a breath-hold after a single vital-capacity inspiration of a xenon–oxygen gas mixture. Image postprocessing by 3-material decomposition generated conventional CT and xenon-enhanced images. Low-attenuation areas on xenon images matched low-attenuation areas on conventional CT in 21 cases but matched normal-attenuation areas in 5 cases. Volumes of Hounsfield unit (HU) histograms of xenon images correlated moderately and highly with vital capacity (VC) and total lung capacity (TLC), respectively (r = 0.68 and 0.85). Means and modes of histograms weakly correlated with VC (r = 0.39 and 0.38), moderately with forced expiratory volume in 1 second (FEV1) (r = 0.59 and 0.56), weakly with the ratio of FEV1 to FVC (r = 0.46 and 0.42), and moderately with the ratio of FEV1 to its predicted value (r = 0.64 and 0.60). Mode and volume of histograms increased in 2 COPD patients after the improvement of FEV1 with bronchodilators. Inhalation of xenon gas caused no adverse effects. Xenon-enhanced CT using a single-breath-hold technique depicted functional abnormalities not detectable on thin-slice CT. Mode, mean, and volume of HU histograms of xenon images

  3. A dual, single detector relaxed eddy accumulation system for long-term measurement of mercury flux

    Directory of Open Access Journals (Sweden)

    S. Osterwalder

    2015-08-01

    Full Text Available The fate of anthropogenic emissions of mercury (Hg to the atmosphere is influenced by the exchange of elemental Hg with the earth surface. This exchange which holds the key to a better understanding of Hg cycling from local to global scales has been difficult to quantify. To advance and facilitate research about land–atmosphere Hg interactions, we developed a dual-intake, single analyzer Relaxed Eddy Accumulation (REA system. REA is an established technique for measuring turbulent fluxes of trace gases and aerosol particles in the atmospheric surface layer. Accurate determination of gaseous elemental mercury (GEM fluxes has proven difficult to technical challenges presented by extremely small concentration differences (typically −3 between updrafts and downdrafts. To address this we present an advanced REA design that uses two inlets and two pair of gold cartridges for semi-continuous monitoring of GEM fluxes. They are then analyzed sequentially on the same detector while another pair of gold cartridges takes over the sample collection. We also added a reference gas module for repeated quality-control measurements. To demonstrate the system performance, we present results from field campaigns in two contrasting environments: an urban setting with a heterogeneous fetch and a boreal mire during snow-melt. The observed emission rates were 15 and 3 ng m−2 h−1. We claim that this dual-inlet, single detector approach is a significant development of the REA system for ultra-trace gases and can help to advance our understanding of long-term land–atmosphere GEM exchange.

  4. Pulmonary fluid overload monitoring in heart failure patients with single and dual chamber defibrillators.

    Science.gov (United States)

    Molon, Giulio; Zanotto, Gabriele; Rahue, Werner; Facchin, Domenico; Leoni, Loira; Morani, Giovanni; Calvi, Valeria; Catanzariti, Domenico; Costa, Alessandro; Zago, Lara; Comisso, Jennifer; Varbaro, Annamaria; Santini, Massimo

    2014-04-01

    Heart failure has a relevant healthcare impact. Monitoring of pulmonary fluid overload (PFO), measured by intrathoracic impedance, has been proposed to alert to heart failure worsening before symptoms become patent. The aim of our research was to evaluate whether PFO diagnostics reduce heart failure hospitalizations in heart failure patients receiving single-chamber or dual-chamber implantable cardioverter-defibrillator (ICD) for primary prevention of sudden death. Twenty-five Italian cardiological centers prospectively followed 221 ICD patients (86% men, 66 ± 11 years, 79% New York Heart Association II and left ventricular ejection fraction 28 ± 5%), of whom 123 received an ICD with PFO monitoring (diagnostics group) and 98 an ICD without such a diagnostics (control group). The association of each patient to a group was assigned a priori, independently of patients' characteristics but based on regional device allocation policies. Patient clinical characteristics and observation period were similar between groups. In a mean follow-up of 17 ± 11 months, heart failure hospitalizations or emergency-room admissions occurred in eight (7%) patients of the diagnostics group and in 16 of the control group (16%; P = 0.02), with an incidence, measured by Kaplan-Meier analysis, of 23% at 2 years and 34% at 3 years in patients of the control group compared with 8% at 2 and 3 years in patients of the diagnostics group (Log rank test P = 0.044). Our data show that in heart failure patients receiving single-chamber or dual-chamber ICD, the use of intrathoracic impedance monitoring is associated with a significant reduction of heart failure hospitalizations. Our results support the hypothesis that PFO diagnostics improve the likelihood of timely detection of heart failure worsening.

  5. Manipulation and Immobilization of a Single Fluorescence Nanosensor for Selective Injection into Cells

    Directory of Open Access Journals (Sweden)

    Hairulazwan Hashim

    2016-12-01

    Full Text Available Manipulation and injection of single nanosensors with high cell viability is an emerging field in cell analysis. We propose a new method using fluorescence nanosensors with a glass nanoprobe and optical control of the zeta potential. The nanosensor is fabricated by encapsulating a fluorescence polystyrene nanobead into a lipid layer with 1,3,3-trimethylindolino-6′-nitrobenzopyrylospiran (SP, which is a photochromic material. The nanobead contains iron oxide nanoparticles and a temperature-sensitive fluorescent dye, Rhodamine B. The zeta potential of the nanosensor switches between negative and positive by photo-isomerization of SP with ultraviolet irradiation. The positively-charged nanosensor easily adheres to a negatively-charged glass nanoprobe, is transported to a target cell, and then adheres to the negatively-charged cell membrane. The nanosensor is then injected into the cytoplasm by heating with a near-infrared (NIR laser. As a demonstration, a single 750 nm nanosensor was picked-up using a glass nanoprobe with optical control of the zeta potential. Then, the nanosensor was transported and immobilized onto a target cell membrane. Finally, it was injected into the cytoplasm using a NIR laser. The success rates of pick-up and cell immobilization of the nanosensor were 75% and 64%, respectively. Cell injection and cell survival rates were 80% and 100%, respectively.

  6. Fluorescent Carbon Quantum Dots as Single Light Converter for White LEDs

    Science.gov (United States)

    Feng, Xiaoting; Zhang, Feng; Wang, Yaling; Zhang, Yi; Yang, Yongzhen; Liu, Xuguang

    2016-06-01

    Synthesis of fluorescent carbon quantum dots (CQDs) as single light converter and their application in white light-emitting diodes (LEDs) are reported. CQDs were prepared by a one-step hydrothermal method using glucose and polyethylene glycol 200 as precursors. The structural and optical properties of the CQDs were investigated. The CQDs with uniform size of 4 nm possessed typical excitation-dependent emission wavelength and quantum yield of 3.5%. Under ultraviolet illumination, the CQDs in deionized water emitted bright blue fluorescence and produced broad visible-light emission with high red, green, and blue spectral component ratio of 63.5% (red-to-blue intensity to total intensity), suggesting great potential as single light converter for white LEDs. To demonstrate their potential, a white LED using CQDs as a single light converter was built. The device exhibited cool white light with corresponding color temperature of 5584 K and color coordinates of (0.32, 0.37), belonging to the white gamut. This research suggests that CQDs could be a promising candidate single light converter for white LEDs.

  7. Effect of Same-dose Single or Dual Field Irradiation on Damage to Miniature Pig Parotid Glands

    Institute of Scientific and Technical Information of China (English)

    Xing Yan; Bo Hai; Zhao-chen Shan; Chang-yu Zheng; Chun-mei Zhang; Song-lin Wang

    2009-01-01

    Aim To evaluate the effect of single or dual field irradiation (IR) with the same dose on damage to miniature pig parotid glands. Methodology Sixteen miniature pigs were divided into two IR groups (n=6) and a control group (n=4). The irradiation groups were subjected to 20 Gy X-radiation to one parotid gland using single-field or dual-field modality by linear accelerator. The dose-volume distributions between two IR groups were compared. Saliva from parotid glands and blood were collected at 0, 4, 8 and 16 weeks after irradiation. Parotid glands were removed at 16 weeks to evaluate tissue morphology. Results The irradiation dose volume distributions were significantly different between single and dual field irradiation groups (t=4.177, P=0.002), although dose volume histogramin (DVH) indicated the equal maximal dose in parotid glands. Saliva flow rates from IR side decreased dramatically at all time points in IR groups, especially in dual field irradiation group. The radiation caused changes of white blood cell count in blood, lactate dehydrogenase and amylase in serum, calcium, potassium and amylase in saliva. Morphologically, more severe radiation damage was found in irradiated parotid glands from dual field irradiation group than that from single field irradiation group. Conclusion Data from this large animal model demonstrated that the radiation damage from the dual field irradiation was more severe than that of the single field irradiation at the same dose, suggesting that dose-volume distribution is an important factor in evaluation of the radiobiology of parotid glands.

  8. Effects of single and dual physical modifications on pinhão starch.

    Science.gov (United States)

    Pinto, Vânia Zanella; Vanier, Nathan Levien; Deon, Vinicius Gonçalves; Moomand, Khalid; El Halal, Shanise Lisie Mello; Zavareze, Elessandra da Rosa; Lim, Loong-Tak; Dias, Alvaro Renato Guerra

    2015-11-15

    Pinhão starch was modified by annealing (ANN), heat-moisture (HMT) or sonication (SNT) treatments. The starch was also modified by a combination of these treatments (ANN-HMT, ANN-SNT, HMT-ANN, HMT-SNT, SNT-ANN, SNT-HMT). Whole starch and debranched starch fractions were analyzed by gel-permeation chromatography. Moreover, crystallinity, morphology, swelling power, solubility, pasting and gelatinization characteristics were evaluated. Native and single ANN and SNT-treated starches exhibited a CA-type crystalline structure while other modified starches showed an A-type structure. The relative crystallinity increased in ANN-treated starches and decreased in single HMT- and SNT-treated starches. The ANN, HMT and SNT did not provide visible cracks, notches or grooves to pinhão starch granule. SNT applied as second treatment was able to increase the peak viscosity of single ANN- and HMT-treated starches. HMT used alone or in dual modifications promoted the strongest effect on gelatinization temperatures and enthalpy.

  9. Endoleak detection using single-acquisition split-bolus dual-energy computer tomography (DECT)

    Energy Technology Data Exchange (ETDEWEB)

    Javor, D.; Wressnegger, A.; Unterhumer, S.; Kollndorfer, K.; Nolz, R.; Beitzke, D.; Loewe, C. [Medical University of Vienna, Department of Biomedical Imaging and Image-guided Therapy, Vienna (Austria)

    2017-04-15

    To assess a single-phase, dual-energy computed tomography (DECT) with a split-bolus technique and reconstruction of virtual non-enhanced images for the detection of endoleaks after endovascular aneurysm repair (EVAR). Fifty patients referred for routine follow-up post-EVAR CT and a history of at least one post-EVAR follow-up CT examination using our standard biphasic (arterial and venous phase) routine protocol (which was used as the reference standard) were included in this prospective trial. An in-patient comparison and an analysis of the split-bolus protocol and the previously used double-phase protocol were performed with regard to differences in diagnostic accuracy, radiation dose, and image quality. The analysis showed a significant reduction of radiation dose of up to 42 %, using the single-acquisition split-bolus protocol, while maintaining a comparable diagnostic accuracy (primary endoleak detection rate of 96 %). Image quality between the two protocols was comparable and only slightly inferior for the split-bolus scan (2.5 vs. 2.4). Using the single-acquisition, split-bolus approach allows for a significant dose reduction while maintaining high image quality, resulting in effective endoleak identification. (orig.)

  10. Dual grafts in adult-to-adult living donor liver transplantation: a single center experience in Taiwan.

    Science.gov (United States)

    Yang, Chin-Hsiang; Chen, Chao-Long; Wang, Chih-Chi; Concejero, Allan M; Wang, Shih-Ho; Liu, Yueh-Wei; Yong, Chee-Chien; Lin, Tsan-Shiun

    2009-02-01

    Volume mismatch is encountered when a single live donor cannot provide adequate graft volume to the recipient with a remnant liver volume which is safe for donation. Our objective is to present our experience in living donor liver transplantation using dual grafts. Record review of 4 dual graft recipients was done. The results were compared with 122 consecutive patients who received a single right lobe. All dual graft recipients were surviving with satisfactory liver function at a median follow-up of 21 months. Two recipients received 1 right and 1 left lobe graft, while the other 2 recipients received 2 left lobe grafts. One donor developed biloma and was managed by percutaneous drainage. The first recipient required re-laparotomy for postoperative bleeding. The second recipient underwent re-laparotomy for bile leak. The third recipient developed grade II decubitus ulcers due to a prolonged sedentary position. When compared with recipients who received a single right lobe, the operative time was prolonged in the dual graft group. There was no apparent increase in the rate of vascular and biliary complications or the incidence of acute cellular rejection. Actuarial patient survivals were comparable in both groups. Dual graft transplantation provides sufficient volume in the recipient without jeopardizing donor safety.

  11. Investigation on utilization of biogas and Karanja oil biodiesel in dual fuel mode in a single cylinder DI diesel engine

    Directory of Open Access Journals (Sweden)

    Bhabani Prasanna Pattanaik, Chandrakanta Nayak, Basanta Kumar Nanda

    2013-01-01

    Full Text Available In this work, experiments were performed on a single cylinder DI diesel engine by using bio-gas as a primary fuel and Karanja oil biodiesel and diesel oil as secondary fuels in dual fuel operation. The experiments were performed to measure performance parameters i.e. (brake specific fuel consumption, brake thermal efficiency and exhaust gas temperature and emission parameters such as carbon monoxide, carbon dioxide, nitrogen oxide unburned hydro carbon and smoke etc. at different load conditions. For the dual-fuel system, the intake system of the test engine was modified to convert into biogas and biodiesel of a dual-fueled combustion engine. Biogas was injected during the intake process by gas injectors. The study showed that, the engine performance parameters like BP, BTE and EGT gradually increase with increase in engine load for all test conditions using both pilot fuels diesel and KOBD. However, the BSFC of the engine showed decreasing slope with increase in engine load for all test conditions. Above 40% engine load the BSFC values for all test fuels are very close to each other. The engine emission analysis showed that the CO2, CO and NOx emissions increase with increase in engine load for both single and dual fuel mode operation using both pilot fuels. The NOx concentration of exhaust gases in dual fuel mode is superior than that of single mode.

  12. Investigation on utilization of biogas and Karanja oil biodiesel in dual fuel mode in a single cylinder DI diesel engine

    Energy Technology Data Exchange (ETDEWEB)

    Prasanna Pattanaik, Bhabani; Nayak, Chandrakanta [Department of Mechanical Eng., Gandhi Institute for Technological Advancement, Madanpur, Bhubaneswar - 752054, Odisha (India); Kumar Nanda, Basanta [Department of Mechanical Eng., Maharaja Institute of Technology, Bhubaneswar, Odisha (India)

    2013-07-01

    In this work, experiments were performed on a single cylinder DI diesel engine by using bio-gas as a primary fuel and Karanja oil biodiesel and diesel oil as secondary fuels in dual fuel operation. The experiments were performed to measure performance parameters i.e. (brake specific fuel consumption, brake thermal efficiency and exhaust gas temperature) and emission parameters such as carbon monoxide, carbon dioxide, nitrogen oxide unburned hydro carbon and smoke etc. at different load conditions. For the dual-fuel system, the intake system of the test engine was modified to convert into biogas and biodiesel of a dual-fueled combustion engine. Biogas was injected during the intake process by gas injectors. The study showed that, the engine performance parameters like BP, BTE and EGT gradually increase with increase in engine load for all test conditions using both pilot fuels diesel and KOBD. However, the BSFC of the engine showed decreasing slope with increase in engine load for all test conditions. Above 40% engine load the BSFC values for all test fuels are very close to each other. The engine emission analysis showed that the CO2, CO and NOx emissions increase with increase in engine load for both single and dual fuel mode operation using both pilot fuels. The NOx concentration of exhaust gases in dual fuel mode is superior than that of single mode.

  13. Single and dual task gait training in people with Parkinson's Disease: A protocol for a randomised controlled trial

    Directory of Open Access Journals (Sweden)

    Silburn Peter

    2011-07-01

    Full Text Available Abstract Background Difficulty performing more than one task at a time (dual tasking is a common and disabling problem experienced by people with Parkinson disease (PD. If asked to perform another task when walking, people with PD often take shorter steps or walk more slowly. Currently there is uncertainty about whether clinicians should teach people with PD to avoid dual tasking or whether they should encourage them to practice dual tasking with the hope that practice will lead to enhanced performance. This study will address this issue by comparing single to dual task gait training. Methods and design A prospective randomised clinical trial is being conducted. Sixty participants with idiopathic PD will be recruited, provided they score I-IV on the modified Hoehn and Yahr (1967 scale, and fulfil other inclusion criteria. Participants will be randomly allocated to either a single or dual task gait training group. Both groups will receive 12 hours of walking training over 4 weeks. The single task group will undertake gait training with cueing strategies to increase step length. The dual task group will train to improve step length when walking and performing a variety of added tasks. Both groups will receive a tailored home program for 6 months. Blinded assessors will conduct four assessments: two baseline assessments, one post intervention and one at 6 months follow-up. The primary outcome measure will be step length when dual tasking over 8 m. Secondary outcome measures include: spatiotemporal gait parameters when walking under single and dual task conditions, measures of executive function, the timed up and go test, measures of community mobility, and quality of life. All analyses will be based on intention to treat principle. Discussion This trial will examine the immediate and longer term effect of dual task walking training as compared to single task training in people with idiopathic PD, at the impairment, activity, and participation

  14. Single and dual task gait training in people with Parkinson's Disease: A protocol for a randomised controlled trial

    Science.gov (United States)

    2011-01-01

    Background Difficulty performing more than one task at a time (dual tasking) is a common and disabling problem experienced by people with Parkinson disease (PD). If asked to perform another task when walking, people with PD often take shorter steps or walk more slowly. Currently there is uncertainty about whether clinicians should teach people with PD to avoid dual tasking or whether they should encourage them to practice dual tasking with the hope that practice will lead to enhanced performance. This study will address this issue by comparing single to dual task gait training. Methods and design A prospective randomised clinical trial is being conducted. Sixty participants with idiopathic PD will be recruited, provided they score I-IV on the modified Hoehn and Yahr (1967) scale, and fulfil other inclusion criteria. Participants will be randomly allocated to either a single or dual task gait training group. Both groups will receive 12 hours of walking training over 4 weeks. The single task group will undertake gait training with cueing strategies to increase step length. The dual task group will train to improve step length when walking and performing a variety of added tasks. Both groups will receive a tailored home program for 6 months. Blinded assessors will conduct four assessments: two baseline assessments, one post intervention and one at 6 months follow-up. The primary outcome measure will be step length when dual tasking over 8 m. Secondary outcome measures include: spatiotemporal gait parameters when walking under single and dual task conditions, measures of executive function, the timed up and go test, measures of community mobility, and quality of life. All analyses will be based on intention to treat principle. Discussion This trial will examine the immediate and longer term effect of dual task walking training as compared to single task training in people with idiopathic PD, at the impairment, activity, and participation levels. It has the potential to

  15. Single-cell analysis reveals gene-expression heterogeneity in syntrophic dual-culture of Desulfovibrio vulgaris with Methanosarcina barkeri

    Science.gov (United States)

    Qi, Zhenhua; Pei, Guangsheng; Chen, Lei; Zhang, Weiwen

    2014-12-01

    Microbial syntrophic metabolism has been well accepted as the heart of how methanogenic and other anaerobic microbial communities function. In this work, we applied a single-cell RT-qPCR approach to reveal gene-expression heterogeneity in a model syntrophic system of Desulfovibrio vulgaris and Methanosarcina barkeri, as compared with the D. vulgaris monoculture. Using the optimized primers and single-cell analytical protocol, we quantitatively determine gene-expression levels of 6 selected target genes in each of the 120 single cells of D. vulgaris isolated from its monoculture and dual-culture with M. barkeri. The results demonstrated very significant cell-to-cell gene-expression heterogeneity for the selected D. vulgaris genes in both the monoculture and the syntrophic dual-culture. Interestingly, no obvious increase in gene-expression heterogeneity for the selected genes was observed for the syntrophic dual-culture when compared with its monoculture, although the community structure and cell-cell interactions have become more complicated in the syntrophic dual-culture. In addition, the single-cell RT-qPCR analysis also provided further evidence that the gene cluster (DVU0148-DVU0150) may be involved syntrophic metabolism between D. vulgaris and M. barkeri. Finally, the study validated that single-cell RT-qPCR analysis could be a valuable tool in deciphering gene functions and metabolism in mixed-cultured microbial communities.

  16. In vivo cancer targeting and fluorescence-CT dual-mode imaging with nanoprobes based on silver sulfide quantum dots and iodinated oil.

    Science.gov (United States)

    Qin, Meng-Yao; Yang, Xiao-Quan; Wang, Kan; Zhang, Xiao-Shuai; Song, Ji-Tao; Yao, Ming-Hao; Yan, Dong-Mei; Liu, Bo; Zhao, Yuan-Di

    2015-12-14

    In this article, a fluorescence-CT dual-mode nanoprobe is successfully synthesized by making use of distearoylphosphatidylethanolamine-poly(ethylene glycol)-folate (DSPE-PEG2000-FA) and other amphiphilic molecules to coat silver sulfide (Ag2S) quantum dots (QDs) and iodinated oil simultaneously. In vitro experiments show that the fluorescence wavelength of the nanoprobe is 1170 nm in the near infrared-II region. Its size is 139.6 nm, it has good dispersibility, and it has low cellular toxicity at concentrations up to 25 μg mL(-1) Ag. In vivo experiments revealed that the probe has a rather long circulation time (blood half-life of 5.7 hours), and the tissue histopathological tests show that it is not obviously harmful to major organs' normal function. Biochemical analysis (glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels) and blood analysis (white blood cell, red blood cell, hemoglobin and blood platelet counts) reveal that it has little influence on blood within 15 days of administration. When injected into HeLa xenograft nude mice by the tail vein, the probe elicited intensely enhanced fluorescence and X-ray computed tomography (CT) signals in the tumors after 24 hours, and the structure, size and position of tumor tissue were shown clearly. In a word, the probe has good tumor targeting capabilities, and it has significant value in fluorescence-CT dual-mode imaging in vivo.

  17. Determination of trace uranium by resonance fluorescence method coupled with photo-catalytic technology and dual cloud point extraction

    Science.gov (United States)

    Li, Jiekang; Li, Guirong; Han, Qian

    2016-12-01

    In this paper, two kinds of salophens (Sal) with different solubilities, Sal1 and Sal2, have been respectively synthesized, and they all can combine with uranyl to form stable complexes: [UO22 +-Sal1] and [UO22 +-Sal2]. Among them, [UO22 +-Sal1] was used as ligand to extract uranium in complex samples by dual cloud point extraction (dCPE), and [UO22 +-Sal2] was used as catalyst for the determination of uranium by photocatalytic resonance fluorescence (RF) method. The photocatalytic characteristic of [UO22 +-Sal2] on the oxidized pyronine Y (PRY) by potassium bromate which leads to the decrease of RF intensity of PRY were studied. The reduced value of RF intensity of reaction system (ΔF) is in proportional to the concentration of uranium (c), and a novel photo-catalytic RF method was developed for the determination of trace uranium (VI) after dCPE. The combination of photo-catalytic RF techniques and dCPE procedure endows the presented methods with enhanced sensitivity and selectivity. Under optimal conditions, the linear calibration curves range for 0.067 to 6.57 ng mL- 1, the linear regression equation was ΔF = 438.0 c (ng mL- 1) + 175.6 with the correlation coefficient r = 0.9981. The limit of detection was 0.066 ng mL- 1. The proposed method was successfully applied for the separation and determination of uranium in real samples with the recoveries of 95.0-103.5%. The mechanisms of the indicator reaction and dCPE are discussed.

  18. Determination of trace uranium by resonance fluorescence method coupled with photo-catalytic technology and dual cloud point extraction.

    Science.gov (United States)

    Li, Jiekang; Li, Guirong; Han, Qian

    2016-12-01

    In this paper, two kinds of salophens (Sal) with different solubilities, Sal1 and Sal2, have been respectively synthesized, and they all can combine with uranyl to form stable complexes: [UO2(2+)-Sal1] and [UO2(2+)-Sal2]. Among them, [UO2(2+)-Sal1] was used as ligand to extract uranium in complex samples by dual cloud point extraction (dCPE), and [UO2(2+)-Sal2] was used as catalyst for the determination of uranium by photocatalytic resonance fluorescence (RF) method. The photocatalytic characteristic of [UO2(2+)-Sal2] on the oxidized pyronine Y (PRY) by potassium bromate which leads to the decrease of RF intensity of PRY were studied. The reduced value of RF intensity of reaction system (ΔF) is in proportional to the concentration of uranium (c), and a novel photo-catalytic RF method was developed for the determination of trace uranium (VI) after dCPE. The combination of photo-catalytic RF techniques and dCPE procedure endows the presented methods with enhanced sensitivity and selectivity. Under optimal conditions, the linear calibration curves range for 0.067 to 6.57ngmL(-1), the linear regression equation was ΔF=438.0 c (ngmL(-1))+175.6 with the correlation coefficient r=0.9981. The limit of detection was 0.066ngmL(-1). The proposed method was successfully applied for the separation and determination of uranium in real samples with the recoveries of 95.0-103.5%. The mechanisms of the indicator reaction and dCPE are discussed.

  19. A tunable and switchable single-longitudinal-mode dual-wavelength fiber laser incorporating a reconfigurable dual-pass Mach-Zehnder interferometer and its application in microwave generation

    Science.gov (United States)

    Wang, Fei; Xu, En-Ming; Dong, Jian-Ji; Zhang, Xin-Liang

    2011-05-01

    A tunable and switchable single-longitudinal-mode (SLM) dual-wavelength fiber laser incorporating a reconfigurable dual-pass Mach-Zehnder interferometer (MZI) filter was proposed and demonstrated, which can be applied in microwave generation. By incorporating a high extinction ratio (ER) dual-pass MZI into an erbium-doped fiber ring cavity, SLM dual-wavelength lasing can be achieved even using a MZI with relatively little free spectrum range (FSR), and by beating the two wavelengths at a photodetector, a 9.76 GHz microwave signal with a 3-dB bandwidth of less than 10 kHz is obtained. Moreover, by direct linking the two outputs of the MZI, the high ER dual-pass MZI is easily reconfigured as a half FSR dual-pass MZI. Using this structure, switchable SLM dual-wavelength lasing can be conveniently realized.

  20. A space- and time-resolved single photon counting detector for fluorescence microscopy and spectroscopy

    Science.gov (United States)

    Michalet, X.; Siegmund, O. H. W.; Vallerga, J. V.; Jelinsky, P.; Millaud, J. E.; Weiss, S.

    2006-02-01

    We have recently developed a wide-field photon-counting detector having high-temporal and high-spatial resolutions and capable of high-throughput (the H33D detector). Its design is based on a 25 mm diameter multi-alkali photocathode producing one photo electron per detected photon, which are then multiplied up to 10 7 times by a 3-microchannel plate stack. The resulting electron cloud is proximity focused on a cross delay line anode, which allows determining the incident photon position with high accuracy. The imaging and fluorescence lifetime measurement performances of the H33D detector installed on a standard epifluorescence microscope will be presented. We compare them to those of standard single-molecule detectors such as single-photon avalanche photodiode (SPAD) or electron-multiplying camera using model samples (fluorescent beads, quantum dots and live cells). Finally, we discuss the design and applications of future generation of H33D detectors for single-molecule imaging and high-throughput study of biomolecular interactions.

  1. Fluorescence-based tools for single-cell approaches in food microbiology.

    Science.gov (United States)

    Bridier, A; Hammes, F; Canette, A; Bouchez, T; Briandet, R

    2015-11-20

    The better understanding of the functioning of microbial communities is a challenging and crucial issue in the field of food microbiology, as it constitutes a prerequisite to the optimization of positive and technological microbial population functioning, as well as for the better control of pathogen contamination of food. Heterogeneity appears now as an intrinsic and multi-origin feature of microbial populations and is a major determinant of their beneficial or detrimental functional properties. The understanding of the molecular and cellular mechanisms behind the behavior of bacteria in microbial communities requires therefore observations at the single-cell level in order to overcome "averaging" effects inherent to traditional global approaches. Recent advances in the development of fluorescence-based approaches dedicated to single-cell analysis provide the opportunity to study microbial communities with an unprecedented level of resolution and to obtain detailed insights on the cell structure, metabolism activity, multicellular behavior and bacterial interactions in complex communities. These methods are now increasingly applied in the field of food microbiology in different areas ranging from research laboratories to industry. In this perspective, we reviewed the main fluorescence-based tools used for single-cell approaches and their concrete applications with specific focus on food microbiology.

  2. Highly sensitive immunoassay of protein molecules based on single nanoparticle fluorescence detection in a nanowell

    Science.gov (United States)

    Han, Jin-Hee; Kim, Hee-Joo; Lakshmana, Sudheendra; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.

    2011-03-01

    A nanoarray based-single molecule detection system was developed for detecting proteins with extremely high sensitivity. The nanoarray was able to effectively trap nanoparticles conjugated with biological sample into nanowells by integrating with an electrophoretic particle entrapment system (EPES). The nanoarray/EPES is superior to other biosensor using immunoassays in terms of saving the amounts of biological solution and enhancing kinetics of antibody binding due to reduced steric hindrance from the neighboring biological molecules. The nanoarray patterned onto a layer of PMMA and LOL on conductive and transparent indium tin oxide (ITO)-glass slide by using e-beam lithography. The suspension of 500 nm-fluorescent (green emission)-carboxylated polystyrene (PS) particles coated with protein-A followed by BDE 47 polyclonal antibody was added to the chip that was connected to the positive voltage. The droplet was covered by another ITO-coated-glass slide and connected to a ground terminal. After trapping the particles into the nanowells, the solution of different concentrations of anti-rabbit- IgG labeled with Alexa 532 was added for an immunoassay. A single molecule detection system could quantify the anti-rabbit IgG down to atto-mole level by counting photons emitted from the fluorescent dye bound to a single nanoparticle in a nanowell.

  3. The reliability of postural balance measures in single and dual tasking in elderly fallers and non-fallers

    NARCIS (Netherlands)

    Swanenburg, Jaap; de Bruin, Eling D.; Favero, Kathrin; Uebelhart, Daniel; Mulder, Theo

    2008-01-01

    Background: The purpose of this study was to determine the reliability of a forceplate postural balance protocol in a group of elderly fallers and non-fallers. The measurements were tested in single and dual-task conditions, with and without vision. Methods: 37 elderly (mean age 73 +/- 6 years)

  4. Age-Related Changes in Brain Activation Underlying Single- and Dual-Task Performance: Visuomanual Drawing and Mental Arithmetic

    Science.gov (United States)

    Van Impe, A.; Coxon, J. P.; Goble, D. J.; Wenderoth, N.; Swinnen, S. P.

    2011-01-01

    Depending on task combination, dual-tasking can either be performed successfully or can lead to performance decrements in one or both tasks. Interference is believed to be caused by limitations in central processing, i.e. structural interference between the neural activation patterns associated with each task. In the present study, single- and…

  5. Burn-back Equations for High Volumetric Loading Single-grain Dual-thrust Rocket Propellant Configuration (Review Paper

    Directory of Open Access Journals (Sweden)

    Himanshu Shekhar

    2011-02-01

    Full Text Available Dual-thrust mode is adopted in solid propellant rocket propulsion through tailoring of burning area, nozzle, rocket motor chamber, propellant type, multiple propellant blocks. In the present study, mathematical formulation has been evolved for generation of burning surface area with web burnt for a simple central blind hole in a solid cylindrical propellant geometry with proper partial inhibition on external and lateral surfaces. The burn-back equation has been validated by static firing and parametric study was conducted to understand effect of various control geometrical parameters. The system is utilised for high volumetric loading, single propellant, single composition, single-chamber, single nozzle dual-thrust mode of burning profiles in rocket application.Defence Science Journal, 2011, 61(2, pp.165-170, DOI:http://dx.doi.org/10.14429/dsj.61.41

  6. Rigid motion correction of dual opposed planar projections in single photon imaging

    Science.gov (United States)

    Angelis, G. I.; Ryder, W. J.; Gillam, J. E.; Boisson, F.; Kyme, A. Z.; Fulton, R. R.; Meikle, S. R.; Kench, P. L.

    2017-05-01

    Awake and/or freely moving small animal single photon emission imaging allows the continuous study of molecules exhibiting slow kinetics without the need to restrain or anaesthetise the animals. Estimating motion free projections in freely moving small animal planar imaging can be considered as a limited angle tomography problem, except that we wish to estimate the 2D planar projections rather than the 3D volume, where the angular sampling in all three axes depends on the rotational motion of the animal. In this study, we hypothesise that the motion corrected planar projections estimated by reconstructing an estimate of the 3D volume using an iterative motion compensating reconstruction algorithm and integrating it along the projection path, will closely match the true, motion-less, planar distribution regardless of the object motion. We tested this hypothesis for the case of rigid motion using Monte-Carlo simulations and experimental phantom data based on a dual opposed detector system, where object motion was modelled with 6 degrees of freedom. In addition, we investigated the quantitative accuracy of the regional activity extracted from the geometric mean of opposing motion corrected planar projections. Results showed that it is feasible to estimate qualitatively accurate motion-corrected projections for a wide range of motions around all 3 axes. Errors in the geometric mean estimates of regional activity were relatively small and within 10% of expected true values. In addition, quantitative regional errors were dependent on the observed motion, as well as on the surrounding activity of overlapping organs. We conclude that both qualitatively and quantitatively accurate motion-free projections of the tracer distribution in a rigidly moving object can be estimated from dual opposed detectors using a correction approach within an iterative reconstruction framework and we expect this approach can be extended to the case of non-rigid motion.

  7. Dual-mode optical microscope based on single-pixel imaging

    Science.gov (United States)

    Rodríguez, A. D.; Clemente, P.; Tajahuerce, E.; Lancis, J.

    2016-07-01

    We demonstrate an inverted microscope that can image specimens in both reflection and transmission modes simultaneously with a single light source. The microscope utilizes a digital micromirror device (DMD) for patterned illumination altogether with two single-pixel photosensors for efficient light detection. The system, a scan-less device with no moving parts, works by sequential projection of a set of binary intensity patterns onto the sample that are codified onto a modified commercial DMD. Data to be displayed are geometrically transformed before written into a memory cell to cancel optical artifacts coming from the diamond-like shaped structure of the micromirror array. The 24-bit color depth of the display is fully exploited to increase the frame rate by a factor of 24, which makes the technique practicable for real samples. Our commercial DMD-based LED-illumination is cost effective and can be easily coupled as an add-on module for already existing inverted microscopes. The reflection and transmission information provided by our dual microscope complement each other and can be useful for imaging non-uniform samples and to prevent self-shadowing effects.

  8. Investigation on Nyquist pulse generation using a single dual-parallel Mach-Zehnder modulator.

    Science.gov (United States)

    Wu, Jian; Zang, Jizhao; Li, Yan; Kong, Deming; Qiu, Jifang; Zhou, Siyuan; Shi, Jindan; Lin, Jintong

    2014-08-25

    The generation of Nyquist pulses with a dual parallel Mach-Zehnder modulator (DPMZM) driven by a single RF signal is demonstrated theoretically and experimentally. A complete theoretical analysis is developed and the limitation of the proposed scheme is also discussed. It is theoretically proved that Nyquist pulses with a spectrum of 5 flat comb lines can be generated using a single DPMZM, which is also verified with simulation. 7 flat comb lines in frequency domain can also be obtained if a large RF driving voltage is applied to DPMZM but the generated waveforms won't present a sinc-shape. This scheme is further investigated experimentally. 40 GHz Nyquist pulses with full-width-at-half-maximum (FWHM) less than 4.65 ps, signal-to-noise ratio (SNR) better than 29.5 dB, and normalized root-mean-square error (NRMSE) less than 2.4% are generated. It is found that a tradeoff exists between the insertion loss of the DPMZM and the deviation of generated pulses. The tunability of repetition rate is experimentally verified by generation of 1 GHz to 40 GHz Nyquist pulses with SNR better than 28.4 dB and NRMSE less than 6.15%.

  9. Experimental investigations of the plasma radial uniformity in single and dual frequency capacitively coupled argon discharges

    Science.gov (United States)

    Zhao, Kai; Liu, Yong-Xin; Gao, Fei; Liu, Gang-Hu; Han, Dao-Man; Wang, You-Nian

    2016-12-01

    In the current work, the radial plasma density has been measured by utilizing a floating double probe in single and dual frequency capacitively coupled argon discharges operated in a cylindrical reactor, aiming at a better understanding of electromagnetic effects and exploring a method of improving the radial uniformity. The experimental results indicate that for single-frequency plasma sustained at low pressure, the plasma density radial profile exhibits a parabolic distribution at 90 MHz, whereas at 180 MHz, the profile evolves into a bimodal distribution, and both cases indicate poor uniformities. With increasing the pressure, the plasma radial uniformity becomes better for both driving frequency cases. By contrast, when discharges are excited by two frequencies (i.e., 90 + 180 MHz), the plasma radial profile is simultaneously influenced by both sources. It is found that by adjusting the low-frequency to high-frequency voltage amplitude ratio β, the radial profile of plasma density could be controlled and optimized for a wide pressure range. To gain a better plasma uniformity, it is necessary to consider the balance between the standing wave effect, which leads to a maximum plasma density at the reactor center, and the edge field effect, which is responsible for a maximum density near the radial electrode edge. This balance can be controlled either by selecting a proper gas pressure or by adjusting the ratio β.

  10. Dual vs. single computer monitor in a Canadian hospital Archiving Department: a study of efficiency and satisfaction.

    Science.gov (United States)

    Poder, Thomas G; Godbout, Sylvie T; Bellemare, Christian

    2011-01-01

    This paper describes a comparative study of clinical coding by Archivists (also known as Clinical Coders in some other countries) using single and dual computer monitors. In the present context, processing a record corresponds to checking the available information; searching for the missing physician information; and finally, performing clinical coding. We collected data for each Archivist during her use of the single monitor for 40 hours and during her use of the dual monitor for 20 hours. During the experimental periods, Archivists did not perform other related duties, so we were able to measure the real-time processing of records. To control for the type of records and their impact on the process time required, we categorised the cases as major or minor, based on whether acute care or day surgery was involved. Overall results show that 1,234 records were processed using a single monitor and 647 records using a dual monitor. The time required to process a record was significantly higher (p= .071) with a single monitor compared to a dual monitor (19.83 vs.18.73 minutes). However, the percentage of major cases was significantly higher (p= .000) in the single monitor group compared to the dual monitor group (78% vs. 69%). As a consequence, we adjusted our results, which reduced the difference in time required to process a record between the two systems from 1.1 to 0.61 minutes. Thus, the net real-time difference was only 37 seconds in favour of the dual monitor system. Extrapolated over a 5-year period, this would represent a time savings of 3.1% and generate a net cost savings of $7,729 CAD (Canadian dollars) for each workstation that devoted 35 hours per week to the processing of records. Finally, satisfaction questionnaire responses indicated a high level of satisfaction and support for the dual-monitor system. The implementation of a dual-monitor system in a hospital archiving department is an efficient option in the context of scarce human resources and has the

  11. Different screening strategies (single or dual for the diagnosis of suspected latent tuberculosis: a cost effectiveness analysis

    Directory of Open Access Journals (Sweden)

    Rook Graham

    2010-02-01

    Full Text Available Abstract Background Previous health economic studies recommend either a dual screening strategy [tuberculin skin test (TST followed by interferon-γ-release assay (IGRA] or a single one [IGRA only] for latent tuberculosis infection (LTBI, the former largely based on claims that it is more cost-effective. We sought to examine that conclusion through the use of a model that accounts for the additional costs of adverse drug reactions and directly compares two commercially available versions of the IGRA: the Quantiferon-TB-Gold-In-Tube (QFT-GIT and T-SPOT.TB. Methods A LTBI screening model directed at screening contacts was used to perform a cost-effectiveness analysis, from a UK healthcare perspective, taking into account the risk of isoniazid-related hepatotoxicity and post-exposure TB (2 years post contact using the TST, QFT-GIT and T-SPOT.TB IGRAs. Results Examining costs alone, the TST/IGRA dual screening strategies (TST/T-SPOT.TB and TST/QFT-GIT; £162,387 and £157,048 per 1000 contacts, respectively cost less than their single strategy counterparts (T-SPOT.TB and QFT-GIT; £203,983 and £202,921 per 1000 contacts which have higher IGRA test costs and greater numbers of persons undergoing LTBI treatment. However, IGRA alone strategies direct healthcare interventions and costs more accurately to those that are truly infected. Subsequently, less contacts need to be treated to prevent an active case of TB (T-SPOT.TB and QFT-GIT; 61.7 and 69.7 contacts in IGRA alone strategies. IGRA single strategies also prevent more cases of post-exposure TB. However, this greater effectiveness does not outweigh the lower incremental costs associated with the dual strategies. Consequently, when these costs are combined with effectiveness, the IGRA dual strategies are more cost-effective than their single strategy counterparts. Comparing between the IGRAs, T-SPOT.TB-based strategies (single and dual; £39,712 and £37,206 per active TB case prevented

  12. Optical mapping of a rice B AC clone using restriction endonuclease and imaging with fluorescent microscopy at single molecule level

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A method of constructing restriction map by optical mapping and single molecule fluorescent microscopy is described. DNA molecules were aligned and adsorbed on a glass coverslip surface by a mbdified "molecular combing"technique, and then the surface-immobilized DNAs were cleaved in situ with a restriction endonuclease. Individual DNA molecules digested by the endonuclease EcoR I were observable with fluorescent microscopy. Using optical mapping, a physical map of a rice bacterial artificial chromosome clone was constructed. This method will facilitate genomic mapping and tracing the dynamic process in real time at a single molecule level with fluorescence microscopy.

  13. 40-Gb/s star 16-QAM transmitter based on single dual-drive Mach-Zehnder modulator

    Institute of Scientific and Technical Information of China (English)

    Junming Gao; Xinyu Xu; Qingjiang Chang; Yikai Su

    2009-01-01

    We propose a 40-Gb/s star 16-ary quadrature amplitude modulation (16-QAM) transmitter using a single dual-drive Mach-Zehnder modulator (DDMZM). This transmitter is demonstrated through experiment and simulation and shows the advantage of simplicity for implementation. Simulation results indicate that error free performance could be achieved for the generated signal after 80-km standard single-mode fiber (SSMF) transmission with coherent detection scheme.

  14. Dynamic fluorescence spectroscopy on single tryptophan mutants of EIImtl in detergent micelles : Effects of substrate binding and phosphorylation on the fluorescence and anisotropy decay

    NARCIS (Netherlands)

    Swaving Dijkstra, Dolf; Broos, J.; Visser, Antonie J.W.G.; van Hoek, A.; Robillard, George

    1997-01-01

    The effects of substrate and substrate analogue binding and phosphorylation on the conformational dynamics of the mannitol permease of Escherichia coli were investigated, using time-resolved fluorescence spectroscopy on mutants containing five single tryptophans situated in the membrane-embedded C d

  15. Single-molecule detection of protein efflux from microorganisms using fluorescent single-walled carbon nanotube sensor arrays

    Science.gov (United States)

    Landry, Markita Patricia; Ando, Hiroki; Chen, Allen Y.; Cao, Jicong; Kottadiel, Vishal Isaac; Chio, Linda; Yang, Darwin; Dong, Juyao; Lu, Timothy K.; Strano, Michael S.

    2017-05-01

    A distinct advantage of nanosensor arrays is their ability to achieve ultralow detection limits in solution by proximity placement to an analyte. Here, we demonstrate label-free detection of individual proteins from Escherichia coli (bacteria) and Pichia pastoris (yeast) immobilized in a microfluidic chamber, measuring protein efflux from single organisms in real time. The array is fabricated using non-covalent conjugation of an aptamer-anchor polynucleotide sequence to near-infrared emissive single-walled carbon nanotubes, using a variable chemical spacer shown to optimize sensor response. Unlabelled RAP1 GTPase and HIV integrase proteins were selectively detected from various cell lines, via large near-infrared fluorescent turn-on responses. We show that the process of E. coli induction, protein synthesis and protein export is highly stochastic, yielding variability in protein secretion, with E. coli cells undergoing division under starved conditions producing 66% fewer secreted protein products than their non-dividing counterparts. We further demonstrate the detection of a unique protein product resulting from T7 bacteriophage infection of E. coli, illustrating that nanosensor arrays can enable real-time, single-cell analysis of a broad range of protein products from various cell types.

  16. Detection of ultra-high energy cosmic ray showers with a single-pixel fluorescence telescope

    CERN Document Server

    Fujii, T; Bertaina, M; Casolino, M; Dawson, B; Horvath, P; Hrabovsky, M; Jiang, J; Mandat, D; Matalon, A; Matthews, J N; Motloch, P; Palatka, M; Pech, M; Privitera, P; Schovanek, P; Takizawa, Y; Thomas, S B; Travnicek, P; Yamazaki, K

    2015-01-01

    We present a concept for large-area, low-cost detection of ultra-high energy cosmic rays (UHECRs) with a Fluorescence detector Array of Single-pixel Tele- scopes (FAST), addressing the requirements for the next generation of UHECR experiments. In the FAST design, a large field of view is covered by a few pixels at the focal plane of a mirror or Fresnel lens. We report first results of a FAST prototype installed at the Telescope Array site, consisting of a single 200 mm photomultiplier tube at the focal plane of a 1 m2 Fresnel lens system taken from the prototype of the JEM-EUSO experiment. The FAST prototype took data for 19 nights, demonstrating remarkable operational stability. We detected laser shots at distances of several kilometres as well as 16 highly significant UHECR shower candidates.

  17. pyFRET: A Python Library for Single Molecule Fluorescence Data Analysis

    CERN Document Server

    Murphy, Rebecca R; Klenerman, David

    2014-01-01

    Single molecule F\\"orster resonance energy transfer (smFRET) is a powerful experimental technique for studying the properties of individual biological molecules in solution. However, as adoption of smFRET techniques becomes more widespread, the lack of available software, whether open source or commercial, for data analysis, is becoming a significant issue. Here, we present pyFRET, an open source Python package for the analysis of data from single-molecule fluorescence experiments from freely diffusing biomolecules. The package provides methods for the complete analysis of a smFRET dataset, from burst selection and denoising, through data visualisation and model fitting. We provide support for both continuous excitation and alternating laser excitation (ALEX) data analysis. pyFRET is available as a package downloadable from the Python Package Index (PyPI) under the open source three-clause BSD licence, together with links to extensive documentation and tutorials, including example usage and test data. Additio...

  18. Single-molecule fluorescence spectroscopy maps the folding landscape of a large protein.

    Science.gov (United States)

    Pirchi, Menahem; Ziv, Guy; Riven, Inbal; Cohen, Sharona Sedghani; Zohar, Nir; Barak, Yoav; Haran, Gilad

    2011-10-11

    Proteins attain their function only after folding into a highly organized three-dimensional structure. Much remains to be learned about the mechanisms of folding of large multidomain proteins, which may populate metastable intermediate states on their energy landscapes. Here we introduce a novel method, based on high-throughput single-molecule fluorescence experiments, which is specifically geared towards tracing the dynamics of folding in the presence of a plethora of intermediates. We employ this method to characterize the folding reaction of a three-domain protein, adenylate kinase. Using thousands of single-molecule trajectories and hidden Markov modelling, we identify six metastable states on adenylate kinase's folding landscape. Remarkably, the connectivity of the intermediates depends on denaturant concentration; at low concentration, multiple intersecting folding pathways co-exist. We anticipate that the methodology introduced here will find broad applicability in the study of folding of large proteins, and will provide a more realistic scenario of their conformational dynamics.

  19. Largely Enhanced Single-molecule Fluorescence in Plasmonic Nanogaps formed by Hybrid Silver Nanostructures

    Science.gov (United States)

    Zhang, Jian; Lakowicz, Joseph R.

    2013-01-01

    It has been suggested that narrow gaps between metallic nanostructures can be practical for producing large field enhancement. We design a hybrid silver nanostructure geometry in which fluorescent emitters are sandwiched between silver nanoparticles and silver island film (SIF). A desired number of polyelectrolyte layers are deposited on the SIF surface before the self-assembly of a second silver nanoparticle layer. Layer-by-layer configuration provides a well-defined dye position. It allows us to study the photophyical behaviors of fluorophores in the resulting gap at the single molecule level. The enhancement factor of a fluorophore located in the gap is much higher than those on silver surfaces alone and on glass. These effects may be used for increased detectability of single molecules bound to surfaces which contain metallic structures for either biophysical studies or high sensitivity assays. PMID:23373787

  20. Subunits of highly Fluorescent Protein R-Phycoerythrin as Probes for Cell Imaging and Single-Molecule Detection

    Energy Technology Data Exchange (ETDEWEB)

    Isailovic, Dragan [Iowa State Univ., Ames, IA (United States)

    2005-01-01

    The purposes of our research were: (1) To characterize subunits of highly fluorescent protein R-Phycoerythrin (R-PE) and check their suitability for single-molecule detection (SMD) and cell imaging, (2) To extend the use of R-PE subunits through design of similar proteins that will be used as probes for microscopy and spectral imaging in a single cell, and (3) To demonstrate a high-throughput spectral imaging method that will rival spectral flow cytometry in the analysis of individual cells. We first demonstrated that R-PE subunits have spectroscopic and structural characteristics that make them suitable for SMD. Subunits were isolated from R-PE by high-performance liquid chromatography (HPLC) and detected as single molecules by total internal reflection fluorescence microscopy (TIRFM). In addition, R-PE subunits and their enzymatic digests were characterized by several separation and detection methods including HPLC, capillary electrophoresis, sodium dodecyl sulfate-polyacrilamide gel electrophoresis (SDS-PAGE) and HPLC-electrospray ionization mass spectrometry (ESI-MS). Favorable absorption and fluorescence of the R-PE subunits and digest peptides originate from phycoerythrobilin (PEB) and phycourobilin (PUB) chromophores that are covalently attached to cysteine residues. High absorption coefficients and strong fluorescence (even under denaturing conditions), broad excitation and emission fluorescence spectra in the visible region of electromagnetic spectrum, and relatively low molecular weights make these molecules suitable for use as fluorescence labels of biomolecules and cells. We further designed fluorescent proteins both in vitro and in vivo (in Escherichia coli) based on the highly specific attachment of PEB chromophore to genetically expressed apo-subunits of R-PE. In one example, apo-alpha and apo-beta R-PE subunits were cloned from red algae Polisiphonia boldii (P. boldii), and expressed in E. coli. Although expressed apo-subunits formed inclusion

  1. Single molecule fluorescence imaging as a technique for barium tagging in neutrinoless double beta decay

    Science.gov (United States)

    Jones, B. J. P.; McDonald, A. D.; Nygren, D. R.

    2016-12-01

    Background rejection is key to success for future neutrinoless double beta decay experiments. To achieve sensitivity to effective Majorana lifetimes of ~ 1028 years, backgrounds must be controlled to better than 0.1 count per ton per year, beyond the reach of any present technology. In this paper we propose a new method to identify the birth of the barium daughter ion in the neutrinoless double beta decay of 136Xe. The method adapts Single Molecule Fluorescent Imaging, a technique from biochemistry research with demonstrated single ion sensitivity. We explore possible SMFI dyes suitable for the problem of barium ion detection in high pressure xenon gas, and develop a fiber-coupled sensing system with which we can detect the presence of bulk Ba++ ions remotely. We show that our sensor produces signal-to-background ratios as high as 85 in response to Ba++ ions when operated in aqueous solution. We then describe the next stage of this R&D program, which will be to demonstrate chelation and fluorescence in xenon gas. If a successful barium ion tag can be developed using SMFI adapted for high pressure xenon gas detectors, the first essentially zero background, ton-scale neutrinoless double beta decay technology could be realized.

  2. Development of a single-cell X-ray fluorescence flow cytometer.

    Science.gov (United States)

    Crawford, Andrew M; Kurecka, Patrick; Yim, Tsz Kwan; Kozemchak, Claire; Deb, Aniruddha; Dostál, Lubomír; Sun, Cheng Jun; Brewe, Dale L; Barrea, Raul; Penner-Hahn, James E

    2016-07-01

    An X-ray fluorescence flow cytometer that can determine the total metal content of single cells has been developed. Capillary action or pressure was used to load cells into hydrophilic or hydrophobic capillaries, respectively. Once loaded, the cells were transported at a fixed vertical velocity past a focused X-ray beam. X-ray fluorescence was then used to determine the mass of metal in each cell. By making single-cell measurements, the population heterogeneity for metals in the µM to mM concentration range on fL sample volumes can be directly measured, a measurement that is difficult using most analytical methods. This approach has been used to determine the metal composition of 936 individual bovine red blood cells (bRBC), 31 individual 3T3 mouse fibroblasts (NIH3T3) and 18 Saccharomyces cerevisiae (yeast) cells with an average measurement frequency of ∼4 cells min(-1). These data show evidence for surprisingly broad metal distributions. Details of the device design, data analysis and opportunities for further sensitivity improvement are described.

  3. Drug transport mechanism of P-glycoprotein monitored by single molecule fluorescence resonance energy transfer

    Science.gov (United States)

    Ernst, S.; Verhalen, B.; Zarrabi, N.; Wilkens, S.; Börsch, M.

    2011-03-01

    In this work we monitor the catalytic mechanism of P-glycoprotein (Pgp) using single-molecule fluorescence resonance energy transfer (FRET). Pgp, a member of the ATP binding cassette family of transport proteins, is found in the plasma membrane of animal cells where it is involved in the ATP hydrolysis driven export of hydrophobic molecules. When expressed in the plasma membrane of cancer cells, the transport activity of Pgp can lead to the failure of chemotherapy by excluding the mostly hydrophobic drugs from the interior of the cell. Despite ongoing effort, the catalytic mechanism by which Pgp couples MgATP binding and hydrolysis to translocation of drug molecules across the lipid bilayer is poorly understood. Using site directed mutagenesis, we have introduced cysteine residues for fluorescence labeling into different regions of the nucleotide binding domains (NBDs) of Pgp. Double-labeled single Pgp molecules showed fluctuating FRET efficiencies during drug stimulated ATP hydrolysis suggesting that the NBDs undergo significant movements during catalysis. Duty cycle-optimized alternating laser excitation (DCO-ALEX) is applied to minimize FRET artifacts and to select the appropriate molecules. The data show that Pgp is a highly dynamic enzyme that appears to fluctuate between at least two major conformations during steady state turnover.

  4. In Vivo Single-Cell Fluorescence and Size Scaling of Phytoplankton Chlorophyll Content.

    Science.gov (United States)

    Álvarez, Eva; Nogueira, Enrique; López-Urrutia, Ángel

    2017-04-01

    In unicellular phytoplankton, the size scaling exponent of chlorophyll content per cell decreases with increasing light limitation. Empirical studies have explored this allometry by combining data from several species, using average values of pigment content and cell size for each species. The resulting allometry thus includes phylogenetic and size scaling effects. The possibility of measuring single-cell fluorescence with imaging-in-flow cytometry devices allows the study of the size scaling of chlorophyll content at both the inter- and intraspecific levels. In this work, the changing allometry of chlorophyll content was estimated for the first time for single phytoplankton populations by using data from a series of incubations with monocultures exposed to different light levels. Interspecifically, our experiments confirm previous modeling and experimental results of increasing size scaling exponents with increasing irradiance. A similar pattern was observed intraspecifically but with a larger variability in size scaling exponents. Our results show that size-based processes and geometrical approaches explain variations in chlorophyll content. We also show that the single-cell fluorescence measurements provided by imaging-in-flow devices can be applied to field samples to understand the changes in the size dependence of chlorophyll content in response to environmental variables affecting primary production.IMPORTANCE The chlorophyll concentrations in phytoplankton register physiological adjustments in cellular pigmentation arising mainly from changes in light conditions. The extent of these adjustments is constrained by the size of the phytoplankton cells, even within single populations. Hence, variations in community chlorophyll derived from photoacclimation are also dependent on the phytoplankton size distribution.

  5. Risk assessment for Helicoverpa zea (Lepidoptera: Noctuidae) resistance on dual-gene versus single-gene corn.

    Science.gov (United States)

    Edwards, Kristine T; Caprio, Michael A; Allen, K Clint; Musser, Fred R

    2013-02-01

    Recent Environmental Protection Agency (EPA) decisions regarding resistance management in Bt-cropping systems have prompted concern in some experts that dual-gene Bt-corn (CrylA.105 and Cry2Ab2 toxins) may result in more rapid selection for resistance in Helicoverpa zea (Boddie) than single-gene Bacillus thuringiensis (Bt)-corn (CrylAb toxin). The concern is that Bt-toxin longevity could be significantly reduced with recent adoption of a natural refuge for dual-gene Bt-cotton (CrylAc and Cry2Ab2 toxins) and concurrent reduction in dual-gene corn refuge from 50 to 20%. A population genetics framework that simulates complex landscapes was applied to risk assessment. Expert opinions on effectiveness of several transgenic corn and cotton varieties were captured and used to assign probabilities to different scenarios in the assessment. At least 350 replicate simulations with randomly drawn parameters were completed for each of four risk assessments. Resistance evolved within 30 yr in 22.5% of simulations with single-gene corn and cotton with no volunteer corn. When volunteer corn was added to this assessment, risk of resistance evolving within 30 yr declined to 13.8%. When dual-gene Bt-cotton planted with a natural refuge and single-gene corn planted with a 50% structured refuge was simulated, simultaneous resistance to both toxins never occurred within 30 yr, but in 38.5% of simulations, resistance evolved to toxin present in single-gene Bt-corn (CrylAb). When both corn and cotton were simulated as dual-gene products, cotton with a natural refuge and corn with a 20% refuge, 3% of simulations evolved resistance to both toxins simultaneously within 30 yr, while 10.4% of simulations evolved resistance to CrylAb/c toxin.

  6. Quality of standing balance in community-dwelling elderly: Age-related differences in single and dual task conditions.

    Science.gov (United States)

    Coelho, Tiago; Fernandes, Ângela; Santos, Rubim; Paúl, Constança; Fernandes, Lia

    2016-01-01

    To examine the relationship between age and quality of standing balance in single and dual task conditions. A cross-sectional study was conducted using a sample of 243 community-dwellers aged ≥65 years. Quality of standing balance was assessed by measuring the center of pressure (COP) sway with a pressure platform. Measurements were performed under single task (orthostatic position) and dual task (orthostatic position while performing a verbal fluency task) conditions. The mean age of the participants was 79.1(±7.3)years and 76.1% were women. Older age was associated with an increased COP sway, mainly in the medial/lateral (ML) direction. Most COP sway parameters were higher under dual task conditions than under single task. After controlling for the effect of the number of words enunciated in dual task conditions, only the differences in COP sway parameters in the ML direction remained significant. There was no significant interaction between age group (65-79; ≥80 years) and condition, which indicates that differences in COP sway caused by performing a secondary task were similar for younger and for older participants. Age did not seem to influence significantly the decline in the quality of standing balance triggered by performing a concurrent cognitive task. However, older age was consistently associated with poorer standing balance, both in single and in dual task conditions. Therefore, performing a secondary task may lead older individuals to reach their postural stability limits and, consequently, to fall. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  7. Applications of capillary electrophoresis and laser-induced fluorescence detection to the analysis of trace species: From single cells to single molecules

    Energy Technology Data Exchange (ETDEWEB)

    Xue, Qifeng [Iowa State Univ., Ames, IA (United States)

    1995-09-26

    This Ph.D. Thesis describes several separation and detection schemes for the analysis of small volume and amount of samples, such as intracellular components and single enzymes developed during research. Indirect Laser-induced fluorescence detection and capillary electrophoresis were used to quantify lactate and pyruvate in single red blood cells. The assay of specific enzyme activities was achieved by monitoring the highly fluorescent enzymatic reaction product, NADH. LDH activity was found not to be a unique marker for diagnosis of leukemia. Reactions of single LDH-1 molecules were investigated by monitoring the reaction product with LIF detection.

  8. Applications of capillary electrophoresis and laser-induced fluorescence detection to the analysis of trace species: From single cells to single molecules

    Energy Technology Data Exchange (ETDEWEB)

    Qifeng, X.

    1995-11-01

    This Ph.D. Thesis describes several separation and detection schemes for the analysis of small volume and amount of samples, such as intracellular components and single enzymes developed during research. Indirect Laser-induced fluorescence detection and capillary electrophoresis were used to quantify lactate and pyruvate in single red blood cells. The assay of specific enzyme activities was achieved by monitoring the highly fluorescent enzymatic reaction product, NADH. LDH activity was found not to be a unique marker for diagnosis of leukemia. Reactions of single LDH-1 molecules were investigated by monitoring the reaction product with LIF detection.

  9. Single- and dual-wavelength switchable erbium-doped fiber ring laser based on intracavity polarization selective tilted fiber gratings.

    Science.gov (United States)

    Mou, Chengbo; Saffari, Pouneh; Fu, Hongyan; Zhou, Kaiming; Zhang, Lin; Bennion, Ian

    2009-06-20

    We propose and demonstrate a single- and dual-wavelength switchable erbium-doped fiber laser (EDFL) by utilizing intracavity polarization selective filters based on tilted fiber gratings (TFGs). In the cavity, one 45 degrees TFG functions as an in-fiber polarizer and the other 77 degrees TFG is used as a fiber polarization dependent loss (PDL) filter. The combined polarization effect from these two TFGs enables the laser to switch between the single- and the dual-wavelength operation with a single-polarization state at room temperature. The laser output at each wavelength shows an optical signal-to-noise ratio (OSNR) of >60 dB, a side mode suppression ratio (SMSR) of >50 dB, and a polarization extinction ratio of approximately 35 dB. The proposed EDFL can give stable output under laboratory conditions.

  10. The temperature dependence of single-event transients in 90-nm CMOS dual-well and triple-well NMOSFETs

    Institute of Scientific and Technical Information of China (English)

    Li Da-Wei; Qin Jun-Rui; Chen Shu-Ming

    2013-01-01

    This paper investigates the temperature dependence of single-event transients (SETs) in 90-nm complementary metat-oxide semiconductor (CMOS) dual-well and triple-well negative metal-oxide semiconductor field-effect transistors (NMOSFETs).Technology computer-aided design (TCAD) three-dimensional (3D) simulations show that the drain current pulse duration increases from 85 ps to 245 ps for triple-well but only increases from 65 ps to 98 ps for dual-well when the temperature increases from-55 ℃C to 125 ℃C,which is closely correlated with the NMOSFET sources.This reveals that the pulse width increases with temperature in dual-well due to the weakening of the anti-amplification bipolar effect while increases with temperature in triple-well due to the enhancement of the bipolar amplification.

  11. Effect of propulsion system characteristics on ascent performance of dual-fueled single-stage earth-to-orbit transports

    Science.gov (United States)

    Rehder, J. J.

    1977-01-01

    The results of a parametric study of ascent performance are presented for a vertical take off, horizontal landing, single stage earth-to-orbit transport vehicle. Two dual fueled concepts, series burn and parallel burn, were investigated, both of which utilized dual position rocket nozzles. The analysis was made by systematically varying a set of propulsion similarity parameters, initial thrust-weight ratio, the proportion of the thrust due to dual position nozzle engines, expansion ratios of the rocket nozzle, and the relative split between the two fuels, hydrogen and hydrocarbon. The data are presented as a series of curves of mass ratio plotted against each of the similarity parameters for various combinations of the other similarity parameters.

  12. Impact of Rayleigh backscattering on single/dual feeder fiber WDM-PON architectures based on array waveguide gratings

    Institute of Scientific and Technical Information of China (English)

    Muhammad Idrees AFRIDI; Jie ZHANG; Yousaf KHAN; Jiawei HAN; Aftab HUSSEIN; Shahab AHMAD

    2013-01-01

    The performance of colorless wavelength- division multiplexing passive optical network (WDM- PON) systems suffers from transmission impairments due to Rayleigh backscattering (RB). A single feeder fiber colorless WDM-PON architecture was modeled, simulated and analyzed at 25 km distance that sustained the noise induced by RB. We analytically compared the performances between single feeder and dual feeder WDM-PON architectures based on array waveguide gratings (AWGs). For single feeder WDM-PON, the high extinction ratios in both return-to-zeros (RZ)-shaped differential phase shift keying (DPSK) downstream and intensity remodulated upstream data signals helped to increase the tolerance to the noise induced by RB. However, a cost effective colorless system in dual feeder WDM-PON architecture was achieved without any optical amplification and dispersion compensation, low power penalty. These results illustrate that single feeder fiber architecture was cost effective in terms of deployment having a power penalty, while dual feeder fiber had lower power penalty thereby with better performance. Simulation results show that downstream and upstream signals achieved error-free performance at 10-Gbps with negligible penalty and enhanced tolerance to the noise induced by RB over 25 km single mode fiber.

  13. Self-assembled dual-modality contrast agents for non-invasive stem cell tracking via near-infrared fluorescence and magnetic resonance imaging.

    Science.gov (United States)

    Liu, Hong; Tan, Yan; Xie, Lisi; Yang, Lei; Zhao, Jing; Bai, Jingxuan; Huang, Ping; Zhan, Wugen; Wan, Qian; Zou, Chao; Han, Yali; Wang, Zhiyong

    2016-09-15

    Stem cells hold great promise for treating various diseases. However, one of the main drawbacks of stem cell therapy is the lack of non-invasive image-tracking technologies. Although magnetic resonance imaging (MRI) and near-infrared fluorescence (NIRF) imaging have been employed to analyse cellular and subcellular events via the assistance of contrast agents, the sensitivity and temporal resolution of MRI and the spatial resolution of NIRF are still shortcomings. In this study, superparamagnetic iron oxide nanocrystals and IR-780 dyes were co-encapsulated in stearic acid-modified polyethylenimine to form a dual-modality contrast agent with nano-size and positive charge. These resulting agents efficiently labelled stem cells and did not influence the cellular viability and differentiation. Moreover, the labelled cells showed the advantages of dual-modality imaging in vivo.

  14. Characteristics of single and dual radio-frequency (RF) plasma sheaths

    Institute of Scientific and Technical Information of China (English)

    DAI Zhong-ling; WANG You-nian

    2006-01-01

    The characteristics of radio-frequency(RF)plasma sheaths have been topics of much scientific study for decades,and have also been of great importance in the manufacture of integrated circuits and fabricating microelectromechanical systems (MEMS),as well as in the study of physical phenomena in dusty plasmas.The sheaths behave special properties under various situations where they can be treated as collisionless or collisional,single- or dual-RF,one- or two-dimensional (1D or 2D) sheaths,etc.This paper reviews our recent progress on the dynamics of RF plasma sheaths using a fluid method that includes the fluid equations and Poission's equation coupled with an equivalent circuit model and a hybrid method in which the fluid model is combined with the Monte-Carlo (MC)method.The structures of RF sheaths behave differently in various situations and plasma parameters such as the ion density,electron temperature,as well as the external parameters such as the applied frequency,power,gas pressure,magnetic field,are crucial for determining the characteristics of plasma sheaths.

  15. Experimental Investigation of Performanec of Single Cylinder 4s Diesel Engine Using Dual Vegetable Oil Blended

    Directory of Open Access Journals (Sweden)

    Prof. C. S. Koli

    2014-03-01

    Full Text Available Over the last two decades there has been a tremendous increase in the number of automobiles and a corresponding increase in the fuel price. In this regard, alternative fuels like vegetable oils play a major role. Use of pure vegetable oil in diesel engines causes some problems due to their high viscosity compared with diesel fuel. To solve the problems due to high viscosity various techniques are used. One such technique is fuel blending. This paper investigated the performance parameters of dual vegetable oil blends (mixture of Mustard oil and Palm oil with diesel on a stationary single cylinder, four stroke direct injection compression ignition engine. The blends of BB 10 (combination of Diesel 90% by volume, Mustard oil 5% by volume and Palm oil 5% by volume and blends of BB 20 (combination of Diesel 80% by volume, Mustard oil 10% by volume and Palm oil 10% by volume gave better brake thermal efficiency, lower total fuel consumption and lower brake specific fuel consumption than other blends (BB 30, BB 40 and BB 50.

  16. Uncertainty characterization of AOD for the AATSR dual and single view retrieval algorithms

    Directory of Open Access Journals (Sweden)

    P. Kolmonen

    2013-04-01

    Full Text Available The uncertainty associated with satellite-retrieved aerosol properties is needed when these data are used to constrain chemical transport or climate models by using data assimilation. Global uncertainty as provided by comparison with independent ground-based observations is usually not adequate for that purpose. Rather the per-pixel uncertainty is needed. In this work we describe how these are determined in the AATSR dual and single view aerosol retrieval algorithms (ADV and ASV which are used to retrieve aerosol optical properties from reflectance measured at the top of the atmosphere. AATSR is the Aerosol Along-Track Scanning Radiometer which flies on the European Space Agency Environmental Satellite ENVISAT. In addition, issues related to multi-year retrievals are described and discussed. The aerosol optical depth (AOD retrieved for the year 2008 is validated versus ground-based AERONET sun photometer measurements with good agreement (r = 0.85. The comparison of the AOD uncertainties with those provided by AERONET shows that they behave well in a statistical sense. Other considerations regarding global multi-year aerosol retrievals are presented and discussed.

  17. Arrhythmia detection in single- and dual-chamber implantable cardioverter defibrillators: the more leads, the better?

    Science.gov (United States)

    Francia, Pietro; Balla, Cristina; Uccellini, Arianna; Cappato, Riccardo

    2009-09-01

    The implantable cardioverter defibrillator (ICD) offers life-saving therapies for primary and secondary prevention of sudden cardiac death in high-risk patients. However, ICD detection algorithms consistently misclassify a substantial proportion of supraventricular rhythms, thus carrying the risk for inappropriate therapies. Although single-chamber ICD (Sc-ICD) discrimination tools have been reported to provide high specificity in rejecting sinus tachycardia and atrial fibrillation with a relatively low ventricular rate, accurate recognition of atrial fibrillation with faster ventricular rates, atrial tachycardias, atrial flutter, and some reentrant tachycardias is still an issue. Dual-chamber ICDs (Dc-ICDs) are supposed to overcome specificity issues by enhancing detection algorithms with information derived from the atrial and ventricular timing relationship. The initial promise of Dc-ICDs was to improve detection specificity without compromising sensitivity, and to translate this advantage over Sc-ICDs in a more selective use of aggressive therapies. Despite this solid background, superiority of Dc- over Sc-ICDs has never been convincingly demonstrated. The present review focuses on the efficacy of contemporary ICD arrhythmia discrimination tools and appraises the so far reported evidence supporting the superiority of Dc-ICDs in preventing inappropriate therapies.

  18. NLOS Signal Detection Based on Single Orthogonal Dual-Polarized GNSS Antenna

    Directory of Open Access Journals (Sweden)

    Ke Zhang

    2017-01-01

    Full Text Available Nowadays users have a high demand for the accuracy of position and velocity, but errors caused by non-line-of-sight (NLOS signals cannot be removed effectively. Since the GNSS signal is right-hand circular polarized (RHCP, the axial ratio of the strong NLOS signal is larger than that of the Line-of-Sight (LOS signal. Based on the difference of the axial ratio, a method for NLOS signal detection using single orthogonal dual-polarized antenna is proposed. The antenna has two channels to receive two orthogonal linear polarized components of the incoming signals. Parallel cross-cancellation is used to remove the LOS signal while maintaining most of the NLOS signals from the receiving signals. The residual NLOS signals are then detected by conventional GNSS digital processor in real time without any prior knowledge of their characteristics. The proposed method makes use of the polarization and spatial information and can detect long delay NLOS signal by miniature and inexpensive receiver GNSS. The effectiveness of the proposed method is confirmed by simulation data.

  19. Optically controllable dual-mode switching in single-mode Fabry-Pérot laser diode subject to one side-mode feedback and external single mode injection

    Science.gov (United States)

    Wu, Jian-Wei; Won, Yong Hyub

    2017-06-01

    In this paper, broadly tunable dual-mode lasing system is presented and demonstrated based on single-mode Fabry-Pérot laser diode subject to the feedback of one side mode amplified by an erbium-doped fiber amplifier in the external feedback cavity. The spacing between two resonance modes in output lasing spectrum is broadly tuned by introducing differently amplified side mode into the single-mode laser via the external cavity consisted of amplifier, filter, and polarization controller so that two difference frequencies of 1 THz and 0.6 THz are given to display the tunable behavior of dual-mode emission in this work. Therefore, under an external injection mode into the laser condition, the power dependent injection locking and optical bistability of generated dual-mode emission are discussed in detail. At different wavelength detunings, the emitted two resonance modes including the dominant and feedback modes are switched to on- or off-state by selecting proper high-low power level of the external injection mode. As a consequence, the maximum value of achieved dual-mode on-off ratio is as high as up to 45 dB.

  20. Hydrophobic fluorescent probes introduce artifacts into single molecule tracking experiments due to non-specific binding.

    Science.gov (United States)

    Zanetti-Domingues, Laura C; Tynan, Christopher J; Rolfe, Daniel J; Clarke, David T; Martin-Fernandez, Marisa

    2013-01-01

    Single-molecule techniques are powerful tools to investigate the structure and dynamics of macromolecular complexes; however, data quality can suffer because of weak specific signal, background noise and dye bleaching and blinking. It is less well-known, but equally important, that non-specific binding of probe to substrates results in a large number of immobile fluorescent molecules, introducing significant artifacts in live cell experiments. Following from our previous work in which we investigated glass coating substrates and demonstrated that the main contribution to this non-specific probe adhesion comes from the dye, we carried out a systematic investigation of how different dye chemistries influence the behaviour of spectrally similar fluorescent probes. Single-molecule brightness, bleaching and probe mobility on the surface of live breast cancer cells cultured on a non-adhesive substrate were assessed for anti-EGFR affibody conjugates with 14 different dyes from 5 different manufacturers, belonging to 3 spectrally homogeneous bands (491 nm, 561 nm and 638 nm laser lines excitation). Our results indicate that, as well as influencing their photophysical properties, dye chemistry has a strong influence on the propensity of dye-protein conjugates to adhere non-specifically to the substrate. In particular, hydrophobicity has a strong influence on interactions with the substrate, with hydrophobic dyes showing much greater levels of binding. Crucially, high levels of non-specific substrate binding result in calculated diffusion coefficients significantly lower than the true values. We conclude that the physic-chemical properties of the dyes should be considered carefully when planning single-molecule experiments. Favourable dye characteristics such as photostability and brightness can be offset by the propensity of a conjugate for non-specific adhesion.

  1. Hydrophobic fluorescent probes introduce artifacts into single molecule tracking experiments due to non-specific binding.

    Directory of Open Access Journals (Sweden)

    Laura C Zanetti-Domingues

    Full Text Available Single-molecule techniques are powerful tools to investigate the structure and dynamics of macromolecular complexes; however, data quality can suffer because of weak specific signal, background noise and dye bleaching and blinking. It is less well-known, but equally important, that non-specific binding of probe to substrates results in a large number of immobile fluorescent molecules, introducing significant artifacts in live cell experiments. Following from our previous work in which we investigated glass coating substrates and demonstrated that the main contribution to this non-specific probe adhesion comes from the dye, we carried out a systematic investigation of how different dye chemistries influence the behaviour of spectrally similar fluorescent probes. Single-molecule brightness, bleaching and probe mobility on the surface of live breast cancer cells cultured on a non-adhesive substrate were assessed for anti-EGFR affibody conjugates with 14 different dyes from 5 different manufacturers, belonging to 3 spectrally homogeneous bands (491 nm, 561 nm and 638 nm laser lines excitation. Our results indicate that, as well as influencing their photophysical properties, dye chemistry has a strong influence on the propensity of dye-protein conjugates to adhere non-specifically to the substrate. In particular, hydrophobicity has a strong influence on interactions with the substrate, with hydrophobic dyes showing much greater levels of binding. Crucially, high levels of non-specific substrate binding result in calculated diffusion coefficients significantly lower than the true values. We conclude that the physic-chemical properties of the dyes should be considered carefully when planning single-molecule experiments. Favourable dye characteristics such as photostability and brightness can be offset by the propensity of a conjugate for non-specific adhesion.

  2. In vivo cancer targeting and fluorescence-CT dual-mode imaging with nanoprobes based on silver sulfide quantum dots and iodinated oil

    Science.gov (United States)

    Qin, Meng-Yao; Yang, Xiao-Quan; Wang, Kan; Zhang, Xiao-Shuai; Song, Ji-Tao; Yao, Ming-Hao; Yan, Dong-Mei; Liu, Bo; Zhao, Yuan-Di

    2015-11-01

    In this article, a fluorescence-CT dual-mode nanoprobe is successfully synthesized by making use of distearoylphosphatidylethanolamine-poly(ethylene glycol)-folate (DSPE-PEG2000-FA) and other amphiphilic molecules to coat silver sulfide (Ag2S) quantum dots (QDs) and iodinated oil simultaneously. In vitro experiments show that the fluorescence wavelength of the nanoprobe is 1170 nm in the near infrared-II region. Its size is 139.6 nm, it has good dispersibility, and it has low cellular toxicity at concentrations up to 25 μg mL-1 Ag. In vivo experiments revealed that the probe has a rather long circulation time (blood half-life of 5.7 hours), and the tissue histopathological tests show that it is not obviously harmful to major organs' normal function. Biochemical analysis (glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels) and blood analysis (white blood cell, red blood cell, hemoglobin and blood platelet counts) reveal that it has little influence on blood within 15 days of administration. When injected into HeLa xenograft nude mice by the tail vein, the probe elicited intensely enhanced fluorescence and X-ray computed tomography (CT) signals in the tumors after 24 hours, and the structure, size and position of tumor tissue were shown clearly. In a word, the probe has good tumor targeting capabilities, and it has significant value in fluorescence-CT dual-mode imaging in vivo.In this article, a fluorescence-CT dual-mode nanoprobe is successfully synthesized by making use of distearoylphosphatidylethanolamine-poly(ethylene glycol)-folate (DSPE-PEG2000-FA) and other amphiphilic molecules to coat silver sulfide (Ag2S) quantum dots (QDs) and iodinated oil simultaneously. In vitro experiments show that the fluorescence wavelength of the nanoprobe is 1170 nm in the near infrared-II region. Its size is 139.6 nm, it has good dispersibility, and it has low cellular toxicity at concentrations up to 25 μg mL-1 Ag. In vivo experiments revealed that the

  3. Single-bead arrays for fluorescence-based immunoassays on capillary-driven microfluidic chips

    Science.gov (United States)

    Temiz, Yuksel; Lim, Michel; Delamarche, Emmanuel

    2016-03-01

    We report a concept for the simple fabrication of easy-to-use chips for immunoassays in the context of point-of-care diagnostics. The chip concept comprises mainly three features: (1) the efficient integration of reagents using beads functionalized with receptors, (2) the generation of capillary-driven liquid flows without using external pumps, and (3) a high-sensitivity detection of analytes using fluorescence microscopy. We fabricated prototype chips using dry etching of Si wafers. 4.5-μm-diameter beads were integrated into hexagonal arrays by sedimentation and removing the excess using a stream of water. We studied the effect of different parameters and showed that array occupancies from 30% to 50% can be achieved by pipetting a 250 nL droplet of 1% bead solution and allowing the beads sediment for 3 min. Chips with integrated beads were sealed using a 50-μm-thick dry-film resist laminated at 45 °C. Liquids pipetted to loading pads were autonomously pulled by capillary pumps at a rate of 0.35 nL s-1 for about 30 min. We studied ligand-receptor interactions and binding kinetics using time-lapse fluorescence microscopy and demonstrated a 5 pM limit of detection (LOD) for an anti-biotin immunoassay. As a clinically-relevant example, we implemented an immunoassay to detect prostate specific antigen (PSA) and showed an LOD of 108 fM (i.e. 3.6 pg mL-1). While a specific implementation is provided here for the detection of PSA, we believe that combining capillary-driven microfluidics with arrays of single beads and fluorescence readout to be very flexible and sufficiently sensitive for the detection of other clinically-relevant analytes.

  4. Fast and single-step immunoassay based on fluorescence quenching within a square glass capillary immobilizing graphene oxide-antibody conjugate and fluorescently labelled antibody.

    Science.gov (United States)

    Shirai, Akihiro; Henares, Terence G; Sueyoshi, Kenji; Endo, Tatsuro; Hisamoto, Hideaki

    2016-05-23

    A single-step, easy-to-use, and fast capillary-type immunoassay device composed of a polyethylene glycol (PEG) coating containing two kinds of antibody-reagents, including an antibody-graphene oxide conjugate and fluorescently labelled antibody, was developed in this study. The working principle involved the spontaneous dissolution of the PEG coating, diffusion of reagents, and subsequent immunoreaction, triggered by the capillary action-mediated introduction of a sample solution. In a sample solution containing the target antigen, two types of antibody reagents form a sandwich-type antigen-antibody complex and fluorescence quenching takes place via fluorescence resonance energy transfer between the labelled fluorescent molecules and graphene oxide. Antigen concentration can be measured based on the decrease in fluorescence intensity. An antigen concentration-dependent response was obtained for the model target protein sample (human IgG, 0.2-10 μg mL(-1)). The present method can shorten the reaction time to within 1 min (approximately 40 s), while conventional methods using the same reagents require reaction times of approximately 20 min because of the large reaction scale. The proposed method is one of the fastest immunoassays ever reported. Finally, the present device was used to measure human IgG in diluted serum samples to demonstrate that this method can be used for fast medical diagnosis.

  5. Quantification of phosphorus in single cells using synchrotron X-ray fluorescence.

    Science.gov (United States)

    Núñez-Milland, Daliángelis R; Baines, Stephen B; Vogt, Stefan; Twining, Benjamin S

    2010-07-01

    Phosphorus is required for numerous cellular compounds and as a result can serve as a useful proxy for total cell biomass in studies of cell elemental composition. Single-cell analysis by synchrotron X-ray fluorescence (SXRF) enables quantitative and qualitative analyses of cell elemental composition with high elemental sensitivity. Element standards are required to convert measured X-ray fluorescence intensities into element concentrations, but few appropriate standards are available, particularly for the biologically important element P. Empirical P conversion factors derived from other elements contained in certified thin-film standards were used to quantify P in the model diatom Thalassiosira pseudonana, and the measured cell quotas were compared with those measured in bulk by spectrophotometry. The mean cellular P quotas quantified with SXRF for cells on Au, Ni and nylon grids using this approach were not significantly different from each other or from those measured spectrophotometrically. Inter-cell variability typical of cell populations was observed. Additionally, the grid substrates were compared for their suitability to P quantification based on the potential for spectral interferences with P. Nylon grids were found to have the lowest background concentrations and limits of detection for P, while background concentrations in Ni and Au grids were 1.8- and 6.3-fold higher. The advantages and disadvantages of each grid type for elemental analysis of individual phytoplankton cells are discussed.

  6. Single Fluorescent Molecules as Nano-Illuminators for Biological Structure and Function

    Science.gov (United States)

    Moerner, W. E.

    2011-03-01

    Since the first optical detection and spectroscopy of a single molecule in a solid (Phys. Rev. Lett. {62}, 2535 (1989)), much has been learned about the ability of single molecules to probe local nanoenvironments and individual behavior in biological and nonbiological materials in the absence of ensemble averaging that can obscure heterogeneity. Because each single fluorophore acts a light source roughly 1 nm in size, microscopic imaging of individual fluorophores leads naturally to superlocalization, or determination of the position of the molecule with precision beyond the optical diffraction limit, simply by digitization of the point-spread function from the single emitter. For example, the shape of single filaments in a living cell can be extracted simply by allowing a single molecule to move through the filament (PNAS {103}, 10929 (2006)). The addition of photoinduced control of single-molecule emission allows imaging beyond the diffraction limit (super-resolution) and a new array of acronyms (PALM, STORM, F-PALM etc.) and advances have appeared. We have used the native blinking and switching of a common yellow-emitting variant of green fluorescent protein (EYFP) reported more than a decade ago (Nature {388}, 355 (1997)) to achieve sub-40 nm super-resolution imaging of several protein structures in the bacterium Caulobacter crescentus: the quasi-helix of the actin-like protein MreB (Nat. Meth. {5}, 947 (2008)), the cellular distribution of the DNA binding protein HU (submitted), and the recently discovered division spindle composed of ParA filaments (Nat. Cell Biol. {12}, 791 (2010)). Even with these advances, better emitters would provide more photons and improved resolution, and a new photoactivatable small-molecule emitter has recently been synthesized and targeted to specific structures in living cells to provide super-resolution images (JACS {132}, 15099 (2010)). Finally, a new optical method for extracting three-dimensional position information based on

  7. Irving Langmuir Prize Talk: Single-Molecule Fluorescence Imaging: Nanoscale Emitters with Photoinduced Switching Enable Superresolution.

    Science.gov (United States)

    Moerner, W. E.

    2009-03-01

    In the two decades since the first optical detection and spectroscopy of a single molecule in a solid (Phys. Rev. Lett. 62, 2535 (1989)), much has been learned about the ability of single molecules to probe local nanoenvironments and individual behavior in biological and nonbiological materials in the absence of ensemble averaging that can obscure heterogeneity. The early years concentrated on high-resolution spectroscopy in solids, which provided observations of lifetime-limited spectra, optical saturation, spectral diffusion, optical switching, vibrational spectra, and magnetic resonance of a single molecular spin. In the mid-1990's, much of the field moved to room temperature, where a wide variety of biophysical effects were subsequently explored, but it is worth noting that several features from the low-temperature studies have analogs at high temperature. For example, in our first studies of yellow-emitting variants of green fluorescent protein (EYFP) in the water-filled pores of a gel (Nature 388, 355 (1997)), optically induced switching of the emission was observed, a room-temperature analog of the earlier low-temperature behavior. Because each single fluorophore acts a light source roughly 1 nm in size, microscopic imaging of individual fluorophores leads naturally to superlocalization, or determination of the position of the molecule with precision beyond the optical diffraction limit, simply by digitization of the point-spread function from the single emitter. Recent work has allowed measurement of the shape of single filaments in a living cell simply by allowing a single molecule to move through the filament (PNAS 103, 10929 (2006)). The additional use of photoinduced control of single-molecule emission allows imaging beyond the diffraction limit (superresolution) by several novel approaches proposed by different researchers. For example, using photoswitchable EYFP, a novel protein superstructure can now be directly imaged in a living bacterial cell at

  8. Utility of single-energy and dual-energy computed tomography in clot characterization: An in-vitro study.

    Science.gov (United States)

    Brinjikji, Waleed; Michalak, Gregory; Kadirvel, Ramanathan; Dai, Daying; Gilvarry, Michael; Duffy, Sharon; Kallmes, David F; McCollough, Cynthia; Leng, Shuai

    2017-06-01

    Background and purpose Because computed tomography (CT) is the most commonly used imaging modality for the evaluation of acute ischemic stroke patients, developing CT-based techniques for improving clot characterization could prove useful. The purpose of this in-vitro study was to determine which single-energy or dual-energy CT techniques provided optimum discrimination between red blood cell (RBC) and fibrin-rich clots. Materials and methods Seven clot types with varying fibrin and RBC densities were made (90% RBC, 99% RBC, 63% RBC, 36% RBC, 18% RBC and 0% RBC with high and low fibrin density) and their composition was verified histologically. Ten of each clot type were created and scanned with a second generation dual source scanner using three single (80 kV, 100 kV, 120 kV) and two dual-energy protocols (80/Sn 140 kV and 100/Sn 140 kV). A region of interest (ROI) was placed over each clot and mean attenuation was measured. Receiver operating characteristic curves were calculated at each energy level to determine the accuracy at differentiating RBC-rich clots from fibrin-rich clots. Results Clot attenuation increased with RBC content at all energy levels. Single-energy at 80 kV and 120 kV and dual-energy 80/Sn 140 kV protocols allowed for distinguishing between all clot types, with the exception of 36% RBC and 18% RBC. On receiver operating characteristic curve analysis, the 80/Sn 140 kV dual-energy protocol had the highest area under the curve for distinguishing between fibrin-rich and RBC-rich clots (area under the curve 0.99). Conclusions Dual-energy CT with 80/Sn 140 kV had the highest accuracy for differentiating RBC-rich and fibrin-rich in-vitro thrombi. Further studies are needed to study the utility of non-contrast dual-energy CT in thrombus characterization in acute ischemic stroke.

  9. Ultrabright fluorescent silica particles with a large number of complex spectra excited with a single wavelength for multiplex applications.

    Science.gov (United States)

    Palantavida, S; Peng, B; Sokolov, I

    2017-02-08

    We report on a novel approach to synthesize ultrabright fluorescent silica particles capable of producing a large number of complex spectra. The spectra can be excited using a single wavelength which is paramount in quantitative fluorescence imaging, flow cytometry and sensing applications. The approach employs the physical encapsulation of organic fluorescent molecules inside a nanoporous silica matrix with no dye leakage. As was recently demonstrated, such an encapsulation allowed for the encapsulation of very high concentrations of organic dyes without quenching their fluorescent efficiency. As a result, dye molecules are distanced within ∼5 nm from each other; it theoretically allows for efficient exchange of excitation energy via Förster resonance energy transfer (FRET). Here we present the first experimental demonstration of the encapsulation of fluorescent dyes in the FRET sequence. Attaining a FRET sequence of up to five different dyes is presented. The number of distinguishable spectra can be further increased by using different relative concentrations of encapsulated dyes. Combining these approaches allows for creating a large number of ultrabright fluorescent particles with substantially different fluorescence spectra. We also demonstrate the utilization of these particles for potential multiplexing applications. Though fluorescence spectra of the obtained multiplex probes are typically overlapping, they can be distinguished by using standard linear decomposition algorithms.

  10. Dual Functional Core-Shell Fluorescent Ag2S@Carbon Nanostructure for Selective Assay of E. coli O157:H7 and Bactericidal Treatment.

    Science.gov (United States)

    Wang, Ning; Wei, Xing; Zheng, An-Qi; Yang, Ting; Chen, Ming-Li; Wang, Jian-Hua

    2017-03-24

    A dual functional fluorescent core-shell Ag2S@Carbon nanostructure is prepared by a hydrothermally assisted multi-amino synthesis approach with folic acid (FA), polyethylenimine (PEI), and mannoses (Mans) as carbon and nitrogen sources (FA-PEI-Mans-Ag2S nanocomposite shortly as Ag2S@C). The nanostructure exhibits strong fluorescent emission at λex/λem = 340/450 nm with a quantum yield of 12.57 ± 0.52%. Ag2S@C is bound to E. coli O157:H7 via strong interaction with the Mans moiety in Ag2S@C with FimH proteins on the fimbriae tip in E. coli O157:H7. Fluorescence emission from Ag2S@C/E. coli conjugate is closely related to the content of E. coli O157:H7. Thus, a novel procedure for fluorescence assay of E. coli O157:H7 is developed, offering a detection limit of 330 cfu mL(-1). Meanwhile, the Ag2S@C nanostructure exhibits excellent antibacterial performance against E. coli O157:H7. A 99.9% sterilization rate can be readily achieved for E. coli O157:H7 at a concentration of 10(6)-10(7) cfu mL(-1) with 3.3 or 10 μg mL(-1) of Ag2S@C with an interaction time of 5 or 0.5 min, respectively.

  11. Dual enzyme-responsive "turn-on" fluorescence sensing systems based on in situ formation of 7-hydroxy-2-iminocoumarin scaffolds.

    Science.gov (United States)

    Debieu, Sylvain; Romieu, Anthony

    2015-11-01

    A new strategy for the simultaneous fluorogenic detection of two distinct enzyme activities namely hydrolase (amidase or esterase) and reductase is described. This innovative biosensing method is based on the powerful "covalent-assembly" principle that involves in situ synthesis of a fluorophore from a non-fluorescent caged precursor and through domino reactions triggered by the two analytes of interest. To establish this approach, penicillin G acylase (PGA) (or pig liver esterase (PLE)) and nitroreductase (NTR) were chosen as model enzymes, and original bis-O-protected 2,4-dihydroxycinnamonitrile derivatives acting as dual-reactive probes readily convertible to highly fluorescent 7-hydroxy-2-iminocoumarin scaffolds upon reacting with the two selected enzymes were synthesised. The two phenolic groups available within the core structure of these probes play a pivotal role in generating iminocoumarin scaffold through an intramolecular cyclisation reaction (hydroxyl group in C-2 position) and in enhancing its push-pull character (hydroxyl group in C-4 position). Their orthogonal and temporary protection with two different enzyme-labile masking groups is the cornerstone in the design of this novel class of fluorogenic "turn-on" probes. Their evaluation using fluorescence-based in vitro assays and HPLC-fluorescence/-MS analyses have enabled us both to demonstrate the claimed activation mechanism (in particular the specific order in which the two enzymes react with the probe) and to highlight the potential utility of these advanced chemical tools in multi-analyte sensing applications.

  12. Visualization of Fluoride Ions In Vivo Using a Gadolinium(III)-Coumarin Complex-Based Fluorescence/MRI Dual-Modal Probe.

    Science.gov (United States)

    Wang, Yue; Song, Renfeng; Feng, Huan; Guo, Ke; Meng, Qingtao; Chi, Haijun; Zhang, Run; Zhang, Zhiqiang

    2016-12-16

    A new Gadolinium(III)-coumarin complex, DO3A-Gd-CA, was designed and prepared as a dual-modal probe for simultaneous fluorescence and relaxivity responses to fluoride ions (F(-)) in aqueous media and mice. DO3A-Gd-CA was designed by using Gd(III) center as an MRI signal output unit and fluoride binding site, and the 4-(diethylamino)-coumarin-3-carboxylic acid (CA) as a fluorescence reporter. Upon the addition of fluoride ions to the solution of DO3A-Gd-CA, the liberation of the coordinated CA ligand led to a 5.7-fold fluorescence enhancement and a 75% increase in the longitudinal relaxivity (r₁). The fluorescent detection limit for fluoride ions was determined to be 8 μM based on a 3σ/slope. The desirable features of the proposed DO3A-Gd-CA, such as high sensitivity and specificity, reliability at physiological pH and low cytotoxicity enable its application in visualization of fluoride ion in mice. The successful in vivo imaging indicates that DO3A-Gd-CA could be potentially used in biomedical diagnosis fields.

  13. Synthesis and application of a N-1' fluorescent biotinyl derivative inducing the specific carboxy-terminal dual labeling of a novel RhoB-selective scFv.

    Science.gov (United States)

    Chaisemartin, L; Chinestra, P; Favre, G; Blonski, C; Faye, J C

    2009-05-20

    The fluorescent site-specific labeling of protein would provide a new, easy-to-use alternative to biochemical and immunochemical methods. We used an intein-mediated strategy for covalent labeling of the carboxy-terminal amino acid of a RhoB-selective scFv previously isolated from a phage display library (a human synthetic V(H) + V(L) scFv phage library). The scFv fused to the Mxe intein was produced in E. coli and purified and was then labeled with a newly synthesized fluorescent biotinyl cysteine derivative capable of inducing scFv-Mxe intein splicing. In this study, we investigated the splicing and labeling properties of various amino acids in the hinge domain between scFv and Mxe under thiol activation. In this dual labeling system, the fluorescein is used for antibody detection and biotin is used for purification, resulting in a high specific activity for fluorescence. We then checked that the purified biotinylated fluorescent scFv retained its selectivity for RhoB without modification of its affinity.

  14. High accuracy microwave frequency measurement based on single-drive dual-parallel Mach-Zehnder modulator

    DEFF Research Database (Denmark)

    Zhao, Ying; Pang, Xiaodan; Deng, Lei

    2011-01-01

    A novel approach for broadband microwave frequency measurement by employing a single-drive dual-parallel Mach-Zehnder modulator is proposed and experimentally demonstrated. Based on bias manipulations of the modulator, conventional frequency-to-power mapping technique is developed by performing a...... 10−3 relative error. This high accuracy frequency measurement technique is a promising candidate for high-speed electronic warfare and defense applications.......A novel approach for broadband microwave frequency measurement by employing a single-drive dual-parallel Mach-Zehnder modulator is proposed and experimentally demonstrated. Based on bias manipulations of the modulator, conventional frequency-to-power mapping technique is developed by performing...... a two-stage frequency measurement cooperating with digital signal processing. In the experiment, 10GHz measurement range is guaranteed and the average uncertainty of estimated microwave frequency is 5.4MHz, which verifies the measurement accuracy is significantly improved by achieving an unprecedented...

  15. Analysis of photon count data from single-molecule fluorescence experiments

    Energy Technology Data Exchange (ETDEWEB)

    Burzykowski, T.; Szubiakowski, J.; Ryden, T

    2003-03-15

    We consider single-molecule fluorescence experiments with data in the form of counts of photons registered over multiple time-intervals. Based on the observation schemes, linking back to works by Dehmelt [Bull. Am. Phys. Soc. 20 (1975) 60] and Cook and Kimble [Phys. Rev. Lett. 54 (1985) 1023], we propose an analytical approach to the data based on the theory of Markov-modulated Poisson processes (MMPP). In particular, we consider maximum-likelihood estimation. The method is illustrated using a real-life dataset. Additionally, the properties of the proposed method are investigated through simulations and compared to two other approaches developed by Yip et al. [J. Phys. Chem. A 102 (1998) 7564] and Molski [Chem. Phys. Lett. 324 (2000) 301].

  16. Single-larger-portion-size and dual-column nutrition labeling may help consumers make more healthful food choices.

    Science.gov (United States)

    Lando, Amy M; Lo, Serena C

    2013-02-01

    The Food and Drug Administration is considering changes to the Nutrition Facts label to help consumers make more healthful choices. To examine the effects of modifications to the Nutrition Facts label on foods that can be listed as having 1 or 2 servings per container, but are reasonably consumed at a single eating occasion. Participants were randomly assigned to study conditions that varied on label format, product, and nutrition profile. Data were collected via an online consumer panel. Adults aged 18 years and older were recruited from Synovate's online household panel. Data were collected during August 2011. A total of 32,897 invitations were sent for a final sample of 9,493 interviews. Participants were randomly assigned to one of 10 label formats classified into three groups: listing 2 servings per container with a single column, listing 2 servings per container with a dual column, and listing a single serving per container. Within these groups there were versions that enlarged the font size for "calories," removed "calories from fat," and changed the wording for serving size declaration. The single product task measured product healthfulness, the amount of calories and various nutrients per serving and per container, and label perceptions. The product comparison task measured ability to identify the healthier product and the product with fewer calories per container and per serving. Analysis of covariance models with Tukey-Kramer tests were used. Covariates included general label use, age, sex, level of education, and race/ethnicity. Single-serving and dual-column formats performed better and scored higher on most outcome measures. For products that contain 2 servings but are customarily consumed at a single eating occasion, using a single-serving or dual-column labeling approach may help consumers make healthier food choices. Published by Elsevier Inc.

  17. The impact of single and dual hydrothermal modifications on the molecular structure and physicochemical properties of normal corn starch.

    Science.gov (United States)

    Chung, Hyun-Jung; Hoover, Ratnajothi; Liu, Qiang

    2009-03-01

    Effect of single and dual hydrothermal modifications with annealing (ANN) and heat-moisture treatment (HMT) on molecular structure and physicochemical properties of corn starch was investigated. Normal corn starch was modified by ANN at 70% moisture at 50 degrees C for 24h and HMT at 30% moisture at 120 degrees C for 24h as well as by the combination of ANN and HMT. The apparent amylose content and swelling factor (SF) decreased on ANN and HMT, but amylose leaching (AML) increased. These changes were more pronounced on dual modification. The crystallinity (determined by X-ray diffraction), the gelatinization enthalpy (determined by differential scanning calorimetry) and ratio of 1047 cm(-1)/1022 cm(-1) (determined by Fourier transform infrared spectroscopy) slightly increased on ANN and decreased on HMT. The ANN and subsequent HMT (ANN-HMT) resulted in the lowest crystallinity, gelatinization enthalpy and ratio of 1047 cm(-1)/1022 cm(-1). The gelatinization temperature range decreased on ANN but increased on HMT. However, the gelatinization range of dually modified starches (ANN-HMT and HMT-ANN) was between ANN starch and HMT starch. Birefringence remained unchanged on ANN but slightly decreased on HMT as well as dual modification. Average chain length and amount of longer branch chains (DP> or =37) remained almost unchanged on ANN but decreased on HMT and dual modifications (ANN-HMT and HMT-ANN). HMT and dual modifications resulted in highly reduced pasting viscosity. ANN and HMT as well as dual modifications increased RDS content and decreased SDS and RS content.

  18. Electrocortical sources related to whole-body surface translations during a single- and dual-task paradigm

    Directory of Open Access Journals (Sweden)

    Mark D Bogost

    2016-10-01

    Full Text Available Appropriate reactive motor responses are essential in maintaining upright balance. However, little is known regarding the potential location of cortical sources that are related to the onset of a perturbation during single- and dual-task paradigms. The purpose of this study was to estimate the location of cortical sources in response to a whole-body surface translation and whether diverted attention decreases the N1 event-related potential (ERP amplitude related to a postural perturbation. This study utilized high-resolution electroencephalography in conjunction with measure projection analysis from ERPs time-locked to backwards surface translation onsets to determine which cortical sources were related to whole-body postural perturbations. Subjects (n = 15 either reacted to whole-body surface translations with (dual task or without (single task performing a visual working memory task. For the single task, four domains were identified that were mainly localized within the frontal and parietal lobes and included sources from the prefrontal, premotor, primary and supplementary motor, somatosensory, and anterior cingulate cortex. Five domains were estimated for the dual task and also included sources within the frontal and parietal lobes, but the sources also shifted to other locations that included areas within the temporal and occipital lobes. Additionally, mean absolute N1 ERP amplitudes representing the activity from similar locations in both tasks were greater for the single than dual task. The present localization results highlight the importance of frontal, parietal and anterior cingulate cortical areas in reactive postural control and suggest a re-allocation or shift of cortical sources related to reactive balance control in the presence of a secondary task. Thus, this study provides novel insight into the underlying neurophysiology and contribution of cortical sources in relation to the neural control of reactive balance.

  19. Electrocortical Sources Related to Whole-Body Surface Translations during a Single- and Dual-Task Paradigm

    Science.gov (United States)

    Bogost, Mark D.; Burgos, Pablo I.; Little, C. Elaine; Woollacott, Marjorie H.; Dalton, Brian H.

    2016-01-01

    Appropriate reactive motor responses are essential in maintaining upright balance. However, little is known regarding the potential location of cortical sources that are related to the onset of a perturbation during single- and dual-task paradigms. The purpose of this study was to estimate the location of cortical sources in response to a whole-body surface translation and whether diverted attention decreases the N1 event-related potential (ERP) amplitude related to a postural perturbation. This study utilized high-resolution electroencephalography in conjunction with measure projection analysis from ERPs time-locked to backwards surface translation onsets to determine which cortical sources were related to whole-body postural perturbations. Subjects (n = 15) either reacted to whole-body surface translations with (dual task) or without (single task) performing a visual working memory task. For the single task, four domains were identified that were mainly localized within the frontal and parietal lobes and included sources from the prefrontal, premotor, primary and supplementary motor, somatosensory and anterior cingulate cortex. Five domains were estimated for the dual task and also included sources within the frontal and parietal lobes, but the sources also shifted to other locations that included areas within the temporal and occipital lobes. Additionally, mean absolute N1 ERP amplitudes representing the activity from similar locations in both tasks were greater for the single than dual task. The present localization results highlight the importance of frontal, parietal and anterior cingulate cortical areas in reactive postural control and suggest a re-allocation or shift of cortical sources related to reactive balance control in the presence of a secondary task. Thus, this study provides novel insight into the underlying neurophysiology and contribution of cortical sources in relation to the neural control of reactive balance.

  20. Signal-to-noise analysis for detection sensitivity of small absorbing heterogeneity in turbid media with single-source and dual-interfering-source.

    Science.gov (United States)

    Chen, Y; Mu, C; Intes, X; Chance, B

    2001-08-13

    Previous studies have suggested that the phased-array detection can achieve high sensitivity in detecting and localizing inhomogeneities embedded in turbid media by illuminating with dual interfering sources. In this paper, we analyze the sensitivity of single-source and dual-interfering-source (phased array) systems with signal-to-noise ratio criteria. Analytical solutions are presented to investigate the sensitivity of detection using different degrees of absorption perturbation by varying the size and contrast of the object under similar configurations for single- and dual-source systems. The results suggest that dual-source configuration can provide higher detection sensitivity. The relation between the amplitude and phase signals for both systems is also analyzed using a vector model. The results can be helpful for optimizing the experimental design by combining the advantages of both single- and dual-source systems in object detection and localization.

  1. Combination of DNA ligase reaction and gold nanoparticle-quenched fluorescent oligonucleotides: a simple and efficient approach for fluorescent assaying of single-nucleotide polymorphisms.

    Science.gov (United States)

    Wang, Hao; Li, Jishan; Wang, Yongxiang; Jin, Jiangyu; Yang, Ronghua; Wang, Kemin; Tan, Weihong

    2010-09-15

    A new fluorescent sensing approach for detection of single-nucleotide polymorphisms (SNPs) is proposed based on the ligase reaction and gold nanoparticle (AuNPs)-quenched fluorescent oligonucleotides. The design exploits the strong fluorescence quenching of AuNPs for organic dyes and the difference in noncovalent interactions of the nanoparticles with single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), where ssDNA can be adsorbed onto the surface of AuNPs while dsDNA cannot be. In the assay, two half primer DNA probes, one being labeled with a dye and the other being phosphorylated, were first incubated with a target DNA template. In the presence of DNA ligase, the two captured ssDNAs are linked for the perfectly matched DNA target to form a stable duplex, but the duplex could not be formed by the single-base mismatched DNA template. After addition of AuNPs, the fluorescence of dye-tagged DNA probe will be efficiently quenched unless the perfectly matched DNA target is present. To demonstrate the feasibility of this design, the performance of SNP detection using two different DNA ligases, T4 DNA ligase and Escherichia coli DNA ligase, were investigated. In the case of T4 DNA ligase, the signal enhancement of the dye-tagged DNA for perfectly matched DNA target is 4.6-fold higher than that for the single-base mismatched DNA. While in the presence of E. coli DNA ligase, the value raises to be 30.2, suggesting excellent capability for SNP discrimination.

  2. Radiation Doses of Dual-Energy CT for Abdominopelvic CT: Comparison with Single-Energy CT

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Young Seo; Jeong, Woo Kyoung; Kim, Yong Soo; Heo, Jeong Nam [Dept. of Radiology, Hanyang University Guro Hospital, Hanyang University College of Medicine, Seoul (Korea, Republic of)

    2011-11-15

    To compare radiation doses of dual-energy CT (DECT) to single-energy CT (SECT) by a phantom experiment, with the application of mean tube currents for abdomino-pelvic CT. This study includes patients who were examined by contrast-enhanced CT for kidney evaluation. We divided the patients into six groups according to sex and body mass index. Each group consisted of five patients and a total of 30 patients were evaluated. We split the body parts (abdomen and pelvis), and calculated the mean tube current of each group as well as investigated the image noise. Applying the mean mAs from a CT scan, we measured the weighted CT dose index (CTDIw) of DECT and SECT. We compared the measured CTDIw to an estimated CTDI value displayed on the CT console. We also compared the radiation dose ratio of DECT to SECT (D/S ratio) for each subgroup. The radiation doses were compared by the student's t-test and analysis of variance. The difference of image noise between DECT and SECT was not statistically significant. Radiation dose of DECT was higher than SECT by about 21.6% (10.69 mGy, 8.79 mGy; p < 0.0001), and the measured CTDI of the DECT was significantly higher than the estimated CTDI by about 6% (p < 0.001). The D/S ratio was not significant between the six groups. The measured CTDIw of abdominopelvic DECT studies were significantly higher than those of SECT.

  3. Dual-isotope single-photon emission computed tomography for dopamine and serotonin transporters in normal and parkinsonian monkey brains

    Energy Technology Data Exchange (ETDEWEB)

    Li, I-H. [Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei 114, Taiwan (China); Huang, W.-S. [Department of Nuclear Medicine, Tri-Service General Hospital, Taipei, 114, Taiwan (China); Yeh, C.-B. [Department of Psychiatry, Tri-Service General Hospital, Taipei, 114, Taiwan (China); Liao, M.-H.; Chen, C.-C.; Shen, L.-H. [Division of Isotope Application, Institute of Nuclear Energy Research, Taoyaun, 325 Taiwan (China); Liu, J.-C. [Department of Biology and Anatomy, National Defense Medical Center, Taipei 114, Taiwan (China); Ma, K.-H. [Department of Biology and Anatomy, National Defense Medical Center, Taipei 114, Taiwan (China)], E-mail: kuohsing91@yahoo.com.tw

    2009-08-15

    Introduction: Parkinson's disease (PD) affects both dopaminergic and serotonergic systems. In this study, we simultaneously evaluated dopamine and serotonin transporters in primates using dual-isotope single-photon emission computed tomography (SPECT) imaging and compared the results with traditional single-isotope imaging. Methods: Four healthy and one 6-OHDA-induced PD monkeys were used for this study. SPECT was performed over 4 h after individual or simultaneous injection of [{sup 99m}Tc]TRODAT-1 (a dopamine transporter imaging agent) and [{sup 123}I]ADAM (a serotonin transporter imaging agent). Results: The results showed that the image quality and uptake ratios in different brain regions were comparable between single- and dual-isotope studies. The striatal [{sup 99m}Tc]TRODAT-1 uptake in the PD monkey was markedly lower than that in normal monkeys. The uptake of [{sup 123}I]ADAM in the midbrain of the PD monkey was comparable to that in the normal monkeys, but there were decreased uptakes in the thalamus and striatum of the PD monkey. Conclusions: Our results suggest that dual-isotope SPECT using [{sup 99m}Tc]TRODAT-1 and [{sup 123}I]ADAM can simultaneously evaluate changes in dopaminergic and serotonergic systems in a PD model.

  4. A novel multiplexed fluorescence polarisation immunoassay based on a recombinant bi-specific single-chain diabody for simultaneous detection of fluoroquinolones and sulfonamides in milk.

    Science.gov (United States)

    Chen, Min; Wen, Kai; Tao, Xiaoqi; Ding, Shuangyang; Xie, Jie; Yu, Xuezhi; Li, Jiancheng; Xia, Xi; Wang, Yang; Xie, Sanlei; Jiang, Haiyang

    2014-01-01

    Major research efforts are focusing on the development of simultaneous multiplexed immunoassays. In this study, a novel dual-binding fluorescence polarisation immunoassay (DB-FPIA) using a broad-specificity bi-specific single-chain diabody (scDb) and two fluorescent-labelled tracers (sulfamethoxypyridazine-fluorescein isothiocyanate (SMP-FITC) and sarafloxacin-Texas Red (SAR-TR)) with different excitation and emission wavelengths was developed for simultaneous and high-throughput detection of 19 fluoroquinolones (FQs) and 13 sulfonamides (SAs) at the maximum residue limits in milk samples. Recoveries for spiked milk samples were from 76.4% to 128.4%, with a relative standard deviation lower than 13.9%. The developed DB-FPIA was then applied to field samples, followed by confirmation by LC-MS/MS. All three instances in which FQs and SAs were present at concentrations near or above the assay limit of detection were identified as positive by the developed DB-FPIA, demonstrating that the method is suitable for rapid screening of FQs and SAs contamination. The novel methodology combines the advantage of the FPIA and the broad sensitivity of scDb and shows great promise for fast multi-analyte screening of low-molecular weight chemical residues in food samples.

  5. Specific solute solvent interactions and dual fluorescence of electron donor substituted bis-pyrazoquinoline in binary mixed solvents

    Science.gov (United States)

    Rotkiewicz, Krystyna; Rettig, Wolfgang; Köhler, Gottfried; Rechthaler, Karl; Danel, Andrzej; Grabka, Danuta

    2004-12-01

    Some extended electron donor and acceptor substituted π-electron systems, composed of a dimethyl- or diethylanilino (DMA or DEA, respectively) group as electron donor and bis-pyrazoloquinoline (DPPQ) or bis-pyrazolopyridine (DMPP) derivatives as acceptors, were studied with the aim to elucidate the contributions of various solute interactions with solvent molecules upon intramolecular charge separation. Steady state and time resolved fluorescence studies were performed for DMA-DPPQ, DMA-DMPP and DEA-DMPP in alcohols and their binary mixtures with heptane. The decay kinetics of the fluorescence of DMA-DPPQ was also studied in neat 1-propanol in dependence of the temperature between 170 and 293 K. Several different fluorescent species, which were ascribed to different complexes with alcohol molecules, contribute to the fluorescence of the various compounds in these binary mixtures and account for the complex fluorescence decay. A simplified consecutive model is proposed to explain the decay behaviour of the excited species.

  6. Highly efficient dual-wavelength mid-infrared CW Laser in diode end-pumped Er:SrF2 single crystals

    Science.gov (United States)

    Ma, Weiwei; Qian, Xiaobo; Wang, Jingya; Liu, Jingjing; Fan, Xiuwei; Liu, Jie; Su, Liangbi; Xu, Jun

    2016-11-01

    The spectral properties and laser performance of Er:SrF2 single crystals were investigated and compared with Er:CaF2. Er:SrF2 crystals have larger absorption cross-sections at the pumping wavelength, larger mid-infrared stimulated emission cross-sections and much longer fluorescence lifetimes of the upper laser level (Er3+:4I11/2 level) than those of Er:CaF2 crystals. Dual-wavelength continuous-wave (CW) lasers around 2.8 μm were demonstrated in both 4at.% and 10at.% Er:SrF2 single crystals under 972 nm laser diode (LD) end pumping. The laser wavelengths are 2789.3 nm and 2791.8 nm in the former, and 2786.4 nm and 2790.7 nm in the latter, respectively. The best laser performance has been demonstrated in lightly doped 4at.% Er:SrF2 with a low threshold of 0.100 W, a high slope efficiency of 22.0%, an maximum output power of 0.483 W.

  7. Single-tube fluorescent product-enhanced reverse transcriptase assay with Ampliwax (STF-PERT) for retrovirus quantitation.

    Science.gov (United States)

    Sears, Johnna F; Khan, Arifa S

    2003-03-01

    A TaqMan fluorescent probe-based product enhanced reverse transcriptase (RT) assay is described in which the RT and polymerase chain reaction (PCR) steps are set-up in a single tube, in two compartments separated by Ampliwax (designated as single-tube fluorescent product-enhanced reverse transcriptase assay (STF-PERT)). This simplification of the two-step method resulted in increased assay reproducibility and handling efficiency while maintaining the sensitivity of the PERT assay (PERT assay can be used to quantitate low amounts of retrovirus in clinical and research materials and to evaluate retrovirus contamination in cell substrates and biological products in human use.

  8. Near-infrared dual-emission quantum dots-gold nanoclusters nanohybrid via co-template synthesis for ratiometric fluorescent detection and bioimaging of ascorbic acid in vitro and in vivo.

    Science.gov (United States)

    Zhao, Peng; He, Kaiyu; Han, Yitao; Zhang, Zhen; Yu, Mengze; Wang, Honghui; Huang, Yan; Nie, Zhou; Yao, Shouzhuo

    2015-10-06

    Near-infrared (NIR) quantum dots (QDs) have emerged as an attractive bioimaging toolkit for exploring biological events because they can provide deep imaging penetration and low fluorescence background. However, the quantitation process of such NIR QDs generally relies on single-emission intensity change, which is susceptible to a variety of environmental factors. Herein, for the first time, we proposed a protein-directed co-template strategy to synthesize a NIR-based, dual-emission fluorescent nanohybrid (DEFN) constructed from far-red gold nanoclusters and NIR PbS QDs (AuNCs-PbS-QDs). The convenient protein-directed co-template synthesis avoids the tedious chemical coupling and modification required in conventional preparation approaches of DEFNs. Additionally, the dual-emission signals of AuNCs-PbS-QDs exhibit two well-resolved emission peaks (640 and 813 nm) separated by 173 nm, which can eliminate environmental interferences by the built-in correction of ratiometric signal, resulting in a more favorable system for bioimaging and biosensing. Next, the target-responsive capability of this NIR-based DEFN to ascorbic acid (AA) was discovered, enabling the proposed DEFN to ratiometrically detect AA with a linear range of 3-40 μM and a detection limit of 1.5 μM. This DEFN sensor possesses high selectivity, rapid response, and excellent photostability. Moreover, the feasibility of this NIR nanosensor has been fully proved by the ratiometric detection of AA for fruit internal quality assessment, in vitro cellular imaging, and in vivo imaging in nude mice.

  9. Attenuation-based characterization of coronary atherosclerotic plaque: Comparison of dual source and dual energy CT with single-source CT and histopathology

    Energy Technology Data Exchange (ETDEWEB)

    Henzler, Thomas, E-mail: Thomas.Henzler@umm.de [Department of Clinical Radiology and Nuclear Medicine, University Medical Center Mannheim, Medical Faculty Mannheim - Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim (Germany); Porubsky, Stefan [Department of Pathology, University Medical Center Mannheim, Medical Faculty Mannheim - Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim (Germany); Kayed, Hany [Department of Clinical Radiology and Nuclear Medicine, University Medical Center Mannheim, Medical Faculty Mannheim - Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim (Germany); Harder, Nils [Department of Clinical Radiology and Nuclear Medicine, University Medical Center Mannheim, Medical Faculty Mannheim - Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim (Germany); Department of Pathology, University Medical Center Mannheim, Medical Faculty Mannheim - Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim (Germany); Krissak, U. Radko; Meyer, Mathias [Department of Clinical Radiology and Nuclear Medicine, University Medical Center Mannheim, Medical Faculty Mannheim - Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim (Germany); Sueselbeck, Tim [1st Department of Medicine University Medical Center Mannheim, Medical Faculty Mannheim - Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim (Germany); Marx, Alexander [Department of Pathology, University Medical Center Mannheim, Medical Faculty Mannheim - Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim (Germany); Michaely, Henrik [Department of Clinical Radiology and Nuclear Medicine, University Medical Center Mannheim, Medical Faculty Mannheim - Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim (Germany); Schoepf, U. Joseph [Department of Radiology and Radiological Science, Medical University of South Carolina, 169 Ashley Avenue, Charleston (United States)

    2011-10-15

    Objective: To compare different CT acquisition techniques regarding for attenuation-based characterization of coronary atherosclerotic plaques using histopathology as the standard of reference. Materials and methods: In a post mortem study 17 human hearts were studied with dual-source CT (DSCT) and dual energy CT (DECT) mode on a DSCT as well as with 16-slice single-source CT (SSCT). At autopsy, atherosclerotic lesions were cut at 5 {mu}m sections. Histopathologic classification of the plaques according to the American Heart Association (AHA) criteria was performed by two pathologists. Attenuation values of all plaques were measured in DSCT, DECT and SSCT studies, respectively and classified based on attenuation according to modified AHA criteria. Results: 58 coronary plaques were identified at autopsy. Regardless of the CT technique only 52/58 plaques were found at CT (sensitivity = 89.6%). There was no significant difference between the mean attenuation values of different plaque types between DSCT, DECT, and SSCT: type IV: 11 HU/8 HU/19 HU; type Va: 44 HU/45 HU/52 HU; type Vb: 1088 HU/966 HU/1079 HU). The sensitivity for correct classification varied depending on the plaque type (type II = 0%, type III = 0%, type IV = 43%, type Va = 58%, Vb = 97%). Conclusion: Independent of the used acquisition technique, SSCT, DSCT and DECT show similar results for attenuation-based characterization of atherosclerotic coronary plaques.

  10. Dual-wavelength imaging of tumor progression by activatable and targeting near-infrared fluorescent probes in a bioluminescent breast cancer model.

    Directory of Open Access Journals (Sweden)

    Bang-Wen Xie

    Full Text Available Bioluminescence imaging (BLI has shown its appeal as a sensitive technique for in vivo whole body optical imaging. However, the development of injectable tumor-specific near-infrared fluorescent (NIRF probes makes fluorescence imaging (FLI a promising alternative to BLI in situations where BLI cannot be used or is unwanted (e.g., spontaneous transgenic tumor models, or syngeneic mice to study immune effects.In this study, we addressed the questions whether it is possible to detect tumor progression using FLI with appropriate sensitivity and how FLI correlates with BLI measurements. In addition, we explored the possibility to simultaneously detect multiple tumor characteristics by dual-wavelength FLI (~700 and ~800 nm in combination with spectral unmixing. Using a luciferase-expressing 4T1-luc2 mouse breast cancer model and combinations of activatable and targeting NIRF probes, we showed that the activatable NIRF probes (ProSense680 and MMPSense680 and the targeting NIRF probes (IRDye 800CW 2-DG and IRDye 800CW EGF were either activated by or bound to 4T1-luc2 cells. In vivo, we implanted 4T1-luc2 cells orthotopically in nude mice and were able to follow tumor progression longitudinally both by BLI and dual-wavelength FLI. We were able to reveal different probe signals within the tumor, which co-localized with immuno-staining. Moreover, we observed a linear correlation between the internal BLI signals and the FLI signals obtained from the NIRF probes. Finally, we could detect pulmonary metastases both by BLI and FLI and confirmed their presence histologically.Taken together, these data suggest that dual-wavelength FLI is a feasible approach to simultaneously detect different features of one tumor and to follow tumor progression with appropriate specificity and sensitivity. This study may open up new perspectives for the detection of tumors and metastases in various experimental models and could also have clinical applications, such as image

  11. Fluorescent metal nanoshell probe to detect single miRNA in lung cancer cell.

    Science.gov (United States)

    Zhang, Jian; Fu, Yi; Mei, Yuping; Jiang, Feng; Lakowicz, Joseph R

    2010-06-01

    In this study, fluorescent metal nanoshells were synthesized as a molecular imaging agent to detect single microRNA (miRNA) molecules in the cells positive to lung cancer. These metal nanoshells were composed of silica spheres with encapsulated Ru(bpy)(3)(2+) complexes as cores and thin silver layers as shells. Compared with the silica spheres in the absence of metal, the metal nanoshells displayed an enhanced emission intensity, shortened lifetime, and extended photostability. The single-stranded probe oligonucleotides were covalently bound on the metal nanoshells to hybridize with the target miRNA-486 molecules in the cells. It was shown that with stronger emission intensity and longer lifetime, the conjugated metal nanoshells were isolated distinctly from the cellular autofluorescence on the cell images. These emission spots on the cell images were counted accurately and analyzed with a pool of cells representing the miRNA-486 expression levels in the cells. The results may reflect a genomic signal change and provide a reference to lung cancer early diagnosis as well as other diseases.

  12. Single Molecule Fluorescence Imaging as a Technique for Barium Tagging in Neutrinoless Double Beta Decay

    CERN Document Server

    Jones, B J P; Nygren, D R

    2016-01-01

    Background rejection is key to success for future neutrinoless double beta decay experiments. To achieve sensitivity to effective Majorana lifetimes of $\\sim10^{28}$ years, backgrounds must be controlled to better than 0.1 count per ton per year, beyond the reach of any present technology. In this paper we propose a new method to identify the birth of the barium daughter ion in the neutrinoless double beta decay of $^{136}$Xe. The method adapts Single Molecule Fluorescent Imaging, a technique from biochemistry research with demonstrated single ion sensitivity. We explore possible SMFI dyes suitable for the problem of barium ion detection in high pressure xenon gas, and develop a fiber-coupled sensing system with which we can detect the presence of bulk Ba$^{++}$ ions remotely. We show that our sensor produces signal-to-background ratios as high as 85 in response to Ba$^{++}$ ions when operated in aqueous solution. We then describe the next stage of this R\\&D program, which will be to demonstrate chelation...

  13. Integration and oligomerization of Bax protein in lipid bilayers characterized by single molecule fluorescence study.

    Science.gov (United States)

    Luo, Lu; Yang, Jun; Liu, Dongxiang

    2014-11-14

    Bax is a pro-apoptotic Bcl-2 family protein. The activated Bax translocates to mitochondria, where it forms pore and permeabilizes the mitochondrial outer membrane. This process requires the BH3-only activator protein (i.e. tBid) and can be inhibited by anti-apoptotic Bcl-2 family proteins such as Bcl-xL. Here by using single molecule fluorescence techniques, we studied the integration and oligomerization of Bax in lipid bilayers. Our study revealed that Bax can bind to lipid membrane spontaneously in the absence of tBid. The Bax pore formation undergoes at least two steps: pre-pore formation and membrane insertion. The activated Bax triggered by tBid or BH3 domain peptide integrates on bilayers and tends to form tetramers, which are termed as pre-pore. Subsequent insertion of the pre-pore into membrane is highly dependent on the composition of cardiolipin in lipid bilayers. Bcl-xL can translocate Bax from membrane to solution and inhibit the pore formation. The study of Bax integration and oligomerization at the single molecule level provides new evidences that may help elucidate the pore formation of Bax and its regulatory mechanism in apoptosis. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Color optimization of single emissive white OLEDs via energy transfer between RGB fluorescent dopants

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Nam Ho; Kim, You-Hyun; Yoon, Ju-An; Lee, Sang Youn [Department of Green Energy and Semiconductor Engineering, Hoseo University, Asan (Korea, Republic of); Ryu, Dae Hyun [Department of Information Technology, Hansei University, Gunpo (Korea, Republic of); Wood, Richard [Department of Engineering Physics, McMaster University, Hamilton, Ontario, Canada L8S 4L7 (Canada); Moon, C.-B. [Department of Green Energy and Semiconductor Engineering, Hoseo University, Asan (Korea, Republic of); Kim, Woo Young, E-mail: wykim@hoseo.edu [Department of Green Energy and Semiconductor Engineering, Hoseo University, Asan (Korea, Republic of); Department of Engineering Physics, McMaster University, Hamilton, Ontario, Canada L8S 4L7 (Canada)

    2013-11-15

    The electroluminescent characteristics of white organic light-emitting diodes (WOLEDs) were investigated including single emitting layer (SEL) with an ADN host and dopants; BCzVBi, C545T, and DCJTB for blue, green and red emission, respectively. The structure of the high efficiency WOLED device was; ITO/NPB(700 Å)/ADN: BCzVBi-7%:C545T-0.05%:DCJTB-0.1%(300 Å)/Bphen(300 Å)/Liq(20 Å)/Al(1200 Å) for mixing three primary colors. Luminous efficiency was 9.08 cd/A at 3.5 V and Commission Intenationale de L’eclairage (CIE{sub x,y}) coordinates of white emission was measured as (0.320, 0.338) at 8 V while simulated CIE{sub x,y} coordinates were (0.336, 0.324) via estimation from each dopant's PL spectrum. -- Highlights: • This paper observes single-emissive-layered white OLED using fluorescent dopants. • Electrical and optical properties are analyzed. • Color stability of white OLED is confirmed for new planar light source.

  15. Quantification of harman alkaloids in sour passion fruit pulp and seeds by a novel dual SBSE-LC/Flu (stir bar sorptive extraction-liquid chromatography with fluorescence detector) method

    OpenAIRE

    PEREIRA, Cíntia A. M.; Rodrigues,Thyago R.; YARIWAKE, Janete H.

    2014-01-01

    A method for the quantification of the alkaloids harmane and harmine in sour passion fruit (Passiflora edulis f. flavicarpa O. Degener, Passifloraceae) pulp and seeds by stir-bar sorptive extraction and high performance liquid chromatography with fluorescence detection (dual SBSE-LC/Flu) is described. The SBSE parameters were optimized using a fractional factorial design, and the dual SBSE-LC/Flu method was validated following the International Conference on Harmonisation of Technical Require...

  16. Dynamic fluorescence spectroscopy on single tryptophan mutants of EII(mtl) in detergent micelles. Effects of substrate binding and phosphorylation on the fluorescence and anisotropy decay.

    Science.gov (United States)

    Dijkstra, D S; Broos, J; Visser, A J; van Hoek, A; Robillard, G T

    1997-04-22

    The effects of substrate and substrate analogue binding and phosphorylation on the conformational dynamics of the mannitol permease of Escherichia coli were investigated, using time-resolved fluorescence spectroscopy on mutants containing five single tryptophans situated in the membrane-embedded C domain of the enzyme [Swaving Dijkstra et al. (1996) Biochemistry 35, 6628-6634]. Since no fluorescent impurities are present in these mutants, the changes in fluorescence and anisotropy could be related with changes in the tryptophan microenvironment. Tryptophans at positions 30 and 42 showed changes in fluorescence intensity decay upon binding mannitol, which were reflected in the changes in lifetime distribution patterns. The disappearance of the shortest-lived decay component in these mutants, as well as in the mutant with a single tryptophan at position 109, indicates a change in the local environment such that quenching via neighboring side chains or solvent is reduced. Phosphorylation at histidine 554 and cysteine 384, located in the cytoplasmatic A and B domains of EII(mtl), respectively, induced an increase in the average fluorescence lifetimes of all of the tryptophans. The effect was most pronounced for tryptophans 30 and 109 which show large increases in the average fluorescence lifetime mainly due to loss of short-lived decay components. A correlation time distribution of the individual tryptophans deduced from an analysis of the anisotropy decay showed that they differed in their rotational mobility with tryptophan 30 showing the least local flexibility. Phosphorylation resulted in immobilization of W109 which, together with changes in the average fluorescence lifetime, is evidence for a conformational coupling between the phosphorylated B domain and the C domain. The influence of mannitol binding on the rotational behavior of the tryptophans is limited; it induces more internal flexibility at all tryptophan positions. A rotational correlation time of 30 ns

  17. Conformational changes of the H+-ATPase from Escherichia coli upon nucleotide binding detected by single molecule fluorescence.

    Science.gov (United States)

    Börsch, M; Turina, P; Eggeling, C; Fries, J R; Seidel, C A; Labahn, A; Gräber, P

    1998-10-23

    Using a confocal fluorescence microscope with an avalanche photodiode as detector, we studied the fluorescence of the tetramethylrhodamine labeled F1 part of the H+-ATPase from Escherichia coli, EF1, carrying the gammaT106-C mutation [Aggeler, J.A. and Capaldi, R.A. (1992) J. Biol. Chem. 267, 21355-21359] in aqueous solution upon excitation with a mode-locked argon ion laser at 528 nm. The diffusion of the labeled EF1 through the confocal volume gives rise to photon bursts, which were analyzed with fluorescence correlation spectroscopy, resulting in a diffusion coefficient of 3.3 x 10(-7) cm2 s(-1). In the presence of nucleotides the diffusion coefficient increases by about 15%. This effect indicates a change of the shape and/or the volume of the enzyme upon binding of nucleotides, i.e. fluorescence correlation spectroscopy with single EF1 molecules allows the detection of conformational changes.

  18. Picosecond wide-field time-correlated single photon counting fluorescence microscopy with a delay line anode detector

    Science.gov (United States)

    Hirvonen, Liisa M.; Becker, Wolfgang; Milnes, James; Conneely, Thomas; Smietana, Stefan; Le Marois, Alix; Jagutzki, Ottmar; Suhling, Klaus

    2016-08-01

    We perform wide-field time-correlated single photon counting-based fluorescence lifetime imaging (FLIM) with a crossed delay line anode image intensifier, where the pulse propagation time yields the photon position. This microchannel plate-based detector was read out with conventional fast timing electronics and mounted on a fluorescence microscope with total internal reflection (TIR) illumination. The picosecond time resolution of this detection system combines low illumination intensity of microwatts with wide-field data collection. This is ideal for fluorescence lifetime imaging of cell membranes using TIR. We show that fluorescence lifetime images of living HeLa cells stained with membrane dye di-4-ANEPPDHQ exhibit a reduced lifetime near the coverslip in TIR compared to epifluorescence FLIM.

  19. CMOS single-photon avalanche diodes and micromachined optical filters for integrated fluorescence sensing

    Science.gov (United States)

    Dandin, Marc Peralte

    This dissertation presents a body of work that addresses the two most pressing challenges in the field of integrated fluorescence sensing, namely, the design of integrated optical sensors and the fabrication of high-rejection micro-scale optical filters. Two novel enabling technologies were introduced. They are: the perimeter-gated single-photon avalanche diode (PGSPAD), for on-chip photon counting, and the benzotriazole (BTA)-doped thin-film polymer filter, for on-chip ultraviolet light rejection. Experimental results revealed that the PGSPAD front-end, fabricated in a 0.5 μm standard mixed-signal CMOS process, had the capability of counting photons in the MHz regime. In addition, it was found that a perimeter gate, a structural feature used to suppress edge breakdown in the diode, also maximized the signal-to-noise-ratio in the high-count rate regime whereas it maximized sensitivity at low count rates. On the other hand, BTA-doped filters were demonstrated utilizing three commonly used polymers as hosts. The filters were patternable, utilizing the same procedures traditionally used to pattern the undoped polymer hosts, a key advantage for integration into microsystems. Filter performance was analyzed using a set of metrics developed for optoelectronic characterization of integrated fluorescence sensors; high rejection levels (nearing -40 dB) of UV light were observed in films of only 5 μm in thickness. Ultimately, BTA-doped filters were integrated into a portable sensor, and their use was demonstrated in two types of bioassays.

  20. Alcohol consumption patterns among Mexican American mothers and among children from single- and dual-headed households: findings from HHANES 1982-84.

    Science.gov (United States)

    Stroup-Benham, C A; Treviño, F M; Treviño, D B

    1990-01-01

    Data from the southwestern United States sample of the Hispanic Health and Nutrition Examination Survey were employed to compare the patterns of alcohol use among Mexican American mothers and children in female-headed households with use patterns among mothers and children in couple-headed households. Single female heads of household drank more alcoholic beverages on more days than females from dual-headed households. As a whole, the children of single heads of household still living at home did not demonstrate significantly different drinking patterns from their dual-headed household counterparts. While male children of single-headed households drank more days and total drinks than their dual-headed household counterparts, female children of dual-headed households drank more days and total drinks than female children from single-headed households. PMID:9187580

  1. Stable dual-wavelength single-longitudinal-mode ring erbium-doped fiber laser for optical generation of microwave frequency

    Science.gov (United States)

    Wang, T.; Liang, G.; Miao, X.; Zhou, X.; Li, Q.

    2012-05-01

    We demonstrate a simple dual-wavelength ring erbium-doped fiber laser operating in single-longitudinal-mode (SLM) at room temperature. A pair of reflection type short-period fiber Bragg gratings (FBGs), which have two different center wavelengths of 1545.072 and 1545.284 nm, are used as the wavelength-selective component of the laser. A segment of unpumped polarization maintaining erbium-doped fiber (PM-EDF) is acted as a narrow multiband filter. By turning the polarization controller (PC) to enhance the polarization hole burning (PHB), the single-wavelength and dual-wavelength laser oscillations are observed at 1545.072 and 1545.284 nm. The output power variation is less than 0.6 dB for both wavelengths over a five-minute period and the optical signal to noise ratio (OSNR) is greater than 50 dB. By beating the dual-wavelengths at a photodetector (PD), a microwave signal at 26.44 GHz is demonstrated.

  2. Characterising coarse PBA dynamics in real-time above and below a tropical rainforest canopy using a dual channel UV fluorescence aerosol spectrometer.

    Science.gov (United States)

    Gabey, A.; Gallagher, M. W.; Burgess, R.; Coe, H.; McFiggans, G.,; Kaye, P. H.; Stanley, W. R.; Davies, F.; Foot, V. E.

    2009-04-01

    single-particle dual channel UV fluorescence spectrometer (Kaye et al., 2008) capable of detecting PBA by inducing fluorescence in two so-called biofluorophores - one present during metabolism and the other an amino acid - in the particle size range 1 m 2 m. In qualitative agreement with measurements of culturable airborne material in a tropical forest's understory (Gilbert, 2005) a diurnal cycle of PBA number concentration is present, reaching a maximum of ~4000 l-1 at local midnight and falling to ~100 l-1 around midday. The role of the planetary boundary layer's collapse and re-establishment in dictating this variation in is also investigated using LIDAR data. Transient PBA concentration spikes lasting several minutes are superposed on the smooth underlying diurnal variation and occur at similar times each day. Nucleopore filter samples were also taken in-situ and analysed under an Environmental scanning electron microscope (ESEM) in Manchester. The images obtained showed the PBA fraction to be dominated by fungal spores of diameter 2-5 m, from various species including ABM. Since such species tend to release spores in bursts at regular times this appears to account for the PBA concentration spikes.

  3. Correlative atomic force and confocal fluorescence microscopy: single molecule imaging and force induced spectral shifts (Conference Presentation)

    Science.gov (United States)

    Basché, Thomas; Hinze, Gerald; Stöttinger, Sven

    2016-09-01

    A grand challenge in nanoscience is to correlate structure or morphology of individual nano-sized objects with their photo-physical properties. An early example have been measurements of the emission spectra and polarization of single semiconductor quantum dots as well as their crystallographic structure by a combination of confocal fluorescence microscopy and transmission electron microscopy.[1] Recently, the simultaneous use of confocal fluorescence and atomic force microscopy (AFM) has allowed for correlating the morphology/conformation of individual nanoparticle oligomers or molecules with their photo-physics.[2, 3] In particular, we have employed the tip of an AFM cantilever to apply compressive stress to single molecules adsorbed on a surface and follow the effect of the impact on the electronic states of the molecule by fluorescence spectroscopy.[3] Quantum mechanical calculations corroborate that the spectral changes induced by the localized force can be associated to transitions among the different possible conformers of the adsorbed molecule.

  4. Analytical 1-D dual-porosity equivalent solutions to 3-D discrete single-continuum models. Application to karstic spring hydrograph modelling

    CERN Document Server

    Cornaton, F

    2011-01-01

    One dimensional analytical porosity-weighted solutions of the dual-porosity model are derived, providing insights on how to relate exchange and storage coefficients to the volumetric density of the high-permeability medium. It is shown that porosity-weighted storage and exchange coefficients are needed when handling highly heterogeneous systems - such as karstic aquifers - using equivalent dual-porosity models. The sensitivity of these coefficients is illustrated by means of numerical experiments with theoretical karst systems. The presented 1-D dual-porosity analytical model is used to reproduce the hydraulic responses of reference 3-D karst aquifers, modelled by a discrete single-continuum approach. Under various stress conditions, simulation results show the relations between the dual-porosity model coefficients and the structural features of the discrete single-continuum model. The calibration of the equivalent 1-D analytical dual-porosity model on reference hydraulic responses confirms the dependence of ...

  5. Solvent tuned single molecule dual emission in protic solvents: effect of polarity and H-bonding.

    Science.gov (United States)

    Chevreux, S; Allain, C; Wilbraham, L; Nakatani, K; Jacques, P; Ciofini, I; Lemercier, G

    2015-01-01

    Phen-PENMe2 has recently been proposed as a promising new molecule displaying solvent-tuned dual emission, highlighting an original and newly-described charge transfer model. The study of the photophysical behaviour of this molecule was extended to include protic solvents. The effects of polarity and hydrogen bonding lead to an even more evident dual emission associated with a large multi-emission band in some solvents like methanol, highlighting Phen-PENMe2 as a promising candidate for white light emission.

  6. Diagnostic accuracy: theoretical models for preimplantation genetic testing of a single nucleus using the fluorescence in situ hybridization technique

    NARCIS (Netherlands)

    P.N. Scriven; P.M.M. Bossuyt

    2010-01-01

    The aim of this study was to develop and use theoretical models to investigate the accuracy of the fluorescence in situ hybridization (FISH) technique in testing a single nucleus from a preimplantation embryo without the complicating effect of mosaicism. Mathematical models were constructed for thre

  7. Ti:sapphire rib waveguides as single-transverse-mode broadband fluorescence sources for optical coherence tomography applications

    NARCIS (Netherlands)

    Grivas, C.; Shepherd, D.P.; May-Smith, T.C.; Eason, R.W.; Pollnau, M.; Crunteanu, A.; Jelinek, M.

    2003-01-01

    Ar+-beam-milled rib waveguides in pulsed-laser-deposited Ti:sapphire layers show broadband single transverse mode fluorescence emission at output powers up to 300 μW and propagation losses comparable to those in unstructured planar waveguide counterparts.

  8. Comparison of single-beam and dual-beam laser welding of Ti-22Al-25Nb/TA15 dissimilar titanium alloys

    Science.gov (United States)

    Shen, Junqi; Li, Bo; Hu, Shengsun; Zhang, Hao; Bu, Xianzheng

    2017-08-01

    Laser beam welding (LBW) was used to join Ti-22Al-25Nb/TA15 dissimilar titanium alloys. The microstructure and mechanical properties of the welded joints under single and dual beam welding were analyzed and compared. In the mode of single laser beam, the fusion zone only consisted of B2 phase because of existence of β-phase stabilizer and rapid cooling rate of LBW. However, O phase was formed in the fusion zone while applying dual-beam laser welding due to decrease of the cooling rate. The microhardness distribution of the welded joint in dual-beam welding mode was consistent with that in single mode, but the hardness of the weld under dual laser beam was higher than that of single laser beam. In room-temperature tensile tests, the fractures all occurred in the weld, but the morphology exhibited a quasi-cleavage feature in single mode while the morphology was dimple fracture in the mode of dual laser beam. The tensile strength and elongation were both increased under dual-beam laser welding compared with those under single-beam laser welding.

  9. Comparative effectiveness of single and dual rapid diagnostic tests for syphilis and HIV in antenatal care services in Colombia.

    Science.gov (United States)

    Gaitán-Duarte, Hernando Guillermo; Newman, Lori; Laverty, Maura; Habib, Ndema Abu; González-Gordon, Lina María; Ángel-Müller, Edith; Abella, Catleya; Barros, Esther Cristina; Rincón, Carlos; Caicedo, Sidia; Gómez, Bertha; Pérez, Freddy

    2016-12-01

    To assess the effectiveness of a dual rapid test compared to a single rapid test for syphilis and HIV screening. A cluster-randomized open-label clinical trial was performed in 12 public antenatal care (ANC) centers in the cities of Bogotá and Cali, Colombia. Pregnant women who were over 14 years of age at their first antenatal visit and who had not been previously tested for HIV and syphilis during the current pregnancy were included. Pregnant women were randomized to single HIV and single syphilis rapid diagnostic tests (Arm A) or to dual HIV and syphilis rapid diagnostic tests (Arm B). The four main outcomes measured were: (1) acceptability of the test, (2) uptake in testing, (3) treatment on the same day (that is, timely treatment), and (4) treatment at any time for positive rapid test cases. Bivariate and multivariate analyses were calculated to adjust for the clustering effect and the period. A total of 1 048 patients were analyzed in Arm A, and 1 166 in Arm B. Acceptability of the rapid tests was 99.8% in Arm A and 99.6% in Arm B. The prevalence of positive rapid tests was 2.21% for syphilis and 0.36% for HIV. Timely treatment was provided to 20 of 29 patients (69%) in Arm A and 16 of 20 patients (80%) in Arm B (relative risk (RR), 1.10; 95% confidence interval (CI): (1.00 -1.20). Treatment at any time was given to 24 of 29 patients (83%) in Arm A and to 20 of 20 (100%) in Arm B (RR, 1.11; 95% CI: 1.01-1.22). There were no differences in patient acceptability, testing and timely treatment between dual rapid tests and single rapid tests for HIV and syphilis screening in the ANC centers. Same-day treatment depends also on the interpretation of and confidence in the results by the health providers.

  10. Subunits of highly Fluorescent Protein R-Phycoerythrin as Probes for Cell Imaging and Single-Molecule Detection

    Energy Technology Data Exchange (ETDEWEB)

    Isailovic, Dragan [Iowa State Univ., Ames, IA (United States)

    2005-01-01

    The purposes of our research were: (1) To characterize subunits of highly fluorescent protein R-Phycoerythrin (R-PE) and check their suitability for single-molecule detection (SMD) and cell imaging, (2) To extend the use of R-PE subunits through design of similar proteins that will be used as probes for microscopy and spectral imaging in a single cell, and (3) To demonstrate a high-throughput spectral imaging method that will rival spectral flow cytometry in the analysis of individual cells. We first demonstrated that R-PE subunits have spectroscopic and structural characteristics that make them suitable for SMD. Subunits were isolated from R-PE by high-performance liquid chromatography (HPLC) and detected as single molecules by total internal reflection fluorescence microscopy (TIRFM). In addition, R-PE subunits and their enzymatic digests were characterized by several separation and detection methods including HPLC, capillary electrophoresis, sodium dodecyl sulfate-polyacrilamide gel electrophoresis (SDS-PAGE) and HPLC-electrospray ionization mass spectrometry (ESI-MS). Favorable absorption and fluorescence of the R-PE subunits and digest peptides originate from phycoerythrobilin (PEB) and phycourobilin (PUB) chromophores that are covalently attached to cysteine residues. High absorption coefficients and strong fluorescence (even under denaturing conditions), broad excitation and emission fluorescence spectra in the visible region of electromagnetic spectrum, and relatively low molecular weights make these molecules suitable for use as fluorescence labels of biomolecules and cells. We further designed fluorescent proteins both in vitro and in vivo (in Escherichia coli) based on the highly specific attachment of PEB chromophore to genetically expressed apo-subunits of R-PE. In one example, apo-alpha and apo-beta R-PE subunits were cloned from red algae Polisiphonia boldii (P. boldii), and expressed in E. coli. Although expressed apo-subunits formed inclusion

  11. Single laser pulse generates dual photoacoustic signals for differential contrast photoacoustic imaging.

    Science.gov (United States)

    Gao, Fei; Feng, Xiaohua; Zhang, Ruochong; Liu, Siyu; Ding, Ran; Kishor, Rahul; Zheng, Yuanjin

    2017-04-04

    Photoacoustic sensing and imaging techniques have been studied widely to explore optical absorption contrast based on nanosecond laser illumination. In this paper, we report a long laser pulse induced dual photoacoustic (LDPA) nonlinear effect, which originates from unsatisfied stress and thermal confinements. Being different from conventional short laser pulse illumination, the proposed method utilizes a long square-profile laser pulse to induce dual photoacoustic signals. Without satisfying the stress confinement, the dual photoacoustic signals are generated following the positive and negative edges of the long laser pulse. More interestingly, the first expansion-induced photoacoustic signal exhibits positive waveform due to the initial sharp rising of temperature. On the contrary, the second contraction-induced photoacoustic signal exhibits exactly negative waveform due to the falling of temperature, as well as pulse-width-dependent signal amplitude. An analytical model is derived to describe the generation of the dual photoacoustic pulses, incorporating Gruneisen saturation and thermal diffusion effect, which is experimentally proved. Lastly, an alternate of LDPA technique using quasi-CW laser excitation is also introduced and demonstrated for both super-contrast in vitro and in vivo imaging. Compared with existing nonlinear PA techniques, the proposed LDPA nonlinear effect could enable a much broader range of potential applications.

  12. Single- and dual energy QCT around acetabular cups in total hip arthroplasty using 3-dimensional segmentation

    DEFF Research Database (Denmark)

    Mussmann, Bo Redder; Andersen, Poul Erik; Torfing, Trine

    Introduction: Bone density measurements around hip implants are challenged by artifacts and the complex anatomy of the acetabulum. We developed 3D segmentation software and used dual energy CT to reduce artifacts. The between-scan agreement and reliability of the software was tested and bone mine...

  13. Reliable dual tensor model estimation in single and crossing fibers based on jeffreys prior

    NARCIS (Netherlands)

    J. Yang (Jianfei); D.H.J. Poot; M.W.A. Caan (Matthan); Su, T. (Tanja); C.B. Majoie (Charles); L.J. van Vliet (Lucas); F. Vos (Frans)

    2016-01-01

    textabstractPurpose This paper presents and studies a framework for reliable modeling of diffusion MRI using a data-acquisition adaptive prior. Methods Automated relevance determination estimates the mean of the posterior distribution of a rank-2 dual tensor model exploiting Jeffreys prior (JARD).

  14. Excited-state structural dynamics of a dual-emission calmodulin-green fluorescent protein sensor for calcium ion imaging

    Science.gov (United States)

    Oscar, Breland G.; Liu, Weimin; Zhao, Yongxin; Tang, Longteng; Wang, Yanli; Campbell, Robert E.; Fang, Chong

    2014-01-01

    Fluorescent proteins (FPs) have played a pivotal role in bioimaging and advancing biomedicine. The versatile fluorescence from engineered, genetically encodable FP variants greatly enhances cellular imaging capabilities, which are dictated by excited-state structural dynamics of the embedded chromophore inside the protein pocket. Visualization of the molecular choreography of the photoexcited chromophore requires a spectroscopic technique capable of resolving atomic motions on the intrinsic timescale of femtosecond to picosecond. We use femtosecond stimulated Raman spectroscopy to study the excited-state conformational dynamics of a recently developed FP-calmodulin biosensor, GEM-GECO1, for calcium ion (Ca2+) sensing. This study reveals that, in the absence of Ca2+, the dominant skeletal motion is a ∼170 cm−1 phenol-ring in-plane rocking that facilitates excited-state proton transfer (ESPT) with a time constant of ∼30 ps (6 times slower than wild-type GFP) to reach the green fluorescent state. The functional relevance of the motion is corroborated by molecular dynamics simulations. Upon Ca2+ binding, this in-plane rocking motion diminishes, and blue emission from a trapped photoexcited neutral chromophore dominates because ESPT is inhibited. Fluorescence properties of site-specific protein mutants lend further support to functional roles of key residues including proline 377 in modulating the H-bonding network and fluorescence outcome. These crucial structural dynamics insights will aid rational design in bioengineering to generate versatile, robust, and more sensitive optical sensors to detect Ca2+ in physiologically relevant environments. PMID:24987121

  15. Fluorescence imaging of local membrane electric fields during the excitation of single neurons in culture.

    Science.gov (United States)

    Gogan, P; Schmiedel-Jakob, I; Chitti, Y; Tyc-Dumont, S

    1995-01-01

    The spatial distribution of depolarized patches of membrane during the excitation of single neurons in culture has been recorded with a high spatial resolution (1 micron2/pixel) imaging system based on a liquid-nitrogen-cooled astronomical camera mounted on an inverted microscope. Images were captured from rat nodose neurons stained with the voltage-sensitive dye RH237. Conventional intracellular microelectrode recordings were made in synchrony with the images. During an action potential the fluorescence changes occurred in localized, unevenly distributed membrane areas, which formed clusters of depolarized sites of different sizes and intensities. When fast conductances were blocked by the addition of tetrodotoxin, a reduction in the number and the intensities of the depolarized sites was observed. The blockade by tetrodotoxin of voltage-clamped neurons also reduced the number of depolarized sites, although the same depolarizing voltage step was applied. Similarly, when a voltage-clamped neuron was depolarized by a constant-amplitude voltage step, the number of depolarized sites varied according to the degree of activation of the voltage-sensitive channels, which was modified by changing the holding potential. These results suggest that the spatial patterns of depolarization observed during excitation are related to the operations of ionic channels in the membrane. Images FIGURE 1 FIGURE 2 FIGURE 4 FIGURE 5 FIGURE 6 FIGURE 7 PMID:8527643

  16. Single-molecule-sensitive fluorescence resonance energy transfer in freely-diffusing attoliter droplets

    Energy Technology Data Exchange (ETDEWEB)

    Rahmanseresht, Sheema; Ramos, Kieran P.; Gamari, Ben D.; Goldner, Lori S., E-mail: lgoldner@physics.umass.edu [Department of Physics, University of Massachusetts, Amherst, Massachusetts 01003 (United States); Milas, Peker [Department of Neuroscience, University of Wisconsin, Madison, Wisconsin 53705 (United States)

    2015-05-11

    Fluorescence resonance energy transfer (FRET) from individual, dye-labeled RNA molecules confined in freely-diffusing attoliter-volume aqueous droplets is carefully compared to FRET from unconfined RNA in solution. The use of freely-diffusing droplets is a remarkably simple and high-throughput technique that facilitates a substantial increase in signal-to-noise for single-molecular-pair FRET measurements. We show that there can be dramatic differences between FRET in solution and in droplets, which we attribute primarily to an altered pH in the confining environment. We also demonstrate that a sufficient concentration of a non-ionic surfactant mitigates this effect and restores FRET to its neutral-pH solution value. At low surfactant levels, even accounting for pH, we observe differences between the distribution of FRET values in solution and in droplets which remain unexplained. Our results will facilitate the use of nanoemulsion droplets as attoliter volume reactors for use in biophysical and biochemical assays, and also in applications such as protein crystallization or nanoparticle synthesis, where careful attention to the pH of the confined phase is required.

  17. Integrating Optical Tweezers, DNA Tightropes, and Single-Molecule Fluorescence Imaging: Pitfalls and Traps.

    Science.gov (United States)

    Wang, J; Barnett, J T; Pollard, M R; Kad, N M

    2017-01-01

    Fluorescence imaging is one of the cornerstone techniques for understanding how single molecules search for their targets on DNA. By tagging individual proteins, it is possible to track their position with high accuracy. However, to understand how proteins search for targets, it is necessary to elongate the DNA to avoid protein localization ambiguities. Such structures known as "DNA tightropes" are tremendously powerful for imaging target location; however, they lack information about how force and load affect protein behavior. The use of optically trapped microstructures offers the means to apply and measure force effects. Here we describe a system that we recently developed to enable individual proteins to be directly manipulated on DNA tightropes. Proteins bound to DNA can be conjugated with Qdot fluorophores for visualization and also directly manipulated by an optically trapped, manufactured microstructure. Together this offers a new approach to understanding the physical environment of molecules, and the combination with DNA tightropes presents opportunities to study complex biological phenomena. © 2017 Elsevier Inc. All rights reserved.

  18. Transforming single DNA molecules into fluorescent magnetic particles for detection and enumeration of genetic variations

    Science.gov (United States)

    Dressman, Devin; Yan, Hai; Traverso, Giovanni; Kinzler, Kenneth W.; Vogelstein, Bert

    2003-01-01

    Many areas of biomedical research depend on the analysis of uncommon variations in individual genes or transcripts. Here we describe a method that can quantify such variation at a scale and ease heretofore unattainable. Each DNA molecule in a collection of such molecules is converted into a single magnetic particle to which thousands of copies of DNA identical in sequence to the original are bound. This population of beads then corresponds to a one-to-one representation of the starting DNA molecules. Variation within the original population of DNA molecules can then be simply assessed by counting fluorescently labeled particles via flow cytometry. This approach is called BEAMing on the basis of four of its principal components (beads, emulsion, amplification, and magnetics). Millions of individual DNA molecules can be assessed in this fashion with standard laboratory equipment. Moreover, specific variants can be isolated by flow sorting and used for further experimentation. BEAMing can be used for the identification and quantification of rare mutations as well as to study variations in gene sequences or transcripts in specific populations or tissues. PMID:12857956

  19. 5-fluoro-D,L-tryptophan as a dual NMR and fluorescent probe of α-synuclein.

    Science.gov (United States)

    Pfefferkorn, Candace M; Lee, Jennifer C

    2012-01-01

    Analysis of conventional proton nuclear magnetic resonance (NMR) experiments on intrinsically disordered proteins (IDPs) is challenging because of the highly flexible and multiple rapidly exchanging conformations typifying this class of proteins. One method to circumvent some of these difficulties is to incorporate nonnative fluorine ((19)F) nuclei at specific sites within the polypeptide. (19)F NMR is particularly suitable for characterization of unfolded structures because (19)F chemical shifts are highly sensitive to local environments and conformations. Furthermore, the incorporation of fluorine analogs of fluorescent amino acids such as 5-fluoro-D: ,L: -tryptophan (5FW) allows for complementary studies of protein microenvironment via fluorescence spectroscopy. Herein, we describe methods to produce, purify, characterize, and perform steady-state fluorescence and 1D NMR experiments on 5FW analogs of the IDP α-synuclein.

  20. Enhancement of single-molecule fluorescence signals by colloidal silver nanoparticles in studies of protein translation.

    Science.gov (United States)

    Bharill, Shashank; Chen, Chunlai; Stevens, Benjamin; Kaur, Jaskiran; Smilansky, Zeev; Mandecki, Wlodek; Gryczynski, Ignacy; Gryczynski, Zygmunt; Cooperman, Barry S; Goldman, Yale E

    2011-01-25

    Metal-enhanced fluorescence (MEF) increased total photon emission of Cy3- and Cy5-labeled ribosomal initiation complexes near 50 nm silver particles 4- and 5.5-fold, respectively. Fluorescence intensity fluctuations above shot noise, at 0.1-5 Hz, were greater on silver particles. Overall signal-to-noise ratio was similar or slightly improved near the particles. Proximity to silver particles did not compromise ribosome function, as measured by codon-dependent binding of fluorescent tRNA, dynamics of fluorescence resonance energy transfer between adjacent tRNAs in the ribosome, and tRNA translocation induced by elongation factor G.

  1. Quenching and enhancement of single-molecule fluorescence under metallic and dielectric tips

    Science.gov (United States)

    Azoulay, J.; Débarre, A.; Richard, A.; Tchénio, P.

    2000-08-01

    We report on fluorescence experiments by apertureless near-field optical microscopy. We develop a simple model that demonstrates the importance of non-radiative transfer and that takes into account the dependence of non-radiative transfer on tip geometry. This process is in competition with field enhancement and it is a key process to understand the observed fluorescence enhancement factors. The analysis of the different factors involved in the global fluorescence enhancement or quenching leads to new strategies to reach resolution down to a few nanometers by apertureless fluorescence microscopy.

  2. Genetic mutations in live infectious bronchitis vaccine viruses following single or dual in vitro infection of tracheal organ cultures.

    Science.gov (United States)

    Ball, Christopher; Bennett, Sarah; Forrester, Anne; Ganapathy, Kannan

    2016-12-01

    Despite regular co-vaccination of two different strains of live infectious bronchitis vaccine viruses, little is known about possible mutations in these viruses following vaccination. As an alternative to chicks, this study used an in vitro infection model to identify single-nucleotide polymorphisms (SNPs) within the part-S1 gene of two live infectious bronchitis virus vaccine strains (793B and Massachusetts) following single or dual inoculation onto tracheal organ cultures. Results indicate that viral titres reduced over the duration of the study; conversely, the amount of detected infectious bronchitis virus genome increased. Results demonstrate a greater number of non-synonymous SNPs in both vaccine strains when they are co-inoculated, compared with the single inoculations. The influence of the increased SNP and hydrophobic properties of the translated proteins on the vaccine viruses' virulence is unknown.

  3. Computed Tomography of the Head and Neck Region for Tumor Staging-Comparison of Dual-Source, Dual-Energy and Low-Kilovolt, Single-Energy Acquisitions.

    Science.gov (United States)

    May, Matthias Stefan; Bruegel, Joscha; Brand, Michael; Wiesmueller, Marco; Krauss, Bernhard; Allmendinger, Thomas; Uder, Michael; Wuest, Wolfgang

    2017-09-01

    The aim of this study was to intra-individually compare the image quality obtained by dual-source, dual-energy (DSDE) computed tomography (CT) examinations and different virtual monoenergetic reconstructions to a low single-energy (SE) scan. Third-generation DSDE-CT was performed in 49 patients with histologically proven malignant disease of the head and neck region. Weighted average images (WAIs) and virtual monoenergetic images (VMIs) for low (40 and 60 keV) and high (120 and 190 keV) energies were reconstructed. A second scan aligned to the jaw, covering the oral cavity, was performed for every patient to reduce artifacts caused by dental hardware using a SE-CT protocol with 70-kV tube voltages and matching radiation dose settings. Objective image quality was evaluated by calculating contrast-to-noise ratios. Subjective image quality was evaluated by experienced radiologists. Highest contrast-to-noise ratios for vessel and tumor attenuation were obtained in 40-keV VMI (all P VMI, WAI, and the 70-kV SE examinations. Overall subjective image quality was also highest for 40-keV, but differences to 60-keV VMI, WAI, and 70-kV SE were nonsignificant (all P > 0.05). High kiloelectron volt VMIs reduce metal artifacts with only limited diagnostic impact because of insufficiency in case of severe dental hardware. CTDIvol did not differ significantly between both examination protocols (DSDE: 18.6 mGy; 70-kV SE: 19.4 mGy; P = 0.10). High overall image quality for tumor delineation in head and neck imaging were obtained with 40-keV VMI. However, 70-kV SE examinations are an alternative and modified projections aligned to the jaw are recommended in case of severe artifacts caused by dental hardware.

  4. Single- and dual-energy CT of the abdomen: comparison of radiation dose and image quality of 2nd and 3rd generation dual-source CT

    Energy Technology Data Exchange (ETDEWEB)

    Wichmann, Julian L. [Medical University of South Carolina, Department of Radiology and Radiological Science, Charleston, SC (United States); University Hospital Frankfurt, Department of Diagnostic and Interventional Radiology, Frankfurt (Germany); Hardie, Andrew D.; Felmly, Lloyd M.; Perry, Jonathan D.; Varga-Szemes, Akos; De Cecco, Carlo N. [Medical University of South Carolina, Department of Radiology and Radiological Science, Charleston, SC (United States); Schoepf, U.J. [Medical University of South Carolina, Department of Radiology and Radiological Science, Charleston, SC (United States); Medical University of South Carolina, Division of Cardiology, Department of Medicine, Charleston, SC (United States); Mangold, Stefanie [University Hospital of Tuebingen, Department of Diagnostic and Interventional Radiology, Tuebingen (Germany); Caruso, Damiano [Medical University of South Carolina, Department of Radiology and Radiological Science, Charleston, SC (United States); University of Rome ' ' Sapienza' ' , Department of Radiological Sciences, Oncological and Pathological Sciences, Latina (Italy); Canstein, Christian [Medical University of South Carolina, Department of Radiology and Radiological Science, Charleston, SC (United States); Siemens Medical Solutions USA, Malvern, PA (United States); Vogl, Thomas J. [University Hospital Frankfurt, Department of Diagnostic and Interventional Radiology, Frankfurt (Germany)

    2017-02-15

    To compare single-energy (SECT) and dual-energy (DECT) abdominal CT examinations in matched patient cohorts regarding differences in radiation dose and image quality performed with second- and third-generation dual-source CT (DSCT). We retrospectively analysed 200 patients (100 male, 100 female; mean age 61.2 ± 13.5 years, mean body mass index 27.5 ± 3.8 kg/m{sup 2}) equally divided into four groups matched by gender and body mass index, who had undergone portal venous phase abdominal CT with second-generation (group A, 120-kV-SECT; group B, 80/140-kV-DECT) and third-generation DSCT (group C, 100-kV-SECT; group D, 90/150-kV-DECT). The radiation dose was normalised for 40-cm scan length. Dose-independent figure-of-merit (FOM) contrast-to-noise ratios (CNRs) were calculated for various organs and vessels. Subjective overall image quality and reader confidence were assessed. The effective normalised radiation dose was significantly lower (P < 0.001) in groups C (6.2 ± 2.0 mSv) and D (5.3 ± 1.9 mSv, P = 0.103) compared to groups A (8.8 ± 2.3 mSv) and B (9.7 ± 2.4 mSv, P = 0.102). Dose-independent FOM-CNR peaked for liver, kidney, and portal vein measurements (all P ≤ 0.0285) in group D. Subjective image quality and reader confidence were consistently rated as excellent in all groups (all ≥1.53 out of 5). With both DSCT generations, abdominal DECT can be routinely performed without radiation dose penalty compared to SECT, while third-generation DSCT shows improved dose efficiency. (orig.)

  5. pH-Responsive Tumor-Targetable Theranostic Nanovectors Based on Core Crosslinked (CCL Micelles with Fluorescence and Magnetic Resonance (MR Dual Imaging Modalities and Drug Delivery Performance

    Directory of Open Access Journals (Sweden)

    Sidan Tian

    2016-06-01

    Full Text Available The development of novel theranostic nanovectors is of particular interest in treating formidable diseases (e.g., cancers. Herein, we report a new tumor-targetable theranostic agent based on core crosslinked (CCL micelles, possessing tumor targetable moieties and fluorescence and magnetic resonance (MR dual imaging modalities. An azide-terminated diblock copolymer, N3-POEGMA-b-P(DPA-co-GMA, was synthesized via consecutive atom transfer radical polymerization (ATRP, where OEGMA, DPA, and GMA are oligo(ethylene glycolmethyl ether methacrylate, 2-(diisopropylaminoethyl methacrylate, and glycidyl methacrylate, respectively. The resulting diblock copolymer was further functionalized with DOTA(Gd (DOTA is 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakisacetic acid or benzaldehyde moieties via copper(I-catalyzed alkyne-azide cycloaddition (CuAAC chemistry, resulting in the formation of DOTA(Gd-POEGMA-b-P(DPA-co-GMA and benzaldehyde-POEGMA-b-P(DPA-co-GMA copolymers. The resultant block copolymers co-assembled into mixed micelles at neutral pH in the presence of tetrakis[4-(2-mercaptoethoxyphenyl]ethylene (TPE-4SH, which underwent spontaneous crosslinking reactions with GMA residues embedded within the micellar cores, simultaneously switching on TPE fluorescence due to the restriction of intramolecular rotation. Moreover, camptothecin (CPT was encapsulated into the crosslinked cores at neutral pH, and tumor-targeting pH low insertion peptide (pHLIP, sequence: AEQNPIYWARYADWLFTTPLLLLDLALLVDADEGTCG moieties were attached to the coronas through the Schiff base chemistry, yielding a theranostic nanovector with fluorescence and MR dual imaging modalities and tumor-targeting capability. The nanovectors can be efficiently taken up by A549 cells, as monitored by TPE fluorescence. After internalization, intracellular acidic pH triggered the release of loaded CPT, killing cancer cells in a selective manner. On the other hand, the nanovectors labeled with DOTA

  6. Noise-tolerance analysis for detection and reconstruction of absorbing inhomogeneities with diffuse optical tomography using single- and phase-correlated dual-source schemes

    Science.gov (United States)

    Kanmani, B.; Vasu, R. M.

    2007-03-01

    An iterative reconstruction procedure is used to invert intensity data from both single- and phase-correlated dual-source illuminations for absorption inhomogeneities. The Jacobian for the dual source is constructed by an algebraic addition of the Jacobians estimated for the two sources separately. By numerical simulations, it is shown that the dual-source scheme performs superior to the single-source system in regard to (i) noise tolerance in data and (ii) ability to reconstruct smaller and lower contrast objects. The quality of reconstructions from single-source data, as indicated by mean-square error at convergence, is markedly poorer compared to their dual-source counterpart, when noise in data was in excess of 2%. With fixed contrast and decreasing inhomogeneity diameter, our simulations showed that, for diameters below 7 mm, the dual-source scheme has a higher percentage contrast recovery compared to the single-source scheme. Similarly, the dual-source scheme reconstructs to a higher percentage contrast recovery from lower contrast inhomogeneity, in comparison to the single-source scheme.

  7. Novel challenges to gait in Parkinson's disease: the effect of concurrent music in single- and dual-task contexts.

    Science.gov (United States)

    Brown, Lesley A; de Bruin, Natalie; Doan, Jon B; Suchowersky, Oksana; Hu, Bin

    2009-09-01

    To investigate the effect of concurrent music on parkinsonian gait in single- and dual-task contexts. A counterbalanced repeated-measure design. A university balance research laboratory. People with idiopathic Parkinson's disease (PD) (n=10) (67+/-7 y) and healthy age-matched (65+/-6 y) control subjects (n=10). Subjects walked at a self-selected pace along an unobstructed walkway in 4 differing test conditions. Test conditions were differentiated by the presence of music accompaniment (no music/music) and the presence of a secondary cognitive task (single/dual). Single- and dual-task conditions were randomized; trials were blocked by the presence of music and counterbalanced between subjects. Music was self-selected by subjects. The cognitive task consisted of serial subtractions (3's). Subjects were not instructed to attend to the music nor were they provided with instructions regarding task prioritization. Mean gait velocity, stride length, and the percentage of the gait cycle spent in double-limb support. Gait among the PD patients was adversely affected by concurrent music. In contrast, gait performance in the control subjects showed no significant difference between no music and music conditions. The added requirement of a cognitive task differentially influenced gait performance in PD patients and control subjects, with PD patients displaying a further decrease in spatiotemporal parameters of gait and control subjects displaying a marginal improvement. Gait impairments associated with PD are exacerbated in the presence of concurrent music, an effect that is further exaggerated by the addition of a cognitive task. These results have implications for patient safety in multitasking situations.

  8. The reliability of postural balance measures in single and dual tasking in elderly fallers and non-fallers

    Directory of Open Access Journals (Sweden)

    Favero Kathrin

    2008-12-01

    Full Text Available Abstract Background The purpose of this study was to determine the reliability of a forceplate postural balance protocol in a group of elderly fallers and non-fallers. The measurements were tested in single and dual-task conditions, with and without vision. Methods 37 elderly (mean age 73 ± 6 years community-dwellers were included in this study. All were tested in a single (two-legged stance and in a dual-task (two-legged stance while counting backwards aloud in steps of 7's condition, with and without vision. A forceplate was used for registering postural variables: the maximal and the root-mean-square amplitude in medio-lateral (Max-ML, RMS-ML and antero-posterior (Max-AP, RMS-AP direction, mean velocity (MV, and the area of the 95% confidence ellipse (AoE. Reliability of the test protocol was expressed with intraclass correlation coefficients (ICC, with 95% limits of agreement (LoA, and with the smallest detectable difference (SDD. Results The ICCs for inter-rater reliability and test-retest reliability of the balance variables were r = 0.70–0.89. For the variables Max-AP and RMS-AP the ICCs were r = 0.52–0.74. The SDD values were for variable Max-ML and Max-AP between 0.37 cm and 0.83 cm, for MV between 0.48 cm/s and 1.2 cm/s and for AoE between 1.48 cm2 and 3.75 cm2. The LoA analysis by Bland-Altman plots showed no systematic differences between test-retest measurements. Conclusion The study showed good reliability results for group assessment and no systematic errors of the measurement protocol in measuring postural balance in the elderly in a single-task and dual-task condition.

  9. Antibiotic transport in resistant bacteria: synchrotron UV fluorescence microscopy to determine antibiotic accumulation with single cell resolution.

    Directory of Open Access Journals (Sweden)

    Slávka Kaščáková

    Full Text Available A molecular definition of the mechanism conferring bacterial multidrug resistance is clinically crucial and today methods for quantitative determination of the uptake of antimicrobial agents with single cell resolution are missing. Using the naturally occurring fluorescence of antibacterial agents after deep ultraviolet (DUV excitation, we developed a method to non-invasively monitor the quinolones uptake in single bacteria. Our approach is based on a DUV fluorescence microscope coupled to a synchrotron beamline providing tuneable excitation from 200 to 600 nm. A full spectrum was acquired at each pixel of the image, to study the DUV excited fluorescence emitted from quinolones within single bacteria. Measuring spectra allowed us to separate the antibiotic fluorescence from the autofluorescence contribution. By performing spectroscopic analysis, the quantification of the antibiotic signal was possible. To our knowledge, this is the first time that the intracellular accumulation of a clinical antibiotic could be determined and discussed in relation with the level of drug susceptibility for a multiresistant strain. This method is especially important to follow the behavior of quinolone molecules at individual cell level, to quantify the intracellular concentration of the antibiotic and develop new strategies to combat the dissemination of MDR-bacteria. In addition, this original approach also indicates the heterogeneity of bacterial population when the same strain is under environmental stress like antibiotic attack.

  10. Linear modeling of single-shot dual-energy x-ray imaging using a sandwich detector

    Science.gov (United States)

    Kim, J.; Kim, D. W.; Kim, S. H.; Yun, S.; Youn, H.; Jeon, H.; Kim, H. K.

    2017-01-01

    For single-shot dual-energy (DE) imaging, a sandwich detector typically consists of a thin front detector and a thick rear detector. Therefore, the spatial-resolution characteristics of the two detectors are different, and as a result, weighted subtraction of the corresponding two images gives rise to edge-enhancement characteristics in the resulting DE images. This is a unique characteristic of single-shot DE imaging compared to the conventional dual-shot DE imaging which uses the same detector to acquire low- and high-energy images. Using a linear-systems theory, in this paper, we show that the modulation-transfer function (MTF) of a sandwich detector is a weighted average of contributions from each MTF characteristic of two detector layers forming the sandwich detector. The MTF results obtained using the developed model are validated with those measured directly from single-shot DE images for an edge-knife phantom. Weighting larger than at least 0.5 in DE reconstruction gives an enhancement in DE MTF at mid and high spatial frequencies compared to the MTFs obtained from each detector layer. The behavior of the linear model as a function of weighting factor used for DE reconstruction is discussed in comparisons with numerical simulations.

  11. Modified low temperature Czochralski growth of xylenol orange doped benzopheone single crystal for fabricating dual band patch antenna

    Science.gov (United States)

    Yadav, Harsh; Sinha, Nidhi; Kumar, Binay

    2016-09-01

    Organic non-linear optical pure and xylenol orange (XO) doped benzophenone (BP) single crystals have been grown by a modified Czochralski technique. A low cost CZ system was designed and fabricated that is suitable for the growth of single crystals of low melting point organic materials. Structural analysis was performed by powder and single crystal XRD. LC-HRMS spectra reveal that the dye molecules are present in the doped crystal. The linear optical characterization was carried out by UV-vis spectroscopy. In the case of the XO doped BP crystal, two absorption peaks were found at 504 nm and 620 nm. The enhancement of photoluminescence intensity of blue emission was observed in the dye doped crystal. Dielectric studies reveal that the XO doped BP has shown improved a dielectric constant with low dielectric loss. A dual band compact circular patch antenna was simulated and fabricated using the XO doped crystal. Resonant frequencies of the dual bands at 4.80 GHz and 9.22 GHz were achieved by introducing a defect ground state in the circular patch antenna. The piezoelectric coefficient (d33) value was increased from 1 to 4 pC/N by XO dye doping, which opens up the possibilities of simultaneous transducer applications.

  12. The role of molecular dipole orientation in single-molecule fluorescence microscopy and implications for super-resolution imaging.

    Science.gov (United States)

    Backlund, Mikael P; Lew, Matthew D; Backer, Adam S; Sahl, Steffen J; Moerner, W E

    2014-03-17

    Numerous methods for determining the orientation of single-molecule transition dipole moments from microscopic images of the molecular fluorescence have been developed in recent years. At the same time, techniques that rely on nanometer-level accuracy in the determination of molecular position, such as single-molecule super-resolution imaging, have proven immensely successful in their ability to access unprecedented levels of detail and resolution previously hidden by the optical diffraction limit. However, the level of accuracy in the determination of position is threatened by insufficient treatment of molecular orientation. Here we review a number of methods for measuring molecular orientation using fluorescence microscopy, focusing on approaches that are most compatible with position estimation and single-molecule super-resolution imaging. We highlight recent methods based on quadrated pupil imaging and on double-helix point spread function microscopy and apply them to the study of fluorophore mobility on immunolabeled microtubules.

  13. Development of a strain visualization system for microstructures using single fluorescent molecule tracking on a three-dimensional orientation microscope

    Science.gov (United States)

    Yoshida, Shintaro; Yoshiki, Keisuke; Namazu, Takahiro; Araki, Nozomu; Hashimoto, Mamoru; Kurihara, Makoto; Hashimoto, Nobuyuki; Inoue, Shozo

    2011-09-01

    We propose a technique that employs single fluorescent molecules for visualizing the distribution of strain induced in microstructures. We sprayed single-molecule tracers on microstructures by ultrasonic atomization and traced the position and orientation of the tracers by a single-molecule detection technique with a three-dimensional (3D) orientation microscope, which consists of a conventional fluorescent microscope and a polarization-mode converter. By using 3D spline interpolation, we visualized the surface geometry of a microelectromechanical (MEMS) device. We tracked the 3D position and orientation of tracers attached to a supporting beam of the MEMS mirror. The surface declination angles calculated from the orientation of the tracers were in agreement with the tilt angle obtained from the 3D position of the tracers.

  14. Single molecule detection of nitric oxide enabled by d(AT)15 DNA adsorbed to near infrared fluorescent single-walled carbon nanotubes.

    Science.gov (United States)

    Zhang, Jingqing; Boghossian, Ardemis A; Barone, Paul W; Rwei, Alina; Kim, Jong-Ho; Lin, Dahua; Heller, Daniel A; Hilmer, Andrew J; Nair, Nitish; Reuel, Nigel F; Strano, Michael S

    2011-01-26

    We report the selective detection of single nitric oxide (NO) molecules using a specific DNA sequence of d(AT)(15) oligonucleotides, adsorbed to an array of near-infrared fluorescent semiconducting single-walled carbon nanotubes (AT(15)-SWNT). While SWNT suspended with eight other variant DNA sequences show fluorescence quenching or enhancement from analytes such as dopamine, NADH, L-ascorbic acid, and riboflavin, d(AT)(15) imparts SWNT with a distinct selectivity toward NO. In contrast, the electrostatically neutral polyvinyl alcohol enables no response to nitric oxide, but exhibits fluorescent enhancement to other molecules in the tested library. For AT(15)-SWNT, a stepwise fluorescence decrease is observed when the nanotubes are exposed to NO, reporting the dynamics of single-molecule NO adsorption via SWNT exciton quenching. We describe these quenching traces using a birth-and-death Markov model, and the maximum likelihood estimator of adsorption and desorption rates of NO is derived. Applying the method to simulated traces indicates that the resulting error in the estimated rate constants is less than 5% under our experimental conditions, allowing for calibration using a series of NO concentrations. As expected, the adsorption rate is found to be linearly proportional to NO concentration, and the intrinsic single-site NO adsorption rate constant is 0.001 s(-1) μM NO(-1). The ability to detect nitric oxide quantitatively at the single-molecule level may find applications in new cellular assays for the study of nitric oxide carcinogenesis and chemical signaling, as well as medical diagnostics for inflammation.

  15. Dual-mode chemical vapor generation for simultaneous determination of hydride-forming and non-hydride-forming elements by atomic fluorescence spectrometry.

    Science.gov (United States)

    Wang, Yu; Xu, Kailai; Jiang, Xiaoming; Hou, Xiandeng; Zheng, Chengbin

    2014-05-21

    A dual-mode chemical vapor generation integrating hydride generation and photochemical vapor generation was developed for simultaneous multi-element analysis of hydride-forming and non-hydride-forming elements by atomic fluorescence spectrometry. Four elements were selected as model elements of hydride-forming (As, Cd) and non-hydride-forming (Ni, Fe) elements to validate this proposed method. Standard or sample solutions were separately pumped to mix with tetrahydroborate, and concentrated formic acid and ammonia, and then directed to a hydride generator and a photochemical reactor to realize simultaneous hydride generation and photochemical vapor generation, respectively. Optimum conditions for dual-mode chemical vapor generation were carefully investigated. Under the optimized conditions, limits of detection of 0.05, 0.008, 0.8 and 0.1 μg L(-1) were obtained for As, Cd, Fe and Ni, respectively. The precisions were 5.0, 5.5, 4.3 and 4.5% (n = 6, RSDs) for 2 μg L(-1) of As, 1 μg L(-1) of Cd, 50 μg L(-1) of Fe and 10 μg L(-1) of Ni, respectively. This method was validated for accuracy with three certified reference water samples and applied to the simultaneous determination of these elements in a tap water sample with spike recoveries in the range of 95-99%.

  16. Photobleaching of asymmetric cyanines used for fluorescence imaging of single DNA molecules.

    Science.gov (United States)

    Kanony, C; Akerman, B; Tuite, E

    2001-08-22

    The photobleaching of the cyanine dyes YO and YOYO has been investigated for both free and DNA-bound dyes, using absorption and fluorescence spectroscopy coupled with fluorescence microscopy. For the free dyes, the nature of the reactive species involved in the photodegradation process is different for the monomer and the dimer, as shown by scavenger studies. For DNA-bound dyes, photoinduced fading of the visible absorption band occurs by different pathways depending on the drug binding mode and can be attenuated by appropriate scavengers. However, none of these scavengers were found to have any significant effect on the photobleaching of dye fluorescence. It appears that the reduction of fluorescence intensity comes from a quenching of the dye fluorescence by modified DNA bases, possibly 8-oxo-7,8-dihydro-2'-deoxyguanosine.

  17. A Novel Real-time Fluorescence Mutant-allele-specific Amplification Method for Rapid Single Nucleotide Polymorphism Analysis

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Current methods for single nucleotide polymorphism (SNP) analysis are timeconsuming and complicated. We aimed at development of one-step real-time fluorescence mutant-allele-specific amplification (MASA) method for rapid SNP analysis. The method is a marriage of two technologies: MASA primers for target DNA and a double-stranded DNA-selective fluorescent dye, SYBR Green I. Genotypes are separated according to the different threshold cycles of the wild-type and mutant primers. K-ras oncogene was used as a target to validate the feasibility of the method. The experimental results showed that the different genotypes can be clearly discriminated by the assay. The real-time fluorescence MASA method will have an enormous potential for fast and reliable SNP analysis due to its simplicity and low cost.

  18. Laser-Induced Fluorescence Detection in High-Throughput Screening of Heterogeneous Catalysts and Single Cells Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Hui Su

    2001-05-25

    Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, the author introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, they demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm{sup 2} for 40-{micro}m wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection. In the second part of this dissertation, the author used laser-induced native fluorescence coupled with capillary electrophoresis (LINF-CE) and microscope imaging to study the single cell degranulation. On the basis of good temporal correlation with events observed through an optical microscope, they have identified individual peaks in the fluorescence electropherograms as serotonin released from the granular core on contact with the surrounding fluid.

  19. Laser-Induced Fluorescence Detection in High-Throughput Screening of Heterogeneous Catalysts and Single Cells Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Su, Hui [Iowa State Univ., Ames, IA (United States)

    2001-01-01

    Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, the author introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, they demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm2 for 40-μm wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection. In the second part of this dissertation, the author used laser-induced native fluorescence coupled with capillary electrophoresis (LINF-CE) and microscope imaging to study the single cell degranulation. On the basis of good temporal correlation with events observed through an optical microscope, they have identified individual peaks in the fluorescence electropherograms as serotonin released from the granular core on contact with the surrounding fluid.

  20. Fluorescence spectra of atmospheric aerosol at Adelphi, Maryland, USA: measurement and classification of single particles containing organic carbon

    Science.gov (United States)

    Pinnick, Ronald G.; Hill, Steven C.; Pan, Yong-Le; Chang, Richard K.

    We measured laser-induced fluorescence spectra from individual supermicron-sized atmospheric particles drawn into our laboratory at Adelphi, MD, an urban site in the Washington, DC metroplex. A virtural impactor concentrator is used along with an aerodynamic-focusing-nozzle which forms, within an optical chamber, a focused aerosol jet where single aerosol particles can be interrogated on-the-fly with a pulsed 266-nm-wavelength laser. Sample rates are a few liter per minute, and are size dependent. Crossed-diode laser beams indicate when a particle is traversing the sample region and are used to trigger the UV laser to fire and the gated intensified CCD to record the fluorescence spectrum. Our breadboard fluorescence particle spectrometer measures particles in the 3-10 μm diameter size range. Typical trigger rates are a few per second. The usable spectral range is from about 295 to 605 nm. The majority of the particles have very weak fluorescence (on average 8 percent of particles have fluorescence signals above noise). The spectra were grouped using a heirarchical cluster analysis, with parameters chosen so that spectra typically cluster into 4-12 main categories. From the set of all cluster spectra we chose 8 template spectra for reanalyzing all the data. On average, 92 percent (81-94 percent) of the spectra were similar to these templates (using the same thresholds used for the cluster analysis). The major emission bands of the most commonly occurring spectra have peaks: near 460 nm (28 percent of fluorescent particles on average), a very broad hump, and may be humic acids or humic like substances; near 317 nm (on average 24 percent of fluorescent particles); near 321 and 460 nm (a double hump, 12 percent of fluorescent particles); and near 341 nm (8 percent of fluorescent particles). Some of the fluorescence in spectra peaking in the 317-341 nm range is probably from dicyclic aromatics and heterocyclics, including the amino acid tryptophan in biological

  1. Magnetic and fluorescent Gd2O3:Yb(3+)/Ln(3+) nanoparticles for simultaneous upconversion luminescence/MR dual modal imaging and NIR-induced photodynamic therapy.

    Science.gov (United States)

    Liu, Jun; Huang, Long; Tian, Xiumei; Chen, Xiaoming; Shao, Yuanzhi; Xie, Fukang; Chen, Dihu; Li, Li

    The development of upconversion nanoparticles (UCNs) for theranostics application is a new strategy toward the accurate diagnosis and efficient treatment of cancer. Here, magnetic and fluorescent lanthanide-doped gadolinium oxide (Gd2O3) UCNs with bright upconversion luminescence (UCL) and high longitudinal relaxivity (r1) are used for simultaneous magnetic resonance imaging (MRI)/UCL dual-modal imaging and photodynamic therapy (PDT). In vitro and in vivo MRI studies show that these products can serve as good MRI contrast agents. The bright upconversion luminescence of the products allows their use as fluorescence nanoprobes for live cells imaging. We also utilized the luminescence-emission capability of the UCNs for the activation of a photosensitizer to achieve significant PDT results. To the best of our knowledge, this study is the first use of lanthanide-doped Gd2O3 UCNs in a theranostics application. This investigation provides a useful platform for the development of Gd2O3-based UCNs for clinical diagnosis, treatment, and imaging-guided therapy of cancer.

  2. Magnetic and fluorescent Gd2O3:Yb3+/Ln3+ nanoparticles for simultaneous upconversion luminescence/MR dual modal imaging and NIR-induced photodynamic therapy

    Science.gov (United States)

    Liu, Jun; Huang, Long; Tian, Xiumei; Chen, Xiaoming; Shao, Yuanzhi; Xie, Fukang; Chen, Dihu; Li, Li

    2017-01-01

    The development of upconversion nanoparticles (UCNs) for theranostics application is a new strategy toward the accurate diagnosis and efficient treatment of cancer. Here, magnetic and fluorescent lanthanide-doped gadolinium oxide (Gd2O3) UCNs with bright upconversion luminescence (UCL) and high longitudinal relaxivity (r1) are used for simultaneous magnetic resonance imaging (MRI)/UCL dual-modal imaging and photodynamic therapy (PDT). In vitro and in vivo MRI studies show that these products can serve as good MRI contrast agents. The bright upconversion luminescence of the products allows their use as fluorescence nanoprobes for live cells imaging. We also utilized the luminescence-emission capability of the UCNs for the activation of a photosensitizer to achieve significant PDT results. To the best of our knowledge, this study is the first use of lanthanide-doped Gd2O3 UCNs in a theranostics application. This investigation provides a useful platform for the development of Gd2O3-based UCNs for clinical diagnosis, treatment, and imaging-guided therapy of cancer. PMID:28031709

  3. Direct and accurate measurement of CAG repeat configuration in the ataxin-1 (ATXN-1) gene by "dual-fluorescence labeled PCR-restriction fragment length analysis".

    Science.gov (United States)

    Lin, Jiang X; Ishikawa, Kinya; Sakamoto, Masaki; Tsunemi, Taiji; Ishiguro, Taro; Amino, Takeshi; Toru, Shuta; Kondo, Ikuko; Mizusawa, Hidehiro

    2008-01-01

    Spinocerebellar ataxia type 1 (SCA1; OMIM: #164400) is an autosomal dominant cerebellar ataxia caused by an expansion of CAG repeat, which encodes polyglutamine, in the ataxin-1 (ATXN1) gene. Length of polyglutamine in the ATXN1 protein is the critical determinant of pathogenesis of this disease. Molecular diagnosis of SCA1 is usually undertaken by assessing the length of CAG repeat configuration using primers spanning this configuration. However, this conventional method may potentially lead to misdiagnosis in assessing polyglutamine-encoding CAG repeat length, since CAT interruptions may be present within the CAG repeat configuration, not only in normal controls but also in neurologically symptomatic subjects. We developed a new method for assessing actual CAG repeat numbers not interrupted by CAT sequences. Polymerase chain reaction using a primer pair labeled with two different fluorescences followed by restriction enzyme digestion with SfaNI which recognizes the sequence "GCATC(N)(5)", lengths of actual CAG repeats that encode polyglutamine were directly detected. We named this method "dual fluorescence labeled PCR-restriction fragment length analysis". We found that numbers of actual CAG repeat encoding polyglutamine do not overlap between our cohorts of normal chromosomes (n=385) and SCA1 chromosomes (n=5). We conclude that the present method is a useful way for molecular diagnosis of SCA1.

  4. Selective Detection and Automated Counting of Fluorescently-Labeled Chrysotile Asbestos Using a Dual-Mode High-Throughput Microscopy (DM-HTM Method

    Directory of Open Access Journals (Sweden)

    Jung Kyung Kim

    2013-05-01

    Full Text Available Phase contrast microscopy (PCM is a widely used analytical method for airborne asbestos, but it is unable to distinguish asbestos from non-asbestos fibers and requires time-consuming and laborious manual counting of fibers. Previously, we developed a high-throughput microscopy (HTM method that could greatly reduce human intervention and analysis time through automated image acquisition and counting of fibers. In this study, we designed a dual-mode HTM (DM-HTM device for the combined reflection and fluorescence imaging of asbestos, and automated a series of built-in image processing commands of ImageJ software to test its capabilities. We used DksA, a chrysotile-adhesive protein, for selective detection of chrysotile fibers in the mixed dust-free suspension of crysotile and amosite prepared in the laboratory. We demonstrate that fluorescently-stained chrysotile and total fibers can be identified and enumerated automatically in a high-throughput manner by the DM-HTM system. Combined with more advanced software that can correctly identify overlapping and branching fibers and distinguish between fibers and elongated dust particles, the DM-HTM method should enable fully automated counting of airborne asbestos.

  5. Ratiometric Molecular Probes Based on Dual Emission of a Blue Fluorescent Coumarin and a Red Phosphorescent Cationic Iridium(III) Complex for Intracellular Oxygen Sensing.

    Science.gov (United States)

    Yoshihara, Toshitada; Murayama, Saori; Tobita, Seiji

    2015-06-09

    Ratiometric molecular probes RP1 and RP2 consisting of a blue fluorescent coumarin and a red phosphorescent cationic iridium complex connected by a tetra- or octaproline linker, respectively, were designed and synthesized for sensing oxygen levels in living cells. These probes exhibited dual emission with good spectral separation in acetonitrile. The photorelaxation processes, including intramolecular energy transfer, were revealed by emission quantum yield and lifetime measurements. The ratios (R(I) = (I(p)/I(f))) between the phosphorescence (I(p)) and fluorescence (I(f)) intensities showed excellent oxygen responses; the ratio of R(I) under degassed and aerated conditions ( R(I)(0) was 20.3 and 19.6 for RP1 and RP2. The introduction of the cationic Ir (III) complex improved the cellular uptake efficiency compared to that of a neutral analogue with a tetraproline linker. The emission spectra of the ratiometric probes internalized into living HeLa or MCF-7 cells could be obtained using a conventional microplate reader. The complex RP2 with an octaproline linker provided ratios comparable to the ratiometric measurements obtained using a microplate reader: the ratio of the R(I)) value of RP2 under hypoxia (2.5% O2) to that under normoxia (21% O2) was 1.5 and 1.7 for HeLa and MCF-7 cells, respectively. Thus, the intracellular oxygen levels of MCF-7 cells could be imaged by ratiometric emission measurements using the complex RP2.

  6. In Vivo Magnetic Resonance and Fluorescence Dual-Modality Imaging of Tumor Angiogenesis in Rats Using GEBP11 Peptide Targeted Magnetic Nanoparticles.

    Science.gov (United States)

    Su, Tao; Wang, Yabin; Wang, Jiinda; Han, Dong; Ma, Sai; Cao, Jianbo; Li, Xiujuan; Zhang, Ran; Qiao, Hongyu; Liang, Jimin; Liu, Gang; Yang, Bo; Liang, Shuhui; Nie, Yongzhan; Wu, Kaichun; Li, Jiayi; Cao, Feng

    2016-05-01

    Angiogenesis is an essential process for tumor progression. Tumor vasculature-targeting peptides have shown great potential for use in cancer imaging and therapy. Our previous studies have shown that GEBP11, a novel vasculature-specific binding peptide that exhibits high affinity and specificity to tumor angiogenesis, is a promising candidate for the diagnosis and targeted radiotherapy of gastric cancer. In the present study, we developed a novel magnetic resonance and fluorescence (MR/Fluo) dual-modality imaging probe by covalently coupling 2,3-dimercaptosuccinnic acid-coated paramagnetic nanoparticles (DMSA-MNPs) and Cy5.5 to the GEBP11 peptide. The probe Cy5.5-GEBP11-DMSA-MNPs (CGD-MNPs), with a hydrodynamic diameter of 82.8 ± 6.5 nm, exhibited good imaging properties, high stability and little cytotoxicity. In vivo MR/Fluo imaging revealed that CGD-MNPs were successfully applied to visualize tumor angiogenesis in SGC-7901 xenograft mouse models. Prussian blue and CD31 immunohistochemical staining confirmed that CGD-MNPs co-localized with tumor blood vessels. In conclusion, CGD-MNPs are promising candidates for use as MR and fluorescence imaging probes for visualizing gastric cancer angiogenesis in vivo.

  7. Stimulus Response of Au-NPs@GMP-Tb Core-Shell Nanoparticles: Toward Colorimetric and Fluorescent Dual-Mode Sensing of Alkaline Phosphatase Activity in Algal Blooms of a Freshwater Lake.

    Science.gov (United States)

    Zhang, Xiaolei; Deng, Jingjing; Xue, Yumeng; Shi, Guoyue; Zhou, Tianshu

    2016-01-19

    In this study, we demonstrate a colorimetric and fluorescent dual-mode method for alkaline phosphatase activity (APA) sensing in freshwater lake with stimuli-responsive gold nanoparticles@terbium-guanosine monophosphate (Au-NPs@GMP-Tb) core-shell nanoparticles. Initially, the core-shell nanoparticles were fabricated based on Au-NPs decorated with a fluorescent GMP-Tb shell. Upon being excited at 290 nm, the as-formed Au-NPs@GMP-Tb core-shell nanoparticles emit green fluorescence, and the decorated GMP-Tb shell causes the aggregation of Au-NPs. However, the addition of ALP destroys GMP-Tb shell, resulting in the release of Au-NPs from the shell into the solvent. As a consequence, the aggregated Au-NPs solubilizes with the changes in the UV-vis spectrum of the dispersion, and in the meantime, the fluorescence of GMP-Tb shell turns off, which constitutes a new mechanism for colorimetric and fluorescent dual-mode sensing of APA. With the method developed here, we could monitor the dynamic change of APA during an algal bloom of a freshwater lake, both by the naked eye and further confirmed by fluorometric determination. This study not only offers a new method for on-site visible detection of APA but also provides a strategy for dual-mode sensing mechanisms by the rational design of the excellent optical properties of Au-NPs and the adaptive inclusion properties of the luminescent infinite coordination polymers.

  8. Multicolor imaging of hydrogen peroxide level in living and apoptotic cells by a single fluorescent probe.

    Science.gov (United States)

    Wen, Ying; Xue, Fengfeng; Lan, Haichuang; Li, Zhenhua; Xiao, Shuzhang; Yi, Tao

    2017-05-15

    To understand the entangled relationship between reactive oxygen species (ROS) and apoptosis, there is urgent need for simultaneous dynamic monitoring of these two important biological events. In this study, we have developed a fluorescent probe, pep4-NP1, which can simultaneously detect H2O2 and caspase 3, the respective markers of ROS and apoptosis. The probe contains a H2O2 fluorescence reporter (NP1) and Cy5 fluorescent chromophore connected by a caspase 3 specific recognition peptide. The detecting strategy was realized through a controllable fluorescence resonance energy transfer (FRET) process between NP1 and Cy5 of pep4-NP1, after reaction with H2O2, which was verified by molecular calculation and in vitro spectral studies. In the absent of caspase 3, the accumulation of H2O2 induces red fluorescence of pep4-NP1 centered at 663nm in living cells due to the existence of FRET. In contrast, FRET is inhibited in apoptotic cells due to cleavage of the peptide spacer of pep4-NP1 by over-expressed caspase 3. Consequently, green fluorescence (555nm) predominated when labelling production of H2O2 in apoptotic cells. Moreover, Pep4-NP1 shows excellent selectivity towards H2O2 and caspase 3 on their respective reaction sites. Therefore, pep4-NP1 can distinguish endogenously generated H2O2 between living cells and apoptotic cells with different fluorescence wavelengths, providing additional information on the ROS production pathways.

  9. Dendrimer-encapsulated naphthalocyanine as a single agent-based theranostic nanoplatform for near-infrared fluorescence imaging and combinatorial anticancer phototherapy

    Science.gov (United States)

    Taratula, Olena; Schumann, Canan; Duong, Tony; Taylor, Karmin L.; Taratula, Oleh

    2015-02-01

    Multifunctional theranostic platforms capable of concurrent near-infrared (NIR) fluorescence imaging and phototherapies are strongly desired for cancer diagnosis and treatment. However, the integration of separate imaging and therapeutic components into nanocarriers results in complex theranostic systems with limited translational potential. A single agent-based theranostic nanoplatform, therefore, was developed for concurrent NIR fluorescence imaging and combinatorial phototherapy with dual photodynamic (PDT) and photothermal (PTT) therapeutic mechanisms. The transformation of a substituted silicon naphthalocyanine (SiNc) into a biocompatible nanoplatform (SiNc-NP) was achieved by SiNc encapsulation into the hydrophobic interior of a generation 5 polypropylenimine dendrimer following surface modification with polyethylene glycol. Encapsulation provides aqueous solubility to SiNc and preserves its NIR fluorescence, PDT and PTT properties. Moreover, an impressive photostability in the dendrimer-encapsulated SiNc has been detected. Under NIR irradiation (785 nm, 1.3 W cm-2), SiNc-NP manifested robust heat generation capability (ΔT = 40 °C) and efficiently produced reactive oxygen species essential for PTT and PDT, respectively, without releasing SiNc from the nanopaltform. By varying the laser power density from 0.3 W cm-2 to 1.3 W cm-2 the therapeutic mechanism of SiNc-NP could be switched from PDT to combinatorial PDT-PTT treatment. In vitro and in vivo studies confirmed that phototherapy mediated by SiNc can efficiently destroy chemotherapy resistant ovarian cancer cells. Remarkably, solid tumors treated with a single dose of SiNc-NP combined with NIR irradiation were completely eradicated without cancer recurrence. Finally, the efficiency of SiNc-NP as an NIR imaging agent was confirmed by recording the strong fluorescence signal in the tumor, which was not photobleached during the phototherapeutic procedure.Multifunctional theranostic platforms capable of

  10. A closed-form method for single-point positioning with six satellites in dual-GNSS constellations

    Science.gov (United States)

    Teng, Yunlong; Huang, Qi; Ao, Yongcai; Li, Yun

    2016-12-01

    With the impact of the Global Navigation Satellite System (GNSS), dual-GNSS constellations are playing an increasingly significant role in positioning, navigation and timing (PNT) applications. Aiming at improving from the existing method, i.e., linearization, of solving the single-point positioning problem under a dual-GNSS, this paper develops a closed-form method for solving PNT problems in the case of six satellites. This method reduces the positioning problem to a simple mathematical problem of finding solutions to a quadratic equation, thereby needing only one receiver clock bias (RCB) as variable. By solving the RCB, the positioning information in three dimensions is obtained by utilizing a linear equation. Compared with the existing method, the closed-form method requires no initial position or iterations. This method thus provides a direct solution to single-point positioning. Further, how to check the uniqueness and the validity of the solutions is also derived. Experimental results verify the validity, applicability and efficiency of the proposed method.

  11. Dual-modal photoacoustic and optical coherence tomography using a single near-infrared supercontinuum laser source

    Science.gov (United States)

    Lee, Changho; Han, Seunghoon; Kim, Sehui; Jeon, Minyoung; Jeon, Mansik; Kim, Chulhong; Kim, Jeehyun

    2013-03-01

    We report the development of a combined dual-modal photoacoustic and optical coherence tomography (PA-OCT) system using a single near-infrared (NIR) supercontinuum laser source which can provide both optical absorption and scattering contrasts simultaneously. By using a small sized pulsed Nd:YAG microchip laser and a photonic crystal fiber, we fabricated a pulsed broadband supercontinuum source from 600 to 1700 nm. Under the same optical hardware system, intrinsically registered PA and OCT images are acquired in a single scanning. In order to demonstrate feasibility of our system, we successfully acquired the PA and OCT images of black and white hairs images at the same time. The black hair was detected in both PA and OCT images, while the white hair appeared only in the OCT image. This result suggests the potential of compact, cost-effective, and simple dual-modal PA-OCT system. Moreover, we believe that this approach will be a key point for commercialization and clinical translation.

  12. Donor-estimated GFR as an appropriate criterion for allocation of ECD kidneys into single or dual kidney transplantation.

    Science.gov (United States)

    Snanoudj, R; Rabant, M; Timsit, M O; Karras, A; Savoye, E; Tricot, L; Loupy, A; Hiesse, C; Zuber, J; Kreis, H; Martinez, F; Thervet, E; Méjean, A; Lebret, T; Legendre, C; Delahousse, M

    2009-11-01

    It has been suggested that dual kidney transplantation (DKT) improves outcomes for expanded criteria donor (ECD) kidneys. However, no criteria for allocation to single or dual transplantation have been assessed prospectively. The strategy of DKT remains underused and potentially eligible kidneys are frequently discarded. We prospectively compared 81 DKT and 70 single kidney transplant (SKT) receiving grafts from ECD donors aged >65 years, allocated according to donor estimated glomerular filtration rate (eGFR): DKT if eGFR between 30 and 60 mL/min, SKT if eGFR greater than 60 mL/min. Patient and graft survival were similar in the two groups. In the DKT group, 13/81 patients lost one of their two kidneys due to hemorrhage, arterial or venous thrombosis. Mean eGFR at month 12 was similar in the DKT and SKT groups (47.8 mL/min and 46.4 mL/min, respectively). Simulated allocation of kidneys according to criteria based on day 0 donor parameters such as those described by Remuzzi et al., Andres et al. and UNOS, did not indicate an improvement in 12-month eGFR compared to our allocation based on donor eGFR.

  13. SISTEM KONTROL OTOMATIK DENGAN MODEL SINGLE-INPUT-DUAL-OUTPUT DALAM KENDALI EFISIENSI UMUR-PEMAKAIAN INSTRUMEN

    Directory of Open Access Journals (Sweden)

    S.N.M.P. Simamora

    2014-10-01

    Full Text Available Efficiency condition occurs when the value of the used outputs compared to the resource total that has been used almost close to the value 1 (absolute environment. An instrument to achieve efficiency if the power output level has decreased significantly in the life of the instrument used, if it compared to the previous condition, when the instrument is not equipped with additional systems (or proposed model improvement. Even more effective if the inputs model that are used in unison to achieve a homogeneous output. On this research has been designed and implemented the automatic control system for models of single input-dual-output, wherein the sampling instruments used are lamp and fan. Source voltage used is AC (alternate-current and tested using quantitative research methods and instrumentation (with measuring instruments are observed. The results obtained demonstrate the efficiency of the instrument experienced a significant current model of single-input-dual-output applied separately instrument trials such as lamp and fan when it compared to the condition or state before. And the result show that the design has been built, can also run well.

  14. Dual-function photonic integrated circuit for frequency octo-tupling or single-side-band modulation.

    Science.gov (United States)

    Hasan, Mehedi; Maldonado-Basilio, Ramón; Hall, Trevor J

    2015-06-01

    A dual-function photonic integrated circuit for microwave photonic applications is proposed. The circuit consists of four linear electro-optic phase modulators connected optically in parallel within a generalized Mach-Zehnder interferometer architecture. The photonic circuit is arranged to have two separate output ports. A first port provides frequency up-conversion of a microwave signal from the electrical to the optical domain; equivalently single-side-band modulation. A second port provides tunable millimeter wave carriers by frequency octo-tupling of an appropriate amplitude RF carrier. The circuit exploits the intrinsic relative phases between the ports of multi-mode interference couplers to provide substantially all the static optical phases needed. The operation of the proposed dual-function photonic integrated circuit is verified by computer simulations. The performance of the frequency octo-tupling and up-conversion functions is analyzed in terms of the electrical signal to harmonic distortion ratio and the optical single side band to unwanted harmonics ratio, respectively.

  15. Single-polarization, switchable dual-wavelength erbium-doped fiber laser with two polarization-maintaining fiber Bragg gratings.

    Science.gov (United States)

    Feng, Suchun; Xu, Ou; Lu, Shaohua; Mao, Xiangqiao; Ning, Tigang; Jian, Shuisheng

    2008-08-04

    An improved erbium-doped fiber laser configuration for achieving single-polarization, switchable dual-wavelength of orthogonal polarizations oscillations at room temperature is proposed. For the first time, two fiber Bragg gratings (FBGs) directly written in a polarization-maintaining (PM) and photosensitive erbium-doped fiber (PMPEDF) as the wavelength-selective component are used in a linear laser cavity. Due to the polarization hole burning (PHB) enhanced by the polarization-maintaining FBG (PMFBG), the laser can be designed to operate in stable dual-wavelength or wavelength-switching modes with a wavelength spacing of 0.336 nm at room temperature by adjusting a polarization controller (PC). Each lasing line shows a single polarization with a polarization extinction ratio of >25 dB under different pump levels. The optical signal-to-noise ratio (OSNR) is greater than 50 dB. The amplitude variation with 16 times scans in nearly one and half an hour is less than 0.5 dB at both operating wavelength.

  16. Medium dependent dual turn on/turn off fluorescence sensing for Cu2 + ions using AMI/SDS assemblies

    Science.gov (United States)

    Gujar, Varsha B.; Ottoor, Divya

    2017-02-01

    Behavior of Amiloride (AMI) as a metal ion sensor in anionic surfactant assemblies of varying concentrations at different pH is depicted in this work. From a non-sensor fluorophore, AMI has been transformed in to a tunable fluorosensor for Cu2 + ions in various SDS concentrations. At premicellar concentration of SDS, ion-pair complex is expected to be formed between AMI and SDS due to electrostatic interactions between them. However at CMC concentrations of SDS, fluorescence intensity of AMI is greatly enhanced with red shift in emission, due to the incorporation of AMI molecule in the hydrophobic micellar interface. The behavior of metal sensing by AMI-SDS assemblies gives rise to several interesting observations. Micellation of SDS has been greatly enhanced by increasing copper ion concentrations, as these counter ions screens the charge on monomers of SDS which lead to the aggregation at premicellar concentrations only. Concentrations and pH dependent discrete trends of interactions between SDS-AMI and SDS-Cu2 + ions, have given tunable fluorescence responses (fluorescence turn on/turn off) of AMI for added Cu2 + ions. The electrostatic interaction between the metal cations and the anionic surfactants is the driving force for bringing the metal ions near to the vicinity of micelle where AMI resides. Thus, a comprehensive understanding of the mechanism related to the 'turn on-turn off' fluorescence response of AMI with respect to pH and SDS concentration for effective Cu2 + ion sensing is illustrated in this work.

  17. A versatile dual spot laser scanning confocal microscopy system for advanced fluorescence correlation spectroscopy analysis in living cell

    CERN Document Server

    Ferrand, P; Kress, A; Aillaud, A; Rigneault, H; Marguet, D

    2009-01-01

    A fluorescence correlation spectroscopy (FCS) system based on two independent measurement volumes is presented. The optical setup and data acquisition hardware are detailed, as well as a complete protocol to control the location, size and shape of the measurement volumes. A method that allows to monitor independently the excitation and collection efficiency distribution is proposed. Finally, a few examples of measurements that exploit the two spots in static and/or scanning schemes, are reported.

  18. Peering into Cells One Molecule at a Time: Single-molecule and plasmon-enhanced fluorescence super-resolution imaging

    Science.gov (United States)

    Biteen, Julie

    2013-03-01

    Single-molecule fluorescence brings the resolution of optical microscopy down to the nanometer scale, allowing us to unlock the mysteries of how biomolecules work together to achieve the complexity that is a cell. This high-resolution, non-destructive method for examining subcellular events has opened up an exciting new frontier: the study of macromolecular localization and dynamics in living cells. We have developed methods for single-molecule investigations of live bacterial cells, and have used these techniques to investigate thee important prokaryotic systems: membrane-bound transcription activation in Vibrio cholerae, carbohydrate catabolism in Bacteroides thetaiotaomicron, and DNA mismatch repair in Bacillus subtilis. Each system presents unique challenges, and we will discuss the important methods developed for each system. Furthermore, we use the plasmon modes of bio-compatible metal nanoparticles to enhance the emissivity of single-molecule fluorophores. The resolution of single-molecule imaging in cells is generally limited to 20-40 nm, far worse than the 1.5-nm localization accuracies which have been attained in vitro. We use plasmonics to improve the brightness and stability of single-molecule probes, and in particular fluorescent proteins, which are widely used for bio-imaging. We find that gold-coupled fluorophores demonstrate brighter, longer-lived emission, yielding an overall enhancement in total photons detected. Ultimately, this results in increased localization accuracy for single-molecule imaging. Furthermore, since fluorescence intensity is proportional to local electromagnetic field intensity, these changes in decay intensity and rate serve as a nm-scale read-out of the field intensity. Our work indicates that plasmonic substrates are uniquely advantageous for super-resolution imaging, and that plasmon-enhanced imaging is a promising technique for improving live cell single-molecule microscopy.

  19. Dual rotor single- stator axial air gap PMSM motor/generator drive for high torque vehicles applications

    Science.gov (United States)

    Tutelea, L. N.; Deaconu, S. I.; Boldea, I.; Popa, G. N.

    2014-03-01

    The actual e - continuously variable transmission (e-CVT) solution for the parallel Hybrid Electric Vehicle (HEV) requires two electric machines, two inverters, and a planetary gear. A distinct electric generator and a propulsion electric motor, both with full power converters, are typical for a series HEV. In an effort to simplify the planetary-geared e-CVT for the parallel HEV or the series HEV we hereby propose to replace the basically two electric machines and their two power converters by a single, axial-air-gap, electric machine central stator, fed from a single PWM converter with dual frequency voltage output and two independent PM rotors, destined for hybrid electric vehicles (HEV) and military vehicles applications. The proposed topologies and the magneto-motive force analysis are the core of the paper.

  20. Single-coil and dual-coil defibrillator leads and association with clinical outcomes in a complete Danish nationwide ICD cohort

    DEFF Research Database (Denmark)

    Larsen, Jacob M; Hjortshøj, Søren P; Nielsen, Jens C;

    2016-01-01

    BACKGROUND: The best choice of defibrillator lead in patients with routine implantable cardioverter-defibrillator (ICD) is not settled. Traditionally, most physicians prefer dual-coil leads but the use of single-coil leads is increasing. OBJECTIVE: The purpose of this study was to compare clinical...... outcomes in patients with single- and dual-coil leads. METHODS: All 4769 Danish patients 18 years or older with first-time ICD implants from 2007 to 2011 were included from the Danish Pacemaker and ICD Register. Defibrillator leads were 38.9% single-coil leads and 61.1% dual-coil leads. The primary end...... point was all-cause mortality. Secondary end points were lowest successful energy at implant defibrillation testing, first shock failure in spontaneous arrhythmias, structural lead failure, and lead extraction outcomes. RESULTS: Single-coil leads were associated with lower all-cause mortality...

  1. Development of a dual-modal tissue diagnostic system combining time-resolved fluorescence spectroscopy and ultrasonic backscatter microscopy

    Science.gov (United States)

    Sun, Yang; Park, Jesung; Stephens, Douglas N.; Jo, Javier A.; Sun, Lei; Cannata, Jonathan M.; Saroufeem, Ramez M. G.; Shung, K. Kirk; Marcu, Laura

    2009-06-01

    We report a tissue diagnostic system which combines two complementary techniques of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) and ultrasonic backscatter microscopy (UBM). TR-LIFS evaluates the biochemical composition of tissue, while UBM provides tissue microanatomy and enables localization of the region of diagnostic interest. The TR-LIFS component consists of an optical fiber-based time-domain apparatus including a spectrometer, gated multichannel plate photomultiplier, and fast digitizer. It records the fluorescence with high sensitivity (nM concentration range) and time resolution as low as 300 ps. The UBM system consists of a transducer, pulser, receiving circuit, and positioning stage. The transducer used here is 45 MHz, unfocused, with axial and lateral resolutions 38 and 200 μm. Validation of the hybrid system and ultrasonic and spectroscopic data coregistration were conducted both in vitro (tissue phantom) and ex vivo (atherosclerotic tissue specimens of human aorta). Standard histopathological analysis of tissue samples was used to validate the UBM-TRLIFS data. Current results have demonstrated that spatially correlated UBM and TR-LIFS data provide complementary characterization of both morphology (necrotic core and calcium deposits) and biochemistry (collagen, elastin, and lipid features) of the atherosclerotic plaques at the same location. Thus, a combination of fluorescence spectroscopy with ultrasound imaging would allow for better identification of features associated with tissue pathologies. Current design and performance of the hybrid system suggests potential applications in clinical diagnosis of atherosclerotic plaque.

  2. Magnetic/upconversion fluorescent NaGdF4:Yb,Er nanoparticle-based dual-modal molecular probes for imaging tiny tumors in vivo.

    Science.gov (United States)

    Liu, Chunyan; Gao, Zhenyu; Zeng, Jianfeng; Hou, Yi; Fang, Fang; Li, Yilin; Qiao, Ruirui; Shen, Lin; Lei, Hao; Yang, Wensheng; Gao, Mingyuan

    2013-08-27

    Detection of early malignant tumors remains clinically difficult; developing ultrasensitive imaging agents is therefore highly demanded. Owing to the unusual magnetic and optical properties associated with f-electrons, rare-earth elements are very suitable for creating functional materials potentially useful for tumor imaging. Nanometer-sized particles offer such a platform with which versatile unique properties of the rare-earth elements can be integrated. Yet the development of rare-earth nanoparticle-based tumor probes suitable for imaging tiny tumors in vivo remains difficult, which challenges not only the physical properties of the nanoparticles but also the rationality of the probe design. Here we report new approaches for size control synthesis of magnetic/upconversion fluorescent NaGdF4:Yb,Er nanocrystals and their applications for imaging tiny tumors in vivo. By independently varying F(-):Ln(3+) and Na(+):Ln(3+) ratios, the size and shape regulation mechanisms were investigated. By replacing the oleic acid ligand with PEG2000 bearing a maleimide group at one end and two phosphate groups at the other end, PEGylated NaGdF4:Yb,Er nanoparticles with optimized size and upconversion fluorescence were obtained. Accordingly, a dual-modality molecular tumor probe was prepared, as a proof of concept, by covalently attaching antitumor antibody to PEGylated NaGdF4:Yb,Er nanoparticles through a "click" reaction. Systematic investigations on tumor detections, through magnetic resonance imaging and upconversion fluorescence imaging, were carried out to image intraperitoneal tumors and subcutaneous tumors in vivo. Owing to the excellent properties of the molecular probes, tumors smaller than 2 mm was successfully imaged in vivo. In addition, pharmacokinetic studies on differently sized particles were performed to disclose the particle size dependent biodistributions and elimination pathways.

  3. Stress and gender effects on prefrontal cortex oxygenation levels assessed during single and dual-task walking conditions.

    Science.gov (United States)

    Holtzer, Roee; Schoen, Chelsea; Demetriou, Eleni; Mahoney, Jeannette R; Izzetoglu, Meltem; Wang, Cuiling; Verghese, Joe

    2017-03-01

    The ability to walk is critical for functional independence and wellbeing. The pre-frontal cortex (PFC) plays a key role in cognitive control of locomotion, notably under attention-demanding conditions. Factors that influence brain responses to cognitive demands of locomotion, however, are poorly understood. Herein, we evaluated the individual and combined effects of gender and perceived stress on stride velocity and PFC Oxygenated Hemoglobin (HbO2 ) assessed during single and dual-task walking conditions. The experimental paradigm included Normal Walk (NW); Cognitive Interference (Alpha); and Walk-While-Talk (WWT) tasks. An instrumented walkway was used to assess stride velocity in NW and WWT conditions. Functional Near-Infrared-Spectroscopy (fNIRS) was used to quantify PFC HbO2 levels during NW, Alpha and WWT. Perceived task-related stress was evaluated with a single 11-point scale item. Participants were community residing older adults (age = 76.8 ± 6.7 years; %female = 56). Results revealed that higher perceived stress was associated with greater decline in stride velocity from single to dual-task conditions among men. Three-way interactions revealed that gender moderated the effect of perceived stress on changes in HbO2 levels comparing WWT to NW and Alpha. Attenuation in the increase in HbO2 levels, in high compared to low perceived stress levels, from the two single task conditions to WWT was observed only in men. Thus, older men may be more vulnerable to the effect of perceived stress on the change in PFC oxygenation levels across walking conditions that vary in terms of cognitive demands. These findings confer important implications for assessment and treatment of individuals at risk of mobility impairments. © 2016 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

  4. Dual-wavelength single-longitudinal-mode erbium-doped fiber laser based on inverse-Gaussian apodized fiber Bragg grating and its application in microwave generation

    Science.gov (United States)

    Lin, Bo; Tjin, Swee Chuan; Zhang, Han; Tang, Dingyuan; Liang, Sheng; Hao, Jianzhong; Dong, Bo

    2011-03-01

    We propose a simple erbium-doped fiber ring laser. It consists of an inverse-Gaussian apodized fiber Bragg grating filter which has two ultra-narrow transmission bands, and an unpumped erbium-doped fiber as a saturable absorber. Stable