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Sample records for counting fluorescence lifetime

  1. Single photon counting fluorescence lifetime detection of pericellular oxygen concentrations.

    Science.gov (United States)

    Hosny, Neveen A; Lee, David A; Knight, Martin M

    2012-01-01

    Fluorescence lifetime imaging microscopy offers a non-invasive method for quantifying local oxygen concentrations. However, existing methods are either invasive, require custom-made systems, or show limited spatial resolution. Therefore, these methods are unsuitable for investigation of pericellular oxygen concentrations. This study describes an adaptation of commercially available equipment which has been optimized for quantitative extracellular oxygen detection with high lifetime accuracy and spatial resolution while avoiding systematic photon pile-up. The oxygen sensitive fluorescent dye, tris(2,2'-bipyridyl)ruthenium(II) chloride hexahydrate [Ru(bipy)(3)](2+), was excited using a two-photon excitation laser. Lifetime was measured using a Becker & Hickl time-correlated single photon counting, which will be referred to as a TCSPC card. [Ru(bipy)(3)](2+) characterization studies quantified the influences of temperature, pH, cellular culture media and oxygen on the fluorescence lifetime measurements. This provided a precisely calibrated and accurate system for quantification of pericellular oxygen concentration based on measured lifetimes. Using this technique, quantification of oxygen concentrations around isolated viable chondrocytes, seeded in three-dimensional agarose gel, revealed a subpopulation of cells that exhibited significant spatial oxygen gradients such that oxygen concentration reduced with increasing proximity to the cell. This technique provides a powerful tool for quantifying spatial oxygen gradients within three-dimensional cellular models.

  2. Gating circuit for single photon-counting fluorescence lifetime instruments using high repetition pulsed light sources

    International Nuclear Information System (INIS)

    Laws, W.R.; Potter, D.W.; Sutherland, J.C.

    1984-01-01

    We have constructed a circuit that permits conventional timing electronics to be used in single photon-counting fluorimeters with high repetition rate excitation sources (synchrotrons and mode-locked lasers). Most commercial time-to-amplitude and time-to-digital converters introduce errors when processing very short time intervals and when subjected to high-frequency signals. This circuit reduces the frequency of signals representing the pulsed light source (stops) to the rate of detected fluorescence events (starts). Precise timing between the start/stop pair is accomplished by using the second stop pulse after a start pulse. Important features of our design are that the circuit is insensitive to the simultaneous occurrence of start and stop signals and that the reduction in the stop frequency allows the start/stop time interval to be placed in linear regions of the response functions of commercial timing electronics

  3. Fluorescence lifetime imaging of skin cancer

    Science.gov (United States)

    Patalay, Rakesh; Talbot, Clifford; Munro, Ian; Breunig, Hans Georg; König, Karsten; Alexandrov, Yuri; Warren, Sean; Neil, Mark A. A.; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Chris

    2011-03-01

    Fluorescence intensity imaging and fluorescence lifetime imaging microscopy (FLIM) using two photon microscopy (TPM) have been used to study tissue autofluorescence in ex vivo skin cancer samples. A commercially available system (DermaInspect®) was modified to collect fluorescence intensity and lifetimes in two spectral channels using time correlated single photon counting and depth-resolved steady state measurements of the fluorescence emission spectrum. Uniquely, image segmentation has been used to allow fluorescence lifetimes to be calculated for each cell. An analysis of lifetime values obtained from a range of pigmented and non-pigmented lesions will be presented.

  4. Fluorescence lifetime based bioassays

    Science.gov (United States)

    Meyer-Almes, Franz-Josef

    2017-12-01

    Fluorescence lifetime (FLT) is a robust intrinsic property and material constant of fluorescent matter. Measuring this important physical indicator has evolved from a laboratory curiosity to a powerful and established technique for a variety of applications in drug discovery, medical diagnostics and basic biological research. This distinct trend was mainly driven by improved and meanwhile affordable laser and detection instrumentation on the one hand, and the development of suitable FLT probes and biological assays on the other. In this process two essential working approaches emerged. The first one is primarily focused on high throughput applications employing biochemical in vitro assays with no requirement for high spatial resolution. The second even more dynamic trend is the significant expansion of assay methods combining highly time and spatially resolved fluorescence data by fluorescence lifetime imaging. The latter approach is currently pursued to enable not only the investigation of immortal tumor cell lines, but also specific tissues or even organs in living animals. This review tries to give an actual overview about the current status of FLT based bioassays and the wide range of application opportunities in biomedical and life science areas. In addition, future trends of FLT technologies will be discussed.

  5. Time variation of fluorescence lifetime in enhanced cyan fluorescence protein

    International Nuclear Information System (INIS)

    Lee, Soonhyouk; Kim, Soo Yong; Park, Kyoungsook; Jeong, Jinyoung; Chung, Bong Hyun; Kim, Sok Won

    2010-01-01

    The lifetime variations of enhanced cyan fluorescence protein (ECFP) in relatively short integration time bins were studied via time-correlated single photon counting (TCSPC) measurement. We observed that minimum photon counts are necessary for the lifetime estimation to achieve a certain range of variance. The conditions to decrease the variance of lifetime were investigated and the channel width of the measurement of TCSPC data was found to be another important factor for the variance of lifetime. Though the lifetime of ECFP is best fit by a double exponential, a mono exponential fit for the same integration time is more stable. The results may be useful in the analysis of photophysical dynamics for ensemble molecules in short measurement time windows.

  6. Laser induced fluorescence lifetime characterization of Bacillus endospore species using time correlated single photon counting analysis with the multi-exponential fit method

    Science.gov (United States)

    Smith, Clint; Edwards, Jarrod; Fisher, Andmorgan

    2010-04-01

    Rapid detection of biological material is critical for determining presence/absence of bacterial endospores within various investigative programs. Even more critical is that if select material tests positive for bacillus endospores then tests should provide data at the species level. Optical detection of microbial endospore formers such as Bacillus sp. can be heavy, cumbersome, and may only identify at the genus level. Data provided from this study will aid in characterization needed by future detection systems for further rapid breakdown analysis to gain insight into a more positive signature collection of Bacillus sp. Literature has shown that fluorescence spectroscopy of endospores could be statistically separated from other vegetative genera, but could not be separated among one another. Results of this study showed endospore species separation is possible using laser-induce fluorescence with lifetime decay analysis for Bacillus endospores. Lifetime decays of B. subtilis, B. megaterium, B. coagulans, and B. anthracis Sterne strain were investigated. Using the Multi-Exponential fit method data showed three distinct lifetimes for each species within the following ranges, 0.2-1.3 ns; 2.5-7.0 ns; 7.5-15.0 ns, when laser induced at 307 nm. The four endospore species were individually separated using principle component analysis (95% CI).

  7. Three-dimensional fluorescence lifetime tomography

    International Nuclear Information System (INIS)

    Godavarty, Anuradha; Sevick-Muraca, Eva M.; Eppstein, Margaret J.

    2005-01-01

    Near-infrared fluorescence tomography using molecularly targeted lifetime-sensitive, fluorescent contrast agents have applications for early-stage cancer diagnostics. Yet, although the measurement of fluorescent lifetime imaging microscopy (FLIM) is extensively used in microscopy and spectroscopy applications, demonstration of fluorescence lifetime tomography for medical imaging is limited to two-dimensional studies. Herein, the feasibility of three-dimensional fluorescence-lifetime tomography on clinically relevant phantom volumes is established, using (i) a gain-modulated intensified charge coupled device (CCD) and modulated laser diode imaging system, (ii) two fluorescent contrast agents, e.g., Indocyanine green and 3-3'-Diethylthiatricarbocyanine iodide differing in their fluorescence lifetime by 0.62 ns, and (iii) a two stage approximate extended Kalman filter reconstruction algorithm. Fluorescence measurements of phase and amplitude were acquired on the phantom surface under different target to background fluorescence absorption (70:1, 100:1) and fluorescence lifetime (1:1, 2.1:1) contrasts at target depths of 1.4-2 cm. The Bayesian tomography algorithm was employed to obtain three-dimensional images of lifetime and absorption owing to the fluorophores

  8. Fluorescence lifetime imaging using light emitting diodes

    Energy Technology Data Exchange (ETDEWEB)

    Kennedy, Gordon T; Munro, Ian; Poher, Vincent; French, Paul M W; Neil, Mark A A [Blackett Laboratory, Imperial College London, South Kensington Campus, London SW7 2AZ (United Kingdom); Elson, Daniel S [Institute of Biomedical Engineering, Imperial College London, South Kensington Campus, London SW7 2AZ (United Kingdom); Hares, Jonathan D [Kentech Instruments Ltd, Unit 9, Hall Farm Workshops, South Moreton, Didcot, Oxfordshire, OX11 9AG (United Kingdom)], E-mail: gordon.kennedy@imperial.ac.uk

    2008-05-07

    We demonstrate flexible use of low cost, high-power light emitting diodes as illumination sources for fluorescence lifetime imaging (FLIM). Both time-domain and frequency-domain techniques have been implemented at wavelengths spanning the range 450-640 nm. Additionally, we demonstrate optically sectioned fluorescence lifetime imaging by combining structured illumination with frequency-domain FLIM.

  9. Clinical results of fluorescence lifetime imaging in ophthalmology

    Science.gov (United States)

    Schweitzer, D.; Quick, S.; Klemm, M.; Hammer, M.; Jentsch, S.; Dawczynski, J.; Becker, W.

    2009-07-01

    A laser scanner ophthalmoscope was developed for in vivo fluorescence lifetime measurements at the human retina. Measurements were performed in 30 degree fundus images. The fundus was excited by pulses of 75 ps (FWHM). The dynamic fluorescence was detected in two spectral channels K1(490-560nm), K2(560-700 nm) by time-correlated single photon counting. The decay of fluorescence was three-exponentially. Local and global alterations in lifetimes were found between healthy subjects and patients suffering from age-related macular degeneration, diabetic retinopathy, and vessel occlusion. The lifetimes T1, T2, and T3 in both channels are changed to longer values in AMD and diabetic retinopathy in comparison with healthy subjects. The lifetime T2 in K1 is most sensitive to metabolic alterations in branch arterial vessel occlusion.

  10. Remote UV Fluorescence Lifetime Spectrometer, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — The goal of this project is to develop, demonstrate, and deliver to NASA an innovative, portable, and power efficient Remote UV Fluorescence Lifetime Spectrometer...

  11. Fluorescence lifetime imaging microscopy using near-infrared contrast agents.

    Science.gov (United States)

    Nothdurft, R; Sarder, P; Bloch, S; Culver, J; Achilefu, S

    2012-08-01

    Although single-photon fluorescence lifetime imaging microscopy (FLIM) is widely used to image molecular processes using a wide range of excitation wavelengths, the captured emission of this technique is confined to the visible spectrum. Here, we explore the feasibility of utilizing near-infrared (NIR) fluorescent molecular probes with emission >700 nm for FLIM of live cells. The confocal microscope is equipped with a 785 nm laser diode, a red-enhanced photomultiplier tube, and a time-correlated single photon counting card. We demonstrate that our system reports the lifetime distributions of NIR fluorescent dyes, cypate and DTTCI, in cells. In cells labelled separately or jointly with these dyes, NIR FLIM successfully distinguishes their lifetimes, providing a method to sort different cell populations. In addition, lifetime distributions of cells co-incubated with these dyes allow estimate of the dyes' relative concentrations in complex cellular microenvironments. With the heightened interest in fluorescence lifetime-based small animal imaging using NIR fluorophores, this technique further serves as a bridge between in vitro spectroscopic characterization of new fluorophore lifetimes and in vivo tissue imaging. © 2012 The Author Journal of Microscopy © 2012 Royal Microscopical Society.

  12. Fluorescence lifetime measurement of radical ions

    International Nuclear Information System (INIS)

    Ichinose, Nobuyuki; Kinugasa, Jun-ichiro; Hagiri, Masahide; Nakayama, Toshihiro; Murakami, Hiroshi; Kishimoto, Maki; Daido, Hiroyuki

    2004-01-01

    One-photonic excitation of a charge transfer complex of hexamethoxybenzene (HMB) and nitrosonium tetrafluoroborate (NO + BF 4 - ) in acetonitrile afforded fluorescences emission from excited radical cation of HMB (HMB + *). Lifetime of the excited radical ion species was measured to be 7 ps by the pump-probe transient absorption technique. The lifetime was much shorter than that of free radical ion (63 ps), indicating the presence of an interaction between HMB + * and NO in the excited complex. (author)

  13. Two-dimensional fluorescence lifetime correlation spectroscopy. 2. Application.

    Science.gov (United States)

    Ishii, Kunihiko; Tahara, Tahei

    2013-10-03

    In the preceding article, we introduced the theoretical framework of two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS). In this article, we report the experimental implementation of 2D FLCS. In this method, two-dimensional emission-delay correlation maps are constructed from the photon data obtained with the time-correlated single photon counting (TCSPC), and then they are converted to 2D lifetime correlation maps by the inverse Laplace transform. We develop a numerical method to realize reliable transformation, employing the maximum entropy method (MEM). We apply the developed actual 2D FLCS to two real systems, a dye mixture and a DNA hairpin. For the dye mixture, we show that 2D FLCS is experimentally feasible and that it can identify different species in an inhomogeneous sample without any prior knowledge. The application to the DNA hairpin demonstrates that 2D FLCS can disclose microsecond spontaneous dynamics of biological molecules in a visually comprehensible manner, through identifying species as unique lifetime distributions. A FRET pair is attached to the both ends of the DNA hairpin, and the different structures of the DNA hairpin are distinguished as different fluorescence lifetimes in 2D FLCS. By constructing the 2D correlation maps of the fluorescence lifetime of the FRET donor, the equilibrium dynamics between the open and the closed forms of the DNA hairpin is clearly observed as the appearance of the cross peaks between the corresponding fluorescence lifetimes. This equilibrium dynamics of the DNA hairpin is clearly separated from the acceptor-missing DNA that appears as an isolated diagonal peak in the 2D maps. The present study clearly shows that newly developed 2D FLCS can disclose spontaneous structural dynamics of biological molecules with microsecond time resolution.

  14. Application of time-correlated single photon counting and stroboscopic detection methods with an evanescent-wave fibre-optic sensor for fluorescence-lifetime-based pH measurements

    International Nuclear Information System (INIS)

    Henning, Paul E; Geissinger, Peter

    2012-01-01

    Quasi-distributed optical fibre sensor arrays containing luminescent sensor molecules can be read out spatially resolved utilizing optical time-of-flight detection (OTOFD) methods, which employ pulsed laser interrogation of the luminosensors and time-resolved detection of the sensor signals. In many cases, sensing is based on a change in sensor luminescence intensity; however, sensing based on luminescence lifetime changes is preferable because it reduces the need for field calibration. Because in OTOFD detection is time-resolved, luminescence-lifetime information is already available through the signal pulses, although in practise applications were restricted to sensors with long luminescence lifetimes (hundreds of ns). To implement lifetime-based sensing in crossed-optical-fibre-sensor arrays for sensor molecules with lifetimes less than 10 ns, two time-domain methods, time-correlated single photon counting and stroboscopic detection, were used to record the pH-dependent emission of a fluorescein derivative covalently attached to a highly-porous polymer. A two-term nonexponential decay function yielded both a good fit for experimental lifetime data during reconvolution and a pH response that matches Henderson–Hasselbalch behaviour, yielding a sensor accuracy of 0.02 pH units. Moreover, strong agreement was obtained for the two lifetime determination methods and with intensity-based measurements taken previously. (paper)

  15. Non-invasive imaging of skin cancer with fluorescence lifetime imaging using two photon tomography

    Science.gov (United States)

    Patalay, Rakesh; Talbot, Clifford; Alexandrov, Yuriy; Munro, Ian; Breunig, Hans Georg; König, Karsten; Warren, Sean; Neil, Mark A. A.; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Christopher

    2011-07-01

    Multispectral fluorescence lifetime imaging (FLIM) using two photon microscopy as a non-invasive technique for the diagnosis of skin lesions is described. Skin contains fluorophores including elastin, keratin, collagen, FAD and NADH. This endogenous contrast allows tissue to be imaged without the addition of exogenous agents and allows the in vivo state of cells and tissues to be studied. A modified DermaInspect® multiphoton tomography system was used to excite autofluorescence at 760 nm in vivo and on freshly excised ex vivo tissue. This instrument simultaneously acquires fluorescence lifetime images in four spectral channels between 360-655 nm using time-correlated single photon counting and can also provide hyperspectral images. The multispectral fluorescence lifetime images were spatially segmented and binned to determine lifetimes for each cell by fitting to a double exponential lifetime model. A comparative analysis between the cellular lifetimes from different diagnoses demonstrates significant diagnostic potential.

  16. A measurement of the neutron lifetime by counting trapped protons

    CERN Document Server

    Snow, W M; Dewey, M S; Fei, X; Gilliam, D M; Greene, G L; Nico, J S; Wietfeldt, F E

    2000-01-01

    A measurement of the neutron lifetime tau sub n performed by trapping and counting decay protons from in-beam neutron decays in a Penning trap is in progress at the National Institute of Standards and Technology (NIST). A description of the measurement technique, the status of the data analysis, and prospects for improvements in the measurement are discussed.

  17. Theoretical lifetimes and fluorescence yields for multiply-ionized fluorine

    International Nuclear Information System (INIS)

    Tunnell, T.W.; Can, C.; Bhalla, C.P.

    1978-01-01

    Theoretical lifetimes and multiplet partial fluorescence yields for various fluorine ions with a single K-shell vacancy were calculated. For few-electron systems, the lifetimes and line fluorescence yields were computed in the intermediate coupling scheme with the inclusion of the effects arising from configuration interactions. 6 references

  18. Measurement of the neutron lifetime by counting trapped protons

    International Nuclear Information System (INIS)

    Byrne, J.; Dawber, P.G.; Spain, J.A.; Williams, A.P.; Dewey, M.S.; Gilliam, D.M.; Greene, G.L.; Lamaze, G.P.; Scott, R.D.; Pauwels, J.; Eykens, R.; Lamberty, A.

    1990-01-01

    The neutron lifetime τ n has been measured by counting decay protons stored in a Penning trap whose magnetic axis coincided with a neutron-beam axis. The result of the measurement is τ n =893.6±5.3 s, which agrees well with the value predicted by precise measurements of the β-decay asymmetry parameter A and the standard model

  19. Towards sensitive, high-throughput, biomolecular assays based on fluorescence lifetime

    Science.gov (United States)

    Ioanna Skilitsi, Anastasia; Turko, Timothé; Cianfarani, Damien; Barre, Sophie; Uhring, Wilfried; Hassiepen, Ulrich; Léonard, Jérémie

    2017-09-01

    Time-resolved fluorescence detection for robust sensing of biomolecular interactions is developed by implementing time-correlated single photon counting in high-throughput conditions. Droplet microfluidics is used as a promising platform for the very fast handling of low-volume samples. We illustrate the potential of this very sensitive and cost-effective technology in the context of an enzymatic activity assay based on fluorescently-labeled biomolecules. Fluorescence lifetime detection by time-correlated single photon counting is shown to enable reliable discrimination between positive and negative control samples at a throughput as high as several hundred samples per second.

  20. Quantitative analysis of fluorescence lifetime measurements of the macula using the fluorescence lifetime imaging ophthalmoscope in healthy subjects.

    Science.gov (United States)

    Dysli, Chantal; Quellec, Gwénolé; Abegg, Mathias; Menke, Marcel N; Wolf-Schnurrbusch, Ute; Kowal, Jens; Blatz, Johannes; La Schiazza, Olivier; Leichtle, Alexander B; Wolf, Sebastian; Zinkernagel, Martin S

    2014-04-03

    Fundus autofluorescence (FAF) cannot only be characterized by the intensity or the emission spectrum, but also by its lifetime. As the lifetime of a fluorescent molecule is sensitive to its local microenvironment, this technique may provide more information than fundus autofluorescence imaging. We report here the characteristics and repeatability of FAF lifetime measurements of the human macula using a new fluorescence lifetime imaging ophthalmoscope (FLIO). A total of 31 healthy phakic subjects were included in this study with an age range from 22 to 61 years. For image acquisition, a fluorescence lifetime ophthalmoscope based on a Heidelberg Engineering Spectralis system was used. Fluorescence lifetime maps of the retina were recorded in a short- (498-560 nm) and a long- (560-720 nm) spectral channel. For quantification of fluorescence lifetimes a standard ETDRS grid was used. Mean fluorescence lifetimes were shortest in the fovea, with 208 picoseconds for the short-spectral channel and 239 picoseconds for the long-spectral channel, respectively. Fluorescence lifetimes increased from the central area to the outer ring of the ETDRS grid. The test-retest reliability of FLIO was very high for all ETDRS areas (Spearman's ρ = 0.80 for the short- and 0.97 for the long-spectral channel, P macula in healthy subjects. By using a custom-built software, we were able to quantify fluorescence lifetimes within the ETDRS grid. Establishing a clinically accessible standard against which to measure FAF lifetimes within the retina is a prerequisite for future studies in retinal disease.

  1. Fluorescence lifetime assays: current advances and applications in drug discovery.

    Science.gov (United States)

    Pritz, Stephan; Doering, Klaus; Woelcke, Julian; Hassiepen, Ulrich

    2011-06-01

    Fluorescence lifetime assays complement the portfolio of established assay formats available in drug discovery, particularly with the recent advances in microplate readers and the commercial availability of novel fluorescent labels. Fluorescence lifetime assists in lowering complexity of compound screening assays, affording a modular, toolbox-like approach to assay development and yielding robust homogeneous assays. To date, materials and procedures have been reported for biochemical assays on proteases, as well as on protein kinases and phosphatases. This article gives an overview of two assay families, distinguished by the origin of the fluorescence signal modulation. The pharmaceutical industry demands techniques with a robust, integrated compound profiling process and short turnaround times. Fluorescence lifetime assays have already helped the drug discovery field, in this sense, by enhancing productivity during the hit-to-lead and lead optimization phases. Future work will focus on covering other biochemical molecular modifications by investigating the detailed photo-physical mechanisms underlying the fluorescence signal.

  2. Fluorescence lifetime imaging of oxygen in dental biofilm

    Science.gov (United States)

    Gerritsen, Hans C.; de Grauw, Cees J.

    2000-12-01

    Dental biofilm consists of micro-colonies of bacteria embedded in a matrix of polysaccharides and salivary proteins. pH and oxygen concentration are of great importance in dental biofilm. Both can be measured using fluorescence techniques. The imaging of dental biofilm is complicated by the thickness of the biofilms that can be up to several hundred micrometers thick. Here, we employed a combination of two-photon excitation microscopy with fluorescence lifetime imaging to quantify the oxygen concentration in dental biofilm. Collisional quenching of fluorescent probes by molecular oxygen leads to a reduction of the fluorescence lifetime of the probe. We employed this mechanism to measure the oxygen concentration distribution in dental biofilm by means of fluorescence lifetime imaging. Here, TRIS Ruthenium chloride hydrate was used as an oxygen probe. A calibration procedure on buffers was use to measure the lifetime response of this Ruthenium probe. The results are in agreement with the Stern-Volmer equation. A linear relation was found between the ratio of the unquenched and the quenched lifetime and the oxygen concentration. The biofilm fluorescence lifetime imaging results show a strong oxygen gradient at the buffer - biofilm interface and the average oxygen concentration in the biofilm amounted to 50 μM.

  3. Refractive index sensing using Fluorescence Lifetime Imaging (FLIM)

    International Nuclear Information System (INIS)

    Jones, Carolyn; Suhling, Klaus

    2006-01-01

    The fluorescence lifetime is a function of the refractive index of the fluorophore's environment, for example in the case of the biologically important green fluorescent protein (GFP). In order to address the question whether this effect can be exploited to image the local environment of specific proteins in cell biology, we need to determine the distance over which the fluorophore's lifetime is sensitive to the refractive index. To this end, we employ Fluorescence Lifetime Imaging (FLIM) of fluorescein in NaOH buffer at an interface. This approach allows us to map the fluorescence lifetime as a function of distance from a buffer/air and buffer/oil interface. Preliminary data show that the fluorescence lifetime of fluorescein increases near a buffer/air interface and decreases near a buffer/oil interface. The range over which this fluorescence lifetime change occurs is found to be of the order several μm which is consistent with a theoretical model based on the full width at half maximum of the emission spectrum proposed by Toptygin

  4. Fluorescence lifetime evaluation of whole soils from the Amazon rainforest.

    Science.gov (United States)

    Nicolodelli, Gustavo; Tadini, Amanda Maria; Nogueira, Marcelo Saito; Pratavieira, Sebastião; Mounier, Stephane; Huaman, Jose Luis Clabel; Dos Santos, Cléber Hilário; Montes, Célia Regina; Milori, Débora Marcondes Bastos Pereira

    2017-08-20

    Time-resolved fluorescence spectroscopy (TRFS) is a new tool that can be used to investigate processes of interaction between metal ions and organic matter (OM) in soils, providing a specific analysis of the structure and dynamics of macromolecules. To the best of our knowledge, there are no studies in the literature reporting the use of this technique applied to whole/non-fractionated soil samples, making it a potential method for use in future studies. This work describes the use of TRFS to evaluate the fluorescence lifetimes of OM of whole soils from the Amazon region. Analysis was made of pellets of soils from an oxisol-spodosol system, collected in São Gabriel da Cachoeira (Amazonas, Brazil). The fluorescence lifetimes in the oxisol-spodosol system were attributed to two different fluorophores. One was related to complexation of an OM fraction with metals, resulting in a shorter fluorophore lifetime. A short fluorescence lifetime (2-12 ns) could be associated with simpler structures of the OM, while a long lifetime (19-66 ns) was associated with more complex OM structures. This new TRFS technique for analysis of the fluorescence lifetime in whole soil samples complies with the principles of green chemistry.

  5. Fluorescence decay data analysis correcting for detector pulse pile-up at very high count rates

    Science.gov (United States)

    Patting, Matthias; Reisch, Paja; Sackrow, Marcus; Dowler, Rhys; Koenig, Marcelle; Wahl, Michael

    2018-03-01

    Using time-correlated single photon counting for the purpose of fluorescence lifetime measurements is usually limited in speed due to pile-up. With modern instrumentation, this limitation can be lifted significantly, but some artifacts due to frequent merging of closely spaced detector pulses (detector pulse pile-up) remain an issue to be addressed. We propose a data analysis method correcting for this type of artifact and the resulting systematic errors. It physically models the photon losses due to detector pulse pile-up and incorporates the loss in the decay fit model employed to obtain fluorescence lifetimes and relative amplitudes of the decay components. Comparison of results with and without this correction shows a significant reduction of systematic errors at count rates approaching the excitation rate. This allows quantitatively accurate fluorescence lifetime imaging at very high frame rates.

  6. Multiphoton fluorescence lifetime imaging of chemotherapy distribution in solid tumors

    Science.gov (United States)

    Carlson, Marjorie; Watson, Adrienne L.; Anderson, Leah; Largaespada, David A.; Provenzano, Paolo P.

    2017-11-01

    Doxorubicin is a commonly used chemotherapeutic employed to treat multiple human cancers, including numerous sarcomas and carcinomas. Furthermore, doxorubicin possesses strong fluorescent properties that make it an ideal reagent for modeling drug delivery by examining its distribution in cells and tissues. However, while doxorubicin fluorescence and lifetime have been imaged in live tissue, its behavior in archival samples that frequently result from drug and treatment studies in human and animal patients, and murine models of human cancer, has to date been largely unexplored. Here, we demonstrate imaging of doxorubicin intensity and lifetimes in archival formalin-fixed paraffin-embedded sections from mouse models of human cancer with multiphoton excitation and multiphoton fluorescence lifetime imaging microscopy (FLIM). Multiphoton excitation imaging reveals robust doxorubicin emission in tissue sections and captures spatial heterogeneity in cells and tissues. However, quantifying the amount of doxorubicin signal in distinct cell compartments, particularly the nucleus, often remains challenging due to strong signals in multiple compartments. The addition of FLIM analysis to display the spatial distribution of excited state lifetimes clearly distinguishes between signals in distinct compartments such as the cell nuclei versus cytoplasm and allows for quantification of doxorubicin signal in each compartment. Furthermore, we observed a shift in lifetime values in the nuclei of transformed cells versus nontransformed cells, suggesting a possible diagnostic role for doxorubicin lifetime imaging to distinguish normal versus transformed cells. Thus, data here demonstrate that multiphoton FLIM is a highly sensitive platform for imaging doxorubicin distribution in normal and diseased archival tissues.

  7. Mesh adaptation technique for Fourier-domain fluorescence lifetime imaging

    International Nuclear Information System (INIS)

    Soloviev, Vadim Y.

    2006-01-01

    A novel adaptive mesh technique in the Fourier domain is introduced for problems in fluorescence lifetime imaging. A dynamical adaptation of the three-dimensional scheme based on the finite volume formulation reduces computational time and balances the ill-posed nature of the inverse problem. Light propagation in the medium is modeled by the telegraph equation, while the lifetime reconstruction algorithm is derived from the Fredholm integral equation of the first kind. Stability and computational efficiency of the method are demonstrated by image reconstruction of two spherical fluorescent objects embedded in a tissue phantom

  8. Measuring and sorting cell populations expressing isospectral fluorescent proteins with different fluorescence lifetimes.

    Directory of Open Access Journals (Sweden)

    Bryan Sands

    Full Text Available Study of signal transduction in live cells benefits from the ability to visualize and quantify light emitted by fluorescent proteins (XFPs fused to different signaling proteins. However, because cell signaling proteins are often present in small numbers, and because the XFPs themselves are poor fluorophores, the amount of emitted light, and the observable signal in these studies, is often small. An XFP's fluorescence lifetime contains additional information about the immediate environment of the fluorophore that can augment the information from its weak light signal. Here, we constructed and expressed in Saccharomyces cerevisiae variants of Teal Fluorescent Protein (TFP and Citrine that were isospectral but had shorter fluorescence lifetimes, ∼ 1.5 ns vs ∼ 3 ns. We modified microscopic and flow cytometric instruments to measure fluorescence lifetimes in live cells. We developed digital hardware and a measure of lifetime called a "pseudophasor" that we could compute quickly enough to permit sorting by lifetime in flow. We used these abilities to sort mixtures of cells expressing TFP and the short-lifetime TFP variant into subpopulations that were respectively 97% and 94% pure. This work demonstrates the feasibility of using information about fluorescence lifetime to help quantify cell signaling in living cells at the high throughput provided by flow cytometry. Moreover, it demonstrates the feasibility of isolating and recovering subpopulations of cells with different XFP lifetimes for subsequent experimentation.

  9. Nanoantenna array-induced fluorescence enhancement and reduced lifetimes

    DEFF Research Database (Denmark)

    Bakker, R. M.; Drachev, V. P.; Liu, Z.

    2008-01-01

    Enhanced fluorescence is observed from dye molecules interacting with optical nanoantenna arrays. Elliptical gold dimers form individual nanoantennae with tunable plasmon resonances depending upon the geometry of the two particles and the size of the gap between them. A fluorescent dye, Rhodamine...... 800, is uniformly embedded in a dielectric host that coats the nanoantennae. The nanoantennae act to enhance the dye absorption. In turn, emission from the dye drives the plasmon resonance of the antennae; the nanoantennae act to enhance the fluorescence signal and change the angular distribution...... of emission. These effects depend upon the overlap of the plasmon resonance with the excitation wavelength and the fluorescence emission band. A decreased fluorescence lifetime is observed along with highly polarized emission that displays the characteristics of the nanoantenna's dipole mode. Being able...

  10. In vivo imaging of cerebral energy metabolism with two-photon fluorescence lifetime microscopy of NADH.

    Science.gov (United States)

    Yaseen, Mohammad A; Sakadžić, Sava; Wu, Weicheng; Becker, Wolfgang; Kasischke, Karl A; Boas, David A

    2013-02-01

    Minimally invasive, specific measurement of cellular energy metabolism is crucial for understanding cerebral pathophysiology. Here, we present high-resolution, in vivo observations of autofluorescence lifetime as a biomarker of cerebral energy metabolism in exposed rat cortices. We describe a customized two-photon imaging system with time correlated single photon counting detection and specialized software for modeling multiple-component fits of fluorescence decay and monitoring their transient behaviors. In vivo cerebral NADH fluorescence suggests the presence of four distinct components, which respond differently to brief periods of anoxia and likely indicate different enzymatic formulations. Individual components show potential as indicators of specific molecular pathways involved in oxidative metabolism.

  11. Fluorescence Lifetime Correlation Spectroscopy (FLCS): Concepts, Applications and Outlook

    Czech Academy of Sciences Publication Activity Database

    Kapusta, Peter; Macháň, Radek; Benda, A.; Hof, Martin

    2012-01-01

    Roč. 13, č. 10 (2012), s. 12890-12910 E-ISSN 1422-0067 R&D Projects: GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : fluorescence correlation spectroscopy (FCS) * time correlated single photon counting (TCSPC) * fluorescence cross-correlation spectroscopy (FCCS) Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.464, year: 2012

  12. Fluorescence detection of single molecules using pulsed near-field optical excitation and time correlated photon counting

    International Nuclear Information System (INIS)

    Ambrose, W.P.; Goodwin, P.M.; Martin, J.C.; Keller, R.A.

    1994-01-01

    Pulsed excitation, time correlated single photon counting and time gated detection are used in near-field optical microscopy to enhance fluorescence images and measure the fluorescence lifetimes of single molecules of Rhodamine 6G on silica surfaces. Time gated detection is used to reject prompt scattered background and to improve the image signal to noise ratio. The excited state lifetime of a single Rhodamine 6G molecule is found to depend on the position of the near-field probe. We attribute the lifetime variations to spontaneous emission rate alterations by the fluorescence reflected from and quenching by the aluminum coated probe

  13. Refractive index sensing of green fluorescent proteins in living cells using fluorescence lifetime imaging microscopy

    NARCIS (Netherlands)

    van Manen, Henk-Jan; Verkuijlen, Paul; Wittendorp, Paul; Subramaniam, Vinod; van den Berg, Timo K; Roos, Dirk; Otto, Cees

    2008-01-01

    We show that fluorescence lifetime imaging microscopy (FLIM) of green fluorescent protein (GFP) molecules in cells can be used to report on the local refractive index of intracellular GFP. We expressed GFP fusion constructs of Rac2 and gp91(phox), which are both subunits of the phagocyte NADPH

  14. Azadioxatriangulenium: exploring the effect of a 20 ns fluorescence lifetime in fluorescence anisotropy measurements

    Science.gov (United States)

    Bogh, Sidsel A.; Bora, Ilkay; Rosenberg, Martin; Thyrhaug, Erling; Laursen, Bo W.; Just Sørensen, Thomas

    2015-12-01

    Azaoxatriangulenium (ADOTA) has been shown to be highly emissive despite a moderate molar absorption coefficient of the primary electronic transition. As a result, the fluorescence lifetime is ~20 ns, longer than all commonly used red fluorescent organic probes. The electronic transitions in ADOTA are highly polarised (r 0  =  0.38), which in combination with the long fluorescence lifetime extents the size-range of biomolecular weights that can be detected in fluorescence polarisation-based experiments. Here, the rotational dynamics of bovine serum albumin (BSA) are monitored with three different ADOTA derivatives, differing only in constitution of the reactive linker. A detailed study of the degree of labelling, the steady-state anisotropy, and the time-resolved anisotropy of the three different ADOTA-BSA conjugates are reported. The fluorescence quantum yields (ϕ fl) of the free dyes in PBS solution are determined to be ~55%, which is reduced to ~20% in the ADOTA-BSA conjugates. Despite the reduction in ϕ fl, a ~20 ns intensity averaged lifetime is maintained, allowing for the rotational dynamics of BSA to be monitored for up to 100 ns. Thus, ADOTA can be used in fluorescence polarisation assays to fill the gap between commonly used organic dyes and the long luminescence lifetime transition metal complexes. This allows for efficient steady-state fluorescence polarisation assays for detecting binding of analytes with molecular weights of up to 100 kDa.

  15. Modulated electron-multiplied fluorescence lifetime imaging microscope: all-solid-state camera for fluorescence lifetime imaging.

    Science.gov (United States)

    Zhao, Qiaole; Schelen, Ben; Schouten, Raymond; van den Oever, Rein; Leenen, René; van Kuijk, Harry; Peters, Inge; Polderdijk, Frank; Bosiers, Jan; Raspe, Marcel; Jalink, Kees; Geert Sander de Jong, Jan; van Geest, Bert; Stoop, Karel; Young, Ian Ted

    2012-12-01

    We have built an all-solid-state camera that is directly modulated at the pixel level for frequency-domain fluorescence lifetime imaging microscopy (FLIM) measurements. This novel camera eliminates the need for an image intensifier through the use of an application-specific charge coupled device design in a frequency-domain FLIM system. The first stage of evaluation for the camera has been carried out. Camera characteristics such as noise distribution, dark current influence, camera gain, sampling density, sensitivity, linearity of photometric response, and optical transfer function have been studied through experiments. We are able to do lifetime measurement using our modulated, electron-multiplied fluorescence lifetime imaging microscope (MEM-FLIM) camera for various objects, e.g., fluorescein solution, fixed green fluorescent protein (GFP) cells, and GFP-actin stained live cells. A detailed comparison of a conventional microchannel plate (MCP)-based FLIM system and the MEM-FLIM system is presented. The MEM-FLIM camera shows higher resolution and a better image quality. The MEM-FLIM camera provides a new opportunity for performing frequency-domain FLIM.

  16. Bessel beam fluorescence lifetime tomography of live embryos (Conference Presentation)

    Science.gov (United States)

    Xu, Dongli; Peng, Leilei

    2016-03-01

    Optical tomography allows isotropic 3D imaging of embryos. Scanning-laser optical tomography (SLOT) has superior light collecting efficiency than wide-field optical tomography, making it ideal for fluorescence imaging of live embryos. We previously reported an imaging system that combines SLOT with a novel Fourier-multiplexed fluorescence lifetime imaging (FmFLIM) technique named FmFLIM-SLOT. FmFLIM-SLOT performs multiplexed FLIM-FRET readout of multiple FRET sensors in live embryos. Here we report a recent effort on improving the spatial resolution of the FmFLIM-SLOT system in order to image complex biochemical processes in live embryos at the cellular level. Optical tomography has to compromise between resolution and the depth of view. In SLOT, the commonly-used focused Gaussian beam diverges quickly from the focal plane, making it impossible to achieve high resolution imaging in a large volume specimen. We thus introduce Bessel beam laser-scanning tomography, which illuminates the sample with a spatial-light-modulator-generated Bessel beam that has an extended focal depth. The Bessel beam is scanned across the whole specimen. Fluorescence projection images are acquired at equal angular intervals as the sample rotates. Reconstruction artifacts due to annular-rings of the Bessel beam are removed by a modified 3D filtered back projection algorithm. Furthermore, in combination of Fourier-multiplexing fluorescence lifetime imaging (FmFLIM) method, the Bessel FmFLIM-SLOT system is capable of perform 3D lifetime imaging of live embryos at cellular resolution. The system is applied to in-vivo imaging of transgenic Zebrafish embryos. Results prove that Bessel FmFLIM-SLOT is a promising imaging method in development biology research.

  17. Picosecond wide-field time-correlated single photon counting fluorescence microscopy with a delay line anode detector

    Energy Technology Data Exchange (ETDEWEB)

    Hirvonen, Liisa M.; Le Marois, Alix; Suhling, Klaus, E-mail: klaus.suhling@kcl.ac.uk [Department of Physics, King' s College London, Strand, London WC2R 2LS (United Kingdom); Becker, Wolfgang; Smietana, Stefan [Becker & Hickl GmbH, Nahmitzer Damm 30, 12277 Berlin (Germany); Milnes, James; Conneely, Thomas [Photek Ltd., 26 Castleham Rd, Saint Leonards-on-Sea TN38 9NS (United Kingdom); Jagutzki, Ottmar [Institut für Kernphysik, Max-von-Laue-Str. 1, 60438 Frankfurt (Germany)

    2016-08-15

    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.

  18. Fluorescence lifetime, dipole orientation and bilayer polymer films

    Science.gov (United States)

    Ho, Xuan Long; Chen, Po-Jui; Woon, Wei-Yen; White, Jonathon David

    2017-10-01

    Bilayer films consisting of the optically transparent polymers, polystyrene (PS) and poly(methyl methacrylate) (PMMA) were spin-cast on glass substrates. The upper 13.5 nm layer (PS) was lightly doped with Rhodamine-6 G (RH6G) or MEH-PPV. While the fluorescence of MEH-PPV was independent of PMMA thickness, the lifetime of RH6G increased 3-fold as the underlying PMMA thickness increased from 0 to 500 nm while the collected flux decreased suggesting a reorientation of the smaller molecule's dipole with respect to the air-polymer interface with PMMA thickness. This suggests lifetime may find application for nondestructive thickness measurements of transparent films with sub-micron lateral resolution and large range.

  19. Fluorescence Lifetime Imaging in Stargardt Disease: Potential Marker for Disease Progression

    OpenAIRE

    Dysli Chantal; Wolf Sebastian; Hatz Katja; Zinkernagel Martin

    2016-01-01

    PURPOSE The purpose of this study was to describe autofluorescence lifetime characteristics in Stargardt disease (STGD) using fluorescence lifetime imaging ophthalmoscopy (FLIO) and to investigate potential prognostic markers for disease activity and progression. METHODS Fluorescence lifetime data of 16 patients with STGD (mean age, 40 years; range, 22-56 years) and 15 age-matched controls were acquired using a fluorescence lifetime imaging ophthalmoscope based on a Heidelberg Eng...

  20. Refractive Index Sensing of Green Fluorescent Proteins in Living Cells Using Fluorescence Lifetime Imaging Microscopy

    Science.gov (United States)

    van Manen, Henk-Jan; Verkuijlen, Paul; Wittendorp, Paul; Subramaniam, Vinod; van den Berg, Timo K.; Roos, Dirk; Otto, Cees

    2008-01-01

    We show that fluorescence lifetime imaging microscopy (FLIM) of green fluorescent protein (GFP) molecules in cells can be used to report on the local refractive index of intracellular GFP. We expressed GFP fusion constructs of Rac2 and gp91phox, which are both subunits of the phagocyte NADPH oxidase enzyme, in human myeloid PLB-985 cells and showed by high-resolution confocal fluorescence microscopy that GFP-Rac2 and GFP-gp91phox are targeted to the cytosol and to membranes, respectively. Frequency-domain FLIM experiments on these PLB-985 cells resulted in average fluorescence lifetimes of 2.70 ns for cytosolic GFP-Rac2 and 2.31 ns for membrane-bound GFP-gp91phox. By comparing these lifetimes with a calibration curve obtained by measuring GFP lifetimes in PBS/glycerol mixtures of known refractive index, we found that the local refractive indices of cytosolic GFP-Rac2 and membrane-targeted GFP-gp91phox are ∼1.38 and ∼1.46, respectively, which is in good correspondence with reported values for the cytosol and plasma membrane measured by other techniques. The ability to measure the local refractive index of proteins in living cells by FLIM may be important in revealing intracellular spatial heterogeneities within organelles such as the plasma and phagosomal membrane. PMID:18223002

  1. The enhanced cyan fluorescent protein: a sensitive pH sensor for fluorescence lifetime imaging.

    Science.gov (United States)

    Poëa-Guyon, Sandrine; Pasquier, Hélène; Mérola, Fabienne; Morel, Nicolas; Erard, Marie

    2013-05-01

    pH is an important parameter that affects many functions of live cells, from protein structure or function to several crucial steps of their metabolism. Genetically encoded pH sensors based on pH-sensitive fluorescent proteins have been developed and used to monitor the pH of intracellular compartments. The quantitative analysis of pH variations can be performed either by ratiometric or fluorescence lifetime detection. However, most available genetically encoded pH sensors are based on green and yellow fluorescent proteins and are not compatible with multicolor approaches. Taking advantage of the strong pH sensitivity of enhanced cyan fluorescent protein (ECFP), we demonstrate here its suitability as a sensitive pH sensor using fluorescence lifetime imaging. The intracellular ECFP lifetime undergoes large changes (32 %) in the pH 5 to pH 7 range, which allows accurate pH measurements to better than 0.2 pH units. By fusion of ECFP with the granular chromogranin A, we successfully measured the pH in secretory granules of PC12 cells, and we performed a kinetic analysis of intragranular pH variations in living cells exposed to ammonium chloride.

  2. Thermally activated delayed fluorescence organic dots for two-photon fluorescence lifetime imaging

    Science.gov (United States)

    He, Tingchao; Ren, Can; Li, Zhuohua; Xiao, Shuyu; Li, Junzi; Lin, Xiaodong; Ye, Chuanxiang; Zhang, Junmin; Guo, Lihong; Hu, Wenbo; Chen, Rui

    2018-05-01

    Autofluorescence is a major challenge in complex tissue imaging when molecules present in the biological tissue compete with the fluorophore. This issue may be resolved by designing organic molecules with long fluorescence lifetimes. The present work reports the two-photon absorption (TPA) properties of a thermally activated delayed fluorescence (TADF) molecule with carbazole as the electron donor and dicyanobenzene as the electron acceptor (i.e., 4CzIPN). The results indicate that 4CzIPN exhibits a moderate TPA cross-section (˜9 × 10-50 cm4 s photon-1), high fluorescence quantum yield, and a long fluorescence lifetime (˜1.47 μs). 4CzIPN was compactly encapsulated into an amphiphilic copolymer via nanoprecipitation to achieve water-soluble organic dots. Interestingly, 4CzIPN organic dots have been utilized in applications involving two-photon fluorescence lifetime imaging (FLIM). Our work aptly demonstrates that TADF molecules are promising candidates of nonlinear optical probes for developing next-generation multiphoton FLIM applications.

  3. Community detection for fluorescent lifetime microscopy image segmentation

    Science.gov (United States)

    Hu, Dandan; Sarder, Pinaki; Ronhovde, Peter; Achilefu, Samuel; Nussinov, Zohar

    2014-03-01

    Multiresolution community detection (CD) method has been suggested in a recent work as an efficient method for performing unsupervised segmentation of fluorescence lifetime (FLT) images of live cell images containing fluorescent molecular probes.1 In the current paper, we further explore this method in FLT images of ex vivo tissue slices. The image processing problem is framed as identifying clusters with respective average FLTs against a background or "solvent" in FLT imaging microscopy (FLIM) images derived using NIR fluorescent dyes. We have identified significant multiresolution structures using replica correlations in these images, where such correlations are manifested by information theoretic overlaps of the independent solutions ("replicas") attained using the multiresolution CD method from different starting points. In this paper, our method is found to be more efficient than a current state-of-the-art image segmentation method based on mixture of Gaussian distributions. It offers more than 1:25 times diversity based on Shannon index than the latter method, in selecting clusters with distinct average FLTs in NIR FLIM images.

  4. Fluorescent Biosensors Based on Single-Molecule Counting.

    Science.gov (United States)

    Ma, Fei; Li, Ying; Tang, Bo; Zhang, Chun-Yang

    2016-09-20

    Biosensors for highly sensitive, selective, and rapid quantification of specific biomolecules make great contributions to biomedical research, especially molecular diagnostics. However, conventional methods for biomolecular assays often suffer from insufficient sensitivity and poor specificity. In some case (e.g., early disease diagnostics), the concentration of target biomolecules is too low to be detected by these routine approaches, and cumbersome procedures are needed to improve the detection sensitivity. Therefore, there is an urgent need for rapid and ultrasensitive analytical tools. In this respect, single-molecule fluorescence approaches may well satisfy the requirement and hold promising potential for the development of ultrasensitive biosensors. Encouragingly, owing to the advances in single-molecule microscopy and spectroscopy over past decades, the detection of single fluorescent molecule comes true, greatly boosting the development of highly sensitive biosensors. By in vitro/in vivo labeling of target biomolecules with proper fluorescent tags, the quantification of certain biomolecule at the single-molecule level is achieved. In comparison with conventional ensemble measurements, single-molecule detection-based analytical methods possess the advantages of ultrahigh sensitivity, good selectivity, rapid analysis time, and low sample consumption. Consequently, single-molecule detection may be potentially employed as an ideal analytical approach to quantify low-abundant biomolecules with rapidity and simplicity. In this Account, we will summarize our efforts for developing a series of ultrasensitive biosensors based on single-molecule counting. Single-molecule counting is a member of single-molecule detection technologies and may be used as a very simple and ultrasensitive method to quantify target molecules by simply counting the individual fluorescent bursts. In the fluorescent sensors, the signals of target biomolecules may be translated to the

  5. A Compact Fluorescence Lifetime Excitation-Emission Spectrometer (FLEXEMS) for Detecting Trace Organics, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — In this Small Business Innovative Research (SBIR) effort, Leiden Measurement Technology (LMT) proposes to design and build the Fluorescence Lifetime Excitation...

  6. A Compact Fluorescence Lifetime Excitation-Emission Spectrometer (FLEXEMS) for Detecting Trace Organics, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — In this Small Business Innovative Research (SBIR) effort, Leiden Measurement Technology (LMT) proposes to design and build the Fluorescence Lifetime Excitation...

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

    Science.gov (United States)

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

    2006-10-01

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

  8. An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy

    International Nuclear Information System (INIS)

    Dunsby, C; Lanigan, P M P; McGinty, J; Elson, D S; Requejo-Isidro, J; Munro, I; Galletly, N; McCann, F; Treanor, B; Oenfelt, B; Davis, D M; Neil, M A A; French, P M W

    2004-01-01

    Fluorescence imaging is used widely in microscopy and macroscopic imaging applications for fields ranging from biomedicine to materials science. A critical component for any fluorescence imaging system is the excitation source. Traditionally, wide-field systems use filtered thermal or arc-generated white light sources, while point scanning confocal microscope systems require spatially coherent (point-like) laser sources. Unfortunately, the limited range of visible wavelengths available from conventional laser sources constrains the design and usefulness of fluorescent probes in confocal microscopy. A 'hands-off' laser-like source, electronically tunable across the visible spectrum, would be invaluable for fluorescence imaging and provide new opportunities, e.g. automated excitation fingerprinting and in situ measurement of excitation cross-sections. Yet more information can be obtained using fluorescence lifetime imaging (FLIM), which requires that the light source be pulsed or rapidly modulated. We show how a white light continuum, generated by injecting femtosecond optical radiation into a micro-structured optical fibre, coupled with a simple prism-based tunable filter arrangement, can fulfil all these roles as a continuously electronically tunable (435-1150 nm) visible ultrafast light source in confocal, wide-field and FLIM systems

  9. Time-resolved laser fluorescence spectroscopy of organic ligands by europium: Fluorescence quenching and lifetime properties

    Science.gov (United States)

    Nouhi, A.; Hajjoul, H.; Redon, R.; Gagné, J. P.; Mounier, S.

    2018-03-01

    Time-resolved Laser Fluorescence Spectroscopy (TRLFS) has proved its usefulness in the fields of biophysics, life science and geochemistry to characterize the fluorescence probe molecule with its chemical environment. The purpose of this study is to demonstrate the applicability of this powerful technique combined with Steady-State (S-S) measurements. A multi-mode factor analysis, in particular CP/PARAFAC, was used to analyze the interaction between Europium (Eu) and Humic substances (HSs) extracted from Saint Lawrence Estuary in Canada. The Saint Lawrence system is a semi-enclosed water stream with connections to the Atlantic Ocean and is an excellent natural laboratory. CP/PARAFAC applied to fluorescence S-S data allows introspecting ligands-metal interactions and the one-site 1:1 modeling gives information about the stability constants. From the spectral signatures and decay lifetimes data given by TRLFS, one can deduce the fluorescence quenching which modifies the fluorescence and discuss its mechanisms. Results indicated a relatively strong binding ability between europium and humic substances samples (Log K value varies from 3.38 to 5.08 at pH 7.00). Using the Stern-Volmer plot, it has been concluded that static and dynamic quenching takes places in the case of salicylic acid and europium interaction while for HSs interaction only a static quenching is observed.

  10. Development of new photon-counting detectors for single-molecule fluorescence microscopy

    Science.gov (United States)

    Michalet, X.; Colyer, R. A.; Scalia, G.; Ingargiola, A.; Lin, R.; Millaud, J. E.; Weiss, S.; Siegmund, Oswald H. W.; Tremsin, Anton S.; Vallerga, John V.; Cheng, A.; Levi, M.; Aharoni, D.; Arisaka, K.; Villa, F.; Guerrieri, F.; Panzeri, F.; Rech, I.; Gulinatti, A.; Zappa, F.; Ghioni, M.; Cova, S.

    2013-01-01

    Two optical configurations are commonly used in single-molecule fluorescence microscopy: point-like excitation and detection to study freely diffusing molecules, and wide field illumination and detection to study surface immobilized or slowly diffusing molecules. Both approaches have common features, but also differ in significant aspects. In particular, they use different detectors, which share some requirements but also have major technical differences. Currently, two types of detectors best fulfil the needs of each approach: single-photon-counting avalanche diodes (SPADs) for point-like detection, and electron-multiplying charge-coupled devices (EMCCDs) for wide field detection. However, there is room for improvements in both cases. The first configuration suffers from low throughput owing to the analysis of data from a single location. The second, on the other hand, is limited to relatively low frame rates and loses the benefit of single-photon-counting approaches. During the past few years, new developments in point-like and wide field detectors have started addressing some of these issues. Here, we describe our recent progresses towards increasing the throughput of single-molecule fluorescence spectroscopy in solution using parallel arrays of SPADs. We also discuss our development of large area photon-counting cameras achieving subnanosecond resolution for fluorescence lifetime imaging applications at the single-molecule level. PMID:23267185

  11. Ns-scaled time-gated fluorescence lifetime imaging for forensic document examination

    Science.gov (United States)

    Zhong, Xin; Wang, Xinwei; Zhou, Yan

    2018-01-01

    A method of ns-scaled time-gated fluorescence lifetime imaging (TFLI) is proposed to distinguish different fluorescent substances in forensic document examination. Compared with Video Spectral Comparator (VSC) which can examine fluorescence intensity images only, TFLI can detect questioned documents like falsification or alteration. TFLI system can enhance weak signal by accumulation method. The two fluorescence intensity images of the interval delay time tg are acquired by ICCD and fitted into fluorescence lifetime image. The lifetimes of fluorescence substances are represented by different colors, which make it easy to detect the fluorescent substances and the sequence of handwritings. It proves that TFLI is a powerful tool for forensic document examination. Furthermore, the advantages of TFLI system are ns-scaled precision preservation and powerful capture capability.

  12. The LB Films of Dansyl Chloride Labeled Octadecylamine and Its Fluorescence Lifetime

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Octadecylamine was derivatized with dansyl chloride (5-dimethylaminonaphthalene-1-sulfonyl chloride) in order to simplify and understand the LB films of fluorescent probe labeling proteins.Its monolayer and multilayers in the absence and presence of stearic acid were deposited by LB technique.Fluorescence spectra and lifetimes of the fluorescent products were studied to elucidate the microenvironment of molecules in the LB films.

  13. Quantitative fluorescence lifetime spectroscopy in turbid media: comparison of theoretical, experimental and computational methods

    International Nuclear Information System (INIS)

    Vishwanath, Karthik; Mycek, Mary-Ann; Pogue, Brian

    2002-01-01

    A Monte Carlo model developed to simulate time-resolved fluorescence propagation in a semi-infinite turbid medium was validated against previously reported theoretical and computational results. Model simulations were compared to experimental measurements of fluorescence spectra and lifetimes on tissue-simulating phantoms for single and dual fibre-optic probe geometries. Experiments and simulations using a single probe revealed that scattering-induced artefacts appeared in fluorescence emission spectra, while fluorescence lifetimes were unchanged. Although fluorescence lifetime measurements are generally more robust to scattering artefacts than are measurements of fluorescence spectra, in the dual-probe geometry scattering-induced changes in apparent lifetime were predicted both from diffusion theory and via Monte Carlo simulation, as well as measured experimentally. In all cases, the recovered apparent lifetime increased with increasing scattering and increasing source-detector separation. Diffusion theory consistently underestimated the magnitude of these increases in apparent lifetime (predicting a maximum increase of ∼15%), while Monte Carlo simulations and experiment were closely matched (showing increases as large as 30%). These results indicate that quantitative simulations of time-resolved fluorescence propagation in turbid media will be important for accurate recovery of fluorophore lifetimes in biological spectroscopy and imaging applications. (author)

  14. Standard reference for instrument response function in fluorescence lifetime measurements in visible and near infrared

    International Nuclear Information System (INIS)

    Chib, Rahul; Shah, Sunil; Gryczynski, Zygmunt; Fudala, Rafal; Borejdo, Julian; Gryczynski, Ignacy; Zelent, Bogumil; Corradini, Maria G; Ludescher, Richard D

    2016-01-01

    Allura red (AR) fluorophore, a common dye in the food industry, displays a broad emission spectrum in water (visible-to-near infrared region of the electromagnetic spectrum) and has a remarkably short fluorescence lifetime of about 10 ps. This short lifetime does not depend on the emission (observation) wavelength. We examined time responses of AR fluorescence across emission wavelengths from 550 nm to 750 nm and found that it is an ideal candidate for impulse response functions in fluorescence lifetime measurements. (technical note)

  15. Chlorophyll fluorescence lifetime imaging provides new insight into the chlorosis induced by plant virus infection.

    Science.gov (United States)

    Lei, Rong; Jiang, Hongshan; Hu, Fan; Yan, Jin; Zhu, Shuifang

    2017-02-01

    Leaf chlorosis induced by plant virus infection has a short fluorescence lifetime, which reflects damaged photosynthetic complexes and degraded chloroplasts. Plant viruses often induce chlorosis and necrosis, which are intimately related to photosynthetic functions. Chlorophyll fluorescence lifetime measurement is a valuable noninvasive tool for analyzing photosynthetic processes and is a sensitive indicator of the environment surrounding the fluorescent molecules. In this study, our central goal was to explore the effect of viral infection on photosynthesis by employing chlorophyll fluorescence lifetime imaging (FLIM), steady-state fluorescence, non-photochemical quenching (NPQ), transmission electron microscopy (TEM), and pigment analysis. The data indicated that the chlorophyll fluorescence lifetime of chlorotic leaves was significantly shorter than that of healthy control leaves, and the fitted short lifetime component of chlorophyll fluorescence of chlorotic leaves was dominant. This dominant short lifetime component may result from damage to the structure of thylakoid, which was confirmed by TEM. The NPQ value of chlorotic leaves was slightly higher than that of healthy green leaves, which can be explained by increased neoxanthin, lutein and violaxanthin content relative to chlorophyll a. The difference in NPQ is slight, but FLIM can provide simple and direct characterization of PSII structure and photosynthetic function. Therefore, this technique shows great potential as a simple and rapid method for studying mechanisms of plant virus infection.

  16. Alterations in cerebral metabolism observed in living rodents using fluorescence lifetime microscopy of intrinsic NADH (Conference Presentation)

    Science.gov (United States)

    Yaseen, Mohammad A.; Sakadžić, Sava; Sutin, Jason; Wu, Weicheng; Fu, Buyin; Boas, David A.

    2017-02-01

    Monitoring cerebral energy metabolism at a cellular level is essential to improve our understanding of healthy brain function and its pathological alterations. In this study, we resolve specific alterations in cerebral metabolism utilizing minimally-invasive 2-Photon fluorescence lifetime imaging (2P-FLIM) measurements of reduced nicotinamide adenine dinucleotide (NADH) fluorescence, collected in vivo from anesthetized rats and mice. Time-resolved lifetime measurements enables distinction of different components contributing to NADH autofluorescence. These components reportedly represent different enzyme-bound formulations of NADH. Our observations from this study confirm the hypothesis that NADH FLIM can identify specific alterations in cerebral metabolism. Using time-correlated single photon counting (TCSPC) equipment and a custom-built multimodal imaging system, 2-photon fluorescence lifetime imaging (FLIM) was performed in cerebral tissue with high spatial and temporal resolution. Multi-exponential fits for NADH fluorescence lifetimes indicate 4 distinct components, or 'species.' We observed distinct variations in the relative proportions of these components before and after pharmacological-induced impairments to several reactions involved in anaerobic glycolysis and aerobic oxidative metabolism. Classification models developed with experimental data correctly predict the metabolic impairments associated with bicuculline-induced focal seizures in separate experiments. Compared to traditional intensity-based NADH measurements, lifetime imaging of NADH is less susceptible to the adverse effects of overlying blood vessels. Evaluating NADH measurements will ultimately lead to a deeper understanding of cerebral energetics and its pathology-related alterations. Such knowledge will likely aid development of therapeutic strategies for neurodegenerative diseases such as Alzheimer's Disease, Parkinson's disease, and stroke.

  17. A Fast Global Fitting Algorithm for Fluorescence Lifetime Imaging Microscopy Based on Image Segmentation

    OpenAIRE

    Pelet, S.; Previte, M.J.R.; Laiho, L.H.; So, P.T. C.

    2004-01-01

    Global fitting algorithms have been shown to improve effectively the accuracy and precision of the analysis of fluorescence lifetime imaging microscopy data. Global analysis performs better than unconstrained data fitting when prior information exists, such as the spatial invariance of the lifetimes of individual fluorescent species. The highly coupled nature of global analysis often results in a significantly slower convergence of the data fitting algorithm as compared with unconstrained ana...

  18. In vivo multiphoton tomography and fluorescence lifetime imaging of human brain tumor tissue.

    Science.gov (United States)

    Kantelhardt, Sven R; Kalasauskas, Darius; König, Karsten; Kim, Ella; Weinigel, Martin; Uchugonova, Aisada; Giese, Alf

    2016-05-01

    High resolution multiphoton tomography and fluorescence lifetime imaging differentiates glioma from adjacent brain in native tissue samples ex vivo. Presently, multiphoton tomography is applied in clinical dermatology and experimentally. We here present the first application of multiphoton and fluorescence lifetime imaging for in vivo imaging on humans during a neurosurgical procedure. We used a MPTflex™ Multiphoton Laser Tomograph (JenLab, Germany). We examined cultured glioma cells in an orthotopic mouse tumor model and native human tissue samples. Finally the multiphoton tomograph was applied to provide optical biopsies during resection of a clinical case of glioblastoma. All tissues imaged by multiphoton tomography were sampled and processed for conventional histopathology. The multiphoton tomograph allowed fluorescence intensity- and fluorescence lifetime imaging with submicron spatial resolution and 200 picosecond temporal resolution. Morphological fluorescence intensity imaging and fluorescence lifetime imaging of tumor-bearing mouse brains and native human tissue samples clearly differentiated tumor and adjacent brain tissue. Intraoperative imaging was found to be technically feasible. Intraoperative image quality was comparable to ex vivo examinations. To our knowledge we here present the first intraoperative application of high resolution multiphoton tomography and fluorescence lifetime imaging of human brain tumors in situ. It allowed in vivo identification and determination of cell density of tumor tissue on a cellular and subcellular level within seconds. The technology shows the potential of rapid intraoperative identification of native glioma tissue without need for tissue processing or staining.

  19. Time gated fluorescence lifetime imaging and micro-volume spectroscopy using two-photon excitation

    NARCIS (Netherlands)

    Sytsma, J.; Vroom, J.M.; de Grauw, C.J.; Gerritsen, H.C.

    A scanning microscope utilizing two-photon excitation in combination with fluorescence lifetime contrast is presented. The microscope makes use of a tunable femtosecond titanium:sapphire laser enabling the two-photon excitation of a broad range of fluorescent molecules, including UV probes.

  20. Counting constituents in molecular complexes by fluorescence photon antibunching

    Energy Technology Data Exchange (ETDEWEB)

    Fore, S; Laurence, T; Hollars, C; Huser, T

    2007-04-17

    Modern single molecule fluorescence microscopy offers new, highly quantitative ways of studying the systems biology of cells while keeping the cells healthy and alive in their natural environment. In this context, a quantum optical technique, photon antibunching, has found a small niche in the continuously growing applications of single molecule techniques to small molecular complexes. Here, we review some of the most recent applications of photon antibunching in biophotonics, and we provide a guide for how to conduct photon antibunching experiments at the single molecule level by applying techniques borrowed from time-correlated single photon counting. We provide a number of new examples for applications of photon antibunching to the study of multichromophoric molecules and small molecular complexes.

  1. Fluorophore:dendrimer ratio impacts cellular uptake and intracellular fluorescence lifetime.

    Science.gov (United States)

    Dougherty, Casey A; Vaidyanathan, Sriram; Orr, Bradford G; Banaszak Holl, Mark M

    2015-02-18

    G5-NH2-TAMRAn (n = 1-4, 5+, and 1.5(avg)) were prepared with n = 1-4 as a precise dye:dendrimer ratio, 5+ as a mixture of dendrimers with 5 or more dye per dendrimer, and 1.5(avg) as a Poisson distribution of dye:dendrimer ratios with a mean of 1.5 dye per dendrimer. The absorption intensity increased sublinearly with n whereas the fluorescence emission and lifetime decreased with an increasing number of dyes per dendrimer. Flow cytometry was employed to quantify uptake into HEK293A cells. Dendrimers with 2-4 dyes were found to have greater uptake than dendrimer with a single dye. Fluorescence lifetime imaging microscopy (FLIM) showed that the different dye:dendrimer ratio alone was sufficient to change the fluorescence lifetime of the material observed inside cells. We also observed that the lifetime of G5-NH2-TAMRA5+ increased when present in the cell as compared to solution. However, cells treated with G5-NH2-TAMRA1.5(avg) did not exhibit the high lifetime components present in G5-NH2-TAMRA1 and G5-NH2-TAMRA5+. In general, the effects of the dye:dendrimer ratio on fluorescence lifetime were of similar magnitude to environmentally induced lifetime shifts.

  2. Miniaturized side-viewing imaging probe for fluorescence lifetime imaging (FLIM): validation with fluorescence dyes, tissue structural proteins and tissue specimens

    OpenAIRE

    Elson, DS; Jo, JA; Marcu, L

    2007-01-01

    We report a side viewing fibre-based endoscope that is compatible with intravascular imaging and fluorescence lifetime imaging microscopy (FLIM). The instrument has been validated through testing with fluorescent dyes and collagen and elastin powders using the Laguerre expansion deconvolution technique to calculate the fluorescence lifetimes. The instrument has also been tested on freshly excised unstained animal vascular tissues.

  3. Development of windows based software to analyze fluorescence decay with time-correlated single photon counting (TCSPC) setup

    International Nuclear Information System (INIS)

    Mallick, M.B.; Ravindranath, S.V.G.; Das, N.C.

    2002-07-01

    A VUV spectroscopic facility for studies in photophysics and photochemistry is being set up at INDUS-I synchrotron source, CAT, Indore. For this purpose, a data acquisition system based on time-correlated single photon counting method is being developed for fluorescence lifetime measurement. To estimate fluorescence lifetime from the data collected with this sytem, a Windows based program has been developed using Visual Basic 5.0. It uses instrument response function (IRF) and observed decay curve and estimates parameters of single exponential decay by least square analysis and Marquardt method as convergence mechanism. Estimation of parameters was performed using data collected with a commercial setup. Goodness of fit was judged by evaluating χR 2 , weighted residuals and autocorrelation function. Performance is compared with two commercial software packages and found to be satisfactory. (author)

  4. Distribution of diffusion times determined by fluorescence (lifetime) correlation spectroscopy

    Czech Academy of Sciences Publication Activity Database

    Pánek, Jiří; Loukotová, Lenka; Hrubý, Martin; Štěpánek, Petr

    2018-01-01

    Roč. 51, č. 8 (2018), s. 2796-2804 ISSN 0024-9297 R&D Projects: GA MŠk(CZ) LO1507 Institutional support: RVO:61389013 Keywords : polymer solution * fluorescence correlation spectroscopy * diffusion time distribution Subject RIV: CD - Macromolecular Chemistry OBOR OECD: Polymer science Impact factor: 5.835, year: 2016

  5. In vivo fluorescence lifetime imaging for monitoring the efficacy of the cancer treatment.

    Science.gov (United States)

    Ardeshirpour, Yasaman; Chernomordik, Victor; Hassan, Moinuddin; Zielinski, Rafal; Capala, Jacek; Gandjbakhche, Amir

    2014-07-01

    Advances in tumor biology created a foundation for targeted therapy aimed at inactivation of specific molecular mechanisms responsible for cell malignancy. In this paper, we used in vivo fluorescence lifetime imaging with HER2-targeted fluorescent probes as an alternative imaging method to investigate the efficacy of targeted therapy with 17-DMAG (an HSP90 inhibitor) on tumors with high expression of HER2 receptors. HER2-specific Affibody, conjugated to Alexafluor 750, was injected into nude mice bearing HER2-positive tumor xenograft. The fluorescence lifetime was measured before treatment and monitored after the probe injections at 12 hours after the last treatment dose, when the response to the 17-DMAG therapy was the most pronounced as well as a week after the last treatment when the tumors grew back almost to their pretreatment size. Imaging results showed significant difference between the fluorescence lifetimes at the tumor and the contralateral site (∼0.13 ns) in the control group (before treatment) and 7 days after the last treatment when the tumors grew back to their pretreatment dimensions. However, at the time frame that the treatment had its maximum effect (12 hours after the last treatment), the difference between the fluorescence lifetime at the tumor and contralateral site decreased to 0.03 ns. The results showed a good correlation between fluorescence lifetime and the efficacy of the treatment. These findings show that in vivo fluorescence lifetime imaging can be used as a promising molecular imaging tool for monitoring the treatment outcome in preclinical models and potentially in patients. ©2014 American Association for Cancer Research.

  6. In-vivo fluorescence lifetime imaging for monitoring the efficacy of the cancer treatment

    Science.gov (United States)

    Ardeshirpour, Yasaman; Chernomordik, Victor; Hassan, Moinuddin; Zielinski, Rafal; Capala, Jacek; Gandjbakhche, Amir

    2015-01-01

    Purpose Advances in tumor biology created a foundation for targeted therapy aimed at inactivation of specific molecular mechanisms responsible for cell malignancy. In this paper, we used in-vivo fluorescence lifetime imaging with HER2 targeted fluorescent probes as an alternative imaging method to investigate the efficacy of targeted therapy with 17-DMAG (an HSP90 inhibitor) on tumors with high expression of HER2 receptors. Experimental Design HER2-specific Affibody, conjugated to Alexafluor 750, was injected into nude mice, bearing HER2-positive tumor xenograft. The fluorescence lifetime was measured before treatment and monitored after the probe injections at 12 hours after the last treatment dose, when the response to the 17-DMAG therapy was the most pronounced as well as a week after the last treatment when the tumors grew back almost to their pre-treatment size. Results Imaging results showed significant difference between the fluorescence lifetimes at the tumor and the contralateral site (~0.13ns) in the control group (before treatment) and 7 days after the last treatment when the tumors grew back to their pretreatment dimensions. However, at the time frame that the treatment had its maximum effect (12 hours after the last treatment) the difference between the fluorescence lifetime at the tumor and contralateral site decreased to 0.03ns. Conclusions The results showed a good correlation between fluorescence lifetime and the efficacy of the treatment. These findings show that in-vivo fluorescence lifetime imaging can be used as a promising molecular imaging tool for monitoring the treatment outcome in preclinical models and potentially in patients. PMID:24671949

  7. Portable fluorescence lifetime spectroscopy system for in-situ interrogation of biological tissues

    Science.gov (United States)

    Saito Nogueira, Marcelo; Cosci, Alessandro; Teixeira Rosa, Ramon Gabriel; Salvio, Ana Gabriela; Pratavieira, Sebastião; Kurachi, Cristina

    2017-12-01

    Fluorescence spectroscopy and lifetime techniques are potential methods for optical diagnosis and characterization of biological tissues with an in-situ, fast, and noninvasive interrogation. Several diseases may be diagnosed due to differences in the fluorescence spectra of targeted fluorophores, when, these spectra are similar, considering steady-state fluorescence, others may be detected by monitoring their fluorescence lifetime. Despite this complementarity, most of the current fluorescence lifetime systems are not robust and portable, and not being feasible for clinical applications. We describe the assembly of a fluorescence lifetime spectroscopy system in a suitcase, its characterization, and validation with clinical measurements of skin lesions. The assembled system is all encased and robust, maintaining its mechanical, electrical, and optical stability during transportation, and is feasible for clinical measurements. The instrument response function measured was about 300 ps, and the system is properly calibrated. At the clinical study, the system showed to be reliable, and the achieved spectroscopy results support its potential use as an auxiliary tool for skin diagnostics.

  8. Portable fluorescence lifetime spectroscopy system for in-situ interrogation of biological tissues.

    Science.gov (United States)

    Saito Nogueira, Marcelo; Cosci, Alessandro; Teixeira Rosa, Ramon Gabriel; Salvio, Ana Gabriela; Pratavieira, Sebastião; Kurachi, Cristina

    2017-10-01

    Fluorescence spectroscopy and lifetime techniques are potential methods for optical diagnosis and characterization of biological tissues with an in-situ, fast, and noninvasive interrogation. Several diseases may be diagnosed due to differences in the fluorescence spectra of targeted fluorophores, when, these spectra are similar, considering steady-state fluorescence, others may be detected by monitoring their fluorescence lifetime. Despite this complementarity, most of the current fluorescence lifetime systems are not robust and portable, and not being feasible for clinical applications. We describe the assembly of a fluorescence lifetime spectroscopy system in a suitcase, its characterization, and validation with clinical measurements of skin lesions. The assembled system is all encased and robust, maintaining its mechanical, electrical, and optical stability during transportation, and is feasible for clinical measurements. The instrument response function measured was about 300 ps, and the system is properly calibrated. At the clinical study, the system showed to be reliable, and the achieved spectroscopy results support its potential use as an auxiliary tool for skin diagnostics. (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

  9. Use of multiphoton tomography and fluorescence lifetime imaging to investigate skin pigmentation in vivo

    Science.gov (United States)

    Dancik, Yuri; Favre, Amandine; Loy, Chong Jin; Zvyagin, Andrei V.; Roberts, Michael S.

    2013-02-01

    There is a growing body of literature showing the usefulness of multiphoton tomography (MPT) and fluorescence lifetime imaging for in situ characterization of skin constituents and the ensuing development of noninvasive diagnostic tools against skin diseases. Melanin and pigmentation-associated skin cancers constitute some of the major applications. We show that MPT and fluorescence lifetime imaging can be used to measure changes in cutaneous melanin concentration and that these can be related to the visible skin color. Melanin in the skin of African, Indian, Caucasian, and Asian volunteers is detected on the basis of its emission wavelength and fluorescence lifetimes in solution and in a melanocyte-keratinocyte cell culture. Fluorescence intensity is used to characterize the melanin content and distribution as a function of skin type and depth into the skin (stratum granulosum and stratum basale). The measured fluorescence intensities in given skin types agree with melanin amounts reported by others using biopsies. Our results suggest that spatial distribution of melanin in skin can be studied using MPT and fluorescence lifetime imaging, but further studies are needed to ascertain that the method can resolve melanin amount in smaller depth intervals.

  10. Plasmonic-based instrument response function for time-resolved fluorescence: toward proper lifetime analysis

    Energy Technology Data Exchange (ETDEWEB)

    Szlazak, Radoslaw; Tutaj, Krzysztof; Grudzinski, Wojciech; Gruszecki, Wieslaw I.; Luchowski, Rafal, E-mail: rafal.luchowski@umcs.pl [Maria Curie-Sklodowska University, Department of Biophysics, Institute of Physics (Poland)

    2013-06-15

    In this report, we investigated the so-called plasmonic platforms prepared to target ultra-short fluorescence and accurate instrumental response function in a time-domain spectroscopy and microscopy. The interaction of metallic nanoparticles with nearby fluorophores results in the increase of the dye fluorescence quantum yield, photostability and decrease of the lifetime parameter. The mentioned properties of platforms were applied to achieve a picosecond fluorescence lifetime (21 ps) of erythrosin B, used later as a better choice for deconvolution of fluorescence decays measured with 'color' sensitive photo-detectors. The ultra-short fluorescence standard based on combination of thin layers of silver film, silver colloidal nanoparticles (about 60 nm in diameter), and top layer of erythrosin B embedded in 0.2 % poly(vinyl) alcohol. The response functions were monitored on two photo-detectors; microchannel plate photomultiplier and single photon avalanche photodiode as a Rayleigh scattering and ultra-short fluorescence. We demonstrated that use of the plasmonic base fluorescence standard as an instrumental response function results in the absence of systematic error in lifetime measurements and analysis.

  11. Fluorescence lifetime correlation spectroscopy combined with lifetime tuning: New perspectives in supported phospholipid bilayer research

    Czech Academy of Sciences Publication Activity Database

    Benda, Aleš; Fagulová, Veronika; Deyneka, Alexander; Enderlain, J.; Hof, Martin

    2006-01-01

    Roč. 22, č. 23 (2006), s. 9580-9585 ISSN 0743-7463 R&D Projects: GA ČR GA203/05/2308; GA MŠk LC06063 Institutional research plan: CEZ:AV0Z40400503; CEZ:AV0Z10100522 Keywords : spectroscopy * fluorescence * FLCS Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.902, year: 2006

  12. Time-Resolved Fluorescence Spectroscopy and Fluorescence Lifetime Imaging Microscopy for Characterization of Dendritic Polymer Nanoparticles and Applications in Nanomedicine

    Directory of Open Access Journals (Sweden)

    Alexander Boreham

    2016-12-01

    Full Text Available The emerging field of nanomedicine provides new approaches for the diagnosis and treatment of diseases, for symptom relief and for monitoring of disease progression. One route of realizing this approach is through carefully constructed nanoparticles. Due to the small size inherent to the nanoparticles a proper characterization is not trivial. This review highlights the application of time-resolved fluorescence spectroscopy and fluorescence lifetime imaging microscopy (FLIM for the analysis of nanoparticles, covering aspects ranging from molecular properties to particle detection in tissue samples. The latter technique is particularly important as FLIM allows for distinguishing of target molecules from the autofluorescent background and, due to the environmental sensitivity of the fluorescence lifetime, also offers insights into the local environment of the nanoparticle or its interactions with other biomolecules. Thus, these techniques offer highly suitable tools in the fields of particle development, such as organic chemistry, and in the fields of particle application, such as in experimental dermatology or pharmaceutical research.

  13. Time-Resolved Fluorescence Spectroscopy and Fluorescence Lifetime Imaging Microscopy for Characterization of Dendritic Polymer Nanoparticles and Applications in Nanomedicine.

    Science.gov (United States)

    Boreham, Alexander; Brodwolf, Robert; Walker, Karolina; Haag, Rainer; Alexiev, Ulrike

    2016-12-24

    The emerging field of nanomedicine provides new approaches for the diagnosis and treatment of diseases, for symptom relief and for monitoring of disease progression. One route of realizing this approach is through carefully constructed nanoparticles. Due to the small size inherent to the nanoparticles a proper characterization is not trivial. This review highlights the application of time-resolved fluorescence spectroscopy and fluorescence lifetime imaging microscopy (FLIM) for the analysis of nanoparticles, covering aspects ranging from molecular properties to particle detection in tissue samples. The latter technique is particularly important as FLIM allows for distinguishing of target molecules from the autofluorescent background and, due to the environmental sensitivity of the fluorescence lifetime, also offers insights into the local environment of the nanoparticle or its interactions with other biomolecules. Thus, these techniques offer highly suitable tools in the fields of particle development, such as organic chemistry, and in the fields of particle application, such as in experimental dermatology or pharmaceutical research.

  14. Fluorescence lifetime imaging of induced pluripotent stem cells

    Science.gov (United States)

    Uchugonova, Aisada; Batista, Ana; König, Karsten

    2014-02-01

    The multiphoton FLIM tomograph MPTflex with its flexible scan head, articulated arm, and the tunable femtosecond laser source was employed to study cell monolayers and 3D cell clusters. FLIM was performed with 250 ps temporal resolution and submicron special resolution using time-correlated single photon counting. The autofluorescence based on NAD(P)H and flavins/flavoproteins has been measured in mouse embryonic fibroblasts, induced pluripotent stem cells (iPS cells) originated from mouse embryonic fibroblasts and non-proliferative mouse embryonic fibroblasts.

  15. Measurements of fluorescence lifetime of group III metalo-8-quinolinolates and their use in analytical chemistry

    Energy Technology Data Exchange (ETDEWEB)

    Nishikawa, Y; Hiraki, K; Morishige, K; Takahashi, K [Kinki Univ., Higashi-Osaka, Osaka (Japan). Faculty of Science and Technology; Shigematsu, T

    1976-07-01

    8-Quinolinolates of aluminum, gallium, and indium in chloroform exhibit strong yellowish green fluorescence with an emission maximum at 510, 526, and 528 nm, respectively. The time resolved fluorescence spectra and the fluorescence lifetime properties of these chelates were measured with a time-resolved spectrofluorometer. The fluorescence intensity of these chelates decays exponentially with time t, and obeys the following equation: F=F/sub 0/e-t/tau=F/sub 0/e-k sub(f).t where F/sub 0/ and F are the fluorescence intensity when the exciting light is irradiating and shut off, respectively; tau and k sub(f) being the lifetime and the rate constant for the process of fluorescence emission. The lifetimes of these chelates in chloroform solution at the ordinary temperature were 17.8, 10.1, and 8.4 ns for Al(C/sub 9/H/sub 6/ON)/sub 3/, Ga(C/sub 9/H/sub 6/ON)/sub 3/, and In(C/sub 9/H/sub 6/ON)/sub 3/, respectively. Thus, 8-quinolinolates of group III metals emit the same type radiation with different lifetimes. Between Al-chelate and In-chelate, there were significant difference in the lifetime by 9.4 ns. Then, the logarithmic plot of the composite fluorescence intensity against time is the overlap of some straight lines with different slopes which indicate k sub(f) of various decay processes. The linear portion of the logarithmic plot of the composite fluorescence intensity corresponded to the longer lifetime component (Al-chelate), and by substracting this component from the whole one, the straight line due to the shorter lifetime component (In-chelate) is obtained. Aluminum and indium contents were then determined by comparing the fluorescence intensity of the sample with that of the standard at a definite time (extrapolated to t=0). By using this composite decay curve, the composition of mixtures of nx10/sup -4/ mol/l of Al and In-chelates in chloroform could be determined.

  16. Multiphoton Laser Microscopy and Fluorescence Lifetime Imaging for the Evaluation of the Skin

    Directory of Open Access Journals (Sweden)

    Stefania Seidenari

    2012-01-01

    Full Text Available Multiphoton laser microscopy is a new, non-invasive technique providing access to the skin at a cellular and subcellular level, which is based both on autofluorescence and fluorescence lifetime imaging. Whereas the former considers fluorescence intensity emitted by epidermal and dermal fluorophores and by the extra-cellular matrix, fluorescence lifetime imaging (FLIM, is generated by the fluorescence decay rate. This innovative technique can be applied to the study of living skin, cell cultures and ex vivo samples. Although still limited to the clinical research field, the development of multiphoton laser microscopy is thought to become suitable for a practical application in the next few years: in this paper, we performed an accurate review of the studies published so far, considering the possible fields of application of this imaging method and providing high quality images acquired in the Department of Dermatology of the University of Modena.

  17. Fluorescence lifetime measurements of native and glycated human serum albumin and bovine serum albumin

    Science.gov (United States)

    Joshi, Narahari V.; Joshi, Virgina O. d.; Contreras, Silvia; Gil, Herminia; Medina, Honorio; Siemiarczuk, Aleksander

    1999-05-01

    Nonenzymatic glycation, also known as Maillard reaction, plays an important role in the secondary complications of the diabetic pathology and aging, therefore, human serum albumin (HSA) and bovine serum albumin (BSA) were glycated by a conventional method in our laboratory using glucose as the glycating agent. Fluorescence lifetime measurements were carried out with a laser strobe fluorometer equipped with a nitrogen/dye laser and a frequency doubler as a pulsed excitation source. The samples were excited at 295 nm and the emission spectra were recorded at 345 nm. The obtained decay curves were tried for double and triple exponential functions. It has been found that the shorter lifetime increases for glycated proteins as compared with that of the native ones. For example, in the case of glycated BSA the lifetime increased from 1.36 ns to 2.30 ns. Similarly, for HSA, the lifetime increases from 1.58 ns to 2.26 ns. Meanwhile, the longer lifetime changed very slightly for both proteins (from 6.52 ns to 6.72 ns). The increase in the lifetime can be associated with the environmental effect; originated from the attachment of glucose to some lysine residues. A good example is Trp 214 which is in the cage of Lys 225, Lys 212, Lys 233, Lys 205, Lys 500, Lys 199 and Lys 195. If fluorescence lifetime technique is calibrated and properly used it could be employed for assessing glycation of proteins.

  18. Fluorescence lifetime imaging of lipids during 3T3-L1 cell differentiation

    Science.gov (United States)

    Song, Young Sik; Won, Young Jae; Lee, Sang-Hak; Kim, Dug Young

    2014-03-01

    Obesity is becoming a big health problem in these days. Since increased body weight is due to increased number and size of the triglyceride-storing adipocytes, many researchers are working on differentiation conditions and processes of adipocytes. Adipocytes also work as regulators of whole-body energy homeostasis by secreting several proteins that regulate processes as diverse as haemostasis, blood pressure, immune function, angiogenesis and energy balance. 3T3-L1 cells are widely used cell line for studying adipogenesis because it can differentiate into an adipocyte-like phenotype under appropriate conditions. In this paper, we propose an effective fluorescence lifetime imaging technique which can easily distinguish lipids in membrane and those in lipid droplets. Nile red dyes are attached to lipids in 3T3-L1 cells. Fluorescence lifetime images were taken for 2 week during differentiation procedure of 3T3-L1 cells into adipocytes. We used 488 nm pulsed laser with 5MHz repetition rate and emission wavelength is 520 nm of Nile Red fluorescent dye. Results clearly show that the lifetime of Nile red in lipid droplets are smaller than those in cell membrane. Our results suggest that fluorescence lifetime imaging can be a very powerful tool to monitor lipid droplet formation in adipocytes from 3T3-L1 cells.

  19. Fluorescence lifetime of emitters with broad homogeneous linewidths modified in opal photonic crystals

    DEFF Research Database (Denmark)

    Nikolaev, Ivan S.; Lodahl, Peter; Vos, Willem L.

    2008-01-01

    We have investigated the dynamics of spontaneous emission from dye molecules embedded in opal photonic crystals. Fluorescence lifetimes of Rhodamine 6G (R6G) dye were measured as a function of both optical frequency and crystal lattice parameter of the polystyrene opals. Due to the broad...

  20. Time-Domain Fluorescence Lifetime Imaging Techniques Suitable for Solid-State Imaging Sensor Arrays

    Directory of Open Access Journals (Sweden)

    Robert K. Henderson

    2012-05-01

    Full Text Available We have successfully demonstrated video-rate CMOS single-photon avalanche diode (SPAD-based cameras for fluorescence lifetime imaging microscopy (FLIM by applying innovative FLIM algorithms. We also review and compare several time-domain techniques and solid-state FLIM systems, and adapt the proposed algorithms for massive CMOS SPAD-based arrays and hardware implementations. The theoretical error equations are derived and their performances are demonstrated on the data obtained from 0.13 μm CMOS SPAD arrays and the multiple-decay data obtained from scanning PMT systems. In vivo two photon fluorescence lifetime imaging data of FITC-albumin labeled vasculature of a P22 rat carcinosarcoma (BD9 rat window chamber are used to test how different algorithms perform on bi-decay data. The proposed techniques are capable of producing lifetime images with enough contrast.

  1. Parallel excitation-emission multiplexed fluorescence lifetime confocal microscopy for live cell imaging.

    Science.gov (United States)

    Zhao, Ming; Li, Yu; Peng, Leilei

    2014-05-05

    We present a novel excitation-emission multiplexed fluorescence lifetime microscopy (FLIM) method that surpasses current FLIM techniques in multiplexing capability. The method employs Fourier multiplexing to simultaneously acquire confocal fluorescence lifetime images of multiple excitation wavelength and emission color combinations at 44,000 pixels/sec. The system is built with low-cost CW laser sources and standard PMTs with versatile spectral configuration, which can be implemented as an add-on to commercial confocal microscopes. The Fourier lifetime confocal method allows fast multiplexed FLIM imaging, which makes it possible to monitor multiple biological processes in live cells. The low cost and compatibility with commercial systems could also make multiplexed FLIM more accessible to biological research community.

  2. Enhancement of early cervical cancer diagnosis with epithelial layer analysis of fluorescence lifetime images.

    Directory of Open Access Journals (Sweden)

    Jun Gu

    Full Text Available This work reports the use of layer analysis to aid the fluorescence lifetime diagnosis of cervical intraepithelial neoplasia (CIN from H&E stained cervical tissue sections. The mean and standard deviation of lifetimes in single region of interest (ROI of cervical epithelium were previously shown to correlate to the gold standard histopathological classification of early cervical cancer. These previously defined single ROIs were evenly divided into layers for analysis. A 10-layer model revealed a steady increase in fluorescence lifetime from the inner to the outer epithelial layers of healthy tissue sections, suggesting a close association with cellular maturity. The shorter lifetime and minimal lifetime increase towards the epithelial surface of CIN-affected regions are in good agreement with the absence of cellular maturation in CIN. Mean layer lifetimes in the top-half cervical epithelium were used as feature vectors for extreme learning machine (ELM classifier discriminations. It was found that the proposed layer analysis technique greatly improves the sensitivity and specificity to 94.6% and 84.3%, respectively, which can better supplement the traditional gold standard cervical histopathological examinations.

  3. An automated cell-counting algorithm for fluorescently-stained cells in migration assays

    Directory of Open Access Journals (Sweden)

    Novielli Nicole M

    2011-10-01

    Full Text Available Abstract A cell-counting algorithm, developed in Matlab®, was created to efficiently count migrated fluorescently-stained cells on membranes from migration assays. At each concentration of cells used (10,000, and 100,000 cells, images were acquired at 2.5 ×, 5 ×, and 10 × objective magnifications. Automated cell counts strongly correlated to manual counts (r2 = 0.99, P

  4. Azadioxatriangulenium: a long fluorescence lifetime fluorophore for large biomolecule binding assay

    International Nuclear Information System (INIS)

    Sørensen, Thomas Just; Thyrhaug, Erling; Szabelski, Mariusz; Gryczynski, Ignacy; Gryczynski, Zygmunt; Luchowski, Rafal; Laursen, Bo W

    2013-01-01

    Of the many optical bioassays available, sensing by fluorescence anisotropy has great advantages as it provides a sensitive, instrumentally simple, ratiometric method of detection. However, it is hampered by a severe limitation, as the emission lifetime of the label needs to be comparable to the correlation lifetime (tumbling time) of the biomolecule which is labelled. For proteins of moderate size this is on the order of 20–200 ns, which due to practical issues currently limits the choice of labels to the dansyl-type dyes and certain aromatic dyes. These have the significant drawback of UV/blue absorption and emission as well as an often significant solvent sensitivity. Here, we report the synthesis and characterization of a new fluorescent label for high molecular weight biomolecule assay based on the azadioxatriangulenium motif. The NHS ester of the long fluorescence lifetime, red-emitting fluorophore: azadioxatriangulenium (ADOTA-NHS) was conjugated to anti-rabbit Immunoglobulin G (antiIgG). The long fluorescence lifetime was exploited to determine the correlation time of the high molecular weight antibody and its complex with rabbit Immunoglobulin G (IgG) with steady-state fluorescence anisotropy and time-resolved methods: solution phase immuno-assay was performed following either steady-state or time-resolved fluorescence anisotropy. By performing a variable temperature experiment it was determined that the binding of the ligand resulted in an increase in correlation time of more than 75%, and an increase in the steady-state anisotropy of 18%. The results show that the triangulenium class of dyes can be used in anisotropy assay to detect binding events involving biomolecules of far larger size than what is possible with most other red-emitting organic dyes. (paper)

  5. Azadioxatriangulenium: a long fluorescence lifetime fluorophore for large biomolecule binding assay

    Science.gov (United States)

    Just Sørensen, Thomas; Thyrhaug, Erling; Szabelski, Mariusz; Luchowski, Rafal; Gryczynski, Ignacy; Gryczynski, Zygmunt; Laursen, Bo W.

    2013-06-01

    Of the many optical bioassays available, sensing by fluorescence anisotropy has great advantages as it provides a sensitive, instrumentally simple, ratiometric method of detection. However, it is hampered by a severe limitation, as the emission lifetime of the label needs to be comparable to the correlation lifetime (tumbling time) of the biomolecule which is labelled. For proteins of moderate size this is on the order of 20-200 ns, which due to practical issues currently limits the choice of labels to the dansyl-type dyes and certain aromatic dyes. These have the significant drawback of UV/blue absorption and emission as well as an often significant solvent sensitivity. Here, we report the synthesis and characterization of a new fluorescent label for high molecular weight biomolecule assay based on the azadioxatriangulenium motif. The NHS ester of the long fluorescence lifetime, red-emitting fluorophore: azadioxatriangulenium (ADOTA-NHS) was conjugated to anti-rabbit Immunoglobulin G (antiIgG). The long fluorescence lifetime was exploited to determine the correlation time of the high molecular weight antibody and its complex with rabbit Immunoglobulin G (IgG) with steady-state fluorescence anisotropy and time-resolved methods: solution phase immuno-assay was performed following either steady-state or time-resolved fluorescence anisotropy. By performing a variable temperature experiment it was determined that the binding of the ligand resulted in an increase in correlation time of more than 75%, and an increase in the steady-state anisotropy of 18%. The results show that the triangulenium class of dyes can be used in anisotropy assay to detect binding events involving biomolecules of far larger size than what is possible with most other red-emitting organic dyes.

  6. Azadioxatriangulenium (ADOTA+): A long fluorescence lifetime fluorophore for large biomolecule binding assay

    Science.gov (United States)

    Sørensen, Thomas Just; Thyrhaug, Erling; Szabelski, Mariusz; Luchowski, Rafal; Gryczynski, Ignacy; Gryczynski, Zygmunt; Laursen, Bo W.

    2013-01-01

    Of the many optical bioassays available, sensing by fluorescence anisotropy have great advantages as it provides a sensitive, instrumentally simple, ratiometric method of detection. However, it is hampered by a severe limitation as the emission lifetime of the label needs to be comparable to the correlation lifetime (tumbling time) of the biomolecule which is labelled. For proteins of moderate size this is in the order of 20–200 ns, which due to practical issues currently limits the choice of labels to the dansyl-type dyes and certain aromatics dyes. These have the significant drawback of UV/blue absorption and emission as well as an often significant solvent sensitivity. Here, we report the synthesis and characterization of a new fluorescent label for high molecular weight biomolecules assay based on the azadioxatriangulenium motif. The NHS ester of the long fluorescence lifetime, red emitting fluorophore: azadioxatriangulenium (ADOTA-NHS) was conjugated to anti-rabbit Immunoglobulin G (antiIgG). The long fluorescence lifetime was exploited to determine the correlation time of the high molecular weight antibody and its complex with rabbit Immuniglobulin G (IgG) with steady-state fluorescence anisotropy and time-resolved methods: solution phase immuno-assay was performed following either steady-state or time-resolved fluorescence anisotropy. By performing a variable temperature experiment it was determined that the binding of the ligand resulted in an increase in correlation time by more than 75 %, and a change in the steady-state anisotropy increase of 18%. The results show that the triangulenium class of dyes can be used in anisotropy assay for detecting binding events involving biomolecules of far larger size than what is possible with the other red emitting organic dyes. PMID:24058730

  7. Time-correlated single-photon counting study of multiple photoluminescence lifetime components of silicon nanoclusters

    Energy Technology Data Exchange (ETDEWEB)

    Diamare, D., E-mail: d.diamare@ee.ucl.ac.uk [Department of Electronic and Electrical Engineering, University College London, Torrington Place, London, WC1E 7JE (United Kingdom); Wojdak, M. [Department of Electronic and Electrical Engineering, University College London, Torrington Place, London, WC1E 7JE (United Kingdom); Lettieri, S. [Institute for Superconductors and Innovative Materials, National Council of Research (CNR-SPIN), Via Cintia 80126, Naples (Italy); Department of Physical Sciences, University of Naples “Federico II”, Via Cintia 80126, Naples (Italy); Kenyon, A.J. [Department of Electronic and Electrical Engineering, University College London, Torrington Place, London, WC1E 7JE (United Kingdom)

    2013-04-15

    We report time-resolved photoluminescence measurements of thin films of silica containing silicon nanoclusters (Si NCs), produced by PECVD and annealed at temperatures between 700 °C and 1150 °C. While the near infrared emission of Si NCs has long been studied, visible light emission has only recently attracted interest due to its very short decay times and its recently-reported redshift with decreasing NCs size. We analyse the PL decay dynamics in the range 450–700 nm with picosecond time resolution using Time Correlated Single Photon Counting. In the resultant multi-exponential decays two dominant components can clearly be distinguished: a very short component, in the range of hundreds of picoseconds, and a nanosecond component. In this wavelength range we do not detect the microsecond component generally associated with excitonic recombination. We associate the nanosecond component to defect relaxation: it decreases in intensity in the sample annealed at higher temperature, suggesting that the contribution from defects decreases with increasing temperature. The origin of the very fast PL component (ps time region) is also discussed. We show that it is consistent with the Auger recombination times of multiple excitons. Further work needs to be done in order to assess the contribution of the Auger-controlled recombinations to the defect-assisted mechanism of photoluminescence. -- Highlights: ► We report time-resolved PL measurements of Si-Ncs embedded in SiO{sub 2} matrix. ► Net decrease of PL with increasing the annealing temperature has been observed. ► Lifetime distribution analysis revealed a multiexponential decay with ns and ps components. ► Ps components are consistent with the lifetime range of the Auger recombination times. ► No evidence for a fast direct transition at the Brillouin zone centre.

  8. Online multispectral fluorescence lifetime values estimation and overlay onto tissue white-light video frames

    Science.gov (United States)

    Gorpas, Dimitris; Ma, Dinglong; Bec, Julien; Yankelevich, Diego R.; Marcu, Laura

    2016-03-01

    Fluorescence lifetime imaging has been shown to be a robust technique for biochemical and functional characterization of tissues and to present great potential for intraoperative tissue diagnosis and guidance of surgical procedures. We report a technique for real-time mapping of fluorescence parameters (i.e. lifetime values) onto the location from where the fluorescence measurements were taken. This is achieved by merging a 450 nm aiming beam generated by a diode laser with the excitation light in a single delivery/collection fiber and by continuously imaging the region of interest with a color CMOS camera. The interrogated locations are then extracted from the acquired frames via color-based segmentation of the aiming beam. Assuming a Gaussian profile of the imaged aiming beam, the segmentation results are fitted to ellipses that are dynamically scaled at the full width of three automatically estimated thresholds (50%, 75%, 90%) of the Gaussian distribution's maximum value. This enables the dynamic augmentation of the white-light video frames with the corresponding fluorescence decay parameters. A fluorescence phantom and fresh tissue samples were used to evaluate this method with motorized and hand-held scanning measurements. At 640x512 pixels resolution the area of interest augmented with fluorescence decay parameters can be imaged at an average 34 frames per second. The developed method has the potential to become a valuable tool for real-time display of optical spectroscopy data during continuous scanning applications that subsequently can be used for tissue characterization and diagnosis.

  9. Rotational multispectral fluorescence lifetime imaging and intravascular ultrasound: bimodal system for intravascular applications

    Science.gov (United States)

    Ma, Dinglong; Bec, Julien; Yankelevich, Diego R.; Gorpas, Dimitris; Fatakdawala, Hussain; Marcu, Laura

    2014-01-01

    Abstract. We report the development and validation of a hybrid intravascular diagnostic system combining multispectral fluorescence lifetime imaging (FLIm) and intravascular ultrasound (IVUS) for cardiovascular imaging applications. A prototype FLIm system based on fluorescence pulse sampling technique providing information on artery biochemical composition was integrated with a commercial IVUS system providing information on artery morphology. A customized 3-Fr bimodal catheter combining a rotational side-view fiberoptic and a 40-MHz IVUS transducer was constructed for sequential helical scanning (rotation and pullback) of tubular structures. Validation of this bimodal approach was conducted in pig heart coronary arteries. Spatial resolution, fluorescence detection efficiency, pulse broadening effect, and lifetime measurement variability of the FLIm system were systematically evaluated. Current results show that this system is capable of temporarily resolving the fluorescence emission simultaneously in multiple spectral channels in a single pullback sequence. Accurate measurements of fluorescence decay characteristics from arterial segments can be obtained rapidly (e.g., 20 mm in 5 s), and accurate co-registration of fluorescence and ultrasound features can be achieved. The current finding demonstrates the compatibility of FLIm instrumentation with in vivo clinical investigations and its potential to complement conventional IVUS during catheterization procedures. PMID:24898604

  10. A fast global fitting algorithm for fluorescence lifetime imaging microscopy based on image segmentation.

    Science.gov (United States)

    Pelet, S; Previte, M J R; Laiho, L H; So, P T C

    2004-10-01

    Global fitting algorithms have been shown to improve effectively the accuracy and precision of the analysis of fluorescence lifetime imaging microscopy data. Global analysis performs better than unconstrained data fitting when prior information exists, such as the spatial invariance of the lifetimes of individual fluorescent species. The highly coupled nature of global analysis often results in a significantly slower convergence of the data fitting algorithm as compared with unconstrained analysis. Convergence speed can be greatly accelerated by providing appropriate initial guesses. Realizing that the image morphology often correlates with fluorophore distribution, a global fitting algorithm has been developed to assign initial guesses throughout an image based on a segmentation analysis. This algorithm was tested on both simulated data sets and time-domain lifetime measurements. We have successfully measured fluorophore distribution in fibroblasts stained with Hoechst and calcein. This method further allows second harmonic generation from collagen and elastin autofluorescence to be differentiated in fluorescence lifetime imaging microscopy images of ex vivo human skin. On our experimental measurement, this algorithm increased convergence speed by over two orders of magnitude and achieved significantly better fits. Copyright 2004 Biophysical Society

  11. Using multiphoton fluorescence lifetime imaging to characterize liver damage and fluorescein disposition in liver in vivo

    Science.gov (United States)

    Thorling, Camilla A.; Studier, Hauke; Crawford, Darrell; Roberts, Michael S.

    2016-03-01

    Liver disease is the fifth most common cause of death and unlike many other major causes of mortality, liver disease rates are increasing rather than decreasing. There is no ideal measurement of liver disease and although biopsies are the gold standard, this only allows for a spot examination and cannot follow dynamic processes of the liver. Intravital imaging has the potential to extract detailed information over a larger sampling area continuously. The aim of this project was to investigate whether multiphoton and fluorescence lifetime imaging microscopy could detect early liver damage and to assess whether it could detect changes in metabolism of fluorescein in normal and diseased livers. Four experimental groups were used in this study: 1) control; 2) ischemia reperfusion injury; 3) steatosis and 4) steatosis with ischemia reperfusion injury. Results showed that multiphoton microscopy could visualize morphological changes such as decreased fluorescence of endogenous fluorophores and the presence of lipid droplets, characteristic of steatosis. Fluorescence lifetime imaging microscopy showed increase in NADPH in steatosis with and without ischemia reperfusion injury and could detect changes in metabolism of fluorescein to fluorescein monoglurcuronide, which was impaired in steatosis with ischemia reperfusion injury. These results concluded that the combination of multiphoton microscopy and fluorescence lifetime imaging is a promising method of assessing early stage liver damage and that it can be used to study changes in drug metabolism in the liver as an indication of liver disease and has the potential to replace the traditional static liver biopsy currently used.

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

    Science.gov (United States)

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

    1999-07-01

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

  13. Fluorescence lifetime imaging of endogenous molecules in live mouse cancer models (Conference Presentation)

    Science.gov (United States)

    Svindrych, Zdenek; Wang, Tianxiong; Hu, Song; Periasamy, Ammasi

    2017-02-01

    NADH and FAD are important endogenous fluorescent coenzymes participating in key enzymatic reactions of cellular metabolism. While fluorescence intensities of NADH and FAD have been used to determine the redox state of cells and tissues, this simple approach breaks down in the case of deep-tissue intravital imaging due to depth- and wavelength-dependent light absorption and scattering. To circumvent this limitation, our research focuses on fluorescence lifetimes of two-photon excited NADH and FAD emission to study the metabolic state of live tissues. In our custom-built scanning microscope we combine tunable femtosecond Ti:sapphire laser (operating at 740 nm for NADH excitation and 890 nm for FAD excitation), two GaAsP hybrid detectors for registering individual fluorescence photons and two Becker and Hickl time correlator boards for high precision lifetime measurements. Together with our rigorous FLIM analysis approach (including image segmentation, multi-exponential decay fitting and detailed statistical analysis) we are able to detect metabolic changes in cancer xenografts (human pancreatic cancer MPanc96 cells injected subcutaneously into the ear of an immunodeficient nude mouse), relative to surrounding healthy tissue. Advantageously, with the same instrumentation we can also take high-resolution and high-contrast images of second harmonic signal (SHG) originating from collagen fibers of both the healthy skin and the growing tumor. The combination of metabolic measurements (NADH and FAD lifetime) and morphological information (collagen SHG) allows us to follow the tumor growth in live mouse model and the changes in tumor microenvironment.

  14. In Vivo Fluorescence Lifetime Imaging Monitors Binding of Specific Probes to Cancer Biomarkers

    Science.gov (United States)

    Ardeshirpour, Yasaman; Chernomordik, Victor; Zielinski, Rafal; Capala, Jacek; Griffiths, Gary; Vasalatiy, Olga; Smirnov, Aleksandr V.; Knutson, Jay R.; Lyakhov, Ilya; Achilefu, Samuel; Gandjbakhche, Amir; Hassan, Moinuddin

    2012-01-01

    One of the most important factors in choosing a treatment strategy for cancer is characterization of biomarkers in cancer cells. Particularly, recent advances in Monoclonal Antibodies (MAB) as primary-specific drugs targeting tumor receptors show that their efficacy depends strongly on characterization of tumor biomarkers. Assessment of their status in individual patients would facilitate selection of an optimal treatment strategy, and the continuous monitoring of those biomarkers and their binding process to the therapy would provide a means for early evaluation of the efficacy of therapeutic intervention. In this study we have demonstrated for the first time in live animals that the fluorescence lifetime can be used to detect the binding of targeted optical probes to the extracellular receptors on tumor cells in vivo. The rationale was that fluorescence lifetime of a specific probe is sensitive to local environment and/or affinity to other molecules. We attached Near-InfraRed (NIR) fluorescent probes to Human Epidermal Growth Factor 2 (HER2/neu)-specific Affibody molecules and used our time-resolved optical system to compare the fluorescence lifetime of the optical probes that were bound and unbound to tumor cells in live mice. Our results show that the fluorescence lifetime changes in our model system delineate HER2 receptor bound from the unbound probe in vivo. Thus, this method is useful as a specific marker of the receptor binding process, which can open a new paradigm in the “image and treat” concept, especially for early evaluation of the efficacy of the therapy. PMID:22384092

  15. In vivo fluorescence lifetime imaging monitors binding of specific probes to cancer biomarkers.

    Directory of Open Access Journals (Sweden)

    Yasaman Ardeshirpour

    Full Text Available One of the most important factors in choosing a treatment strategy for cancer is characterization of biomarkers in cancer cells. Particularly, recent advances in Monoclonal Antibodies (MAB as primary-specific drugs targeting tumor receptors show that their efficacy depends strongly on characterization of tumor biomarkers. Assessment of their status in individual patients would facilitate selection of an optimal treatment strategy, and the continuous monitoring of those biomarkers and their binding process to the therapy would provide a means for early evaluation of the efficacy of therapeutic intervention. In this study we have demonstrated for the first time in live animals that the fluorescence lifetime can be used to detect the binding of targeted optical probes to the extracellular receptors on tumor cells in vivo. The rationale was that fluorescence lifetime of a specific probe is sensitive to local environment and/or affinity to other molecules. We attached Near-InfraRed (NIR fluorescent probes to Human Epidermal Growth Factor 2 (HER2/neu-specific Affibody molecules and used our time-resolved optical system to compare the fluorescence lifetime of the optical probes that were bound and unbound to tumor cells in live mice. Our results show that the fluorescence lifetime changes in our model system delineate HER2 receptor bound from the unbound probe in vivo. Thus, this method is useful as a specific marker of the receptor binding process, which can open a new paradigm in the "image and treat" concept, especially for early evaluation of the efficacy of the therapy.

  16. Efficacy of photodynamic therapy against larvae of Aedes aegypti: confocal microscopy and fluorescence-lifetime imaging

    Science.gov (United States)

    de Souza, L. M.; Pratavieira, S.; Inada, N. M.; Kurachi, C.; Corbi, J.; Guimarães, F. E. G.; Bagnato, V. S.

    2014-03-01

    Recently a few demonstration on the use of Photodynamic Reaction as possibility to eliminate larvae that transmit diseases for men has been successfully demonstrated. This promising tool cannot be vastly used due to many problems, including the lake of investigation concerning the mechanisms of larvae killing as well as security concerning the use of photosensitizers in open environment. In this study, we investigate some of the mechanisms in which porphyrin (Photogem) is incorporated on the Aedes aegypti larvae previously to illumination and killing. Larvae at second instar were exposed to the photosensitizer and after 30 minutes imaged by a confocal fluorescence microscope. It was observed the presence of photosensitizer in the gut and at the digestive tract of the larva. Fluorescence-Lifetime Imaging showed greater photosensitizer concentration in the intestinal wall of the samples, which produces a strong decrease of the Photogem fluorescence lifetime. For Photodynamic Therapy exposition to different light doses and concentrations of porphyrin were employed. Three different light sources (LED, Fluorescent lamp, Sun light) also were tested. Sun light and fluorescent lamp shows close to 100% of mortality after 24 hrs. of illumination. These results indicate the potential use of photodynamic effect against the LARVAE of Aedes aegypti.

  17. Fluorescence lifetime measurement with confocal endomicroscopy for direct analysis of tissue biochemistry in vivo

    Directory of Open Access Journals (Sweden)

    Youngjae Won

    2016-08-01

    Full Text Available Confocal endomicroscopy is a powerful tool for in vivo real-time imaging at cellular resolution inside a living body without tissue resection. Microscopic fluorescence lifetime measurement can provide information about localized biochemical conditions such as pH and the concentrations of oxygen and calcium. We hypothesized that combining these techniques could assist accurate cancer discrimination by providing both biochemical and morphological information. We designed a dual-mode experimental setup for confocal endomicroscopic imaging and fluorescence lifetime measurement and applied it to a mouse xenograft model of activated human pancreatic cancer generated by subcutaneous injection of AsPC-1 tumor cells. Using this method with pH-sensitive sodium fluorescein injection, we demonstrated discrimination between normal and cancerous tissues in a living mouse. With further development, this method may be useful for clinical cancer detection.

  18. Label-free identification of macrophage phenotype by fluorescence lifetime imaging microscopy

    Science.gov (United States)

    Alfonso-García, Alba; Smith, Tim D.; Datta, Rupsa; Luu, Thuy U.; Gratton, Enrico; Potma, Eric O.; Liu, Wendy F.

    2016-04-01

    Macrophages adopt a variety of phenotypes that are a reflection of the many functions they perform as part of the immune system. In particular, metabolism is a phenotypic trait that differs between classically activated, proinflammatory macrophages, and alternatively activated, prohealing macrophages. Inflammatory macrophages have a metabolism based on glycolysis while alternatively activated macrophages generally rely on oxidative phosphorylation to generate chemical energy. We employ this shift in metabolism as an endogenous marker to identify the phenotype of individual macrophages via live-cell fluorescence lifetime imaging microscopy (FLIM). We demonstrate that polarized macrophages can be readily discriminated with the aid of a phasor approach to FLIM, which provides a fast and model-free method for analyzing fluorescence lifetime images.

  19. Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate.

    Science.gov (United States)

    Eibl, Matthias; Karpf, Sebastian; Weng, Daniel; Hakert, Hubertus; Pfeiffer, Tom; Kolb, Jan Philip; Huber, Robert

    2017-07-01

    Two-photon-excited fluorescence lifetime imaging microscopy (FLIM) is a chemically specific 3-D sensing modality providing valuable information about the microstructure, composition and function of a sample. However, a more widespread application of this technique is hindered by the need for a sophisticated ultra-short pulse laser source and by speed limitations of current FLIM detection systems. To overcome these limitations, we combined a robust sub-nanosecond fiber laser as the excitation source with high analog bandwidth detection. Due to the long pulse length in our configuration, more fluorescence photons are generated per pulse, which allows us to derive the lifetime with a single excitation pulse only. In this paper, we show high quality FLIM images acquired at a pixel rate of 1 MHz. This approach is a promising candidate for an easy-to-use and benchtop FLIM system to make this technique available to a wider research community.

  20. Exciton-polaron quenching in organic thin-film transistors studied by fluorescence lifetime imaging microscopy

    DEFF Research Database (Denmark)

    Jensen, Per Baunegaard With; Leißner, Till; Osadnik, Andreas

    Organic semiconductors show great potential in electronic and optical applications. However, a major challenge is the degradation of the semiconductor materials that cause a reduction in device performance. Here, we present our investigations of Organic Thin Film Transistors (OTFT) based...... that correlates with the local charge density indicates a pronounced exciton quenching by the injected charges. Subsequent FLIM measurements on previously biased OTFT devices show a general decrease in fluorescence lifetime suggesting degradation of the organic semiconductor. This is correlated with the results...

  1. Diagnosis of basal cell carcinoma by two photon excited fluorescence combined with lifetime imaging

    Science.gov (United States)

    Fan, Shunping; Peng, Xiao; Liu, Lixin; Liu, Shaoxiong; Lu, Yuan; Qu, Junle

    2014-02-01

    Basal cell carcinoma (BCC) is the most common type of human skin cancer. The traditional diagnostic procedure of BCC is histological examination with haematoxylin and eosin staining of the tissue biopsy. In order to reduce complexity of the diagnosis procedure, a number of noninvasive optical methods have been applied in skin examination, for example, multiphoton tomography (MPT) and fluorescence lifetime imaging microscopy (FLIM). In this study, we explored two-photon optical tomography of human skin specimens using two-photon excited autofluorescence imaging and FLIM. There are a number of naturally endogenous fluorophores in skin sample, such as keratin, melanin, collagen, elastin, flavin and porphyrin. Confocal microscopy was used to obtain structures of the sample. Properties of epidermic and cancer cells were characterized by fluorescence emission spectra, as well as fluorescence lifetime imaging. Our results show that two-photon autofluorescence lifetime imaging can provide accurate optical biopsies with subcellular resolution and is potentially a quantitative optical diagnostic method in skin cancer diagnosis.

  2. Assessment of post-implantation integration of engineered tissues using fluorescence lifetime spectroscopy

    Science.gov (United States)

    Elahi, Sakib F.; Lee, Seung Y.; Lloyd, William R.; Chen, Leng-Chun; Kuo, Shiuhyang; Zhou, Ying; Kim, Hyungjin M.; Kennedy, Robert; Marcelo, Cynthia; Feinberg, Stephen E.; Mycek, Mary-Ann

    2018-02-01

    Clinical translation of engineered tissue constructs requires noninvasive methods to assess construct health and viability after implantation in patients. However, current practices to monitor post-implantation construct integration are either qualitative (visual assessment) or destructive (tissue histology). As label-free fluorescence lifetime sensing can noninvasively characterize pre-implantation construct viability, we employed a handheld fluorescence lifetime spectroscopy probe to quantitatively and noninvasively assess tissue constructs that were implanted in a murine model. We designed the system to be suitable for intravital measurements: portability, localization with precise maneuverability, and rapid data acquisition. Our model tissue constructs were manufactured from primary human cells to simulate patient variability and were stressed to create a range of health states. Secreted amounts of three cytokines that relate to cellular viability were measured in vitro to assess pre-implantation construct health. In vivo optical sensing assessed tissue integration of constructs at one-week and three-weeks post-implantation. At one-week post-implantation, optical parameters correlated with in vitro pre-implantation secretion levels of all three cytokines (p clinical optical diagnostic tools based on label-free fluorescence lifetime sensing of endogenous tissue fluorophores could noninvasively monitor post-implantation integration of engineered tissues.

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

    International Nuclear Information System (INIS)

    Badugu, Ramachandram; Lakowicz, Joseph R.; Geddes, Chris D.

    2004-01-01

    We characterize six new fluorescent probes that show both intensity and lifetime changes in the presence of free uncomplexed aqueous cyanide, allowing for fluorescence based cyanide sensing up to physiological safeguard levels, i.e. 2 to the anionic R-B - (CN) 3 form, a new cyanide binding mechanism which we have recently reported. The presence of an electron deficient quaternary heterocyclic nitrogen nucleus, and the electron rich cyanide bound form, provides for the intensity changes observed. We have determined the disassociation constants of the probes to be in the range ∼15-84 μM 3 . In addition we have synthesized control compounds which do not contain the boronic acid moiety, allowing for a rationale of the cyanide responses between the probe isomers to be made. The lifetime of the cyanide bound probes are significantly shorter than the free R-B(OH) 2 probe forms, providing for the opportunity of lifetime based cyanide sensing up to physiologically lethal levels. Finally, while fluorescent probes containing the boronic acid moiety have earned a well-deserved reputation for monosaccharide sensing, we show that strong bases such as CN - and OH - preferentially bind as compared to glucose, enabling the potential use of these probes for cyanide safeguard and determination in physiological fluids, especially given that physiologies do not experience any notable changes in pH

  4. Fluorescence Lifetime Readouts of Troponin-C-Based Calcium FRET Sensors: A Quantitative Comparison of CFP and mTFP1 as Donor Fluorophores

    Science.gov (United States)

    Laine, Romain; Stuckey, Daniel W.; Manning, Hugh; Warren, Sean C.; Kennedy, Gordon; Carling, David

    2012-01-01

    We have compared the performance of two Troponin-C-based calcium FRET sensors using fluorescence lifetime read-outs. The first sensor, TN-L15, consists of a Troponin-C fragment inserted between CFP and Citrine while the second sensor, called mTFP-TnC-Cit, was realized by replacing CFP in TN-L15 with monomeric Teal Fluorescent Protein (mTFP1). Using cytosol preparations of transiently transfected mammalian cells, we have measured the fluorescence decay profiles of these sensors at controlled concentrations of calcium using time-correlated single photon counting. These data were fitted to discrete exponential decay models using global analysis to determine the FRET efficiency, fraction of donor molecules undergoing FRET and calcium affinity of these sensors. We have also studied the decay profiles of the donor fluorescent proteins alone and determined the sensitivity of the donor lifetime to temperature and emission wavelength. Live-cell fluorescence lifetime imaging (FLIM) of HEK293T cells expressing each of these sensors was also undertaken. We confirmed that donor fluorescence of mTFP-TnC-Cit fits well to a two-component decay model, while the TN-L15 lifetime data was best fitted to a constrained four-component model, which was supported by phasor analysis of the measured lifetime data. If the constrained global fitting is employed, the TN-L15 sensor can provide a larger dynamic range of lifetime readout than the mTFP-TnC-Cit sensor but the CFP donor is significantly more sensitive to changes in temperature and emission wavelength compared to mTFP and, while the mTFP-TnC-Cit solution phase data broadly agreed with measurements in live cells, this was not the case for the TN-L15 sensor. Our titration experiment also indicates that a similar precision in determination of calcium concentration can be achieved with both FRET biosensors when fitting a single exponential donor fluorescence decay model to the fluorescence decay profiles. We therefore suggest that m

  5. Multimodal optical coherence tomography and fluorescence lifetime imaging with interleaved excitation sources for simultaneous endogenous and exogenous fluorescence.

    Science.gov (United States)

    Shrestha, Sebina; Serafino, Michael J; Rico-Jimenez, Jesus; Park, Jesung; Chen, Xi; Zhaorigetu, Siqin; Walton, Brian L; Jo, Javier A; Applegate, Brian E

    2016-09-01

    Multimodal imaging probes a variety of tissue properties in a single image acquisition by merging complimentary imaging technologies. Exploiting synergies amongst the data, algorithms can be developed that lead to better tissue characterization than could be accomplished by the constituent imaging modalities taken alone. The combination of optical coherence tomography (OCT) with fluorescence lifetime imaging microscopy (FLIM) provides access to detailed tissue morphology and local biochemistry. The optical system described here merges 1310 nm swept-source OCT with time-domain FLIM having excitation at 355 and 532 nm. The pulses from 355 and 532 nm lasers have been interleaved to enable simultaneous acquisition of endogenous and exogenous fluorescence signals, respectively. The multimodal imaging system was validated using tissue phantoms. Nonspecific tagging with Alexa Flour 532 in a Watanbe rabbit aorta and active tagging of the LOX-1 receptor in human coronary artery, demonstrate the capacity of the system for simultaneous acquisition of OCT, endogenous FLIM, and exogenous FLIM in tissues.

  6. Fluorescence lifetime microscopy for monitoring cell adhesion using metal induced energy transfer

    Science.gov (United States)

    Hwang, Wonsang; Seo, JinWon; Song, Jun ho; Kim, DongEun; Won, YoungJae; Choi, In-Hong; Yoo, Kyung-Hwa; Kim, Dug Young

    2018-02-01

    A precise control and a reliable monitoring tool for the adhesion properties of a cell are very important in atherosclerosis studies. If endothelial cells in contact with the intracellular membrane are not attached securely, low-density lipoprotein (LDL) particles can enter into the inner membrane. It is therefore necessary to measure conditions under which endothelial cell detachment occurs. When a cell is attached to a metal thin film, the lifetime of a fluorescence probe attached to the membrane of the cell is reduced by the metal-induced energy transfer (MIET). Fluorescence lifetime imaging microscopy (FLIM) is used to monitor the attachment condition of a cell to a metal surface using FRET. However, this requires high numerical aperture (NA) objective lens because axial confocal resolution must be smaller than the cell thickness. This requirement limits the field of view of the measurement specimen. In this study we provides a new method which can measure adhesion properties of endothelial cells even with a low NA objective lens by resolving two lifetime components in FLIM.

  7. Xanthophyll cycle-dependent quenching of photosystem II chlorophyll a fluorescence: Formation of a quenching complex with a short fluorescence lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Gilmore, A.M.; Hazlett, T.L.; Govindjee [Univ. of Illinois, Urbana, IL (United States)

    1995-03-14

    Excess light triggers protective nonradiative dissipation of excitation energy in photosystem II through the formation of a trans-thylakoid pH gradient that in turn stimulates formation of zeaxanthin and antheraxanthin. These xanthophylls when combined with protonation of antenna pigment-protein complexes may increase nonradiative dissipation and, thus, quench chlorophyll a fluorescence. Here we measured, in parallel, the chlorophyll a fluorescence lifetime and intensity to understand the mechanism of this process. Increasing the xanthophyll concentration in the presence of a pH gradient (quenched conditions) decreases the fractional intensity of a fluorescence lifetime component centered at {approx}2 ns and increases a component at {approx}0.4 ns. Uncoupling the pH gradient (unquenched conditions) eliminates the 0.4-ns component. Changes in the xanthophyll concentration do not significantly affect the fluorescence lifetimes in either the quenched or unquenched sample conditions. However, there are differences in fluorescence lifetimes between the quenched and unquenched states that are due to pH-related, but nonxanthophyll-related, processes. Quenching of the maximal fluorescence intensity correlates with both the xanthophyll concentration and the fractional intensity of the 0.4-ns component. The unchanged fluorescence lifetimes and the proportional quenching of the maximal and dark-level fluorescence intensities indicate that the xanthophyllact on antenna, not reaction center processes. Further, the fluorescence quenching is interpreted as the combined effect of the pH gradient and xanthophyll concentration, resulting in the formation of a quenching complex with a short ({approx}0.4 ns) fluorescence lifetime. 33 refs., 6 figs., 2 tabs.

  8. Fluorescence lifetime microscopy of NADH distinguishes alterations in cerebral metabolism in vivo.

    Science.gov (United States)

    Yaseen, Mohammad A; Sutin, Jason; Wu, Weicheng; Fu, Buyin; Uhlirova, Hana; Devor, Anna; Boas, David A; Sakadžić, Sava

    2017-05-01

    Evaluating cerebral energy metabolism at microscopic resolution is important for comprehensively understanding healthy brain function and its pathological alterations. Here, we resolve specific alterations in cerebral metabolism in vivo in Sprague Dawley rats utilizing minimally-invasive 2-photon fluorescence lifetime imaging (2P-FLIM) measurements of reduced nicotinamide adenine dinucleotide (NADH) fluorescence. Time-resolved fluorescence lifetime measurements enable distinction of different components contributing to NADH autofluorescence. Ostensibly, these components indicate different enzyme-bound formulations of NADH. We observed distinct variations in the relative proportions of these components before and after pharmacological-induced impairments to several reactions involved in glycolytic and oxidative metabolism. Classification models were developed with the experimental data and used to predict the metabolic impairments induced during separate experiments involving bicuculline-induced seizures. The models consistently predicted that prolonged focal seizure activity results in impaired activity in the electron transport chain, likely the consequence of inadequate oxygen supply. 2P-FLIM observations of cerebral NADH will help advance our understanding of cerebral energetics at a microscopic scale. Such knowledge will aid in our evaluation of healthy and diseased cerebral physiology and guide diagnostic and therapeutic strategies that target cerebral energetics.

  9. Carrier Lifetimes in Fluorescent 6H-SiC for LEDs Application

    DEFF Research Database (Denmark)

    Grivickas, Vytautas; Gulbinas, Karolis; Jokubavičius, Valdas

    Recently it was shown a new approach based on all-semiconductor material technology which is composed with a near ultra-violet GaN LED excitation source and fluorescent silicon carbide (f-6H-SiC) substrate which generates a visible broad spectral light by N and B dopants and an efficient donor...... to acceptor pair recombination [1,2]. This combination can achieve higher electric-light conversion efficiency and high color rendering in comparison with today’s used blue GaN LED based and phosphors. The devices are promising candidates for general lightning applications and may obtain stability...... under co-linear and orthogonal probe geometry was used to measure carrier lifetimes in the layers under variable injection conditions. Same results are shown in Fig. 1 exaggerating the fact that longer electron lifetime responsible for higher emission and n-type doping should prevail the p-type doping...

  10. Monitoring macular pigment changes in macular holes using fluorescence lifetime imaging ophthalmoscopy.

    Science.gov (United States)

    Sauer, Lydia; Peters, Sven; Schmidt, Johanna; Schweitzer, Dietrich; Klemm, Matthias; Ramm, Lisa; Augsten, Regine; Hammer, Martin

    2017-08-01

    To investigate the impact of macular pigment (MP) on fundus autofluorescence (FAF) lifetimes in vivo by characterizing full-thickness idiopathic macular holes (MH) and macular pseudo-holes (MPH). A total of 37 patients with MH and 52 with MPH were included. Using the fluorescence lifetime imaging ophthalmoscope (FLIO), based on a Heidelberg Engineering Spectralis system, a 30° retinal field was investigated. FAF decays were detected in a short (498-560 nm; ch1) and long (560-720 nm; ch2) wavelength channel. τ m , the mean fluorescence lifetime, was calculated from a three-exponential approximation of the FAF decays. Macular coherence tomography scans were recorded, and macular pigment's optical density (MPOD) was measured (one-wavelength reflectometry). Two MH subgroups were analysed according to the presence or absence of an operculum above the MH. A total of 17 healthy fellow eyes were included. A longitudinal FAF decay examination was conducted in nine patients, which were followed up after surgery and showed a closed MH. In MH without opercula, significant τ m differences (p hole area (MHa) and surrounding areas (MHb) (ch1: MHa 238 ± 64 ps, MHb 181 ± 78 ps; ch2: MHa 275 ± 49 ps, MHb 223 ± 48 ps), as well as between MHa and healthy eyes or closed MH. Shorter τ m , adjacent to the hole, can be assigned to areas with equivalently higher MPOD. Opercula containing MP also show short τ m . In MPH, the intactness of the Hele fibre layer is associated with shortest τ m . Shortest τ m originates from MP-containing retinal layers, especially from the Henle fibre layer. Fluorescence lifetime imaging ophthalmoscope (FLIO) provides information on the MP distribution, the pathogenesis and topology of MH. Macular pigment (MP) fluorescence may provide a biomarker for monitoring pathological changes in retinal diseases. © 2016 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

  11. Fluorescence lifetime imaging of microviscosity changes during ER autophagy in live cells

    Science.gov (United States)

    He, Ying; Samanta, Soham; Gong, Wanjun; Liu, Wufan; Pan, Wenhui; Yang, Zhigang; Qu, Junle

    2018-02-01

    Unfolded or misfolded protein accumulation inside Endoplasmic Reticulum (ER) will cause ER stress and subsequently will activate cellular autophagy to release ER stress, which would ultimately result in microviscosity changes. However, even though, it is highly significant to gain a quantitative assessment of microviscosity changes during ER autophagy to study ER stress and autophagy behaviors related diseases, it has rarely been reported yet. In this work, we have reported a BODIPY based fluorescent molecular rotor that can covalently bind with vicinal dithiols containing nascent proteins in ER and hence can result in ER stress through the inhibition of the folding of nascent proteins. The change in local viscosity, caused by the release of the stress in cells through autophagy, was quantified by the probe using fluorescence lifetime imaging. This work basically demonstrates the possibility of introducing synthetic chemical probe as a promising tool to diagnose ER-viscosity-related diseases.

  12. Fluorescence lifetime components reveal kinetic intermediate states upon equilibrium denaturation of carbonic anhydrase II.

    Science.gov (United States)

    Nemtseva, Elena V; Lashchuk, Olesya O; Gerasimova, Marina A; Melnik, Tatiana N; Nagibina, Galina S; Melnik, Bogdan S

    2017-12-21

    In most cases, intermediate states of multistage folding proteins are not 'visible' under equilibrium conditions but are revealed in kinetic experiments. Time-resolved fluorescence spectroscopy was used in equilibrium denaturation studies. The technique allows for detecting changes in the conformation and environment of tryptophan residues in different structural elements of carbonic anhydrase II which in its turn has made it possible to study the intermediate states of carbonic anhydrase II under equilibrium conditions. The results of equilibrium and kinetic experiments using wild-type bovine carbonic anhydrase II and its mutant form with the substitution of leucine for alanine at position 139 (L139A) were compared. The obtained lifetime components of intrinsic tryptophan fluorescence allowed for revealing that, the same as in kinetic experiments, under equilibrium conditions the unfolding of carbonic anhydrase II ensues through formation of intermediate states.

  13. CVD grown 2D MoS{sub 2} layers: A photoluminescence and fluorescence lifetime imaging study

    Energy Technology Data Exchange (ETDEWEB)

    Oezden, Ayberk; Madenoglu, Buesra [Department of Materials Science and Engineering, Faculty of Engineering, Anadolu University, Eskisehir (Turkey); Sar, Hueseyin; Ay, Feridun; Perkgoez, Nihan Kosku [Department of Electrical and Electronics Engineering, Faculty of Engineering, Anadolu University, Eskisehir (Turkey); Yeltik, Aydan [Department of Physics, UNAM Institute of Materials Science and Nanotechnology, Bilkent University, Ankara (Turkey); Sevik, Cem [Department of Mechanical Engineering, Faculty of Engineering, Anadolu University, Eskisehir (Turkey)

    2016-11-15

    In this letter, we report on the fluorescence lifetime imaging and accompanying photoluminescence properties of a chemical vapour deposition (CVD) grown atomically thin material, MoS{sub 2}. μ-Raman, μ-photoluminescence (PL) and fluorescence lifetime imaging microscopy (FLIM) are utilized to probe the fluorescence lifetime and photoluminescence properties of individual flakes of MoS{sub 2} films. Usage of these three techniques allows identification of the grown layers, grain boundaries, structural defects and their relative effects on the PL and fluorescence lifetime spectra. Our investigation on individual monolayer flakes reveals a clear increase of the fluorescence lifetime from 0.3 ns to 0.45 ns at the edges with respect to interior region. On the other hand, investigation of the film layer reveals quenching of PL intensity and lifetime at the grain boundaries. These results could be important for applications where the activity of edges is important such as in photocatalytic water splitting. Finally, it has been demonstrated that PL mapping and FLIM are viable techniques for the investigation of the grain-boundaries. (copyright 2016 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  14. Fluorescence single-molecule counting assays for protein quantification using epi-fluorescence microscopy with quantum dots labeling

    International Nuclear Information System (INIS)

    Jiang Dafeng; Liu Chunxia; Wang Lei; Jiang Wei

    2010-01-01

    A single-molecule counting approach for quantifying the antibody affixed to a surface using quantum dots and epi-fluorescence microscopy is presented. Modifying the glass substrates with carboxyl groups provides a hydrophilic surface that reacts with amine groups of an antibody to allow covalent immobilization of the antibody. Nonspecific adsorption of single molecules on the modified surfaces was first investigated. Then, quantum dots were employed to form complexes with surface-immobilized antibody molecules and used as fluorescent probes for single-molecule imaging. Epi-fluorescence microscopy was chosen as the tool for single-molecule fluorescence detection here. The generated fluorescence signals were taken by an electron multiplying charge-coupled device and were found to be proportional to the sample concentrations. Under optimal conditions, a linear response range of 5.0 x 10 -14 -3.0 x 10 -12 mol L -1 was obtained between the number of single molecules and sample concentration via a single-molecule counting approach.

  15. Conversion of Beckman DK-2A spectrophotometer into an automatic single-photon counting fluorescence spectrophotometer

    International Nuclear Information System (INIS)

    Chikkur, G.C.; Lagare, M.T.; Umakantha, N.

    1981-01-01

    Details of how a DK-2A spectrophotometer can be modified into an automatic single-photon counting fluorescence spectrophotometer for recording a low intensity spectrum, are reported. The single-photon count-rate converted into a DC voltage is applied at the appropriate stage in the sample channel amplifier circuit of a DK-2A to get the pen deflection proportional to the count-rate. A high intensity spectrum may be recorded in the usual way by merely turning the shaft of the mirror motor by 180 degrees. (author)

  16. Measurement of pH micro-heterogeneity in natural cheese matrices by fluorescence lifetime imaging

    Directory of Open Access Journals (Sweden)

    Zuzana eBurdikova

    2015-03-01

    Full Text Available Cheese, a product of microbial fermentation may be defined as a protein matrix entrapping fat, moisture, minerals and solutes as well as dispersed bacterial colonies. The growth and physiology of bacterial cells in these colonies may be influenced by the microenvironment around the colony, or alternatively the cells within the colony may modify the microenvironment (e.g. pH, redox potential due to their metabolic activity. While cheese pH may be measured at macro level there remains a significant knowledge gap relating to the degree of micro-heterogeneity of pH within the cheese matrix and its relationship with microbial, enzymatic and physiochemical parameters and ultimately with cheese quality, consistency and ripening patterns. The pH of cheese samples was monitored both at macroscopic scale and at microscopic scale, using a non-destructive microscopic technique employing C-SNARF-4 and Oregon Green 488 fluorescent probes. The objectives of this work were to evaluate the suitability of these dyes for microscale pH measurements in natural cheese matrices and to enhance the sensitivity and extend the useful pH range of these probes using fluorescence lifetime imaging (FLIM. In particular, fluorescence lifetime of Oregon Green 488 proved to be sensitive probe to map pH micro heterogeneity within cheese matrices. Good agreement was observed between macroscopic scale pH measurement by FLIM and by traditional pH methods, but in addition considerable localized microheterogeneity in pH was evident within the curd matrix with pH range between 4.0 and 5.5. This technique provides significant potential to further investigate the relationship between cheese matrix physico-chemistry and bacterial metabolism during cheese manufacture and ripening.

  17. Measurement of pH micro-heterogeneity in natural cheese matrices by fluorescence lifetime imaging.

    Science.gov (United States)

    Burdikova, Zuzana; Svindrych, Zdenek; Pala, Jan; Hickey, Cian D; Wilkinson, Martin G; Panek, Jiri; Auty, Mark A E; Periasamy, Ammasi; Sheehan, Jeremiah J

    2015-01-01

    Cheese, a product of microbial fermentation may be defined as a protein matrix entrapping fat, moisture, minerals and solutes as well as dispersed bacterial colonies. The growth and physiology of bacterial cells in these colonies may be influenced by the microenvironment around the colony, or alternatively the cells within the colony may modify the microenvironment (e.g., pH, redox potential) due to their metabolic activity. While cheese pH may be measured at macro level there remains a significant knowledge gap relating to the degree of micro-heterogeneity of pH within the cheese matrix and its relationship with microbial, enzymatic and physiochemical parameters and ultimately with cheese quality, consistency and ripening patterns. The pH of cheese samples was monitored both at macroscopic scale and at microscopic scale, using a non-destructive microscopic technique employing C-SNARF-4 and Oregon Green 488 fluorescent probes. The objectives of this work were to evaluate the suitability of these dyes for microscale pH measurements in natural cheese matrices and to enhance the sensitivity and extend the useful pH range of these probes using fluorescence lifetime imaging (FLIM). In particular, fluorescence lifetime of Oregon Green 488 proved to be sensitive probe to map pH micro heterogeneity within cheese matrices. Good agreement was observed between macroscopic scale pH measurement by FLIM and by traditional pH methods, but in addition considerable localized microheterogeneity in pH was evident within the curd matrix with pH range between 4.0 and 5.5. This technique provides significant potential to further investigate the relationship between cheese matrix physico-chemistry and bacterial metabolism during cheese manufacture and ripening.

  18. Generation of a new spectral format, the lifetime synchronous spectrum (LiSS), using phase-resolved fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Shaver, J.M.; McGown, L.B.

    1994-01-01

    A new fluorescence spectral format is introduced in which fluorescence lifetime is shown as a function of synchronously scanned wavelength to generate a Lifetime Synchronous Spectrum (LiSS). Lifetimes are determined in the frequency domain with the use of Phase-Resolved Fluorescence Spectroscopy (PRFS) to obtain the phase of the fluorescence signal. Theory and construction of the LiSS are presented and experimental results are shown for solutions of single components and simple binary and ternary mixtures. These results show how the lifetime information in the LiSS augments the steady-state intensity information of a standard synchronous spectrum, providing unique information for identification of components and resolution of overlapping spectral peaks. The LiSS technique takes advantage of noise reduction inherent in the extraction of lifetime from PRFS in addition to standard spectral smoothing techniques. The precision of phase determination through PRFS is found to be comparable to that of direct phase measurements at normal fluorescence intensities and superior for low-intensity signals

  19. Characterization of energy response for photon-counting detectors using x-ray fluorescence

    International Nuclear Information System (INIS)

    Ding, Huanjun; Cho, Hyo-Min; Molloi, Sabee; Barber, William C.; Iwanczyk, Jan S.

    2014-01-01

    Purpose: To investigate the feasibility of characterizing a Si strip photon-counting detector using x-ray fluorescence. Methods: X-ray fluorescence was generated by using a pencil beam from a tungsten anode x-ray tube with 2 mm Al filtration. Spectra were acquired at 90° from the primary beam direction with an energy-resolved photon-counting detector based on an edge illuminated Si strip detector. The distances from the source to target and the target to detector were approximately 19 and 11 cm, respectively. Four different materials, containing silver (Ag), iodine (I), barium (Ba), and gadolinium (Gd), were placed in small plastic containers with a diameter of approximately 0.7 cm for x-ray fluorescence measurements. Linear regression analysis was performed to derive the gain and offset values for the correlation between the measured fluorescence peak center and the known fluorescence energies. The energy resolutions and charge-sharing fractions were also obtained from analytical fittings of the recorded fluorescence spectra. An analytical model, which employed four parameters that can be determined from the fluorescence calibration, was used to estimate the detector response function. Results: Strong fluorescence signals of all four target materials were recorded with the investigated geometry for the Si strip detector. The average gain and offset of all pixels for detector energy calibration were determined to be 6.95 mV/keV and −66.33 mV, respectively. The detector’s energy resolution remained at approximately 2.7 keV for low energies, and increased slightly at 45 keV. The average charge-sharing fraction was estimated to be 36% within the investigated energy range of 20–45 keV. The simulated detector output based on the proposed response function agreed well with the experimental measurement. Conclusions: The performance of a spectral imaging system using energy-resolved photon-counting detectors is very dependent on the energy calibration of the

  20. GPU accelerated real-time confocal fluorescence lifetime imaging microscopy (FLIM) based on the analog mean-delay (AMD) method

    Science.gov (United States)

    Kim, Byungyeon; Park, Byungjun; Lee, Seungrag; Won, Youngjae

    2016-01-01

    We demonstrated GPU accelerated real-time confocal fluorescence lifetime imaging microscopy (FLIM) based on the analog mean-delay (AMD) method. Our algorithm was verified for various fluorescence lifetimes and photon numbers. The GPU processing time was faster than the physical scanning time for images up to 800 × 800, and more than 149 times faster than a single core CPU. The frame rate of our system was demonstrated to be 13 fps for a 200 × 200 pixel image when observing maize vascular tissue. This system can be utilized for observing dynamic biological reactions, medical diagnosis, and real-time industrial inspection. PMID:28018724

  1. Direct Vpr-Vpr Interaction in Cells monitored by two Photon Fluorescence Correlation Spectroscopy and Fluorescence Lifetime Imaging

    Directory of Open Access Journals (Sweden)

    Mély Yves

    2008-09-01

    Full Text Available Abstract Background The human immunodeficiency virus type 1 (HIV-1 encodes several regulatory proteins, notably Vpr which influences the survival of the infected cells by causing a G2/M arrest and apoptosis. Such an important role of Vpr in HIV-1 disease progression has fuelled a large number of studies, from its 3D structure to the characterization of specific cellular partners. However, no direct imaging and quantification of Vpr-Vpr interaction in living cells has yet been reported. To address this issue, eGFP- and mCherry proteins were tagged by Vpr, expressed in HeLa cells and their interaction was studied by two photon fluorescence lifetime imaging microscopy and fluorescence correlation spectroscopy. Results Results show that Vpr forms homo-oligomers at or close to the nuclear envelope. Moreover, Vpr dimers and trimers were found in the cytoplasm and in the nucleus. Point mutations in the three α helices of Vpr drastically impaired Vpr oligomerization and localization at the nuclear envelope while point mutations outside the helical regions had no effect. Theoretical structures of Vpr mutants reveal that mutations within the α-helices could perturb the leucine zipper like motifs. The ΔQ44 mutation has the most drastic effect since it likely disrupts the second helix. Finally, all Vpr point mutants caused cell apoptosis suggesting that Vpr-mediated apoptosis functions independently from Vpr oligomerization. Conclusion We report that Vpr oligomerization in HeLa cells relies on the hydrophobic core formed by the three α helices. This oligomerization is required for Vpr localization at the nuclear envelope but not for Vpr-mediated apoptosis.

  2. Second-harmonic generation and fluorescence lifetime imaging microscopy through a rodent mammary imaging window

    Science.gov (United States)

    Young, Pamela A.; Nazir, Muhammad; Szulczewski, Michael J.; Keely, Patricia J.; Eliceiri, Kevin W.

    2012-03-01

    Tumor-Associated Collagen Signatures (TACS) have been identified that manifest in specific ways during breast tumor progression and that correspond to patient outcome. There are also compelling metabolic changes associated with carcinoma invasion and progression. We have characterized the difference in the autofluorescent properties of metabolic co-factors, NADH and FAD, between normal and carcinoma breast cell lines. Also, we have shown in vitro that increased collagen density alters metabolic genes which are associated with glycolysis and leads to a more invasive phenotype. Establishing the relationship between collagen density, cellular metabolism, and metastasis in physiologically relevant cancer models is crucial for developing cancer therapies. To study cellular metabolism with respect to collagen density in vivo, we use multiphoton fluorescence excitation microscopy (MPM) in conjunction with a rodent mammary imaging window implanted in defined mouse cancer models. These models are ideal for the study of collagen changes in vivo, allowing determination of corresponding metabolic changes in breast cancer invasion and progression. To measure cellular metabolism, we collect fluorescence lifetime (FLIM) signatures of NADH and FAD, which are known to change based on the microenvironment of the cells. Additionally, MPM systems are capable of collecting second harmonic generation (SHG) signals which are a nonlinear optical property of collagen. Therefore, MPM, SHG, and FLIM are powerful tools with great potential for characterizing key features of breast carcinoma in vivo. Below we present the current efforts of our collaborative group to develop intravital approaches based on these imaging techniques to look at defined mouse mammary models.

  3. In vivo multiphoton and fluorescence lifetime imaging microscopy of the healthy and cholestatic liver

    Science.gov (United States)

    Kuznetsova, Daria S.; Dudenkova, Varvara V.; Rodimova, Svetlana A.; Bobrov, Nikolai V.; Zagainov, Vladimir E.; Zagaynova, Elena V.

    2018-02-01

    A cholestatic liver disease presents one of the most common liver diseases and can potentially progress to cirrhosis or even cholangiocarcinoma. Conventional techniques are insufficient to precisely describe the complex internal structure, heterogeneous cell populations and the dynamics of biological processes of the liver. Currently, the methods of multiphoton and fluorescence lifetime imaging microscopy are actively introducing to biomedical research. Those methods are extremely informative and non-destructive that allows studying of a large number of processes occurring inside cells and tissues, analyzing molecular cellular composition, as well as evaluating the state of connective tissue fibers due to their ability to generate a second optical harmonic. Multiphoton and FLIM microscopy do not need additional staining of samples or the incorporation of any markers to study metabolism, lipid composition, microstructure analysis, evaluation of fibrous structures. These parameters have pronounced changes in hepatocytes of liver with common pathological diseases. Thereby in this study we investigated metabolic changes in the healthy and cholestatic liver based on the fluorescence of the metabolic co-factors NAD(P)H and FAD by multiphoton microscopy combined with FLIM. To estimate the contribution of energy metabolism and lipogenesis in the observed changes of the metabolic profile, a separate analysis of NADH and NADPH was presented. The data can be used to develop new criteria for the identification of hepatic pathology at the level of hepatocyte changes directed to personalized medicine in the future.

  4. Two-photon excitation with pico-second fluorescence lifetime imaging to detect nuclear association of flavanols

    Energy Technology Data Exchange (ETDEWEB)

    Mueller-Harvey, Irene, E-mail: i.mueller-harvey@reading.ac.uk [Chemistry and Biochemistry Laboratory, Food Production and Quality Research Division, School of Agriculture, Policy and Development, University of Reading, P O Box 236, Reading RG6 6AT (United Kingdom); Feucht, Walter, E-mail: walter.feucht@gmail.com [Department of Plant Sciences, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Polster, Juergen, E-mail: j.polster@wzw.tum.de [Department of Physical Biochemistry, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Trnkova, Lucie, E-mail: lucie.trnkova@uhk.cz [University of Hradec Kralove, Faculty of Science, Department of Chemistry, Rokitanskeho 62, 50003 Hradec Kralove (Czech Republic); Burgos, Pierre, E-mail: pierre.burgos@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Parker, Anthony W., E-mail: tony.parker@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Botchway, Stanley W., E-mail: stan.botchway@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom)

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer This fluorescence lifetime imaging microscopy (FLIM) technique for flavanols overcomes autofluorescence interference in cells. Black-Right-Pointing-Pointer Plant flavanols differed in their lifetimes. Black-Right-Pointing-Pointer Dissolved and bound flavanols revealed contrasting lifetime changes. Black-Right-Pointing-Pointer This technique will allow studying of flavanol trafficking in live cells. - Abstract: Two-photon excitation enabled for the first time the observation and measurement of excited state fluorescence lifetimes from three flavanols in solution, which were {approx}1.0 ns for catechin and epicatechin, but <45 ps for epigallocatechin gallate (EGCG). The shorter lifetime for EGCG is in line with a lower fluorescence quantum yield of 0.003 compared to catechin (0.015) and epicatechin (0.018). In vivo experiments with onion cells demonstrated that tryptophan and quercetin, which tend to be major contributors of background fluorescence in plant cells, have sufficiently low cross sections for two-photon excitation at 630 nm and therefore do not interfere with detection of externally added or endogenous flavanols in Allium cepa or Taxus baccata cells. Applying two-photon excitation to flavanols enabled 3-D fluorescence lifetime imaging microscopy and showed that added EGCG penetrated the whole nucleus of onion cells. Interestingly, EGCG and catechin showed different lifetime behaviour when bound to the nucleus: EGCG lifetime increased from <45 to 200 ps, whilst catechin lifetime decreased from 1.0 ns to 500 ps. Semi-quantitative measurements revealed that the relative ratios of EGCG concentrations in nucleoli associated vesicles: nucleus: cytoplasm were ca. 100:10:1. Solution experiments with catechin, epicatechin and histone proteins provided preliminary evidence, via the appearance of a second lifetime ({tau}{sub 2} = 1.9-3.1 ns), that both flavanols may be interacting with histone proteins. We conclude that there

  5. Probing the photoluminescence properties of gold nanoclusters by fluorescence lifetime correlation spectroscopy

    International Nuclear Information System (INIS)

    Yuan, C. T.; Lin, T. N.; Shen, J. L.; Lin, C. A.; Chang, W. H.; Cheng, H. W.; Tang, J.

    2013-01-01

    Gold nanoclusters (Au NCs) have attracted much attention for promising applications in biological imaging owing to their tiny sizes and biocompatibility. So far, most efforts have been focused on the strategies for fabricating high-quality Au NCs and then characterized by conventional ensemble measurement. Here, a fusion single-molecule technique combining fluorescence correlation spectroscopy and time-correlated single-photon counting can be successfully applied to probe the photoluminescence (PL) properties for sparse Au NCs. In this case, the triplet-state dynamics and diffusion process can be observed simultaneously and the relevant time constants can be derived. This work provides a complementary insight into the PL mechanism at the molecular levels for Au NCs in solution

  6. Development of an automated asbestos counting software based on fluorescence microscopy.

    Science.gov (United States)

    Alexandrov, Maxym; Ichida, Etsuko; Nishimura, Tomoki; Aoki, Kousuke; Ishida, Takenori; Hirota, Ryuichi; Ikeda, Takeshi; Kawasaki, Tetsuo; Kuroda, Akio

    2015-01-01

    An emerging alternative to the commonly used analytical methods for asbestos analysis is fluorescence microscopy (FM), which relies on highly specific asbestos-binding probes to distinguish asbestos from interfering non-asbestos fibers. However, all types of microscopic asbestos analysis require laborious examination of large number of fields of view and are prone to subjective errors and large variability between asbestos counts by different analysts and laboratories. A possible solution to these problems is automated counting of asbestos fibers by image analysis software, which would lower the cost and increase the reliability of asbestos testing. This study seeks to develop a fiber recognition and counting software for FM-based asbestos analysis. We discuss the main features of the developed software and the results of its testing. Software testing showed good correlation between automated and manual counts for the samples with medium and high fiber concentrations. At low fiber concentrations, the automated counts were less accurate, leading us to implement correction mode for automated counts. While the full automation of asbestos analysis would require further improvements in accuracy of fiber identification, the developed software could already assist professional asbestos analysts and record detailed fiber dimensions for the use in epidemiological research.

  7. Long-term fluorescence lifetime imaging of a genetically encoded sensor for caspase-3 activity in mouse tumor xenografts

    Science.gov (United States)

    Zherdeva, Victoria; Kazachkina, Natalia I.; Shcheslavskiy, Vladislav; Savitsky, Alexander P.

    2018-03-01

    Caspase-3 is known for its role in apoptosis and programmed cell death regulation. We detected caspase-3 activation in vivo in tumor xenografts via shift of mean fluorescence lifetimes of a caspase-3 sensor. We used the genetically encoded sensor TR23K based on the red fluorescent protein TagRFP and chromoprotein KFP linked by 23 amino acid residues (TagRFP-23-KFP) containing a specific caspase cleavage DEVD motif to monitor the activity of caspase-3 in tumor xenografts by means of fluorescence lifetime imaging-Forster resonance energy transfer. Apoptosis was induced by injection of paclitaxel for A549 lung adenocarcinoma and etoposide and cisplatin for HEp-2 pharynx adenocarcinoma. We observed a shift in lifetime distribution from 1.6 to 1.9 ns to 2.1 to 2.4 ns, which indicated the activation of caspase-3. Even within the same tumor, the lifetime varied presumably due to the tumor heterogeneity and the different depth of tumor invasion. Thus, processing time-resolved fluorescence images allows detection of both the cleaved and noncleaved states of the TR23K sensor in real-time mode during the course of several weeks noninvasively. This approach can be used in drug screening, facilitating the development of new anticancer agents as well as improvement of chemotherapy efficiency and its adaptation for personal treatment.

  8. Using non-empirically tuned range-separated functionals with simulated emission bands to model fluorescence lifetimes.

    Science.gov (United States)

    Wong, Z C; Fan, W Y; Chwee, T S; Sullivan, Michael B

    2017-08-09

    Fluorescence lifetimes were evaluated using TD-DFT under different approximations for the emitting molecule and various exchange-correlation functionals, such as B3LYP, BMK, CAM-B3LYP, LC-BLYP, M06, M06-2X, M11, PBE0, ωB97, ωB97X, LC-BLYP*, and ωB97X* where the range-separation parameters in the last two functionals were tuned in a non-empirical fashion. Changes in the optimised molecular geometries between the ground and electronically excited states were found to affect the quality of the calculated lifetimes significantly, while the inclusion of vibronic features led to further improvements over the assumption of a vertical electronic transition. The LC-BLYP* functional was found to return the most accurate fluorescence lifetimes with unsigned errors that are mostly within 1.5 ns of experimental values.

  9. FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye.

    Directory of Open Access Journals (Sweden)

    Matthias Klemm

    Full Text Available Fluorescence lifetime imaging ophthalmoscopy (FLIO is a new technique for measuring the in vivo autofluorescence intensity decays generated by endogenous fluorophores in the ocular fundus. Here, we present a software package called FLIM eXplorer (FLIMX for analyzing FLIO data. Specifically, we introduce a new adaptive binning approach as an optimal tradeoff between the spatial resolution and the number of photons required per pixel. We also expand existing decay models (multi-exponential, stretched exponential, spectral global analysis, incomplete decay to account for the layered structure of the eye and present a method to correct for the influence of the crystalline lens fluorescence on the retina fluorescence. Subsequently, the Holm-Bonferroni method is applied to FLIO measurements to allow for group comparisons between patients and controls on the basis of fluorescence lifetime parameters. The performance of the new approaches was evaluated in five experiments. Specifically, we evaluated static and adaptive binning in a diabetes mellitus patient, we compared the different decay models in a healthy volunteer and performed a group comparison between diabetes patients and controls. An overview of the visualization capabilities and a comparison of static and adaptive binning is shown for a patient with macular hole. FLIMX's applicability to fluorescence lifetime imaging microscopy is shown in the ganglion cell layer of a porcine retina sample, obtained by a laser scanning microscope using two-photon excitation.

  10. FLIMX: A Software Package to Determine and Analyze the Fluorescence Lifetime in Time-Resolved Fluorescence Data from the Human Eye.

    Science.gov (United States)

    Klemm, Matthias; Schweitzer, Dietrich; Peters, Sven; Sauer, Lydia; Hammer, Martin; Haueisen, Jens

    2015-01-01

    Fluorescence lifetime imaging ophthalmoscopy (FLIO) is a new technique for measuring the in vivo autofluorescence intensity decays generated by endogenous fluorophores in the ocular fundus. Here, we present a software package called FLIM eXplorer (FLIMX) for analyzing FLIO data. Specifically, we introduce a new adaptive binning approach as an optimal tradeoff between the spatial resolution and the number of photons required per pixel. We also expand existing decay models (multi-exponential, stretched exponential, spectral global analysis, incomplete decay) to account for the layered structure of the eye and present a method to correct for the influence of the crystalline lens fluorescence on the retina fluorescence. Subsequently, the Holm-Bonferroni method is applied to FLIO measurements to allow for group comparisons between patients and controls on the basis of fluorescence lifetime parameters. The performance of the new approaches was evaluated in five experiments. Specifically, we evaluated static and adaptive binning in a diabetes mellitus patient, we compared the different decay models in a healthy volunteer and performed a group comparison between diabetes patients and controls. An overview of the visualization capabilities and a comparison of static and adaptive binning is shown for a patient with macular hole. FLIMX's applicability to fluorescence lifetime imaging microscopy is shown in the ganglion cell layer of a porcine retina sample, obtained by a laser scanning microscope using two-photon excitation.

  11. Actin cytoskeleton-dependent Rab GTPase-regulated angiotensin type I receptor lysosomal degradation studied by fluorescence lifetime imaging microscopy

    Science.gov (United States)

    Li, Hewang; Yu, Peiying; Sun, Yuansheng; Felder, Robin A.; Periasamy, Ammasi; Jose, Pedro A.

    2010-09-01

    The dynamic regulation of the cellular trafficking of human angiotensin (Ang) type 1 receptor (AT1R) is not well understood. Therefore, we investigated the cellular trafficking of AT1R-enhanced green fluorescent protein (EGFP) (AT1R-EGFP) heterologously expressed in HEK293 cells by determining the change in donor lifetime (AT1R-EGFP) in the presence or absence of acceptor(s) using fluorescence lifetime imaging-fluorescence resonance energy transfer (FRET) microscopy. The average lifetime of AT1R-EGFP in our donor-alone samples was ~2.33 ns. The basal state lifetime was shortened slightly in the presence of Rab5 (2.01+/-0.10 ns) or Rab7 (2.11+/-0.11 ns) labeled with Alexa 555, as the acceptor fluorophore. A 5-min Ang II treatment markedly shortened the lifetime of AT1R-EGFP in the presence of Rab5-Alexa 555 (1.78+/-0.31 ns) but was affected minimally in the presence of Rab7-Alexa 555 (2.09+/-0.37 ns). A 30-min Ang II treatment further decreased the AT1R-EGFP lifetime in the presence of both Rab5- and Rab7-Alexa 555. Latrunculin A but not nocodazole pretreatment blocked the ability of Ang II to shorten the AT1R-EGFP lifetime. The occurrence of FRET between AT1R-EGFP (donor) and LAMP1-Alexa 555 (acceptor) with Ang II stimulation was impaired by photobleaching the acceptor. These studies demonstrate that Ang II-induced AT1R lysosomal degradation through its association with LAMP1 is regulated by Rab5/7 via mechanisms that are dependent on intact actin cytoskeletons.

  12. On the performance of bioanalytical fluorescence correlation spectroscopy measurements in a multiparameter photon-counting microscope

    Energy Technology Data Exchange (ETDEWEB)

    Mazouchi, Amir; Liu Baoxu; Bahram, Abdullah [Department of Physics, Institute for Optical Sciences, University of Toronto, Toronto (Canada); Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Rd. N., Mississauga, ON, L5L 1C6 (Canada); Gradinaru, Claudiu C., E-mail: claudiu.gradinaru@utoronto.ca [Department of Physics, Institute for Optical Sciences, University of Toronto, Toronto (Canada); Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Rd. N., Mississauga, ON, L5L 1C6 (Canada)

    2011-02-28

    Fluorescence correlation spectroscopy (FCS) data acquisition and analysis routines were developed and implemented in a home-built, multiparameter photon-counting microscope. Laser excitation conditions were investigated for two representative fluorescent probes, Rhodamine110 and enhanced green fluorescent protein (EGFP). Reliable local concentrations and diffusion constants were obtained by fitting measured FCS curves, provided that the excitation intensity did not exceed 20% of the saturation level for each fluorophore. Accurate results were obtained from FCS measurements for sample concentrations varying from pM to {mu}M range, as well as for conditions of high background signals. These experimental constraints were found to be determined by characteristics of the detection system and by the saturation behavior of the fluorescent probes. These factors actually limit the average number of photons that can be collected from a single fluorophore passing through the detection volume. The versatility of our setup and the data analysis capabilities were tested by measuring the mobility of EGFP in the nucleus of Drosophila cells under conditions of high concentration and molecular crowding. As a bioanalytical application, we studied by FCS the binding affinity of a novel peptide-based drug to the cancer-regulating STAT3 protein and corroborated the results with fluorescence polarization analysis derived from the same photon data.

  13. Study on the effect of deposition rate and concentration of Eu on the fluorescent lifetime of CsI: Tl thin film

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Yijun; Guo, Lina [University of Electronic Science and Technology of China, State Key Laboratory of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, Chengdu 610054 (China); Liu, Shuang, E-mail: shuangliu@uestc.edu.cn [University of Electronic Science and Technology of China, State Key Laboratory of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, Chengdu 610054 (China); Wang, Qianfeng; Zhang, Shangjian; Liu, Yong [University of Electronic Science and Technology of China, State Key Laboratory of Electronic Thin Films and Integrated Devices, School of Optoelectronic Information, Chengdu 610054 (China); Zhong, Zhiyong [University of Electronic Science and Technology of China, State Key Laboratory of Electronic Thin Films and Integrated Devices, Chengdu 610054 (China)

    2017-06-21

    Although there are many new scintillators being developed recently, CsI: Tl is still very efficient among them. The fluorescent lifetime is a very important parameter of CsI: Tl thin film and two series of experiments have been conducted to learn about it. Our experiments, however, have demonstrated that the deposition rate and the codoping of Eu{sup 2+} will significantly influence its fluorescent lifetime. In order to increase the efficiency of the imaging system, we intend to obtain a higher fluorescent lifetime for CsI: Tl thin film by controlling these two conditions. - Highlights: • We used vacuum vapor deposition method to grow the high-quality thin films. • The relationship between the deposition rate and the fluorescent lifetime of CsI: Tl thin film was tested. • Concentration of Eu on fluorescent lifetime of the CsI: Tl thin film was studied.

  14. Automated detection of breast cancer in resected specimens with fluorescence lifetime imaging

    Science.gov (United States)

    Phipps, Jennifer E.; Gorpas, Dimitris; Unger, Jakob; Darrow, Morgan; Bold, Richard J.; Marcu, Laura

    2018-01-01

    Re-excision rates for breast cancer lumpectomy procedures are currently nearly 25% due to surgeons relying on inaccurate or incomplete methods of evaluating specimen margins. The objective of this study was to determine if cancer could be automatically detected in breast specimens from mastectomy and lumpectomy procedures by a classification algorithm that incorporated parameters derived from fluorescence lifetime imaging (FLIm). This study generated a database of co-registered histologic sections and FLIm data from breast cancer specimens (N  =  20) and a support vector machine (SVM) classification algorithm able to automatically detect cancerous, fibrous, and adipose breast tissue. Classification accuracies were greater than 97% for automated detection of cancerous, fibrous, and adipose tissue from breast cancer specimens. The classification worked equally well for specimens scanned by hand or with a mechanical stage, demonstrating that the system could be used during surgery or on excised specimens. The ability of this technique to simply discriminate between cancerous and normal breast tissue, in particular to distinguish fibrous breast tissue from tumor, which is notoriously challenging for optical techniques, leads to the conclusion that FLIm has great potential to assess breast cancer margins. Identification of positive margins before waiting for complete histologic analysis could significantly reduce breast cancer re-excision rates.

  15. From morphology to biochemical state - intravital multiphoton fluorescence lifetime imaging of inflamed human skin

    Science.gov (United States)

    Huck, Volker; Gorzelanny, Christian; Thomas, Kai; Getova, Valentina; Niemeyer, Verena; Zens, Katharina; Unnerstall, Tim R.; Feger, Julia S.; Fallah, Mohammad A.; Metze, Dieter; Ständer, Sonja; Luger, Thomas A.; Koenig, Karsten; Mess, Christian; Schneider, Stefan W.

    2016-03-01

    The application of multiphoton microscopy in the field of biomedical research and advanced diagnostics promises unique insights into the pathophysiology of inflammatory skin diseases. In the present study, we combined multiphoton-based intravital tomography (MPT) and fluorescence lifetime imaging (MPT-FLIM) within the scope of a clinical trial of atopic dermatitis with the aim of providing personalised data on the aetiopathology of inflammation in a non-invasive manner at patients’ bedsides. These ‘optical biopsies’ generated via MPT were morphologically analysed and aligned with classical skin histology. Because of its subcellular resolution, MPT provided evidence of a redistribution of mitochondria in keratinocytes, indicating an altered cellular metabolism. Two independent morphometric algorithms reliably showed an even distribution in healthy skin and a perinuclear accumulation in inflamed skin. Moreover, using MPT-FLIM, detection of the onset and progression of inflammatory processes could be achieved. In conclusion, the change in the distribution of mitochondria upon inflammation and the verification of an altered cellular metabolism facilitate a better understanding of inflammatory skin diseases and may permit early diagnosis and therapy.

  16. Characterization of a pulsed x-ray source for fluorescent lifetime measurements

    International Nuclear Information System (INIS)

    Blankespoor, S.C.; Derenzo, S.E.; Moses, W.W.; Rossington, C.S.; Ito, M.; Oba, K.

    1994-01-01

    To search for new, fast, inorganic scintillators, the authors have developed a bench-top pulsed x-ray source for determining fluorescent lifetimes and wavelengths of compounds in crystal or powdered form. This source uses a light-excited x-ray tube which produces x-rays when light from a laser diode strikes its photocathode. The x-ray tube has a tungsten anode, a beryllium exit window, a 30 kV maximum tube bias, and a 50 μA maximum average cathode current. The laser produces 3 x 10 7 photons at 650 nm per ∼100 ps pulse, with up to 10 7 pulses/sec. The time spread for the laser diode, x-ray tube, and a microchannel plate photomultiplier tube is less than 120 ps fwhm. The mean x-ray energy at tube biases of 20, 25, and 30 kV is 9.4, 10.3, and 11.1 keV, respectively. The authors measured 140, 230, and 330 x-ray photons per laser diode pulse per steradian, at tube biases of 20, 25, and 30 kV, respectively. Background x-rays due to dark current occur at a rate of 1 x 10 6 and 3 x 10 6 photons/sec/steradian at biases of 25 and 30 kV, respectively. Data characterizing the x-ray output with an aluminum filter in the x-ray beam are also presented

  17. CONSTRAINING THE LIFETIME AND OPENING ANGLE OF QUASARS USING FLUORESCENT Ly α EMISSION: THE CASE OF Q0420–388

    International Nuclear Information System (INIS)

    Borisova, Elena; Lilly, Simon J.; Cantalupo, Sebastiano; Prochaska, J. Xavier; Rakic, Olivera; Worseck, Gabor

    2016-01-01

    A toy model is developed to understand how the spatial distribution of fluorescent emitters in the vicinity of bright quasars could be affected by the geometry of the quasar bi-conical radiation field and by its lifetime. The model is then applied to the distribution of high-equivalent-width Ly α emitters (with rest-frame equivalent widths above 100 Å, threshold used in, e.g., Trainor and Steidel) identified in a deep narrow-band 36 × 36 arcmin 2 image centered on the luminous quasar Q0420–388. These emitters are found near the edge of the field and show some evidence of an azimuthal asymmetry on the sky of the type expected if the quasar is radiating in a bipolar cone. If these sources are being fluorescently illuminated by the quasar, the two most distant objects require a lifetime of at least 15 Myr for an opening angle of 60° or more, increasing to more than 40 Myr if the opening angle is reduced to a minimum of 30°. However, some other expected signatures of boosted fluorescence are not seen at the current survey limits, e.g., a fall off in Ly α brightness, or equivalent width, with distance. Furthermore, to have most of the Ly α emission of the two distant sources to be fluorescently boosted would require the quasar to have been significantly brighter in the past. This suggests that these particular sources may not be fluorescent, invalidating the above lifetime constraints. This would cast doubt on the use of this relatively low equivalent width threshold and thus also on the lifetime analysis in Trainor and Steidel.

  18. CONSTRAINING THE LIFETIME AND OPENING ANGLE OF QUASARS USING FLUORESCENT Ly α EMISSION: THE CASE OF Q0420–388

    Energy Technology Data Exchange (ETDEWEB)

    Borisova, Elena; Lilly, Simon J.; Cantalupo, Sebastiano [Institute for Astronomy, ETH Zurich, Zurich, CH-8093 (Switzerland); Prochaska, J. Xavier [UCO/Lick Observatory, UC Santa Cruz, Santa Cruz, CA 95064 (United States); Rakic, Olivera; Worseck, Gabor, E-mail: borisova@phys.ethz.ch [Max-Planck-Institut für Astronomie, Heidelberg, D-69117 (Germany)

    2016-10-20

    A toy model is developed to understand how the spatial distribution of fluorescent emitters in the vicinity of bright quasars could be affected by the geometry of the quasar bi-conical radiation field and by its lifetime. The model is then applied to the distribution of high-equivalent-width Ly α emitters (with rest-frame equivalent widths above 100 Å, threshold used in, e.g., Trainor and Steidel) identified in a deep narrow-band 36 × 36 arcmin{sup 2} image centered on the luminous quasar Q0420–388. These emitters are found near the edge of the field and show some evidence of an azimuthal asymmetry on the sky of the type expected if the quasar is radiating in a bipolar cone. If these sources are being fluorescently illuminated by the quasar, the two most distant objects require a lifetime of at least 15 Myr for an opening angle of 60° or more, increasing to more than 40 Myr if the opening angle is reduced to a minimum of 30°. However, some other expected signatures of boosted fluorescence are not seen at the current survey limits, e.g., a fall off in Ly α brightness, or equivalent width, with distance. Furthermore, to have most of the Ly α emission of the two distant sources to be fluorescently boosted would require the quasar to have been significantly brighter in the past. This suggests that these particular sources may not be fluorescent, invalidating the above lifetime constraints. This would cast doubt on the use of this relatively low equivalent width threshold and thus also on the lifetime analysis in Trainor and Steidel.

  19. Automated parasite faecal egg counting using fluorescence labelling, smartphone image capture and computational image analysis.

    Science.gov (United States)

    Slusarewicz, Paul; Pagano, Stefanie; Mills, Christopher; Popa, Gabriel; Chow, K Martin; Mendenhall, Michael; Rodgers, David W; Nielsen, Martin K

    2016-07-01

    Intestinal parasites are a concern in veterinary medicine worldwide and for human health in the developing world. Infections are identified by microscopic visualisation of parasite eggs in faeces, which is time-consuming, requires technical expertise and is impractical for use on-site. For these reasons, recommendations for parasite surveillance are not widely adopted and parasite control is based on administration of rote prophylactic treatments with anthelmintic drugs. This approach is known to promote anthelmintic resistance, so there is a pronounced need for a convenient egg counting assay to promote good clinical practice. Using a fluorescent chitin-binding protein, we show that this structural carbohydrate is present and accessible in shells of ova of strongyle, ascarid, trichurid and coccidian parasites. Furthermore, we show that a cellular smartphone can be used as an inexpensive device to image fluorescent eggs and, by harnessing the computational power of the phone, to perform image analysis to count the eggs. Strongyle egg counts generated by the smartphone system had a significant linear correlation with manual McMaster counts (R(2)=0.98), but with a significantly lower coefficient of variation (P=0.0177). Furthermore, the system was capable of differentiating equine strongyle and ascarid eggs similar to the McMaster method, but with significantly lower coefficients of variation (P<0.0001). This demonstrates the feasibility of a simple, automated on-site test to detect and/or enumerate parasite eggs in mammalian faeces without the need for a laboratory microscope, and highlights the potential of smartphones as relatively sophisticated, inexpensive and portable medical diagnostic devices. Copyright © 2016 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.

  20. Fluorescence life-time imaging and steady state polarization for examining binding of fluorophores to gold nanoparticles.

    Science.gov (United States)

    Schwartz, Shmulik; Fixler, Dror; Popovtzer, Rachela; Shefi, Orit

    2015-11-01

    Nanocomposites as multifunctional agents are capable of combing imaging and cell biology technologies. The conventional methods used for validation of the conjugation process of nanoparticles (NPs) to fluorescent molecules such as spectroscopy analysis and surface potential measurements, are not sufficient. In this paper we present a new and highly sensitive procedure that uses the combination of (1) fluorescence spectrum, (2) fluorescence lifetime, and (3) steady state fluorescence polarization measurements. We characterize and analyze gold NPs with Lucifer yellow (LY) surface coating as a model. We demonstrate the ability to differentiate between LY-GNP (the conjugated complex) and a mixture of coated NP and free dyes. We suggest the approach for neuroscience applications where LY is used for detecting and labeling cells, studying morphology and intracellular communications. Histograms of Fluorescence lifetime imaging (FLIM) of free LY dye (Left) in comparison to the conjugated dye to gold nanoparticles, LY-GNP (Middle) enable the differentiation between LY-GNP (the conjugated complex) and a mixture of coated NP and free dyes (Right). © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. From morphology to clinical pathophysiology: multiphoton fluorescence lifetime imaging at patients' bedside

    Science.gov (United States)

    Mess, Christian; Zens, Katharina; Gorzelanny, Christian; Metze, Dieter; Luger, Thomas A.; König, Karsten; Schneider, Stefan W.; Huck, Volker

    2017-02-01

    Application of multiphoton microscopy in the field of biomedical research and advanced diagnostics promises unique insights into the pathophysiology of skin diseases. By means of multiphoton excitation, endogenous biomolecules like NADH, collagen or elastin show autofluorescence or second harmonic generation. Thus, these molecules provide information about the subcellular morphology, epidermal architecture and physiological condition of the skin. To gain a deeper understanding of the linkage between cellular structure and physiological processes, non-invasive multiphotonbased intravital tomography (MPT) and fluorescence lifetime imaging (FLIM) were combined within the scopes of inflammatory skin, chronic wounds and drug delivery in clinical application. The optical biopsies generated via MPT were morphologically analyzed and aligned with classical skin histology. Because of its subcellular resolution, MPT provided evidence of a redistribution of mitochondria in keratinocytes, indicating an altered cellular metabolism. Independent morphometric algorithms reliably showed a perinuclear accumulation in lesional skin in contrast to an even distribution in healthy skin. Confirmatively, MPT-FLIM showed an obvious metabolic shift in lesions. Moreover, detection of the onset and progression of inflammatory processes could be achieved. The feasibility of primary in vivo tracking of applied therapeutic agents further broadened our scope: We examined the permeation and subsequent distribution of agents directly visualized in patientś skin in short-term repetitive measurements. Furthermore, we performed MPT-FLIM follow-up investigations in the long-term course of therapy. Therefore, clinical MPT-FLIM application offers new insights into the pathophysiology and the individual therapeutic course of skin diseases, facilitating a better understanding of the processes of inflammation and wound healing.

  2. Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging.

    Science.gov (United States)

    Nyström, Sofie; Bäck, Marcus; Nilsson, K Peter R; Hammarström, Per

    2017-10-20

    Proteins that deposit as amyloid in tissues throughout the body can be the cause or consequence of a large number of diseases. Among these we find neurodegenerative diseases such as Alzheimer's and Parkinson's disease afflicting primarily the central nervous system, and systemic amyloidosis where serum amyloid A, transthyretin and IgG light chains deposit as amyloid in liver, carpal tunnel, spleen, kidney, heart, and other peripheral tissues. Amyloid has been known and studied for more than a century, often using amyloid specific dyes such as Congo red and Thioflavin T (ThT) or Thioflavin (ThS). In this paper, we present heptamer-formyl thiophene acetic acid (hFTAA) as an example of recently developed complements to these dyes called luminescent conjugated oligothiophenes (LCOs). hFTAA is easy to use and is compatible with co-staining in immunofluorescence or with other cellular markers. Extensive research has proven that hFTAA detects a wider range of disease associated protein aggregates than conventional amyloid dyes. In addition, hFTAA can also be applied for optical assignment of distinct aggregated morphotypes to allow studies of amyloid fibril polymorphism. While the imaging methodology applied is optional, we here demonstrate hyperspectral imaging (HIS), laser scanning confocal microscopy and fluorescence lifetime imaging (FLIM). These examples show some of the imaging techniques where LCOs can be used as tools to gain more detailed knowledge of the formation and structural properties of amyloids. An important limitation to the technique is, as for all conventional optical microscopy techniques, the requirement for microscopic size of aggregates to allow detection. Furthermore, the aggregate should comprise a repetitive β-sheet structure to allow for hFTAA binding. Excessive fixation and/or epitope exposure that modify the aggregate structure or conformation can render poor hFTAA binding and hence pose limitations to accurate imaging.

  3. Magnetic field and temperature dependence of the fluorescence lifetime of Cr sup(3+) in GdA103

    International Nuclear Information System (INIS)

    Helman, J.S.; Caride, A.O.; Basso, H.C.; Terrile, M.C.; Carvalho, R.A.

    1991-01-01

    The fluorescence lifetime of Cr sup(3+) in GdA10 sub(3) was measured in the range 1.8 - 4.2 K in magnetic fields up to 6 T. The results show a remarkable dependence of the transition probabilities on magnetic order. A model based on the exchange interaction between Cr sup(3+) in highly excited states and the Gd sup(3+) ions is proposed. (author)

  4. Gentamicin differentially alters cellular metabolism of cochlear hair cells as revealed by NAD(P)H fluorescence lifetime imaging

    Science.gov (United States)

    Zholudeva, Lyandysha V.; Ward, Kristina G.; Nichols, Michael G.; Smith, Heather Jensen

    2015-05-01

    Aminoglycoside antibiotics are implicated as culprits of hearing loss in more than 120,000 individuals annually. Research has shown that the sensory cells, but not supporting cells, of the cochlea are readily damaged and/or lost after use of such antibiotics. High-frequency outer hair cells (OHCs) show a greater sensitivity to antibiotics than high- and low-frequency inner hair cells (IHCs). We hypothesize that variations in mitochondrial metabolism account for differences in susceptibility. Fluorescence lifetime microscopy was used to quantify changes in NAD(P)H in sensory and supporting cells from explanted murine cochleae exposed to mitochondrial uncouplers, inhibitors, and an ototoxic antibiotic, gentamicin (GM). Changes in metabolic state resulted in a redistribution of NAD(P)H between subcellular fluorescence lifetime pools. Supporting cells had a significantly longer lifetime than sensory cells. Pretreatment with GM increased NAD(P)H intensity in high-frequency sensory cells, as well as the NAD(P)H lifetime within IHCs. GM specifically increased NAD(P)H concentration in high-frequency OHCs, but not in IHCs or pillar cells. Variations in NAD(P)H intensity in response to mitochondrial toxins and GM were greatest in high-frequency OHCs. These results demonstrate that GM rapidly alters mitochondrial metabolism, differentially modulates cell metabolism, and provides evidence that GM-induced changes in metabolism are significant and greatest in high-frequency OHCs.

  5. Fluorescence lifetime FRET imaging of receptor-ligand complexes in tumor cells in vitro and in vivo

    Science.gov (United States)

    Rudkouskaya, Alena; Sinsuebphon, Nattawut; Intes, Xavier; Mazurkiewicz, Joseph E.; Barroso, Margarida

    2017-02-01

    To guide the development of targeted therapies with improved efficacy and accelerated clinical acceptance, novel imaging methodologies need to be established. Toward this goal, fluorescence lifetime Förster resonance energy transfer (FLIM-FRET) imaging assays capitalize on the ability of antibodies or protein ligands to bind dimerized membrane bound receptors to measure their target engagement levels in cancer cells. Conventional FLIM FRET microscopy has been widely applied at visible wavelengths to detect protein-protein interactions in vitro. However, operation at these wavelengths restricts imaging quality and ability to quantitate lifetime changes in in vivo small animal optical imaging due to high auto-fluorescence and light scattering. Here, we have analyzed the uptake of iron-bound transferrin (Tf) probes into human breast cancer cells using FLIM-FRET microscopy in the visible and near-infrared (NIR) range. The development of NIR FLIM FRET microscopy allows for the use of quantitative lifetime-based molecular assays to measure drug-target engagement levels at multiple scales: from in vitro microscopy to in vivo small animal optical imaging (macroscopy). This novel approach can be extended to other receptors, currently targeted in oncology. Hence, lifetime-based molecular imaging can find numerous applications in drug delivery and targeted therapy assessment and optimization.

  6. A high-throughput direct fluorescence resonance energy transfer-based assay for analyzing apoptotic proteases using flow cytometry and fluorescence lifetime measurements.

    Science.gov (United States)

    Suzuki, Miho; Sakata, Ichiro; Sakai, Takafumi; Tomioka, Hiroaki; Nishigaki, Koichi; Tramier, Marc; Coppey-Moisan, Maïté

    2015-12-15

    Cytometry is a versatile and powerful method applicable to different fields, particularly pharmacology and biomedical studies. Based on the data obtained, cytometric studies are classified into high-throughput (HTP) or high-content screening (HCS) groups. However, assays combining the advantages of both are required to facilitate research. In this study, we developed a high-throughput system to profile cellular populations in terms of time- or dose-dependent responses to apoptotic stimulations because apoptotic inducers are potent anticancer drugs. We previously established assay systems involving protease to monitor live cells for apoptosis using tunable fluorescence resonance energy transfer (FRET)-based bioprobes. These assays can be used for microscopic analyses or fluorescence-activated cell sorting. In this study, we developed FRET-based bioprobes to detect the activity of the apoptotic markers caspase-3 and caspase-9 via changes in bioprobe fluorescence lifetimes using a flow cytometer for direct estimation of FRET efficiencies. Different patterns of changes in the fluorescence lifetimes of these markers during apoptosis were observed, indicating a relationship between discrete steps in the apoptosis process. The findings demonstrate the feasibility of evaluating collective cellular dynamics during apoptosis. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Multiplexed Detection of Attomoles of Nucleic Acids Using Fluorescent Nanoparticle Counting Platform.

    Science.gov (United States)

    Pei, Xiaojing; Yin, Haoyan; Lai, Tiancheng; Zhang, Junlong; Liu, Feng; Xu, Xiao; Li, Na

    2018-01-16

    The sensitive multiplexed detection of nucleic acids in a single sample by a simple manner is of pivotal importance for the diagnosis and therapy of human diseases. Herein, we constructed an automatic fluorescent nanoparticle (FNP) counting platform with a common fluorescence microscopic imaging setup for nonamplification multiplexed detection of attomoles of nucleic acids. Taking the advantages of the highly bright, multicolor emitting FNPs and magnetic separation, the platform enables sensitive multiplexed detection without the need for extra fluorescent labels. Quantification for multiplex DNAs, multiplex microRNAs (miRNA), as well as a DNA and miRNA mixture was achieved with a similar dynamic range, a limit of detection down to 5 amol (5 μL detection volume), and a 81-115% spike recovery from different biological sample matrices. In particular, the sensitivity for multiplex miRNA is by far among the highest without using amplification or the lock nucleic acid hybridization enhancement strategy. Results regarding miRNA-141 from four different cell lines were agreeable with those of the quantitative reverse transcription polymerase chain reaction. Simultaneous detection of miRNA-141 and miRNA-21 in four different cell lines yielded consistent results with publications, indicating the potential for monitoring multiplex miRNA expression associated with the collaborative regulation of important cellular events. This work expands the rule set of multiplex nucleic acid detection strategies and shows promising potential application in clinical diagnosis.

  8. Analysis of photon count data from single-molecule fluorescence experiments

    Science.gov (United States)

    Burzykowski, T.; Szubiakowski, J.; Rydén, T.

    2003-03-01

    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].

  9. Design, construction, and validation of a rotary multifunctional intravascular diagnostic catheter combining multispectral fluorescence lifetime imaging and intravascular ultrasound.

    Science.gov (United States)

    Bec, Julien; Xie, Hongtao; Yankelevich, Diego R; Zhou, Feifei; Sun, Yang; Ghata, Narugopal; Aldredge, Ralph; Marcu, Laura

    2012-10-01

    We report the development and validation of an intravascular rotary catheter for bimodal interrogation of arterial pathologies. This is based on a point-spectroscopy scanning time-resolved fluorescence spectroscopy technique enabling reconstruction of fluorescence lifetime images (FLIm) and providing information on arterial intima composition and intravascular ultrasound (IVUS) providing information on arterial wall morphology. The catheter design allows for independent rotation of the ultrasonic and optical channels within an 8 Fr outer diameter catheter sheath and integrates a low volume flushing channel for blood removal in the optical pathways. In the current configuration, the two channels consist of (a) a standard 3 Fr IVUS catheter with single element transducer (40 MHz) and (b) a side-viewing fiber optic (400 μm core). Experiments conducted in tissue phantoms showed the ability of the catheter to operate in an intraluminal setting and to generate coregistered FLIm and IVUS in one pull-back scan. Current results demonstrate the feasibility of the catheter for simultaneous bimodal interrogation of arterial lumen and for generation of robust fluorescence lifetime data under IVUS guidance. These results facilitate further development of a FLIm-IVUS technique for intravascular diagnosis of atherosclerotic cardiovascular diseases including vulnerable plaques.

  10. Effect of calcinations temperature on the luminescence intensity and fluorescent lifetime of Tb3+-doped hydroxyapatite (Tb-HA nanocrystallines

    Directory of Open Access Journals (Sweden)

    Hairong Yin

    2017-06-01

    Full Text Available Hydroxyapatite luminescent nanocrystallines doped with 6 mol.% Tb3+ (Tb-HA were prepared via chemical deposition method and calcined at different temperature, and the effects of calcinations temperature on the luminescence intensity and fluorescent lifetime were studied. TEM image of Tb-HA revealed that the shape of nanocrystallines changed from needle-like to short rod-like and sphere-like with the increase of calcinations temperature; while the particles sizes decreased from 190 nm to 110 nm. The crystallinity degree increased. The typical emission peaks attributed to Tb3+ ions were observed in emission spectra of 6 mol.% Tb-HA under 378 nm excitation. The luminescent intensity of Tb-HA, which showed the fluorescence quenching, firstly enhanced and then decreased at 700 °C; while the fluorescent lifetime increased firstly and then decreased after 600 °C. Furthermore, the ratio of intensity between 545 nm and 490 nm corresponding to electric-dipole and magnetic-dipole transition (IR: IO increases firstly and then decreases, which revealed that the proportion of substitute type and site of Ca2+ ions by Tb3+ ions were helpful to realize the substitute process and functional structure design.

  11. Segmented frequency-domain fluorescence lifetime measurements: minimizing the effects of photobleaching within a multi-component system.

    Science.gov (United States)

    Marwani, Hadi M; Lowry, Mark; Keating, Patrick; Warner, Isiah M; Cook, Robert L

    2007-11-01

    This study introduces a newly developed frequency segmentation and recombination method for frequency-domain fluorescence lifetime measurements to address the effects of changing fractional contributions over time and minimize the effects of photobleaching within multi-component systems. Frequency segmentation and recombination experiments were evaluated using a two component system consisting of fluorescein and rhodamine B. Comparison of experimental data collected in traditional and segmented fashion with simulated data, generated using different changing fractional contributions, demonstrated the validity of the technique. Frequency segmentation and recombination was also applied to a more complex system consisting of pyrene with Suwannee River fulvic acid reference and was shown to improve recovered lifetimes and fractional intensity contributions. It was observed that photobleaching in both systems led to errors in recovered lifetimes which can complicate the interpretation of lifetime results. Results showed clear evidence that the frequency segmentation and recombination method reduced errors resulting from a changing fractional contribution in a multi-component system, and allowed photobleaching issues to be addressed by commercially available instrumentation.

  12. Silicon photon-counting avalanche diodes for single-molecule fluorescence spectroscopy

    Science.gov (United States)

    Michalet, Xavier; Ingargiola, Antonino; Colyer, Ryan A.; Scalia, Giuseppe; Weiss, Shimon; Maccagnani, Piera; Gulinatti, Angelo; Rech, Ivan; Ghioni, Massimo

    2014-01-01

    Solution-based single-molecule fluorescence spectroscopy is a powerful experimental tool with applications in cell biology, biochemistry and biophysics. The basic feature of this technique is to excite and collect light from a very small volume and work in a low concentration regime resulting in rare burst-like events corresponding to the transit of a single molecule. Detecting photon bursts is a challenging task: the small number of emitted photons in each burst calls for high detector sensitivity. Bursts are very brief, requiring detectors with fast response time and capable of sustaining high count rates. Finally, many bursts need to be accumulated to achieve proper statistical accuracy, resulting in long measurement time unless parallelization strategies are implemented to speed up data acquisition. In this paper we will show that silicon single-photon avalanche diodes (SPADs) best meet the needs of single-molecule detection. We will review the key SPAD parameters and highlight the issues to be addressed in their design, fabrication and operation. After surveying the state-of-the-art SPAD technologies, we will describe our recent progress towards increasing the throughput of single-molecule fluorescence spectroscopy in solution using parallel arrays of SPADs. The potential of this approach is illustrated with single-molecule Förster resonance energy transfer measurements. PMID:25309114

  13. Measurement of apoptosis by SCAN©, a system for counting and analysis of fluorescently labelled nuclei

    Directory of Open Access Journals (Sweden)

    Neta Shlezinger

    2014-11-01

    Full Text Available Apoptosis-like programmed cell death (A-PCD is a universal process common to all types of eukaryotic organisms. Because A-PCD-associated processes are conserved, it is possible to define A-PCD by a standard set of markers. Many of the popular methods to measure A-PCD make use of fluorescent ligands that change in intensity or cellular localization during A-PCD. In single cell organisms, it is possible to quantify levels of A-PCD by scoring the number of apoptotic cells using flow cytometry instruments. In a multicellular organism, quantification of A-PCD is more problematic due to the complex nature of the tissue. The situation is further complicated in filamentous fungi, in which nuclei are divided between compartments, each containing a number of nuclei, which can also migrate between the compartments. We developed SCAN©, a System for Counting and Analysis of Nuclei, and used it to measure A-PCD according to two markers – chromatin condensation and DNA strand breaks. The package includes three modules designed for counting the number of nuclei in multi-nucleated domains, scoring the relative number of nuclei with condensed chromatin, and calculating the relative number of nuclei with DNA strand breaks. The method provides equal or better results compared with manual counting, the analysis is fast and can be applied on large data sets. While we demonstrated the utility of the software for measurement of A-PCD in fungi, the method is readily adopted for measurement of A-PCD in other types of multicellular specimens.

  14. The fluorescence lifetime of BRI1-GFP as probe for the noninvasive determination of the membrane potential in living cells

    Science.gov (United States)

    Elgass, K.; Caesar, K.; Schleifenbaum, F.; Meixner, A. J.; Harter, K.

    2010-02-01

    As the excited state lifetime of a fluorescent molecule depends on its environment, it is possible to use it as a probe for physico-chemical parameters of the surrounding medium. Whereas this is well known for many solid guest/host systems, only few reports of quantitative, temporal resolved in vivo studies to monitor the nano-environment for a protein-coupled chromophore such as GFP are known from literature. Here we present a novel approach to determine the membrane potential of living (plant) cells based on the fluorescence lifetime (FLT) analysis of membrane-located GFP. By using confocal sample scanning microscopy (CSSM) combined with fluorescence lifetime imaging microscopy, we recently showed that the phytohormone brassinolide (BL) induces cell wall expansion and a decrease in the FLT of the BRI1-GFP in living cells of Arabidopsis thaliana seedlings. BRI1 is the dominant functional receptor for BL in Arabidopsis and locates to the plasma membrane. Although the dependence of the FLT of GFP on its physico-chemical environment such as pH-value, refractive index and pressure has been reported, the observed FLT decrease of BRI1-GFP in response to BL application could not be explained by these parameters. However, our in vivo FLT and CSSM analyses indicate that the BLinduced change in the FLT of BRI1-GFP is caused by hyperpolarisation of the plasma membrane (Em). Thus, our results indicate that BRI1-GFP serves as sensitive and non-invasive probe for recording the Em of the plasma membrane in living plant cells with high spatio-temporal resolution.

  15. Morphological spot counting from stacked images for automated analysis of gene copy numbers by fluorescence in situ hybridization.

    Science.gov (United States)

    Grigoryan, Artyom M; Dougherty, Edward R; Kononen, Juha; Bubendorf, Lukas; Hostetter, Galen; Kallioniemi, Olli

    2002-01-01

    Fluorescence in situ hybridization (FISH) is a molecular diagnostic technique in which a fluorescent labeled probe hybridizes to a target nucleotide sequence of deoxyribose nucleic acid. Upon excitation, each chromosome containing the target sequence produces a fluorescent signal (spot). Because fluorescent spot counting is tedious and often subjective, automated digital algorithms to count spots are desirable. New technology provides a stack of images on multiple focal planes throughout a tissue sample. Multiple-focal-plane imaging helps overcome the biases and imprecision inherent in single-focal-plane methods. This paper proposes an algorithm for global spot counting in stacked three-dimensional slice FISH images without the necessity of nuclei segmentation. It is designed to work in complex backgrounds, when there are agglomerated nuclei, and in the presence of illumination gradients. It is based on the morphological top-hat transform, which locates intensity spikes on irregular backgrounds. After finding signals in the slice images, the algorithm groups these together to form three-dimensional spots. Filters are employed to separate legitimate spots from fluorescent noise. The algorithm is set in a comprehensive toolbox that provides visualization and analytic facilities. It includes simulation software that allows examination of algorithm performance for various image and algorithm parameter settings, including signal size, signal density, and the number of slices.

  16. Lifetime-based optical sensor for high-level pCO2 detection employing fluorescence resonance energy transfer

    International Nuclear Information System (INIS)

    Bueltzingsloewen, Christoph von; McEvoy, Aisling K.; McDonagh, Colette; MacCraith, Brian D.

    2003-01-01

    An optical sensor for the measurement of high levels of carbon dioxide in gas phase has been developed. It is based on fluorescence resonance energy transfer (FRET) between a long-lifetime ruthenium polypyridyl complex and the pH-active disazo dye Sudan III. The donor luminophore and the acceptor dye are both immobilised in a hydrophobic silica sol-gel/ethyl cellulose hybrid matrix material. Tetraoctylammonium hydroxide (TOA-OH) is used as an internal buffering system. Fluorescence lifetime is measured in the frequency domain, using low-cost phase modulation measurement technology. The use of Sudan III as an acceptor dye has enabled the sensor to have a dynamic range up to 100% carbon dioxide. The sensor displays 11.2 deg. phase shift between the limit of detection (LOD) of 0.06 and 100% CO 2 with a resolution of better than 2%. The encapsulation in the silica/polymer hybrid material has provided the sensor with good mechanical and chemical stability. The effect of molecular oxygen, humidity and temperature on the sensor performance was studied in detail

  17. Mercury effects on Thalassiosira weissflogii: Applications of two-photon excitation chlorophyll fluorescence lifetime imaging and flow cytometry

    International Nuclear Information System (INIS)

    Wu Yun; Zeng Yan; Qu, Jianan Y.; Wang Wenxiong

    2012-01-01

    The toxic effects of inorganic mercury [Hg(II)] and methylmercury (MeHg) on the photosynthesis and population growth in a marine diatom Thalassiosira weissflogii were investigated using two methods: two-photon excitation fluorescence lifetime imaging (FLIM) and flow cytometry (FCM). For photosynthesis, Hg(II) exposure increased the average chlorophyll fluorescence lifetime, whereas such increment was not found under MeHg stress. This may be caused by the inhibitory effect of Hg(II) instead of MeHg on the electron transport chain. For population growth, modeled specific growth rate data showed that the reduction in population growth by Hg(II) mainly resulted from an increased number of injured cells, while the live cells divided at the normal rates. However, MeHg inhibitory effects on population growth were contributed by the reduced division rates of all cells. Furthermore, the cell images and the FCM data reflected the morphological changes of diatom cells under Hg(II)/MeHg exposure vividly and quantitatively. Our results demonstrated that the toxigenicity mechanisms between Hg(II) and MeHg were different in the algal cells.

  18. Fluorescence lifetime studies of MeV erbium implanted silica glass

    International Nuclear Information System (INIS)

    Lidgard, A.; Polman, A.; Jacobsen, D.C.; Blonder, G.E.; Kistler, R.; Poate, J.M.; Becker, P.C.

    1991-01-01

    MeV erbium ion implantation into various SiO 2 glasses has been studied with the aim of incorporating the rare-earth dopant as an optically active ion in the silica network. The lifetime of the excited state ranges from 1.6 to 12.8 ms, depending on base material and implantation fluence. These results have positive implications for silica-based integrated optical technology. (Author)

  19. Fluorescence lifetime studies of MeV erbium implanted silica glass

    Energy Technology Data Exchange (ETDEWEB)

    Lidgard, A.; Polman, A.; Jacobsen, D.C.; Blonder, G.E.; Kistler, R.; Poate, J.M.; Becker, P.C. (AT and T Bell Labs., Murray Hill, NJ (USA))

    1991-05-23

    MeV erbium ion implantation into various SiO{sub 2} glasses has been studied with the aim of incorporating the rare-earth dopant as an optically active ion in the silica network. The lifetime of the excited state ranges from 1.6 to 12.8 ms, depending on base material and implantation fluence. These results have positive implications for silica-based integrated optical technology. (Author).

  20. A fusion-spliced near-field optical fiber probe using photonic crystal fiber for nanoscale thermometry based on fluorescence-lifetime measurement of quantum dots.

    Science.gov (United States)

    Fujii, Takuro; Taguchi, Yoshihiro; Saiki, Toshiharu; Nagasaka, Yuji

    2011-01-01

    We have developed a novel nanoscale temperature-measurement method using fluorescence in the near-field called fluorescence near-field optics thermal nanoscopy (Fluor-NOTN). Fluor-NOTN enables the temperature distributions of nanoscale materials to be measured in vivo/in situ. The proposed method measures temperature by detecting the temperature dependent fluorescence lifetimes of Cd/Se quantum dots (QDs). For a high-sensitivity temperature measurement, the auto-fluorescence generated from a fiber probe should be reduced. In order to decrease the noise, we have fabricated a novel near-field optical-fiber probe by fusion-splicing a photonic crystal fiber (PCF) and a conventional single-mode fiber (SMF). The validity of the novel fiber probe was assessed experimentally by evaluating the auto-fluorescence spectra of the PCF. Due to the decrease of auto-fluorescence, a six- to ten-fold increase of S/N in the near-field fluorescence lifetime detection was achieved with the newly fabricated fusion-spliced near-field optical fiber probe. Additionally, the near-field fluorescence lifetime of the quantum dots was successfully measured by the fabricated fusion-spliced near-field optical fiber probe at room temperature, and was estimated to be 10.0 ns.

  1. Time-resolved pulse-counting lock-in detection of laser induced fluorescence in the presence of a strong background emission

    International Nuclear Information System (INIS)

    Pelissier, B.; Sadeghi, N.

    1996-01-01

    We describe a time-resolved pulse-counting system well adapted for the detection of continuous laser induced fluorescence (LIF) signals in repetitive phenomena, when a strong background emission is present. It consists of 256 channels coupled to a first in first out memory and interfaced to a 486 DX 33 PC, for data storage. It accepts time-averaged count rates up to 450 kcount/s. Time between channels can be set from 12.5 ns to several μs and the dead time between two consecutive cycles of the physical phenomena is less than 20 ns. In phase with a chopper, which modulates the laser beam, it adds the observed photon signal to the channel memories when the beam is on and substracts it when the beam is stopped, acting like a lock-in amplifier which detect only the modulated part of the signal. The minimum detectivity on the LIF signal is only limited by the shot noise of the plasma induced emission signal. As an application, we studied the time variation of the Ar + *( 2 G 9/2 ) metastable ions, detected by LIF, in two types of plasmas. Their radiative lifetime and collisional quenching frequencies were deduced from their decay rate in the afterglow of a pulsed Helicon reactor. We also observed the evolution of their density in a 455 kHz capacitively coupled argon discharge. copyright 1996 American Institute of Physics

  2. Time-resolved pulse-counting lock-in detection of laser induced fluorescence in the presence of a strong background emission

    Science.gov (United States)

    Pelissier, B.; Sadeghi, N.

    1996-10-01

    We describe a time-resolved pulse-counting system well adapted for the detection of continuous laser induced fluorescence (LIF) signals in repetitive phenomena, when a strong background emission is present. It consists of 256 channels coupled to a first in first out memory and interfaced to a 486 DX 33 PC, for data storage. It accepts time-averaged count rates up to 450 kcount/s. Time between channels can be set from 12.5 ns to several μs and the dead time between two consecutive cycles of the physical phenomena is less than 20 ns. In phase with a chopper, which modulates the laser beam, it adds the observed photon signal to the channel memories when the beam is on and substracts it when the beam is stopped, acting like a lock-in amplifier which detect only the modulated part of the signal. The minimum detectivity on the LIF signal is only limited by the shot noise of the plasma induced emission signal. As an application, we studied the time variation of the Ar+*(2G9/2) metastable ions, detected by LIF, in two types of plasmas. Their radiative lifetime and collisional quenching frequencies were deduced from their decay rate in the afterglow of a pulsed Helicon reactor. We also observed the evolution of their density in a 455 kHz capacitively coupled argon discharge.

  3. Automatic choroid cells segmentation and counting based on approximate convexity and concavity of chain code in fluorescence microscopic image

    Science.gov (United States)

    Lu, Weihua; Chen, Xinjian; Zhu, Weifang; Yang, Lei; Cao, Zhaoyuan; Chen, Haoyu

    2015-03-01

    In this paper, we proposed a method based on the Freeman chain code to segment and count rhesus choroid-retinal vascular endothelial cells (RF/6A) automatically for fluorescence microscopy images. The proposed method consists of four main steps. First, a threshold filter and morphological transform were applied to reduce the noise. Second, the boundary information was used to generate the Freeman chain codes. Third, the concave points were found based on the relationship between the difference of the chain code and the curvature. Finally, cells segmentation and counting were completed based on the characteristics of the number of the concave points, the area and shape of the cells. The proposed method was tested on 100 fluorescence microscopic cell images, and the average true positive rate (TPR) is 98.13% and the average false positive rate (FPR) is 4.47%, respectively. The preliminary results showed the feasibility and efficiency of the proposed method.

  4. The increase of NADH fluorescence lifetime is associated with the metabolic change during osteogenic differentiation of human mesenchymal stem cells (hMSCs)

    Science.gov (United States)

    Guo, Han Wen; Yu, Jia Sin; Hsu, Shu Han; Wei, Yau Huei; Lee, Oscar K.; Wang, Hsing Wen

    2011-03-01

    Fluorescence lifetime of NADH had been used as an optical marker for monitoring cellular metabolism. In our pervious studies, we have demonstrated that NADH lifetime of hMSCs increase gradually with time of osteogenic differentiation. In this study, we measured NADH lifetime of hMSCs from a different donor as well as the corresponding metabolic indices such as ATP level, oxygen consumption and lactate release. We also measure the quantity of Complex I, III, IV and V. The results show that during differentiation more oxygen consumed, higher ATP level expressed and less lactate released, and the increase of NADH lifetime was associated with ATP level. Higher expression of the total Complex protein was observed at 3 and 4 weeks after differentiation than controls. However, Complex I expression did not show significant correlation with the increase of NADH fluorescence lifetime. In summary, we demonstrated that the change of NADH lifetime was associated with the metabolic change during osteogenic differentiation of hMSCs. The increase of NADH lifetime was in part due to the increased Complex protein interaction in mitochondria after differentiation.

  5. Quantification of protein based on single-molecule counting by total internal reflection fluorescence microscopy with adsorption equilibrium

    International Nuclear Information System (INIS)

    Wang Lei; Xu Guang; Shi Zhikun; Jiang Wei; Jin Wenrui

    2007-01-01

    We developed a sensitive single-molecule imaging method for quantification of protein by total internal reflection fluorescence microscopy with adsorption equilibrium. In this method, the adsorption equilibrium of protein was achieved between solution and glass substrate. Then, fluorescence images of protein molecules in a evanescent wave field were taken by a highly sensitive electron multiplying charge coupled device. Finally, the number of fluorescent spots corresponding to the protein molecules in the images was counted. Alexa Fluor 488-labeled goat anti-rat IgG(H + L) was chosen as the model protein. The spot number showed an excellent linear relationship with protein concentration. The concentration linear range was 5.4 x 10 -11 to 8.1 x 10 -10 mol L -1

  6. Fluorescence lifetime imaging ophthalmoscopy in type 2 diabetic patients who have no signs of diabetic retinopathy

    Science.gov (United States)

    Schweitzer, Dietrich; Deutsch, Lydia; Klemm, Matthias; Jentsch, Susanne; Hammer, Martin; Peters, Sven; Haueisen, Jens; Müller, Ulrich A.; Dawczynski, Jens

    2015-06-01

    The time-resolved autofluorescence of the eye is used for the detection of metabolic alteration in diabetic patients who have no signs of diabetic retinopathy. One eye from 37 phakic and 11 pseudophakic patients with type 2 diabetes, and one eye from 25 phakic and 23 pseudophakic healthy subjects were included in the study. After a three-exponential fit of the decay of autofluorescence, histograms of lifetimes τi, amplitudes αi, and relative contributions Qi were statistically compared between corresponding groups in two spectral channels (490diabetic patients and age-matched controls (p450 ps, and the shift of τ3 from ˜3000 to 3700 ps in ch1 of diabetic patients when compared with healthy subjects indicate an increased production of free flavin adenine dinucleotide, accumulation of advanced glycation end products (AGE), and, probably, a change from free to protein-bound reduced nicotinamide adenine dinucleotide at the fundus. AGE also accumulated in the crystalline lens.

  7. Comparitive study of fluorescence lifetime quenching of rhodamine 6G by MoS2 and Au-MoS2

    Science.gov (United States)

    Shakya, Jyoti; Kasana, Parath; Mohanty, T.

    2018-04-01

    Time resolved fluorescence study of Rhodamine 6G (R6G) in the presence of Molybdenum disulfide (MoS2) nanosheets and gold doped MoS2 (Au-MoS2) have been carried out and discussed. We have analyzed the fluorescence decay curves of R6G and it is observed that Au-MoS2 is a better fluorescence lifetime quencher as compare to MoS2 nanosheets. Also, the energy transfer efficiency and energy transfer rate from R6G to MoS2 and Au-MoS2 has been calculated and found higher for Au-MoS2.

  8. Preparation and properties of Nd{sup 3+}:SrAlF{sub 5} nanocrystals embedded fluorophosphate transparent glass-ceramic with long fluorescence lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Ruilin; Wang, Jinlong; Zhang, Liaolin; Liu, Chunxiao; Wei, Wei [Nanjing University of Posts and Telecommunications, School of Optoelectronic Engineering, Nanjing (China)

    2016-07-15

    Nd{sup 3+}:SrAlF{sub 5} nanocrystals embedded fluorophosphate glass-ceramics were prepared by the melt quenching and subsequent thermal treatment method. The formation of SrAlF{sub 5} nanocrystals in the glass was confirmed by X-ray diffraction and scanning electron microscope. The fluorescence intensity and lifetime of the glass-ceramics increased with the increase of size of nanocrystals. Importantly, by controlling growth of nanocrystals, an obvious enhancement of lifetime (725 μs) emerged in the glass-ceramics heat-treated at 510 C and the transmittance can reach to 72.2 % at 1049 nm. The enhanced fluorescence intensity and lifetime were ascribed to the comfortable local environment to the Nd{sup 3+} ion and scattering of the nanoparticle embedded into the glass matrix. (orig.)

  9. Fluorescence lifetime measurements to determine the core-shell nanostructure of FITC-doped silica nanoparticles: An optical approach to evaluate nanoparticle photostability

    International Nuclear Information System (INIS)

    Santra, Swadeshmukul; Liesenfeld, Bernd; Bertolino, Chiara; Dutta, Debamitra; Cao Zehui; Tan Weihong; Moudgil, Brij M.; Mericle, Robert A.

    2006-01-01

    In this paper, we described a novel fluorescence lifetime-based approach to determine the core-shell nanostructure of FITC-(fluorescein isothiocyanate, isomer I) doped fluorescent silica nanoparticles (FSNPs). Because of phase homogeneity between the core and the shell, electron microscopic technique could not be used to characterize such core-shell nanostructure. Our optical approach not only revealed the core-shell nanostructure of FSNPs but also evaluated photobleaching of FSNPs both in the solvated and non-solvated (dry) states. The FSNPs were produced via Stoeber's method by hydrolysis and co-condensation reaction of tetraethylorthosilicate (TEOS) and fluorescein linked (3-aminopropyl)triethoxysilane (FITC-APTS conjugate) in the presence of ammonium hydroxide catalyst. To obtain a pure silica surface coating, FSNPs were then post-coated with TEOS. The average particle size was 135 nm as determined by TEM (transmission electron microscope) measurements. Fluorescence excitation and emission spectral data demonstrated successful doping of FITC dye molecules in FSNPs. Fluorescence lifetime data revealed that approximately 62% of dye molecules remained in the solvated silica shell, while 38% of dye molecules remained in the non-solvated (dry) silica core. Photobleaching experiments of FSNPs were conducted both in DI water (solution state) and in air (dry state). Severe photobleaching of FSNPs was observed in air. However, FSNPs were moderately photostable in the solution state. Photostability of FSNPs in both solution and dry states was explained on the basis of fluorescence lifetime data

  10. Automatic Segmentation of Fluorescence Lifetime Microscopy Images of Cells Using Multi-Resolution Community Detection -A First Study

    Science.gov (United States)

    Hu, Dandan; Sarder, Pinaki; Ronhovde, Peter; Orthaus, Sandra; Achilefu, Samuel; Nussinov, Zohar

    2014-01-01

    Inspired by a multi-resolution community detection (MCD) based network segmentation method, we suggest an automatic method for segmenting fluorescence lifetime (FLT) imaging microscopy (FLIM) images of cells in a first pilot investigation on two selected images. The image processing problem is framed as identifying segments with respective average FLTs against the background in FLIM images. The proposed method segments a FLIM image for a given resolution of the network defined using image pixels as the nodes and similarity between the FLTs of the pixels as the edges. In the resulting segmentation, low network resolution leads to larger segments, and high network resolution leads to smaller segments. Further, using the proposed method, the mean-square error (MSE) in estimating the FLT segments in a FLIM image was found to consistently decrease with increasing resolution of the corresponding network. The MCD method appeared to perform better than a popular spectral clustering based method in performing FLIM image segmentation. At high resolution, the spectral segmentation method introduced noisy segments in its output, and it was unable to achieve a consistent decrease in MSE with increasing resolution. PMID:24251410

  11. Automatic segmentation of fluorescence lifetime microscopy images of cells using multiresolution community detection--a first study.

    Science.gov (United States)

    Hu, D; Sarder, P; Ronhovde, P; Orthaus, S; Achilefu, S; Nussinov, Z

    2014-01-01

    Inspired by a multiresolution community detection based network segmentation method, we suggest an automatic method for segmenting fluorescence lifetime (FLT) imaging microscopy (FLIM) images of cells in a first pilot investigation on two selected images. The image processing problem is framed as identifying segments with respective average FLTs against the background in FLIM images. The proposed method segments a FLIM image for a given resolution of the network defined using image pixels as the nodes and similarity between the FLTs of the pixels as the edges. In the resulting segmentation, low network resolution leads to larger segments, and high network resolution leads to smaller segments. Furthermore, using the proposed method, the mean-square error in estimating the FLT segments in a FLIM image was found to consistently decrease with increasing resolution of the corresponding network. The multiresolution community detection method appeared to perform better than a popular spectral clustering-based method in performing FLIM image segmentation. At high resolution, the spectral segmentation method introduced noisy segments in its output, and it was unable to achieve a consistent decrease in mean-square error with increasing resolution. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

  12. In vivo detection of oral epithelial cancer using endogenous fluorescence lifetime imaging: a pilot human study (Conference Presentation)

    Science.gov (United States)

    Jo, Javier A.; Hwang, Dae Yon; Palma, Jorge; Cheng, Shuna; Cuenca, Rodrigo; Malik, Bilal; Jabbour, Joey; Cheng, Lisa; Wright, John; Maitland, Kristen

    2016-03-01

    Endogenous fluorescence lifetime imaging (FLIM) provides direct access to the concomitant functional and biochemical changes accompanying tissue transition from benign to precancerous and cancerous. Since FLIM can noninvasively measure different and complementary biomarkers of precancer and cancer, we hypothesize that it will aid in clinically detecting early oral epithelial cancer. Our group has recently demonstrated the detection of benign from premalignant and malignant lesions based on endogenous multispectral FLIM in the hamster cheek-pouch model. Encouraged by these positive preliminary results, we have developed a handheld endoscope capable of acquiring multispectral FLIM images in real time from the oral mucosa. This novel FLIM endoscope is being used for imaging clinically suspicious pre-malignant and malignant lesions from patients before undergoing tissue biopsy for histopathological diagnosis of oral epithelial cancer. Our preliminary results thus far are already suggesting the potential of endogenous FLIM for distinguishing a variety of benign lesions from advanced dysplasia and squamous cell carcinoma (SCC). To the best of out knowledge, this is the first in vivo human study aiming to demonstrate the ability to predict the true malignancy of clinically suspicious lesions using endogenous FLIM. If successful, the resulting clinical tool will allow noninvasive real-time detection of epithelial precancerous and cancerous lesions in the oral mucosa and could potentially be used to assist at every step involved on the clinical management of oral cancer patients, from early screening and diagnosis, to treatment and monitoring of recurrence.

  13. The modifier effects of chymotrypsin and trypsin enzymes on fluorescence lifetime distribution of "N-(1-pyrenyl)maleimide-bovine serum albumin" complex.

    Science.gov (United States)

    Özyiğit, İbrahim Ethem; Karakuş, Emine; Pekcan, Önder

    2016-02-05

    Chymotrypsin and trypsin are the well known proteolytic enzymes, both of which are synthesized in the pancreas as their precursors - the inactive forms; chymotrypsinogen and trypsinogen - and then are released into the duodenum to cut proteins into smaller peptides. In this paper, the effects of activities of chymotrypsin and trypsin enzymes on fluorescence lifetime distributions of the substrat bovine serum albumin (BSA) modified with N-(1-pyrenyl)maleimide (PM) were examined. In the labeling study of BSA with PM, it is aimed to attach PM to the single free thiol (Cys34) and to all the free amine groups in accessible positions in order to produce excimers of pyrene planes of the possible highest amount to form the lifetime distributions in the widest range, that may show specifically distinguishing changes resulting from the activities of the proteases. The time resolved spectrofluorometer was used to monitor fluorescence decays, which were analyzed by using the exponential series method (ESM) to obtain the changes of lifetime distributions. After the exposure of the synthesized substrat PM-BSA to the enzymes, the fluorescence lifetime distributions exhibited different structures which were attributed to the different activities of the proteases. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. The modifier effects of chymotrypsin and trypsin enzymes on fluorescence lifetime distribution of "N-(1-pyrenyl)maleimide-bovine serum albumin" complex

    Science.gov (United States)

    Özyiğit, İbrahim Ethem; Karakuş, Emine; Pekcan, Önder

    2016-02-01

    Chymotrypsin and trypsin are the well known proteolytic enzymes, both of which are synthesized in the pancreas as their precursors - the inactive forms; chymotrypsinogen and trypsinogen - and then are released into the duodenum to cut proteins into smaller peptides. In this paper, the effects of activities of chymotrypsin and trypsin enzymes on fluorescence lifetime distributions of the substrat bovine serum albumin (BSA) modified with N-(1-pyrenyl)maleimide (PM) were examined. In the labeling study of BSA with PM, it is aimed to attach PM to the single free thiol (Cys34) and to all the free amine groups in accessible positions in order to produce excimers of pyrene planes of the possible highest amount to form the lifetime distributions in the widest range, that may show specifically distinguishing changes resulting from the activities of the proteases. The time resolved spectrofluorometer was used to monitor fluorescence decays, which were analyzed by using the exponential series method (ESM) to obtain the changes of lifetime distributions. After the exposure of the synthesized substrat PM-BSA to the enzymes, the fluorescence lifetime distributions exhibited different structures which were attributed to the different activities of the proteases.

  15. Subsurface microbial ecology. Epi fluorescence direct counts; Ecologia microbica del sottosuolo: metodo di conta diretta in epifluorescenza

    Energy Technology Data Exchange (ETDEWEB)

    Barra Caracciolo, A.; Silvestri, C.; Creo, C.; Izzo, G. [ENEA, Centro Ricerche Casaccia, Rome (Italy). Dipt. Ambiente

    1998-07-01

    To the aim of recognize the importance of microorganisms in affecting or even determining the fate of xenobiotics in the subsurface environment evaluating bacteria concentration in a subsurface ecosystem, the report discusses a soil sample treatment method which has been developed for epi fluorescence direct counting with DAPI. [Italian] Lo studio discute un metodo di trattamento del campione per la conta diretta in epifluorescenza con un marcatore selettivo per il DNA, il DAPI, al fine di quantificare la concentrazione batterica del sottosuolo e studiare il ruolo dei microrganismi nella biodegradazione delle molecole esogene, ancora poco indagato.

  16. Selective plane illumination microscopy (SPIM) with time-domain fluorescence lifetime imaging microscopy (FLIM) for volumetric measurement of cleared mouse brain samples

    Science.gov (United States)

    Funane, Tsukasa; Hou, Steven S.; Zoltowska, Katarzyna Marta; van Veluw, Susanne J.; Berezovska, Oksana; Kumar, Anand T. N.; Bacskai, Brian J.

    2018-05-01

    We have developed an imaging technique which combines selective plane illumination microscopy with time-domain fluorescence lifetime imaging microscopy (SPIM-FLIM) for three-dimensional volumetric imaging of cleared mouse brains with micro- to mesoscopic resolution. The main features of the microscope include a wavelength-adjustable pulsed laser source (Ti:sapphire) (near-infrared) laser, a BiBO frequency-doubling photonic crystal, a liquid chamber, an electrically focus-tunable lens, a cuvette based sample holder, and an air (dry) objective lens. The performance of the system was evaluated with a lifetime reference dye and micro-bead phantom measurements. Intensity and lifetime maps of three-dimensional human embryonic kidney (HEK) cell culture samples and cleared mouse brain samples expressing green fluorescent protein (GFP) (donor only) and green and red fluorescent protein [positive Förster (fluorescence) resonance energy transfer] were acquired. The results show that the SPIM-FLIM system can be used for sample sizes ranging from single cells to whole mouse organs and can serve as a powerful tool for medical and biological research.

  17. Spot counting on fluorescence in situ hybridization in suspension images using Gaussian mixture model

    Science.gov (United States)

    Liu, Sijia; Sa, Ruhan; Maguire, Orla; Minderman, Hans; Chaudhary, Vipin

    2015-03-01

    Cytogenetic abnormalities are important diagnostic and prognostic criteria for acute myeloid leukemia (AML). A flow cytometry-based imaging approach for FISH in suspension (FISH-IS) was established that enables the automated analysis of several log-magnitude higher number of cells compared to the microscopy-based approaches. The rotational positioning can occur leading to discordance between spot count. As a solution of counting error from overlapping spots, in this study, a Gaussian Mixture Model based classification method is proposed. The Akaike information criterion (AIC) and Bayesian information criterion (BIC) of GMM are used as global image features of this classification method. Via Random Forest classifier, the result shows that the proposed method is able to detect closely overlapping spots which cannot be separated by existing image segmentation based spot detection methods. The experiment results show that by the proposed method we can obtain a significant improvement in spot counting accuracy.

  18. Wide Field-of-View Fluorescence Imaging with Optical-Quality Curved Microfluidic Chamber for Absolute Cell Counting

    Directory of Open Access Journals (Sweden)

    Mohiuddin Khan Shourav

    2016-07-01

    Full Text Available Field curvature and other aberrations are encountered inevitably when designing a compact fluorescence imaging system with a simple lens. Although multiple lens elements can be used to correct most such aberrations, doing so increases system cost and complexity. Herein, we propose a wide field-of-view (FOV fluorescence imaging method with an unconventional optical-quality curved sample chamber that corrects the field curvature caused by a simple lens. Our optics simulations and proof-of-concept experiments demonstrate that a curved substrate with lens-dependent curvature can reduce greatly the distortion in an image taken with a conventional planar detector. Following the validation study, we designed a curved sample chamber that can contain a known amount of sample volume and fabricated it at reasonable cost using plastic injection molding. At a magnification factor of approximately 0.6, the curved chamber provides a clear view of approximately 119 mm2, which is approximately two times larger than the aberration-free area of a planar chamber. Remarkably, a fluorescence image of microbeads in the curved chamber exhibits almost uniform intensity over the entire field even with a simple lens imaging system, whereas the distorted boundary region has much lower brightness than the central area in the planar chamber. The absolute count of white blood cells stained with a fluorescence dye was in good agreement with that obtained by a commercially available conventional microscopy system. Hence, a wide FOV imaging system with the proposed curved sample chamber would enable us to acquire an undistorted image of a large sample volume without requiring a time-consuming scanning process in point-of-care diagnostic applications.

  19. Fluorescence lifetime spectroscopy: potential for in-vivo estimation of skin fluorophores changes after low power laser treatment

    Science.gov (United States)

    Ferulova, Inesa; Lihachev, Alexey; Spigulis, Janis

    2013-11-01

    The impact of visible cwlaser irradiation on skin autofluorescence lifetimes was investigated in spectral range from 450 nm to 600 nm. Skin optical provocations were performed during 1 min by 405 nm low power cw laser with power density up to 20 mW/cm2. Autofluorescence lifetimes were measured before and immediately after the optical provocation.

  20. Non-Euclidean phasor analysis for quantification of oxidative stress in ex vivo human skin exposed to sun filters using fluorescence lifetime imaging microscopy

    Science.gov (United States)

    Osseiran, Sam; Roider, Elisabeth M.; Wang, Hequn; Suita, Yusuke; Murphy, Michael; Fisher, David E.; Evans, Conor L.

    2017-12-01

    Chemical sun filters are commonly used as active ingredients in sunscreens due to their efficient absorption of ultraviolet (UV) radiation. Yet, it is known that these compounds can photochemically react with UV light and generate reactive oxygen species and oxidative stress in vitro, though this has yet to be validated in vivo. One label-free approach to probe oxidative stress is to measure and compare the relative endogenous fluorescence generated by cellular coenzymes nicotinamide adenine dinucleotides and flavin adenine dinucleotides. However, chemical sun filters are fluorescent, with emissive properties that contaminate endogenous fluorescent signals. To accurately distinguish the source of fluorescence in ex vivo skin samples treated with chemical sun filters, fluorescence lifetime imaging microscopy data were processed on a pixel-by-pixel basis using a non-Euclidean separation algorithm based on Mahalanobis distance and validated on simulated data. Applying this method, ex vivo samples exhibited a small oxidative shift when exposed to sun filters alone, though this shift was much smaller than that imparted by UV irradiation. Given the need for investigative tools to further study the clinical impact of chemical sun filters in patients, the reported methodology may be applied to visualize chemical sun filters and measure oxidative stress in patients' skin.

  1. Size-controlled fluorescent nanodiamonds: A facile method of fabrication and color-center counting

    KAUST Repository

    Mahfouz, Remi

    2013-01-01

    We present a facile method for the production of fluorescent diamond nanocrystals (DNCs) of different sizes and efficiently quantify the concentration of emitting defect color centers (DCCs) of each DNC size. We prepared the DNCs by ball-milling commercially available micrometer-sized synthetic (high pressure, high temperature (HPHT)) diamonds and then separated the as-produced DNCs by density gradient ultracentrifugation (DGU) into size-controlled fractions. A protocol to enhance the uniformity of the nitrogen-vacancy (NV) centers in the diamonds was devised by depositing the DNCs as a dense monolayer on amino-silanized silicon substrates and then subjecting the monolayer to He+ beam irradiation. Using a standard confocal setup, we analyzed the average number of NV centers per crystal, and obtained a quantitative relationship between the DNC particle size and the NV number per crystal. This relationship was in good agreement with results from previous studies that used more elaborate setups. Our findings suggest that nanocrystal size separation by DGU may be used to control the number of defects per nanocrystal. The efficient approaches described herein to control and quantify DCCs are valuable to researchers as they explore applications for color centers and new strategies to create them. © 2013 The Royal Society of Chemistry.

  2. Quantitative time domain analysis of lifetime-based Förster resonant energy transfer measurements with fluorescent proteins: Static random isotropic fluorophore orientation distributions

    DEFF Research Database (Denmark)

    Alexandrov, Yuriy; Nikolic, Dino Solar; Dunsby, Christopher

    2018-01-01

    Förster resonant energy transfer (FRET) measurements are widely used to obtain information about molecular interactions and conformations through the dependence of FRET efficiency on the proximity of donor and acceptor fluorophores. Fluorescence lifetime measurements can provide quantitative...... into new software for fitting donor emission decay profiles. Calculated FRET parameters, including molar population fractions, are compared for the analysis of simulated and experimental FRET data under the assumption of static and dynamic fluorophores and the intermediate regimes between fully dynamic...... analysis of FRET efficiency and interacting population fraction. Many FRET experiments exploit the highly specific labelling of genetically expressed fluorescent proteins, applicable in live cells and organisms. Unfortunately, the typical assumption of fast randomization of fluorophore orientations...

  3. Time-resolved fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Gustavsson, Thomas; Mialocq, Jean-Claude

    2007-01-01

    This article addresses the evolution in time of light emitted by a molecular system after a brief photo-excitation. The authors first describe fluorescence from a photo-physical point of view and discuss the characterization of the excited state. Then, they explain some basic notions related to fluorescence characterization (lifetime and decays, quantum efficiency, so on). They present the different experimental methods and techniques currently used to study time-resolved fluorescence. They discuss basic notions of time resolution and spectral reconstruction. They briefly present some conventional methods: intensified Ccd cameras, photo-multipliers and photodiodes associated with a fast oscilloscope, and phase modulation. Other methods and techniques are more precisely presented: time-correlated single photon counting (principle, examples, and fluorescence lifetime imagery), streak camera (principle, examples), and optical methods like the Kerr optical effect (principle and examples) and fluorescence up-conversion (principle and theoretical considerations, examples of application)

  4. Study of lifetimes of fluorescence levels of tetravalent uranium in the incommensurate phase of thorium tetrabromide and tetrachloride

    International Nuclear Information System (INIS)

    Milicic, A.

    1989-01-01

    The lifetimes of radiative levels of tetravalent uranium in the incommensurate phase of thorium tetrahalides have been measured as a function of different parameters: site symmetry, temperature and concentration. The incommensurate phase of thorium tetrabromide and tetrachloride is characterized by a continuous distribution of site symmetries induced by a continuous and weak displacement of the halides around the thorium (uranium) ions. At low temperature, 4.2 K, the lifetime variation as a function of excited classes of symmetry is governed by the radiative process probability as well as the energy transfer between uranium ions in different sites. At higher temperature, a model based on a Boltzmann equilibrium between closed energy levels is able to reproduce the experimental lifetime variation as a function of the temperature, for a given class of symmetry. For the variation of lifetime as a function of uranium ion concentrations, at high dilution and in the case of U 4+ : ThBr 4 , there is a competition between the energy transfer and thermal population of excited states [fr

  5. A rapid and universal bacteria-counting approach using CdSe/ZnS/SiO2 composite nanoparticles as fluorescence probe.

    Science.gov (United States)

    Fu, Xin; Huang, Kelong; Liu, Suqin

    2010-02-01

    In this paper, a rapid, simple, and sensitive method was described for detection of the total bacterial count using SiO(2)-coated CdSe/ZnS quantum dots (QDs) as a fluorescence marker that covalently coupled with bacteria using glutaraldehyde as the crosslinker. Highly luminescent CdSe/ZnS were prepared by applying cadmium oxide and zinc stearate as precursors instead of pyrophoric organometallic precursors. A reverse-microemulsion technique was used to synthesize CdSe/ZnS/SiO(2) composite nanoparticles with a SiO(2) surface coating. Our results showed that CdSe/ZnS/SiO(2) composite nanoparticles prepared with this method possessed highly luminescent, biologically functional, and monodispersive characteristics, and could successfully be covalently conjugated with the bacteria. As a demonstration, it was found that the method had higher sensitivity and could count bacteria in 3 x 10(2) CFU/mL, lower than the conventional plate counting and organic dye-based method. A linear relationship of the fluorescence peak intensity (Y) and the total bacterial count (X) was established in the range of 3 x 10(2)-10(7) CFU/mL using the equation Y = 374.82X-938.27 (R = 0.99574). The results of the determination for the total count of bacteria in seven real samples were identical with the conventional plate count method, and the standard deviation was satisfactory.

  6. Fluorescence resonance energy transfer imaging of CFP/YFP labeled NDH in cyanobacterium cell

    International Nuclear Information System (INIS)

    Ji Dongmei; Lv Wei; Huang Zhengxi; Xia Andong; Xu Min; Ma Weimin; Mi Hualing; Ogawa Teruo

    2007-01-01

    The laser confocal scanning microscopy combined with time-correlated single photon counting imaging technique to obtain fluorescence intensity and fluorescence lifetime images for fluorescence resonance energy transfer measurement is reported. Both the fluorescence lifetime imaging microscopy (FLIM) and intensity images show inhomogeneous cyan fluorescent protein and yellow fluorescent protein (CFP /YFP) expression or inhomogeneous energy transfer between CFP and YFP over whole cell. The results presented in this work show that FLIM could be a potential method to reveal the structure-function behavior of NAD(P)H dehydrogenase complexes in living cell

  7. Total Count of Salmonella typhimurium Coupled on Water Soluble CdSe Quantum Dots by Fluorescence Detection

    Science.gov (United States)

    Feliciano Crespo, Raquel; Perales Perez, Oscar Juan; Ramirez, C.

    2018-05-01

    Health diseases due to the ingestion of water or food contaminated with pathogenic microorganisms are a main health problem around the world. The traditional methods for detecting foodborne pathogens are time-consuming (on the order of days). The development of methods that can help to detect and identify foodborne pathogens with high sensitivity and specificity have been proposed to overcome the limitations of traditional methods. Accordingly, this research is focused on the development of an experimental protocol for a high-sensitivity detection and quantification of bacterial pathogens with reduced detection times. This will lead to the development of a portable and low-cost technology with the opportunity to make onsite detection of pathogenic species. The proposed approach has modified the route reported in the literature; the method proposed is expected to be sensitive enough to detect a low limit of 102 CFU/mL counts of bacteria. The fluorescence-based method was tested in presence of Salmonella typhimurium (ATCC 14020) and Escherichia coli (ATCC 25922). CdSe water-soluble quantum dots (QDs) were synthesized in aqueous phase in presence of thioglycolic acid (TGA) as a capping agent. As-synthesized QDs were characterized by x-ray diffraction, near infrared and Fourier transform infrared spectroscopy, UV-Vis and photoluminescence techniques. Results of the CdSe/TGA-bacteria coupling and the determination of the corresponding quantification profiles (calibration curves) will be presented and discussed.

  8. QUANTIFYING THE SHORT LIFETIME WITH TCSPC-FLIM: FIRST MOMENT VERSUS FITTING METHODS

    Directory of Open Access Journals (Sweden)

    LINGLING XU

    2013-10-01

    Full Text Available Combing the time-correlated single photon counting (TCSPC with fluorescence lifetime imaging microscopy (FLIM provides promising opportunities in revealing important information on the microenvironment of cells and tissues, but the applications are thus far mainly limited by the accuracy and precision of the TCSPC-FLIM technique. Here we present a comprehensive investigation on the performance of two data analysis methods, the first moment (M1 method and the conventional least squares (Fitting method, in quantifying fluorescence lifetime. We found that the M1 method is more superior than the Fitting method when the lifetime is short (70 ~ 400 ps or the signal intensity is weak (<103 photons.

  9. Demonstration of the lack of cytotoxicity of unmodified and folic acid modified graphene oxide quantum dots, and their application to fluorescence lifetime imaging of HaCaT cells.

    Science.gov (United States)

    Goreham, Renee V; Schroeder, Kathryn L; Holmes, Amy; Bradley, Siobhan J; Nann, Thomas

    2018-01-24

    The authors describe the synthesis of water-soluble and fluorescent graphene oxide quantum dots via acid exfoliation of graphite nanoparticles. The resultant graphene oxide quantum dots (GoQDs) were then modified with folic acid. Folic acid receptors are overexpressed in cancer cells and hence can bind to functionalized graphene oxide quantum dots. On excitation at 305 nm, the GoQDs display green fluorescence with a peak wavelength at ~520 nm. The modified GoQDs are non-toxic to macrophage cells even after prolonged exposure and high concentrations. Fluorescence lifetime imaging and multiphoton microscopy was used (in combination) to image HeCaT cells exposed to GoQDs, resulting in a superior method for bioimaging. Graphical abstract Schematic representation of graphene oxide quantum dots, folic acid modified graphene oxide quantum dots (red), and the use of fluorescence lifetime to discriminate against green auto-fluorescence of HeCaT cells.

  10. An Automated System for the Control of, and Data Acquisition from Multiphoton Ionization and Fluorescence Lifetime Measurements.

    Science.gov (United States)

    1986-09-01

    Quanta- Ray company , which also supplied the laser used for the multiphoton work. The, burner was mounted on a translator stage from Velmex, Inc...and no longer exists as a process in the system. When the user analysis program has completed, the lifetime program is again automatically re-started...KCHAR) RETURN 100 FORMAT(I3) 101 FORMAT(F7.2) END SUBROUTINE LAB4 FODA SE"oteD C This routine puts the label "INTEGRAL FROM DATA SET" on the MDP C screen

  11. Upgrading the GSI beamline microscope with a confocal fluorescence lifetime scanner to monitor charged particle induced chromatin decondensation in living cells

    Energy Technology Data Exchange (ETDEWEB)

    Abdollahi, Elham; Taucher-Scholz, Gisela [Department of Biophysics, GSI Helmholtz Center for Heavy Ion Research, Planckstrasse 1, 64291 Darmstadt (Germany); Durante, Marco [Department of Biophysics, GSI Helmholtz Center for Heavy Ion Research, Planckstrasse 1, 64291 Darmstadt (Germany); Institute for Condensed Matter Physics, Darmstadt University of Technology, 64289 Darmstadt (Germany); Jakob, Burkhard, E-mail: B.Jakob@gsi.de [Department of Biophysics, GSI Helmholtz Center for Heavy Ion Research, Planckstrasse 1, 64291 Darmstadt (Germany)

    2015-12-15

    We report the upgrade of the GSI beamline microscope coupled to the linear accelerator UNILAC by a confocal FLIM scanner utilizing time correlated single photon counting technique (TCSPC). The system can now be used to address the radiation induced chromatin decondensation in more detail and with higher sensitivity compared to intensity based methods. This decondensation of heterochromatic areas is one of the early DNA damage responses observed after charged particle irradiation and might facilitate the further processing of the induced lesions. We describe here the establishment of different DNA dyes as chromatin compaction probes usable for quantification of the DNA condensation status in living cells utilizing lifetime imaging. In addition, we find an evidence of heterochromatic chromatin decondensation in ion irradiated murine chromocenters detected after subsequent fixation using FLIM measurements.

  12. Distinct roles of the photosystem II protein PsbS and zeaxanthin in the regulation of light harvesting in plants revealed by fluorescence lifetime snapshots.

    Science.gov (United States)

    Sylak-Glassman, Emily J; Malnoë, Alizée; De Re, Eleonora; Brooks, Matthew D; Fischer, Alexandra Lee; Niyogi, Krishna K; Fleming, Graham R

    2014-12-09

    The photosystem II (PSII) protein PsbS and the enzyme violaxanthin deepoxidase (VDE) are known to influence the dynamics of energy-dependent quenching (qE), the component of nonphotochemical quenching (NPQ) that allows plants to respond to fast fluctuations in light intensity. Although the absence of PsbS and VDE has been shown to change the amount of quenching, there have not been any measurements that can detect whether the presence of these proteins alters the type of quenching that occurs. The chlorophyll fluorescence lifetime probes the excited-state chlorophyll relaxation dynamics and can be used to determine the amount of quenching as well as whether two different genotypes with the same amount of NPQ have similar dynamics of excited-state chlorophyll relaxation. We measured the fluorescence lifetimes on whole leaves of Arabidopsis thaliana throughout the induction and relaxation of NPQ for wild type and the qE mutants, npq4, which lacks PsbS; npq1, which lacks VDE and cannot convert violaxanthin to zeaxanthin; and npq1 npq4, which lacks both VDE and PsbS. These measurements show that although PsbS changes the amount of quenching and the rate at which quenching turns on, it does not affect the relaxation dynamics of excited chlorophyll during quenching. In addition, the data suggest that PsbS responds not only to ΔpH but also to the Δψ across the thylakoid membrane. In contrast, the presence of VDE, which is necessary for the accumulation of zeaxanthin, affects the excited-state chlorophyll relaxation dynamics.

  13. Fluorescence lifetime selectivity in excitation-emission matrices for qualitative analysis of a two-component system

    International Nuclear Information System (INIS)

    Millican, D.W.; McGown, L.B.

    1989-01-01

    Steady-state fluorescence excitation-emission matrices (EEMs), and phase-resolved EEMs (PREEMs) collected at modulation frequencies of 6, 18, and 30 MHz, were used for qualitative analysis of mixtures of benzo[k]fluoranthene (τ = 8 ns) and benzo[b]fluoranthene (τ = 29 ns) in ethanol. The EEMs of the individual components were extracted from mixture EEMs by means of wavelength component vector-gram (WCV) analysis. Phase resolution was found to be superior to steady-state measurements for extraction of the component spectra, for mixtures in which the intensity contributions from the two components are unequal

  14. Anthracene Fluorescence Quenching by a Tetrakis (Ketocarboxamide Cavitand

    Directory of Open Access Journals (Sweden)

    Tibor Zoltan Janosi

    2014-01-01

    Full Text Available Quenching of both fluorescence lifetime and fluorescence intensity of anthracene was investigated in the presence of a newly derived tetrakis (ketocarboxamide cavitand at various concentrations. Time-correlated single photon counting method was applied for the lifetime measurements. A clear correlation between the fluorescence lifetime of anthracene as a function of cavitand concentration in dimethylformamide solution was observed. The bimolecular collisional quenching constant was derived from the decrease of lifetime. Fluorescence intensity was measured in the emission wavelength region around 400 nm as a result of excitation at 280 nm. Effective quenching was observed in the presence of the cavitand. The obtained Stern-Volmer plot displayed upward curvature. The results did not follow even extended Stern-Volmer behavior, often used to describe deviations from static bimolecular quenching. To explain our results we adopted the Smoluchowski model and obtained a reasonable estimate for the molecular radius of the cavitand in solution.

  15. Lifetime measurement of the 8s level in francium

    International Nuclear Information System (INIS)

    Gomez, E.; Sprouse, G.D.; Orozco, L.A.; Galvan, A. Perez

    2005-01-01

    We measure the lifetime of the 8s level of 210 Fr atoms on a magneto-optically trapped sample with time-correlated single-photon counting. The 7P 1/2 state serves as the resonant intermediate level for two-step excitation of the 8s level completed with a 1.3-μm laser. Analysis of the fluorescence decay through the 7P 3/2 level gives 53.30±0.44 ns for the 8s level lifetime

  16. The effect of a chlorhexidine-fluoride varnish on mutans streptococci counts and laser fluorescence readings in occlusal fissures of permanent teeth

    DEFF Research Database (Denmark)

    Lipták, Lídia; Bársony, Nóra; Twetman, Svante

    2016-01-01

    Objective: To evaluate the effect of a new chlorhexidinefluor ide varnish on mutans streptococci (MS) counts and laser fluorescence (LF) readings in fissures of permanent molars. Method and Materials: The study group consisted of 57 healthy schoolchildren (7 to 14 yrs) that volunteered after...... informed consent. A double-blind split-mouth design was employed and 87 pairs of non-cavitated permanent molars were randomly assigned to treatments with either chlorhexidine- fluoride varnish (CHX-F) or chlorhexidine-thymol varnish (CHX-T, Cervitec Plus) as active control. The varnishes were topically...

  17. Simultaneous resolution of spectral and temporal properties of UV and visible fluorescence using single-photon counting with a position-sensitive detector

    International Nuclear Information System (INIS)

    Kelly, L.A.; Trunk, J.G.; Polewski, K.; Sutherland, J.C.

    1995-01-01

    A new fluorescence spectrometer has been assembled at the U9B beamline of the National Synchrotron Light Source to allow simultaneous multiwavelength and time-resolved fluorescence detection, as well as spatial imaging of the sample fluorescence. The spectrometer employs monochromatized, tunable UV and visible excitation light from a synchrotron bending magnet and an imaging spectrograph equipped with a single-photon sensitive emission detector. The detector is comprised of microchannel plates in series, with a resistive anode for encoding the position of the photon-derived current. The centroid position of the photon-induced electron cascade is derived in a position analyzer from the four signals measured at the corners of the resistive anode. Spectral information is obtained by dispersing the fluorescence spectrum across one dimension of the detector photocathode. Timing information is obtained by monitoring the voltage divider circuit at the last MCP detector. The signal from the MCP is used as a ''start'' signal to perform a time-correlated single photon counting experiment. The analog signal representing the position, and hence wavelength, is digitized concomitantly with the start/stop time difference and stored in the two-dimensional histogramming memory of a multiparameter analyzer

  18. New approach to the dosimetry of ionizing radiations by fluorescence measurement, according to the single photon counting technique, correlated in time at the nanosecond scale

    International Nuclear Information System (INIS)

    Sohier, Till

    2011-01-01

    This research thesis reports the first fundamental study of the dosimetry of charged and gamma radiations by measurement of fluorescence resolved in time at a nanosecond scale, in organic matter. This method allows an in-depth and real-time analysis of the deposited dose, while taking ionisation as well as excitation processes into account. The author describes mechanisms of interaction and deposition of energy on dense matter, reports the detailed study of the ion-matter interaction, and the interaction of secondary electrons produced within traces. He addresses mechanisms of energy relaxation, and more particularly the study or organic scintillators. Then, he presents the adopted experimental approach: experimental observation with a statistic reconstitution of the curve representing the intensity of the emitted fluorescence in time and with a nanosecond resolution by using a scintillating sensor for time correlated single photon counting (TCSPC). The next part reports the development of an experimental multi-modal platform for dosimetry by TCSPC aimed at the measurement of fluorescence decays under pulsed excitation (nanosecond pulsed ion beams) and continuous flow excitation (non pulsed beams and radioactive sources). Experimental results are then presented for fluorescence measurements, and compared with measurements obtained by using an ionization chamber under the same irradiation conditions: dose deposited by hellions and carbon ions within polyvinyl toluene and polyethylene terephthalate, use of scintillating optic fibers under gamma irradiation of Caesium 137 and Cobalt 60. A new experimental approach is finally presented to perform dosimetry measurements while experimentally ignoring luminescence produced by Cerenkov effect [fr

  19. Penetration of silver nanoparticles into porcine skin ex vivo using fluorescence lifetime imaging microscopy, Raman microscopy, and surface-enhanced Raman scattering microscopy.

    Science.gov (United States)

    Zhu, Yongjian; Choe, Chun-Sik; Ahlberg, Sebastian; Meinke, Martina C; Alexiev, Ulrike; Lademann, Juergen; Darvin, Maxim E

    2015-05-01

    In order to investigate the penetration depth of silver nanoparticles (Ag NPs) inside the skin, porcine ears treated with Ag NPs are measured by two-photon tomography with a fluorescence lifetime imaging microscopy (TPT-FLIM) technique, confocal Raman microscopy (CRM), and surface-enhanced Raman scattering (SERS) microscopy. Ag NPs are coated with poly-N-vinylpyrrolidone and dispersed in pure water solutions. After the application of Ag NPs, porcine ears are stored in the incubator for 24 h at a temperature of 37°C. The TPT-FLIM measurement results show a dramatic decrease of the Ag NPs' signal intensity from the skin surface to a depth of 4 μm. Below 4 μm, the Ag NPs' signal continues to decline, having completely disappeared at 12 to 14 μm depth. CRM shows that the penetration depth of Ag NPs is 11.1 ± 2.1 μm. The penetration depth measured with a highly sensitive SERS microscopy reaches 15.6 ± 8.3 μm. Several results obtained with SERS show that the penetration depth of Ag NPs can exceed the stratum corneum (SC) thickness, which can be explained by both penetration of trace amounts of Ag NPs through the SC barrier and by the measurements inside the hair follicle, which cannot be excluded in the experiment.

  20. Advanced photon counting applications, methods, instrumentation

    CERN Document Server

    Kapusta, Peter; Erdmann, Rainer

    2015-01-01

    This volume focuses on Time-Correlated Single Photon Counting (TCSPC), a powerful tool allowing luminescence lifetime measurements to be made with high temporal resolution, even on single molecules. Combining spectrum and lifetime provides a "fingerprint" for identifying such molecules in the presence of a background. Used together with confocal detection, this permits single-molecule spectroscopy and microscopy in addition to ensemble measurements, opening up an enormous range of hot life science applications such as fluorescence lifetime imaging (FLIM) and measurement of Förster Resonant Energy Transfer (FRET) for the investigation of protein folding and interaction. Several technology-related chapters present both the basics and current state-of-the-art, in particular of TCSPC electronics, photon detectors and lasers. The remaining chapters cover a broad range of applications and methodologies for experiments and data analysis, including the life sciences, defect centers in diamonds, super-resolution micr...

  1. A 32-channel photon counting module with embedded auto/cross-correlators for real-time parallel fluorescence correlation spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Gong, S.; Labanca, I.; Rech, I.; Ghioni, M. [Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano (Italy)

    2014-10-15

    Fluorescence correlation spectroscopy (FCS) is a well-established technique to study binding interactions or the diffusion of fluorescently labeled biomolecules in vitro and in vivo. Fast FCS experiments require parallel data acquisition and analysis which can be achieved by exploiting a multi-channel Single Photon Avalanche Diode (SPAD) array and a corresponding multi-input correlator. This paper reports a 32-channel FPGA based correlator able to perform 32 auto/cross-correlations simultaneously over a lag-time ranging from 10 ns up to 150 ms. The correlator is included in a 32 × 1 SPAD array module, providing a compact and flexible instrument for high throughput FCS experiments. However, some inherent features of SPAD arrays, namely afterpulsing and optical crosstalk effects, may introduce distortions in the measurement of auto- and cross-correlation functions. We investigated these limitations to assess their impact on the module and evaluate possible workarounds.

  2. A 32-channel photon counting module with embedded auto/cross-correlators for real-time parallel fluorescence correlation spectroscopy

    International Nuclear Information System (INIS)

    Gong, S.; Labanca, I.; Rech, I.; Ghioni, M.

    2014-01-01

    Fluorescence correlation spectroscopy (FCS) is a well-established technique to study binding interactions or the diffusion of fluorescently labeled biomolecules in vitro and in vivo. Fast FCS experiments require parallel data acquisition and analysis which can be achieved by exploiting a multi-channel Single Photon Avalanche Diode (SPAD) array and a corresponding multi-input correlator. This paper reports a 32-channel FPGA based correlator able to perform 32 auto/cross-correlations simultaneously over a lag-time ranging from 10 ns up to 150 ms. The correlator is included in a 32 × 1 SPAD array module, providing a compact and flexible instrument for high throughput FCS experiments. However, some inherent features of SPAD arrays, namely afterpulsing and optical crosstalk effects, may introduce distortions in the measurement of auto- and cross-correlation functions. We investigated these limitations to assess their impact on the module and evaluate possible workarounds

  3. High-resolution imaging of basal cell carcinoma: a comparison between multiphoton microscopy with fluorescence lifetime imaging and reflectance confocal microscopy.

    Science.gov (United States)

    Manfredini, Marco; Arginelli, Federica; Dunsby, Christopher; French, Paul; Talbot, Clifford; König, Karsten; Pellacani, Giovanni; Ponti, Giovanni; Seidenari, Stefania

    2013-02-01

    The aim of this study was to compare morphological aspects of basal cell carcinoma (BCC) as assessed by two different imaging methods: in vivo reflectance confocal microscopy (RCM) and multiphoton tomography with fluorescence lifetime imaging implementation (MPT-FLIM). The study comprised 16 BCCs for which a complete set of RCM and MPT-FLIM images were available. The presence of seven MPT-FLIM descriptors was evaluated. The presence of seven RCM equivalent parameters was scored in accordance to their extension. Chi-squared test with Fisher's exact test and Spearman's rank correlation coefficient were determined between MPT-FLIM scores and adjusted-RCM scores. MPT-FLIM and RCM descriptors of BCC were coupled to match the descriptors that define the same pathological structures. The comparison included: Streaming and Aligned elongated cells, Streaming with multiple directions and Double alignment, Palisading (RCM) and Palisading (MPT-FLIM), Typical tumor islands, and Cell islands surrounded by fibers, Dark silhouettes and Phantom islands, Plump bright cells and Melanophages, Vessels (RCM), and Vessels (MPT-FLIM). The parameters that were significantly correlated were Melanophages/Plump Bright Cells, Aligned elongated cells/Streaming, Double alignment/Streaming with multiple directions, and Palisading (MPT-FLIM)/Palisading (RCM). According to our data, both methods are suitable to image BCC's features. The concordance between MPT-FLIM and RCM is high, with some limitations due to the technical differences between the two devices. The hardest difficulty when comparing the images generated by the two imaging modalities is represented by their different field of view. © 2012 John Wiley & Sons A/S.

  4. Nuclear lifetimes

    International Nuclear Information System (INIS)

    Caraca, J.M.G.

    1976-01-01

    The importance of the results obtained in experiments of measurement of lifetimes for a detailed knowledge of nuclear structure is referred. Direct methods of measurement of nuclear lifetimes are described, namely, electronic methods, recoil-distance method, doppler shift atenuation method and blocking-method. A brief reference is made to indirect methods for measurement of life-times

  5. Detection of X-ray fluorescence of light elements by electron counting in a low-pressure gaseous electron multiplier

    International Nuclear Information System (INIS)

    Pansky, A.; Breskin, A.; Chechik, R.; Malamud, G.

    1992-12-01

    Ionization electrons deposited by soft X-rays in a low pressure (10 Torr) gas medium are efficiently counted by a multistage electron multiplier, providing an accurate measurement of the X-ray photon energy. Energy resolution of 56-28% FWHM were measured for X-rays of 110-676 eV, recording electrical induced charges or visible photons emitted during the avalanche process. It is demonstrated that a combined analysis of the number of electron trail length of an event, provides a powerful and competitive way of resolving ultra soft X-rays. We present the experimental technique, discuss the advantages and limitations of the Primary Electron Counter, and suggest ways to improve its performances. (authors)

  6. Multiphoton autofluorescence lifetime imaging of induced pluripotent stem cells

    Science.gov (United States)

    Uchugonova, Aisada

    2017-06-01

    The multiphoton fluorescence lifetime imaging tomograph MPTflex with its flexible 360-deg scan head, articulated arm, and tunable femtosecond laser source was employed to study induced pluripotent stem cell (iPS) cultures. Autofluorescence (AF) lifetime imaging was performed with 250-ps temporal resolution and submicron spatial resolution using time-correlated single-photon counting. The two-photon excited AF was based on the metabolic coenzymes NAD(P)H and flavin adenine dinucleotide/flavoproteins. iPS cells generated from mouse embryonic fibroblasts (MEFs) and cocultured with growth-arrested MEFs as feeder cells have been studied. Significant differences on AF lifetime signatures were identified between iPS and feeder cells as well as between their differentiating counterparts.

  7. A combination of direct viable count and fluorescence in situ hybridization for specific enumeration of viable Lactobacillus delbrueckii subsp.bulgaricus and Streptococcus thermophilus.

    Science.gov (United States)

    García-Hernández, J; Moreno, Y; Amorocho, C M; Hernández, M

    2012-03-01

    We have developed a direct viable count (DVC)-FISH procedure for quickly and easily discriminating between viable and nonviable cells of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains, the traditional yogurt bacteria. direct viable count method has been modified and adapted for Lact. delbrueckii subsp. bulgaricus and Strep. thermophilus analysis by testing different times of incubation and concentrations of DNA-gyrase inhibitors. DVC procedure has been combined with fluorescent in situ hybridization (FISH) for the specific detection of viable cells of both bacteria with specific rRNA oligonucleotide probes (DVC-FISH). Of the four antibiotics tested (novobiocin, nalidixic acid, pipemidic acid and ciprofloxacin), novobiocin was the most effective for DVC method and the optimum incubation time was 7 h for both bacteria. The number of viable cells was obtained by the enumeration of specific hybridized cells that were elongated at least twice their original length for Lactobacillus and twice their original size for Streptococcus. This technique was successfully applied to detect viable cells in inoculated faeces. Results showed that this DVC-FISH procedure is a quick and culture-independent useful method to specifically detect viable Lact. delbrueckii subsp. bulgaricus and Strep. thermophilus in different samples, being applied for the first time to lactic acid bacteria. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

  8. Fluctuation localization imaging-based fluorescence in situ hybridization (fliFISH) for accurate detection and counting of RNA copies in single cells

    Energy Technology Data Exchange (ETDEWEB)

    Cui, Yi; Hu, Dehong; Markillie, Lye Meng; Chrisler, William B.; Gaffrey, Matthew J.; Ansong, Charles; Sussel, Lori; Orr, Galya

    2017-10-04

    Quantitative gene expression analysis in intact single cells can be achieved using single molecule- based fluorescence in situ hybridization (smFISH). This approach relies on fluorescence intensity to distinguish between true signals, emitted from an RNA copy hybridized with multiple FISH sub-probes, and background noise. Thus, the precision in smFISH is often compromised by partial or nonspecific binding of sub-probes and tissue autofluorescence, limiting its accuracy. Here we provide an accurate approach for setting quantitative thresholds between true and false signals, which relies on blinking frequencies of photoswitchable dyes. This fluctuation localization imaging-based FISH (fliFISH) uses blinking frequency patterns, emitted from a transcript bound to multiple sub-probes, which are distinct from blinking patterns emitted from partial or nonspecifically bound sub-probes and autofluorescence. Using multicolor fliFISH, we identified radial gene expression patterns in mouse pancreatic islets for insulin, the transcription factor, NKX2-2, and their ratio (Nkx2-2/Ins2). These radial patterns, showing higher values in β cells at the islet core and lower values in peripheral cells, were lost in diabetic mouse islets. In summary, fliFISH provides an accurate, quantitative approach for detecting and counting true RNA copies and rejecting false signals by their distinct blinking frequency patterns, laying the foundation for reliable single-cell transcriptomics.

  9. Lifetime measurements

    International Nuclear Information System (INIS)

    Fossan, D.B.; Warburton, E.K.

    1974-01-01

    Lifetime measurements are discussed, concentrating on the electronic technique, the recoil distance method (RDM), and the Doppler shift attenuation method (DSAM). A brief review of several indirect timing techniques is given, and their specific advantages and applicability are considered. The relationship between lifetimes of nuclear states and the nuclear structure information obtained from them is examined. A short discussion of channeling and microwave methods of lifetime measurement is presented. (23 figures, 171 references) (U.S.)

  10. Optimization of time-correlated single photon counting spectrometer

    International Nuclear Information System (INIS)

    Zhang Xiufeng; Du Haiying; Sun Jinsheng

    2011-01-01

    The paper proposes a performance improving scheme for the conventional time-correlated single photon counting spectrometer and develops a high speed data acquisition card based on PCI bus and FPGA technologies. The card is used to replace the multi-channel analyzer to improve the capability and decrease the volume of the spectrometer. The process of operation is introduced along with the integration of the spectrometer system. Many standard samples are measured. The experimental results show that the sensitivity of the spectrometer is single photon counting, and the time resolution of fluorescence lifetime measurement can be picosecond level. The instrument could measure the time-resolved spectroscopy. (authors)

  11. Fluorescence Decay Dynamics of Ethidium Bromide in Polymers

    International Nuclear Information System (INIS)

    Jee, Ah Young; Min Yung

    2010-01-01

    The fluorescence lifetimes of EB in five polymers covering LDPE, HDPE, PC, PS, and PAA were measured by picosecond time-correlated single photon counting. The lifetime change of EB has been previously described by hydrogen bonding ability. In this work, we have observed that the lifetime of EB depends strongly on the Young's modulus of medium. Thus, it is possible that the fluorescence decay dynamics of EB could be influenced by medium rigidity rather than hydrogen bonding ability in polymer. The medium influence on the fluorescence decay dynamics of ethidium bromide (EB) has been investigated in various environments. For example, Ohmstead and Kearns related the fluorescence lifetime of EB to the excited-state proton transfer process. In addition, they reported that the solvent viscosity plays a minor role in the excited state decay process of EB. Chirico et al. measured the fluorescence decay of EB as 1.7 ns in water and 6.5 ns in ethanol and concluded that hydrogen bonding ability is a key factor for the nonradiative relaxation. Pal et al. measured the fluorescence decay time of EB in acetone, acetonitrile, and their mixtures. They observed that the fluorescence decay processes were independent on the solvent polarity. These results show that the EB lifetime does not depend much on polarity or viscosity, but is mainly influenced by hydrogen bonding. Overall, EB is one of most widely used dyes for probing DNA. When EB is intercalated into the helical structure of DNA, a large increase in the fluorescence lifetime has been observed in comparison with water environment, and the fluorescence enhancement was attributed to the blocking of the excited-state proton transfer

  12. Comparative analysis of dose rates in bricks determined by neutron activation analysis, alpha counting and X-ray fluorescence analysis for the thermoluminescence fine grain dating method

    Science.gov (United States)

    Bártová, H.; Kučera, J.; Musílek, L.; Trojek, T.

    2014-11-01

    In order to evaluate the age from the equivalent dose and to obtain an optimized and efficient procedure for thermoluminescence (TL) dating, it is necessary to obtain the values of both the internal and the external dose rates from dated samples and from their environment. The measurements described and compared in this paper refer to bricks from historic buildings and a fine-grain dating method. The external doses are therefore negligible, if the samples are taken from a sufficient depth in the wall. However, both the alpha dose rate and the beta and gamma dose rates must be taken into account in the internal dose. The internal dose rate to fine-grain samples is caused by the concentrations of natural radionuclides 238U, 235U, 232Th and members of their decay chains, and by 40K concentrations. Various methods can be used for determining trace concentrations of these natural radionuclides and their contributions to the dose rate. The dose rate fraction from 238U and 232Th can be calculated, e.g., from the alpha count rate, or from the concentrations of 238U and 232Th, measured by neutron activation analysis (NAA). The dose rate fraction from 40K can be calculated from the concentration of potassium measured, e.g., by X-ray fluorescence analysis (XRF) or by NAA. Alpha counting and XRF are relatively simple and are accessible for an ordinary laboratory. NAA can be considered as a more accurate method, but it is more demanding regarding time and costs, since it needs a nuclear reactor as a neutron source. A comparison of these methods allows us to decide whether the time- and cost-saving simpler techniques introduce uncertainty that is still acceptable.

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

    Directory of Open Access Journals (Sweden)

    Jiayao Li

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

  14. A Rotational BODIPY Nucleotide: An Environment-Sensitive Fluorescence-Lifetime Probe for DNA Interactions and Applications in Live-Cell Microscopy

    Czech Academy of Sciences Publication Activity Database

    Dziuba, Dmytro; Jurkiewicz, Piotr; Cebecauer, Marek; Hof, Martin; Hocek, Michal

    2016-01-01

    Roč. 55, č. 1 (2016), s. 174-178 ISSN 1433-7851 R&D Projects: GA ČR GBP206/12/G151; GA ČR(CZ) GC14-03141J Institutional support: RVO:61388963 ; RVO:61388955 Keywords : DNA * fluorescence spectroscopy * fluorescent probes * nucleosides * time-resolved spectroscopy Subject RIV: CC - Organic Chemistry ; BO - Biophysics (UFCH-W) Impact factor: 11.994, year: 2016

  15. Lifetime measurements

    International Nuclear Information System (INIS)

    Poletti, A.R.

    1976-01-01

    Recent developments in experimental methods of measuring the lifetimes of excited nuclear states is reviewed in three main areas. (a) Doppler Shift Attenuation Measurements (DSAM) Times: 10 -14 - 10 -11 sec.; (b) Recoil Distance Measurements (RDM) Times: 10 -9 - 10 -12 sec.; (c) Direct Electronic Timing Times: down to 10 -10 sec.; A measurement of an excited state lifetime can answer a large number of different questions. Two examples are discussed: (a) The determination of the lifetime of an isomeric transition in 93 Tc and its use in determining an upper limit for the magnitude of the parity non-conserving matrix element - /Hsub(PN)/17/2 + >. (b) The dependence of the strength of M2 transitions on isospin in nuclei in the 1dsub(3/2) -1fsub(7/2) region. (author)

  16. In-situ fluorescence hybridization applied to biological dosimetry: contribution of automation to the counting of radio-induced chromosome aberrations

    International Nuclear Information System (INIS)

    Germain Thomas Roy, Laurence

    1999-01-01

    The frequency of chromosome aberrations on peripheral blood lymphocytes is a dose indicator in the case of ionizing radiations over-exposure. Stable chromosome aberrations (translocations, insertions) are visualized after labelling of some chromosomes using the fluorescence in-situ hybridization (FISH). The study of the use of the FISH technique in biological dosimetry is done with dose-effect curves. It seems that a bias is introduced during the observation of chromosome aberrations involving only 3 pairs of chromosomes. In order to avoid this bias, it would be useful to test the feasibility of using the multi-FISH technique in biological dosimetry. Moreover, this type of chromosome aberration changes with the type of irradiation. It is thus important to define the aberrations to be considered when the FISH technique is used. In order to reduce the time of image analysis, the CYTOGEN system, developed by IMSTAR company (Paris, France) has been adapted to the needs of biological dosimetry. This system allows to localize automatically the metaphases on the slide, which reduces the observation time by 2 or 4. An automatic detection protocol for chromosome aberrations has been implemented. It comprises the image capture, the contours detection and the classification of some chromosome aberrations. The different steps of this protocol have been tested in order to check that no bias is introduced by the automation. However, because radio-induced aberrations are rare events, it seems that a totally automatic system is not foreseeable. A semi-automatic analysis is more suitable. The use of the Slit-Scan technology (Laboratory of applied physics, Heidelberg, Germany) in biological dosimetry has been studied too. This technique allows to analyze rapidly a huge number of chromosomes. A good correlation has been observed between the dicentric frequency measured automatically and by manual counting. The system is under development and should be adapted to the detection of

  17. Fully time-resolved near-field scanning optical microscopy fluorescence imaging

    International Nuclear Information System (INIS)

    Kwak, Eun-Soo; Vanden Bout, David A.

    2003-01-01

    Time-correlated single photon counting has been coupled with near-field scanning optical microscopy (NSOM) to record complete fluorescence lifetime decays at each pixel in an NSOM image. The resulting three-dimensional data sets can be binned in the time dimension to create images of photons at particular time delays or images of the fluorescence lifetime. Alternatively, regions of interest identified in the topography and fluorescence images can be used to bin the data in the spatial dimensions resulting in high signal to noise fluorescence decays of particular regions of the sample. The technique has been demonstrated on films of poly(vinylalcohol), doped with the fluorescent dye, cascade blue (CB). The CB segregates into small circular regions of high concentration within the films during the drying process. The lifetime imaging shows that the spots have slightly faster excited state decays due to quenching of the luminescence as a result of the higher concentration. The technique is also used to image the fluorescence lifetime of an annealed film of poly(dihexylfluorene). The samples show high contrast in the total intensity fluorescence image, but the lifetime image reveals the sample to be extremely uniform

  18. Fluorescence kinetics of Trp-Trp dipeptide and its derivatives in water via ultrafast fluorescence spectroscopy.

    Science.gov (United States)

    Jia, Menghui; Yi, Hua; Chang, Mengfang; Cao, Xiaodan; Li, Lei; Zhou, Zhongneng; Pan, Haifeng; Chen, Yan; Zhang, Sanjun; Xu, Jianhua

    2015-08-01

    Ultrafast fluorescence dynamics of Tryptophan-Tryptophan (Trp-Trp/Trp2) dipeptide and its derivatives in water have been investigated using a picosecond resolved time correlated single photon counting (TCSPC) apparatus together with a femtosecond resolved upconversion spectrophotofluorometer. The fluorescence decay profiles at multiple wavelengths were fitted by a global analysis technique. Nanosecond fluorescence kinetics of Trp2, N-tert-butyl carbonyl oxygen-N'-aldehyde group-l-tryptophan-l-tryptophan (NBTrp2), l-tryptophan-l-tryptophan methyl ester (Trp2Me), and N-acetyl-l-tryptophan-l-tryptophan methyl ester (NATrp2Me) exhibit multi-exponential decays with the average lifetimes of 1.99, 3.04, 0.72 and 1.22ns, respectively. Due to the intramolecular interaction between two Trp residues, the "water relaxation" lifetime was observed around 4ps, and it is noticed that Trp2 and its derivatives also exhibit a new decay with a lifetime of ∼100ps, while single-Trp fluorescence decay in dipeptides/proteins shows 20-30ps. The intramolecular interaction lifetime constants of Trp2, NBTrp2, Trp2Me and NATrp2Me were then calculated to be 3.64, 0.93, 11.52 and 2.40ns, respectively. Candidate mechanisms (including heterogeneity, solvent relaxation, quasi static self-quenching or ET/PT quenching) have been discussed. Copyright © 2015. Published by Elsevier B.V.

  19. Precision lifetime measurements

    International Nuclear Information System (INIS)

    Tanner, C.E.

    1994-01-01

    Precision measurements of atomic lifetimes provide important information necessary for testing atomic theory. The authors employ resonant laser excitation of a fast atomic beam to measure excited state lifetimes by observing the decay-in-flight of the emitted fluorescence. A similar technique was used by Gaupp, et al., who reported measurements with precisions of less than 0.2%. Their program includes lifetime measurements of the low lying p states in alkali and alkali like systems. Motivation for this work comes from a need to test the atomic many-body-perturbation theory (MBPT) that is necessary for interpretation of parity nonconservation experiments in atomic cesium. The authors have measured the cesium 6p 2 P 1/2 and 6p 2 P 3/2 state lifetimes to be 34.934±0.094 ns and 30.499±0.070 ns respectively. With minor changes to the apparatus, they have extended their measurements to include the lithium 2p 2 P 1/2 and 2p 2 P 3/2 states

  20. FUNDUS AUTOFLUORESCENCE LIFETIMES AND CENTRAL SEROUS CHORIORETINOPATHY.

    Science.gov (United States)

    Dysli, Chantal; Berger, Lieselotte; Wolf, Sebastian; Zinkernagel, Martin S

    2017-11-01

    To quantify retinal fluorescence lifetimes in patients with central serous chorioretinopathy (CSC) and to identify disease specific lifetime characteristics over the course of disease. Forty-seven participants were included in this study. Patients with central serous chorioretinopathy were imaged with fundus photography, fundus autofluorescence, optical coherence tomography, and fluorescence lifetime imaging ophthalmoscopy (FLIO) and compared with age-matched controls. Retinal autofluorescence was excited using a 473-nm blue laser light and emitted fluorescence light was detected in 2 distinct wavelengths channels (498-560 nm and 560-720 nm). Clinical features, mean retinal autofluorescence lifetimes, autofluorescence intensity, and corresponding optical coherence tomography (OCT) images were further analyzed. Thirty-five central serous chorioretinopathy patients with a mean visual acuity of 78 ETDRS letters (range, 50-90; mean Snellen equivalent: 20/32) and 12 age-matched controls were included. In the acute stage of central serous chorioretinopathy, retinal fluorescence lifetimes were shortened by 15% and 17% in the respective wavelength channels. Multiple linear regression analysis showed that fluorescence lifetimes were significantly influenced by the disease duration (P autofluorescence lifetimes, particularly in eyes with retinal pigment epithelial atrophy, were associated with poor visual acuity. This study establishes that autofluorescence lifetime changes occurring in central serous chorioretinopathy exhibit explicit patterns which can be used to estimate perturbations of the outer retinal layers with a high degree of statistical significance.

  1. An automated wide-field time-gated optically sectioning fluorescence lifetime imaging multiwell plate reader for high-content analysis of protein-protein interactions

    Science.gov (United States)

    Alibhai, Dominic; Kumar, Sunil; Kelly, Douglas; Warren, Sean; Alexandrov, Yuriy; Munro, Ian; McGinty, James; Talbot, Clifford; Murray, Edward J.; Stuhmeier, Frank; Neil, Mark A. A.; Dunsby, Chris; French, Paul M. W.

    2011-03-01

    We describe an optically-sectioned FLIM multiwell plate reader that combines Nipkow microscopy with wide-field time-gated FLIM, and its application to high content analysis of FRET. The system acquires sectioned FLIM images in fluorescent protein. It has been applied to study the formation of immature HIV virus like particles (VLPs) in live cells by monitoring Gag-Gag protein interactions using FLIM FRET of HIV-1 Gag transfected with CFP or YFP. VLP formation results in FRET between closely packed Gag proteins, as confirmed by our FLIM analysis that includes automatic image segmentation.

  2. RBC count

    Science.gov (United States)

    ... by kidney disease) RBC destruction ( hemolysis ) due to transfusion, blood vessel injury, or other cause Leukemia Malnutrition Bone ... slight risk any time the skin is broken) Alternative Names Erythrocyte count; Red blood cell count; Anemia - RBC count Images Blood test ...

  3. Highly efficient exciplex formation via radical ion pair recombination in X-irradiated alkane solutions for luminophores with short fluorescence lifetimes.

    Science.gov (United States)

    Melnikov, Anatoly R; Kalneus, Evgeny V; Korolev, Valeri V; Dranov, Igor G; Kruppa, Alexander I; Stass, Dmitri V

    2014-08-01

    X-irradiation of alkane solutions of N,N-dimethylaniline with various organic luminophores produces characteristic emission bands ascribed to the corresponding exciplexes. In contrast to optical generation, which requires diffusion-controlled quenching of excited states, an additional channel of exciplex formation via irreversible recombination of radical ion pairs is operative here, which produces exciplexes in solution with high efficiency even for p-terphenyl and diphenylacetylene having fluorescence decay times of 0.95 ns and 8 ps, respectively. The exciplex emission band is sensitive to an external magnetic field and exerts a very large observed magnetic field effect of up to 20%, the maximum possible value under the conditions of the described experiment.

  4. Thermally activated delayed fluorescence of fluorescein derivative for time-resolved and confocal fluorescence imaging.

    Science.gov (United States)

    Xiong, Xiaoqing; Song, Fengling; Wang, Jingyun; Zhang, Yukang; Xue, Yingying; Sun, Liangliang; Jiang, Na; Gao, Pan; Tian, Lu; Peng, Xiaojun

    2014-07-09

    Compared with fluorescence imaging utilizing fluorophores whose lifetimes are in the order of nanoseconds, time-resolved fluorescence microscopy has more advantages in monitoring target fluorescence. In this work, compound DCF-MPYM, which is based on a fluorescein derivative, showed long-lived luminescence (22.11 μs in deaerated ethanol) and was used in time-resolved fluorescence imaging in living cells. Both nanosecond time-resolved transient difference absorption spectra and time-correlated single-photon counting (TCSPC) were employed to explain the long lifetime of the compound, which is rare in pure organic fluorophores without rare earth metals and heavy atoms. A mechanism of thermally activated delayed fluorescence (TADF) that considers the long wavelength fluorescence, large Stokes shift, and long-lived triplet state of DCF-MPYM was proposed. The energy gap (ΔEST) of DCF-MPYM between the singlet and triplet state was determined to be 28.36 meV by the decay rate of DF as a function of temperature. The ΔE(ST) was small enough to allow efficient intersystem crossing (ISC) and reverse ISC, leading to efficient TADF at room temperature. The straightforward synthesis of DCF-MPYM and wide availability of its starting materials contribute to the excellent potential of the compound to replace luminescent lanthanide complexes in future time-resolved imaging technologies.

  5. Counting carbohydrates

    Science.gov (United States)

    Carb counting; Carbohydrate-controlled diet; Diabetic diet; Diabetes-counting carbohydrates ... Many foods contain carbohydrates (carbs), including: Fruit and fruit juice Cereal, bread, pasta, and rice Milk and milk products, soy milk Beans, legumes, ...

  6. Improving fluorescence diagnosis of cancer by SLIM

    Science.gov (United States)

    Rück, Angelika; Dolp, Frank; Kinzler, Ingrid; Hauser, Carmen; Scalfi-Happ, Claudia

    2006-02-01

    Although during the last years, significant progress was made in cancer diagnosis, using either intrinsic or specially designed fluorophores, still problems exist, due to difficulties in spectral separation of highly overlapping probes or in lack of specificity. Many of the problems could be circumvented by focusing on time-resolved methods. In combination with spectral resolved detection (spectral fluorescence lifetime imaging, SLIM) highly sophisticated fluorescence lifetime imaging can be performed which might improve specificity of cell diagnosis. To record lifetime images (τ-mapping) with spectral resolution a setup was realized consisting of a laser scanning microscope equipped with a 16 channel array for time-correlated single photon counting (TCSPC) and a spectrograph in front of the array. A Ti:Saphir laser can be used for excitation or alternatively ps diode lasers. With this system the time- and spectral-resolved fluorescence characteristics of different fluorophores were investigated in solution and in cell culture. As an example, not only the mitochondria staining dye rhodamine 123 could be easily distinguished from DAPI, which intercalates into nucleic acids, but also different binding sites of DAPI. This was proved by the appearance of different lifetime components within different spectral channels. Another example is Photofrin, a photosensitizer which is approved for bladder cancer and for palliative lung and esophageal cancer in 20 countries, including the United States, Canada and many European countries. Photofrin is a complex mixture of different monomeric and aggregated porphyrins. The phototoxic efficiency during photodynamic therapy (PDT) seems to be correlated with the relative amounts of monomers and aggregates. With SLIM different lifetimes could be attributed to various, spectrally highly overlapping compounds. In addition, a detailed analysis of the autofluorescence by SLIM could explain changes of mitochondrial metabolism during

  7. Principles of fluorescence techniques

    CERN Document Server

    2016-01-01

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

  8. Comparative analysis of dose rates in bricks determined by neutron activation analysis, alpha counting and X-ray fluorescence analysis for the thermoluminescence fine grain dating method

    Czech Academy of Sciences Publication Activity Database

    Bártová, H.; Kučera, Jan; Musílek, L.; Trojek, T.

    2014-01-01

    Roč. 104, NOV (2014), s. 393-397 ISSN 0969-806X. [1st International Conference on Dosimetry and its Applications (ICDA). Prague, 23.6.2013-28.6.2013] R&D Projects: GA MŠk(XE) LM2011019 Institutional support: RVO:61389005 Keywords : Alpha coutning * neutron activation analysis * X-ray fluorescence * thermoluminescence dating * dose rate Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 1.380, year: 2014

  9. FastFLIM, the all-in-one engine for measuring photoluminescence lifetime of 100 picoseconds to 100 milliseconds

    Science.gov (United States)

    Sun, Yuansheng; Coskun, Ulas; Liao, Shih-Chu Jeff; Barbieri, Beniamino

    2018-02-01

    Photoluminescence (PL) refers to light emission initiated by any form of photon excitation. PL spectroscopy and microscopy imaging has been widely applied in material, chemical and life sciences. Measuring its lifetime yields a new dimension of the PL imaging and opens new opportunities for many PL applications. In solar cell research, quantification of the PL lifetime has become an important evaluation for the characteristics of the Perovskite thin film. Depending upon the PL process (fluorescence, phosphorescence, photon upconversion, etc.), the PL lifetimes to be measured can vary in a wide timescale range (e.g. from sub-nanoseconds to microseconds or even milliseconds) - it is challenging to cover this wide range of lifetime measurements by a single technique efficiently. Here, we present a novel digital frequency domain (DFD) technique named FastFLIM, capable of measuring the PL lifetime from 100 ps to 100 ms at the high data collection efficiency (up to 140-million counts per second). Other than the traditional nonlinear leastsquare fitting analysis, the raw data acquired by FastFLIM can be directly processed by the model-free phasor plots approach for instant and unbiased lifetime results, providing the ideal routine for the PL lifetime microscopy imaging.

  10. Using the fluorescence of DBO to study the aggregation of asphaltenes

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Zixin; Bohne, Cornelia [Department of Chemistry, University of Victoria (Canada)], email: xyang@uvic.ca

    2010-07-01

    Asphaltene, operationally defined as the fraction of bitumen that is insoluble in heptane but soluble in toluene, is the least characterized component of crude oil. It can aggregate at low concentrations, causing problems in the reservoir and during transport and processing. Fluorescence techniques have been employed to characterize asphaltenes, e.g. by adding external probes. DBO (2,3-diazabicyclo [2.2.2]oct-2-ene), a fluorescence probe molecule with a long fluorescence lifetime, was made sensitive to the presence of aliphatic C-H bonds. DBO was used as an external fluorescent probe to characterize the aggregation of Athabasca asphaltene. The lifetime of DBO was measured using single photon counting. Preliminary lifetime measurements show that AA-5 quenches the emission of DBO, leading to a shortening of the DBO lifetime. The abrupt decrease in lifetime may be related to the interaction of DBO with the AA-5 aggregate; further studies are being performed to test this hypothesis. In conclusion, DBO interacts with asphaltene components and has the potential for being used as a probe to study the asphaltene aggregation.

  11. Improving the counting efficiency in time-correlated single photon counting experiments by dead-time optimization

    Energy Technology Data Exchange (ETDEWEB)

    Peronio, P.; Acconcia, G.; Rech, I.; Ghioni, M. [Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano (Italy)

    2015-11-15

    Time-Correlated Single Photon Counting (TCSPC) has been long recognized as the most sensitive method for fluorescence lifetime measurements, but often requiring “long” data acquisition times. This drawback is related to the limited counting capability of the TCSPC technique, due to pile-up and counting loss effects. In recent years, multi-module TCSPC systems have been introduced to overcome this issue. Splitting the light into several detectors connected to independent TCSPC modules proportionally increases the counting capability. Of course, multi-module operation also increases the system cost and can cause space and power supply problems. In this paper, we propose an alternative approach based on a new detector and processing electronics designed to reduce the overall system dead time, thus enabling efficient photon collection at high excitation rate. We present a fast active quenching circuit for single-photon avalanche diodes which features a minimum dead time of 12.4 ns. We also introduce a new Time-to-Amplitude Converter (TAC) able to attain extra-short dead time thanks to the combination of a scalable array of monolithically integrated TACs and a sequential router. The fast TAC (F-TAC) makes it possible to operate the system towards the upper limit of detector count rate capability (∼80 Mcps) with reduced pile-up losses, addressing one of the historic criticisms of TCSPC. Preliminary measurements on the F-TAC are presented and discussed.

  12. Counting cormorants

    DEFF Research Database (Denmark)

    Bregnballe, Thomas; Carss, David N; Lorentsen, Svein-Håkon

    2013-01-01

    This chapter focuses on Cormorant population counts for both summer (i.e. breeding) and winter (i.e. migration, winter roosts) seasons. It also explains differences in the data collected from undertaking ‘day’ versus ‘roost’ counts, gives some definitions of the term ‘numbers’, and presents two...

  13. Enumeration of Vibrio parahaemolyticus in the viable but nonculturable state using direct plate counts and recognition of individual gene fluorescence in situ hybridization.

    Science.gov (United States)

    Griffitt, Kimberly J; Noriea, Nicholas F; Johnson, Crystal N; Grimes, D Jay

    2011-05-01

    Vibrio parahaemolyticus is a gram-negative, halophilic bacterium indigenous to marine and estuarine environments and it is capable of causing food and water-borne illness in humans. It can also cause disease in marine animals, including cultured species. Currently, culture-based techniques are used for quantification of V. parahaemolyticus in environmental samples; however, these can be misleading as they fail to detect V. parahaemolyticus in a viable but nonculturable (VBNC) state which leads to an underestimation of the population density. In this study, we used a novel fluorescence visualization technique, called recognition of individual gene fluorescence in situ hybridization (RING-FISH), which targets chromosomal DNA for enumeration. A polynucleotide probe labeled with Cyanine 3 (Cy3) was created corresponding to the ubiquitous V. parahaemolyticus gene that codes for thermolabile hemolysin (tlh). When coupled with the Kogure method to distinguish viable from dead cells, RING-FISH probes reliably enumerated total, viable V. parahaemolyticus. The probe was tested for sensitivity and specificity against a pure culture of tlh(+), tdh(-), trh(-)V. parahaemolyticus, pure cultures of Vibrio vulnificus, Vibrio harveyi, Vibrio alginolyticus and Vibrio fischeri, and a mixed environmental sample. This research will provide additional tools for a better understanding of the risk these environmental organisms pose to human health. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. Multimodal fluorescence imaging spectroscopy

    NARCIS (Netherlands)

    Stopel, Martijn H W; Blum, Christian; Subramaniam, Vinod; Engelborghs, Yves; Visser, Anthonie J.W.G.

    2014-01-01

    Multimodal fluorescence imaging is a versatile method that has a wide application range from biological studies to materials science. Typical observables in multimodal fluorescence imaging are intensity, lifetime, excitation, and emission spectra which are recorded at chosen locations at the sample.

  15. Statistical filtering in fluorescence microscopy and fluorescence correlation spectroscopy

    Czech Academy of Sciences Publication Activity Database

    Macháň, Radek; Kapusta, Peter; Hof, Martin

    Roč. 406 , č. 20 (2014), s. 4797-4813 ISSN 1618-2642 R&D Projects: GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : Filtered fluorescence correlation spectroscopy * Fluorescence lifetime correlation spectroscopy * Fluorescence spectral correlation spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.436, year: 2014

  16. Tower counts

    Science.gov (United States)

    Woody, Carol Ann; Johnson, D.H.; Shrier, Brianna M.; O'Neal, Jennifer S.; Knutzen, John A.; Augerot, Xanthippe; O'Neal, Thomas A.; Pearsons, Todd N.

    2007-01-01

    Counting towers provide an accurate, low-cost, low-maintenance, low-technology, and easily mobilized escapement estimation program compared to other methods (e.g., weirs, hydroacoustics, mark-recapture, and aerial surveys) (Thompson 1962; Siebel 1967; Cousens et al. 1982; Symons and Waldichuk 1984; Anderson 2000; Alaska Department of Fish and Game 2003). Counting tower data has been found to be consistent with that of digital video counts (Edwards 2005). Counting towers do not interfere with natural fish migration patterns, nor are fish handled or stressed; however, their use is generally limited to clear rivers that meet specific site selection criteria. The data provided by counting tower sampling allow fishery managers to determine reproductive population size, estimate total return (escapement + catch) and its uncertainty, evaluate population productivity and trends, set harvest rates, determine spawning escapement goals, and forecast future returns (Alaska Department of Fish and Game 1974-2000 and 1975-2004). The number of spawning fish is determined by subtracting subsistence, sport-caught fish, and prespawn mortality from the total estimated escapement. The methods outlined in this protocol for tower counts can be used to provide reasonable estimates ( plus or minus 6%-10%) of reproductive salmon population size and run timing in clear rivers. 

  17. Lifetime measurement of excited atomic and ionic states of some

    Indian Academy of Sciences (India)

    High-frequency deflection (HFD) technique with a delayed coincidence single photon counting arrangement is an efficient technique for radiative lifetime measurement. An apparatus for measurement of the radiative lifetime of atoms and molecules has been developed in our laboratory and measurements have been ...

  18. Counting probe

    International Nuclear Information System (INIS)

    Matsumoto, Haruya; Kaya, Nobuyuki; Yuasa, Kazuhiro; Hayashi, Tomoaki

    1976-01-01

    Electron counting method has been devised and experimented for the purpose of measuring electron temperature and density, the most fundamental quantities to represent plasma conditions. Electron counting is a method to count the electrons in plasma directly by equipping a probe with the secondary electron multiplier. It has three advantages of adjustable sensitivity, high sensitivity of the secondary electron multiplier, and directional property. Sensitivity adjustment is performed by changing the size of collecting hole (pin hole) on the incident front of the multiplier. The probe is usable as a direct reading thermometer of electron temperature because it requires to collect very small amount of electrons, thus it doesn't disturb the surrounding plasma, and the narrow sweep width of the probe voltage is enough. Therefore it can measure anisotropy more sensitively than a Langmuir probe, and it can be used for very low density plasma. Though many problems remain on anisotropy, computer simulation has been carried out. Also it is planned to provide a Helmholtz coil in the vacuum chamber to eliminate the effect of earth magnetic field. In practical experiments, the measurement with a Langmuir probe and an emission probe mounted to the movable structure, the comparison with the results obtained in reverse magnetic field by using a Helmholtz coil, and the measurement of ionic sound wave are scheduled. (Wakatsuki, Y.)

  19. Photon-counting 1.0 GHz-phase-modulation fluorometer

    Energy Technology Data Exchange (ETDEWEB)

    Mizuno, T.; Nakao, S.; Mizutani, Y.; Iwata, T., E-mail: iwata@tokushima-u.ac.jp [Division of Energy System, Institute of Technology and Science, Tokushima University, 2-1 Minami-Jyosanjima, Tokushima 770-8506 (Japan)

    2015-04-15

    We have constructed an improved version of a photon-counting phase-modulation fluorometer (PC-PMF) with a maximum modulation frequency of 1.0 GHz, where a phase domain measurement is conducted with a time-correlated single-photon-counting electronics. While the basic concept of the PC-PMF has been reported previously by one of the authors, little attention has been paid to its significance, other than its weak fluorescence measurement capability. Recently, we have recognized the importance of the PC-PMF and its potential for fluorescence lifetime measurements. One important aspect of the PC-PMF is that it enables us to perform high-speed measurements that exceed the frequency bandwidths of the photomultiplier tubes that are commonly used as fluorescence detectors. We describe the advantages of the PC-PMF and demonstrate its usefulness based on fundamental performance tests. In our new version of the PC-PMF, we have used a laser diode (LD) as an excitation light source rather than the light-emitting diode that was used in the primary version. We have also designed a simple and stable LD driver to modulate the device. Additionally, we have obtained a sinusoidal histogram waveform that has multiple cycles within a time span to be measured, which is indispensable for precise phase measurements. With focus on the fluorescence intensity and the resolution time, we have compared the performance of the PC-PMF with that of a conventional PMF using the analogue light detection method.

  20. Photon-counting 1.0 GHz-phase-modulation fluorometer

    International Nuclear Information System (INIS)

    Mizuno, T.; Nakao, S.; Mizutani, Y.; Iwata, T.

    2015-01-01

    We have constructed an improved version of a photon-counting phase-modulation fluorometer (PC-PMF) with a maximum modulation frequency of 1.0 GHz, where a phase domain measurement is conducted with a time-correlated single-photon-counting electronics. While the basic concept of the PC-PMF has been reported previously by one of the authors, little attention has been paid to its significance, other than its weak fluorescence measurement capability. Recently, we have recognized the importance of the PC-PMF and its potential for fluorescence lifetime measurements. One important aspect of the PC-PMF is that it enables us to perform high-speed measurements that exceed the frequency bandwidths of the photomultiplier tubes that are commonly used as fluorescence detectors. We describe the advantages of the PC-PMF and demonstrate its usefulness based on fundamental performance tests. In our new version of the PC-PMF, we have used a laser diode (LD) as an excitation light source rather than the light-emitting diode that was used in the primary version. We have also designed a simple and stable LD driver to modulate the device. Additionally, we have obtained a sinusoidal histogram waveform that has multiple cycles within a time span to be measured, which is indispensable for precise phase measurements. With focus on the fluorescence intensity and the resolution time, we have compared the performance of the PC-PMF with that of a conventional PMF using the analogue light detection method

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

    International Nuclear Information System (INIS)

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

    2011-01-01

    Highlights: → Novel metal nanoshell as fluorescence imaging agent. → Fluorescent mAb-metal complex with enhanced intensity and shortened lifetime. → Immuno-interactions of mAb-metal complexes with CK19 molecules on CNCAP and HeLa cell surfaces. → 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.

  2. Nuclear lifetime measurement

    International Nuclear Information System (INIS)

    Guillaume, Georges

    Three direct techniques of lifetime measurement are emphasized: electronic methods and two methods based on the Doppler effect (the recoil distance methods or RDM, the Doppler shift attenuation methods or DSAM). Said direct methods are concerned with the direct measurement of the radioactive decay constants of nuclear excited states. They allow lifetimes of nucleus bound states whose deexcitations occur by electromagnetic transitions, to be determined. Other methods for measuring lifetimes are also examined: microwave techniques and those involving the blocking effect in crystals (direct methods) and also various indirect methods of obtaining lifetimes (γ resonance scattering, capture reactions, inelastic electron and nucleus scattering, and Coulomb deexcitation) [fr

  3. Counting Possibilia

    Directory of Open Access Journals (Sweden)

    Alfredo Tomasetta

    2010-06-01

    Full Text Available Timothy Williamson supports the thesis that every possible entity necessarily exists and so he needs to explain how a possible son of Wittgenstein’s, for example, exists in our world:he exists as a merely possible object (MPO, a pure locus of potential. Williamson presents a short argument for the existence of MPOs: how many knives can be made by fitting together two blades and two handles? Four: at the most two are concrete objects, the others being merely possible knives and merely possible objects. This paper defends the idea that one can avoid reference and ontological commitment to MPOs. My proposal is that MPOs can be dispensed with by using the notion of rules of knife-making. I first present a solution according to which we count lists of instructions - selected by the rules - describing physical combinations between components. This account, however, has its own difficulties and I eventually suggest that one can find a way out by admitting possible worlds, entities which are more commonly accepted - at least by philosophers - than MPOs. I maintain that, in answering Williamson’s questions, we count classes of physically possible worlds in which the same instance of a general rule is applied.

  4. Lifetimes for some excited states of sodium

    International Nuclear Information System (INIS)

    Thomas, P.; Campos, J.

    1979-01-01

    The lifetimes of some s,p and d levels of sodium have been measured by the delayed coincidence method, using a single-photon counting technique. The results are compared with the calculated values of the present work, and with other results. The lifetimes of the ns, np, and levels up to n10; of the nf levels up to n-9;and of the ng, nh,n1 and nk levels up to n-8, have been calculated and the transition probabilities of lines with origin in these levels are given. (Author) 38 refs

  5. Lifetime of organic photovoltaics

    DEFF Research Database (Denmark)

    Corazza, Michael; Krebs, Frederik C; Gevorgyan, Suren A.

    2015-01-01

    tests. Comparison of the indoor and outdoor lifetimes was performed by means of the o-diagram, which constitutes the initial steps towards establishing a method for predicting the lifetime of an organic photovoltaic device under real operational conditions based on a selection of accelerated indoor...

  6. Hadronization, spin and lifetimes

    International Nuclear Information System (INIS)

    Grossman, Yuval; Nachshon, Itay

    2008-01-01

    Measurements of lifetimes can be done in two ways. For very short lived particles, the width can be measured. For long lived ones, the lifetime can be directly measured, for example, using a displaced vertex. Practically, the lifetime cannot be extracted for particles with intermediate lifetimes. We show that for such cases information about the lifetime can be extracted for heavy colored particles that can be produced with known polarization. For example, a t-like particle with intermediate lifetime hadronizes into a superposition of the lowest two hadronic states, T* and T (the equivalent of B* and B). Depolarization effects are governed by time scales that are much longer than the hadronization time scale, Λ QCD -1 . After a time of order 1/Δm, with Δm≡m(T*)-m(T), half of the initial polarization is lost. The polarization is totally lost after a time of order 1/Γ γ , with Γ γ = Γ(T* → Tγ). Thus, by comparing the initial and final polarization, we get information on the particle's lifetime.

  7. Temperature and time dependence of free volume in bisphenol A polycarbonate studied by positron lifetime spectroscopy

    NARCIS (Netherlands)

    Kluin, J.E.; Yu, Z.; Vleeshouwers, S.M.; McGervey, J.D.; Jamieson, A.M.; Simha, R.

    1992-01-01

    New positron lifetime expts. were carried out for Bisphenol-A polycarbonate. The influence of unavoidable pos. charged positron irradn. on the lifetime and intensity of o-positronium (o-Ps) annihilation was studied. Results obtained using a state-of-the-art lifetime spectrometer (count rate 670 cps

  8. Categorical counting.

    Science.gov (United States)

    Fetterman, J Gregor; Killeen, P Richard

    2010-09-01

    Pigeons pecked on three keys, responses to one of which could be reinforced after a few pecks, to a second key after a somewhat larger number of pecks, and to a third key after the maximum pecking requirement. The values of the pecking requirements and the proportion of trials ending with reinforcement were varied. Transits among the keys were an orderly function of peck number, and showed approximately proportional changes with changes in the pecking requirements, consistent with Weber's law. Standard deviations of the switch points between successive keys increased more slowly within a condition than across conditions. Changes in reinforcement probability produced changes in the location of the psychometric functions that were consistent with models of timing. Analyses of the number of pecks emitted and the duration of the pecking sequences demonstrated that peck number was the primary determinant of choice, but that passage of time also played some role. We capture the basic results with a standard model of counting, which we qualify to account for the secondary experiments. Copyright 2010 Elsevier B.V. All rights reserved.

  9. The neutron lifetime experiment PENeLOPE

    Energy Technology Data Exchange (ETDEWEB)

    Schreyer, Wolfgang [Technische Universitaet Muenchen (Germany); Collaboration: PENeLOPE-Collaboration

    2015-07-01

    The neutron lifetime τ{sub n}=880.3±1.1 s is an important parameter in the Standard Model of particle physics and in Big Bang cosmology. Several systematic corrections of previously published results reduced the PDG world average by several σ in the last years and call for a new experiment with complementary systematics. The experiment PENeLOPE, currently under construction at the Physik-Department of Technische Universitaet Muenchen, aims to determine the neutron lifetime with a precision of 0.1 s. It will trap ultra-cold neutrons in a magneto-gravitational trap using a large superconducting magnet and will measure their lifetime by both neutron counting and online proton detection. This presentation gives an overview over the latest developments of the experiment.

  10. Gated Detection Measurements of Phosphorescence Lifetimes

    Directory of Open Access Journals (Sweden)

    Yordan Kostov

    2004-10-01

    Full Text Available A low-cost, gated system for measurements of phosphorescence lifetimes is presented. An extensive description of the system operating principles and metrological characteristics is given. Remarkably, the system operates without optical filtering of the LED excitation source. A description of a practical system is also given and its performance is discussed. Because the device effectively suppresses high-level background fluorescence and scattered light, it is expected to find wide-spread application in bioprocess, environmental and biomedical fields.

  11. [Corrected count].

    Science.gov (United States)

    1991-11-27

    The data of the 1991 census indicated that the population count of Brazil fell short of a former estimate by 3 million people. The population reached 150 million people with an annual increase of 2%, while projections in the previous decade expected an increase of 2.48% to 153 million people. This reduction indicates more widespread use of family planning (FP) and control of fertility among families of lower social status as more information is being provided to them. However, the Ministry of Health ordered an investigation of foreign family planning organizations because it was suspected that women were forced to undergo tubal ligation during vaccination campaigns. A strange alliance of left wing politicians and the Roman Catholic Church alleges a conspiracy of international FP organizations receiving foreign funds. The FP strategies of Bemfam and Pro-Pater offer women who have little alternative the opportunity to undergo tubal ligation or to receive oral contraceptives to control fertility. The ongoing government program of distributing booklets on FP is feeble and is not backed up by an education campaign. Charges of foreign interference are leveled while the government hypocritically ignores the grave problem of 4 million abortions a year. The population is expected to continue to grow until the year 2040 and then to stabilize at a low growth rate of .4%. In 1980, the number of children per woman was 4.4 whereas the 1991 census figures indicate this has dropped to 3.5. The excess population is associated with poverty and a forsaken caste in the interior. The population actually has decreased in the interior and in cities with 15,000 people. The phenomenon of the drop of fertility associated with rural exodus is contrasted with cities and villages where the population is 20% less than expected.

  12. Charmed particle lifetimes

    International Nuclear Information System (INIS)

    Rosner, J.L.

    1979-01-01

    Conventional estimates are reviewed for charmed particle lifetimes. Free-quark models give values of (a few) x 10 -13 sec to (a few) x 10 -12 sec. The shorter of these values also follows from an extrapolation based on D → Ke/sup nu/. Possible differences among the lifetimes and production rates of D 0 , D + , F + , C 0 + , the heavy lepton tau, and the fifth quark b are discussed. Extreme values of mixing angles in a six-quark model could extend charmed particle lifetimes by a factor of at most three from the above estimates, while shorter lifetimes than those predicted could occur for some species like D 0 or F + if their nonleptonic decays were enhanced. The predictions are discussed in the light of some current experimental results, and it is estimated that sigma(pp → charm) approx. = 10 μb at 400 GeV/c. 95 references

  13. Lifetime of the metastable 23S1 state in stored Li+ ions

    International Nuclear Information System (INIS)

    Knight, R.D.

    1979-04-01

    A laser-induced fluorescence technique combined with the observation of spontaneous magnetic dipole photons from the highly metastable 2 3 S 1 state of Li + was used to measure the radiative lifetime of this state. The ions are created by electron impact on a lithium atomic beam and are subsequently stored for periods of many seconds in an RF-quadrupole ion trap. A tunable dye laser excites the 2 3 S--2 3 P, transition at 5485A, and the intercombination electric dipole transition 2 3 P 1 --1 1 S 0 at 202A is observed. This process depletes the metastable population in a time tau/sub d/ 3 S 1 / and provides a measure of the total number of metastables. Comparison with the rate of 210A spontaneously emitted photons yields a measured value for the 2 3 S 1 radiative lifetime of tau/sub rad/ = 58.6 +- 12.9 sec, where the quoted error represents 95% confidence levels. The theoretical lifetime is tau/sub theory/ = 49.0 sec. The measured value includes data taken with both 6 Li + and 7 Li + isotopes and was corrected for the slightly different detector efficiencies at 202A and 210A. A careful study of nonradiative quenching of the metastable state was necessary to understand observed differences between tau/sub rad/ and tau/sub 3 S 1 /, the total metastable lifetime. Spatial density profiles of the ions within the trap, useful for determining the ion temperature, were obtained by scanning the laser beam horizontally across the ion trap while storing 2 3 P 1 -- 1 S 0 photon counts as a function of the laser beam's position. Agreement with a simple equilibrium model, including space charge effects, is satisfactory. A study of the optical pumping process is necessary to understand the laser-ion interaction, and observational and theoretical data are presented. 47 references

  14. A study of pile-up in integrated time-correlated single photon counting systems.

    Science.gov (United States)

    Arlt, Jochen; Tyndall, David; Rae, Bruce R; Li, David D-U; Richardson, Justin A; Henderson, Robert K

    2013-10-01

    Recent demonstration of highly integrated, solid-state, time-correlated single photon counting (TCSPC) systems in CMOS technology is set to provide significant increases in performance over existing bulky, expensive hardware. Arrays of single photon single photon avalanche diode (SPAD) detectors, timing channels, and signal processing can be integrated on a single silicon chip with a degree of parallelism and computational speed that is unattainable by discrete photomultiplier tube and photon counting card solutions. New multi-channel, multi-detector TCSPC sensor architectures with greatly enhanced throughput due to minimal detector transit (dead) time or timing channel dead time are now feasible. In this paper, we study the potential for future integrated, solid-state TCSPC sensors to exceed the photon pile-up limit through analytic formula and simulation. The results are validated using a 10% fill factor SPAD array and an 8-channel, 52 ps resolution time-to-digital conversion architecture with embedded lifetime estimation. It is demonstrated that pile-up insensitive acquisition is attainable at greater than 10 times the pulse repetition rate providing over 60 dB of extended dynamic range to the TCSPC technique. Our results predict future CMOS TCSPC sensors capable of live-cell transient observations in confocal scanning microscopy, improved resolution of near-infrared optical tomography systems, and fluorescence lifetime activated cell sorting.

  15. Lifetime measurement of trapped staus using ATLAS

    CERN Document Server

    Sibley, Logan

    I study the creation of long-lived staus at a 14 TeV centre of mass energy in proton-proton collisions at the LHC using both the ATLAS and ACME detectors. The ATLAS overburden or underburden, or even ATLAS itself, may trap the semi-stable staus at that place where they will remain until the time at which they decay, where the stau lifetime ranges between seven days and one year. Using a novel method, one may count the number of muons and pions originating from the stau decay using the standard ATLAS cosmic ray trigger. Using an idealized detector model, I find that this method can lead to measurements of the stau lifetime and SUSY cross-section to within statistical uncertainties of 6% and 1% of their actual values, respectively.

  16. Filter replacement lifetime prediction

    Science.gov (United States)

    Hamann, Hendrik F.; Klein, Levente I.; Manzer, Dennis G.; Marianno, Fernando J.

    2017-10-25

    Methods and systems for predicting a filter lifetime include building a filter effectiveness history based on contaminant sensor information associated with a filter; determining a rate of filter consumption with a processor based on the filter effectiveness history; and determining a remaining filter lifetime based on the determined rate of filter consumption. Methods and systems for increasing filter economy include measuring contaminants in an internal and an external environment; determining a cost of a corrosion rate increase if unfiltered external air intake is increased for cooling; determining a cost of increased air pressure to filter external air; and if the cost of filtering external air exceeds the cost of the corrosion rate increase, increasing an intake of unfiltered external air.

  17. Effects of short term changes in the blood glucose level on the autofluorescence lifetime of the human retina in healthy volunteers

    Science.gov (United States)

    Klemm, Matthias; Nagel, Edgar; Schweitzer, Dietrich; Schramm, Stefan; Haueisen, Jens

    2016-03-01

    Purpose: Fluorescence lifetime imaging ophthalmoscopy (FLIO) provides in vivo metabolic mapping of the ocular fundus. Changes in FLIO have been found in e.g. diabetes patients. The influence of short term metabolic changes caused by blood glucose level changes on is unknown. Aim of this work is the detection of short-term changes in fundus autofluorescence lifetime during an oral glucose tolerance test. Methods: FLIO was performed in 10 healthy volunteers (29+/-4 years, fasting for 12h) using a scanning laser ophthalmoscope (30° fundus, 34μm resolution, excitation with 473nm diode laser with 70 ps pulses at 80 MHz repetition rate, detection in two spectral channels 500-560nm (ch1) and 560-720nm (ch2) using the timecorrelated single photon counting method). The blood glucose level (BGL) was measured by an Accu-Chek® Aviva self-monitoring device. Before and after a glucose drink (300ml solution, containing 75g of glucose (Accu-Chek® Dextrose O.G.T.), BGL and FLIO were measured every 15min. The FLIMX software package was applied to compute the average fluorescence lifetime τ on the inner ring of the ETDRS grid using a modified 3-exponential approach. Results: The results are given as mean +/- standard deviation over all volunteers in ch1. Baseline measurement: BGL: 5.3+/-0.4 mmol/l, τ1: 49+/-6ps. A significant reduction (α=5% Wilcoxon rank-sum test) in τ1 is detected after 15min (BGL: 8.4+/-1.1 mmol/l, τ1: 44+/-5ps) and after 90min (BGL: 6.3+/-1.4 mmol/l, τ1: 41+/-5ps). Results of ch2 show smaller reductions in the fluorescence lifetimes over time.

  18. Determining random counts in liquid scintillation counting

    International Nuclear Information System (INIS)

    Horrocks, D.L.

    1979-01-01

    During measurements involving coincidence counting techniques, errors can arise due to the detection of chance or random coincidences in the multiple detectors used. A method and the electronic circuits necessary are here described for eliminating this source of error in liquid scintillation detectors used in coincidence counting. (UK)

  19. B Lifetimes and Mixing

    International Nuclear Information System (INIS)

    Evans, Harold G.

    2009-01-01

    The Tevatron experiments, CDF and D0, have produced a wealth of new B-physics results since the start of Run II in 2001. We've observed new B-hadrons, seen new effects, and increased many-fold the precision with which we know the properties of b-quark systems. In these proceedings, we will discuss two of the most fruitful areas in the Tevatron B-physics program: lifetimes and mixing. We'll examine the experimental issues driving these analyses, present a summary of the latest results, and discuss prospects for the future.

  20. Lifetimes of heavy flavour particles

    International Nuclear Information System (INIS)

    Forty, R.

    1994-01-01

    The lifetimes of heavy-flavour hadrons are reviewed. After a brief discussion of the theoretical predictions, the problem of averaging lifetime measurements is discussed. The various experimental measurements are then presented and suitable averages performed. Charmed meson lifetimes are now measured to the few percent level, better that theory can predict, whilst for charmed baryons the lifetime hierarchy has been established for the first time. For beauty hadrons the lifetimes are measured at the 6-10 % level, and are in reasonable agreement with theoretical expectations. Beauty baryon studies ar just beginning. (author)

  1. Absorbance and fluorescence studies on porphyrin Nanostructures ...

    African Journals Online (AJOL)

    The aim of this work was to study some photophysical properties of PNR for application as light harvester in dye sensitized solar cells. These properties included absorbance, fluorescence, and fluorescence quantum yield and lifetime. The results of Transmission Electron Microscope (TEM) images showed the formation of ...

  2. Nanosecond fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Leskovar, B.

    1985-03-01

    This article is a summary of a short course lecture given in conjunction with the 1984 Nuclear Science Symposium. Measuring systems for nanosecond fluorescence spectroscopy using single-photon counting techniques are presented. These involve systems based on relaxation-type spark gap light pulser and synchronously pumped mode-locked dye lasers. Furthermore, typical characteristics and optimization of operating conditions of the critical components responsible for the system time resolution are discussed. A short comparison of the most important deconvolution methods for numerical analysis of experimental data is given particularly with respect to the signal-to-noise ratio of the fluorescence signal. 22 refs., 8 figs

  3. Using Fluorescence Intensity of Enhanced Green Fluorescent Protein to Quantify Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Erin Wilson

    2018-05-01

    Full Text Available A variety of direct and indirect methods have been used to quantify planktonic and biofilm bacterial cells. Direct counting methods to determine the total number of cells include plate counts, microscopic cell counts, Coulter cell counting, flow cytometry, and fluorescence microscopy. However, indirect methods are often used to supplement direct cell counting, as they are often more convenient, less time-consuming, and require less material, while providing a number that can be related to the direct cell count. Herein, an indirect method is presented that uses fluorescence emission intensity as a proxy marker for studying bacterial accumulation. A clinical strain of Pseudomonas aeruginosa was genetically modified to express a green fluorescent protein (PA14/EGFP. The fluorescence intensity of EGFP in live cells was used as an indirect measure of live cell density, and was compared with the traditional cell counting methods of optical density (OD600 and plate counting (colony-forming units (CFUs. While both OD600 and CFUs are well-established methods, the use of fluorescence spectroscopy to quantify bacteria is less common. This study demonstrates that EGFP intensity is a convenient reporter for bacterial quantification. In addition, we demonstrate the potential for fluorescence spectroscopy to be used to measure the quantity of PA14/EGFP biofilms, which have important human health implications due to their antimicrobial resistance. Therefore, fluorescence spectroscopy could serve as an alternative or complementary quick assay to quantify bacteria in planktonic cultures and biofilms.

  4. Metabolic Mapping of Breast Cancer with Multiphoton Spectral and Lifetime Imaging

    Science.gov (United States)

    2007-03-01

    2002. Spectrally resolved fluorescence lifetime imaging microscopy. Appl. Spec- trosc. 56 :155-166. 38. Becker, W., A. Bergmann, E. Haustein , Z...photon fluores- cence lifetime imaging microscopy of macrophage-mediated antigen processing. J. Microsc. 185 :339-353. 45. Lin, H.J., P. Herman , and

  5. Autofluorescence Lifetimes in Geographic Atrophy in Patients With Age-Related Macular Degeneration.

    Science.gov (United States)

    Dysli, Chantal; Wolf, Sebastian; Zinkernagel, Martin S

    2016-05-01

    To investigate fluorescence lifetime characteristics in patients with geographic atrophy (GA) in eyes with age-related macular degeneration and to correlate the measurements with clinical data and optical coherence tomography (OCT) findings. Patients with GA were imaged with a fluorescence lifetime imaging ophthalmoscope. Retinal autofluorescence lifetimes were measured in a short and a long spectral channel (498-560 nm and 560-720 nm). Mean retinal fluorescence lifetimes were analyzed within GA and the surrounding retina, and data were correlated with best corrected visual acuity and OCT measurements. Fluorescence lifetime maps of 41 eyes of 41 patients (80 ± 7 years) with GA were analyzed. Mean lifetimes within areas of atrophy were prolonged by 624 ± 276 ps (+152%) in the short spectral channel and 418 ± 186 ps (+83%) in the long spectral channel compared to the surrounding tissue. Autofluorescence lifetime abnormalities in GA occurred with particular patterns, similar to those seen in fundus autofluorescence intensity images. Within the fovea short mean autofluorescence lifetimes were observed, presumably representing macular pigment. Short lifetimes were preserved even in the absence of foveal sparing but were decreased in patients with advanced retinal atrophy in OCT. Short lifetimes in the fovea correlated with better best corrected visual acuity in both spectral channels. This study established that autofluorescence lifetime changes in GA present with explicit patterns. We hypothesize that the short lifetimes seen within the atrophy may be used to estimate damage induced by atrophy and to monitor disease progression in the context of natural history or interventional therapeutic studies.

  6. Clean Hands Count

    Medline Plus

    Full Text Available ... Like this video? Sign in to make your opinion count. Sign in 131 2 Don't like this video? Sign in to make your opinion count. Sign in 3 Loading... Loading... Transcript The ...

  7. The Big Pumpkin Count.

    Science.gov (United States)

    Coplestone-Loomis, Lenny

    1981-01-01

    Pumpkin seeds are counted after students convert pumpkins to jack-o-lanterns. Among the activities involved, pupils learn to count by 10s, make estimates, and to construct a visual representation of 1,000. (MP)

  8. Luminescence and transient lifetime studies for energy transfer of PbS QD films

    Science.gov (United States)

    Wang, Joanna S.; Ullrich, Bruno; Dass, Chandriker K.; Das, Anirban; Wai, Chien M.; Brown, Gail J.; Hendrickson, Joshua R.

    2017-08-01

    Quantum confined semiconductor materials in colloidal form have drawn great attention in scientific communities due to the size-tunability, which controls their optical properties. PbS quantum dots (QDs) are exciting candidates for quantum optics, particularly due to the control of the QD sizes during the synthetic process enabling the realization of precisely tunable emission properties in the near-infrared region. Differently sized pairs of PbS QDs were deposited onto glass substrates to form thin films using supercritical CO2 (sc-CO2) deposition and solvent deposition methods (SDM). The fluorescence and photoluminescence (PL) spectra obtained from these closely packed films prepared by the sc-CO2 method reveal effective Förster resonance energy transfer (FRET) between two different sized dots, while the films composed of three different QD sizes show an even more effective FRET from the smallest to the largest ones. Energy transfer can be observed more directly by temporally resolved PL decay of mixed dots. By means of transient lifetime measurements, a mixed PbS film with 3.1 and 4.7 nm QDs was studied for FRET by time correlated single photon counting. The PL peak of the 3.1 nm QDs is quenched with respect to the emission of the 4.7 nm QDs and decays faster, and the best fit for the lifetime (decay constant)-1 is a biexponential decay mode. The long wavelength decay (4.7 nm QDs) is best fit by a mono-exponential equation. More theoretical and experimental work is required for a thorough understanding of the radiative lifetimes of PbS QDs in mixed QD systems.

  9. B meson lifetime measurement

    International Nuclear Information System (INIS)

    Piccolo, M.

    1989-01-01

    The lifetime of hadrons containing b-quark has been the subject of extensive experimental work and theoretical speculation; its importance is due to implications on some of the fundamental parameters of the Standard Model, such as the top quark mass and the mixing angles. Since the pioneer measurements of the MAC and MARK II collaborations at PEP in 1983 the progress has been impressive; but many issues still remain open and await further study. In this paper the field's present status is discussed. An overview of the theoretical motivations for this measurements in the Standard Model framework is done. Then the experimental techniques used are reviewed, emphasizing the most recent measurements. A comparison of the results obtained is done and systematic errors are discussed. In conclusion there are some remarks on the further developments foreseen in the near future

  10. Radiation counting statistics

    Energy Technology Data Exchange (ETDEWEB)

    Suh, M. Y.; Jee, K. Y.; Park, K. K.; Park, Y. J.; Kim, W. H

    1999-08-01

    This report is intended to describe the statistical methods necessary to design and conduct radiation counting experiments and evaluate the data from the experiment. The methods are described for the evaluation of the stability of a counting system and the estimation of the precision of counting data by application of probability distribution models. The methods for the determination of the uncertainty of the results calculated from the number of counts, as well as various statistical methods for the reduction of counting error are also described. (Author). 11 refs., 8 tabs., 8 figs.

  11. Radiation counting statistics

    Energy Technology Data Exchange (ETDEWEB)

    Suh, M. Y.; Jee, K. Y.; Park, K. K. [Korea Atomic Energy Research Institute, Taejon (Korea)

    1999-08-01

    This report is intended to describe the statistical methods necessary to design and conduct radiation counting experiments and evaluate the data from the experiments. The methods are described for the evaluation of the stability of a counting system and the estimation of the precision of counting data by application of probability distribution models. The methods for the determination of the uncertainty of the results calculated from the number of counts, as well as various statistical methods for the reduction of counting error are also described. 11 refs., 6 figs., 8 tabs. (Author)

  12. Radiation counting statistics

    International Nuclear Information System (INIS)

    Suh, M. Y.; Jee, K. Y.; Park, K. K.; Park, Y. J.; Kim, W. H.

    1999-08-01

    This report is intended to describe the statistical methods necessary to design and conduct radiation counting experiments and evaluate the data from the experiment. The methods are described for the evaluation of the stability of a counting system and the estimation of the precision of counting data by application of probability distribution models. The methods for the determination of the uncertainty of the results calculated from the number of counts, as well as various statistical methods for the reduction of counting error are also described. (Author). 11 refs., 8 tabs., 8 figs

  13. Solvent isotope effect on the fluorescence of azoalkanes

    International Nuclear Information System (INIS)

    Mirbach, M.J.; Mirbach, M.F.; Cherry, W.R.; Turro, N.J.; Engel, P.

    1977-01-01

    A study of fluorescence quantum yields and fluorescence lifetimes of two cyclic azoalkanes reveal a striking dependence of phisub(F) and tausub(F) on solvent and on isotopic substitution (OH → OD). A mechanism involving specific deactivation of the fluorescent state from a hydrogen bonded complex is proposed to rationalize the data. (orig./HK) [de

  14. Quantitative frequency-domain fluorescence spectroscopy in tissues and tissue-like media

    Science.gov (United States)

    Cerussi, Albert Edward

    1999-09-01

    In the never-ending quest for improved medical technology at lower cost, modern near-infrared optical spectroscopy offers the possibility of inexpensive technology for quantitative and non-invasive diagnoses. Hemoglobin is the dominant chromophore in the 700-900 nm spectral region and as such it allows for the optical assessment of hemoglobin concentration and tissue oxygenation by absorption spectroscopy. However, there are many other important physiologically relevant compounds or physiological states that cannot be effectively sensed via optical methods because of poor optical contrast. In such cases, contrast enhancements are required. Fluorescence spectroscopy is an attractive component of optical tissue spectroscopy. Exogenous fluorophores, as well as some endogenous ones, may furnish the desperately needed sensitivity and specificity that is lacking in near-infrared optical tissue spectroscopy. The main focus of this thesis was to investigate the generation and propagation of fluorescence photons inside tissues and tissue-like media (i.e., scattering dominated media). The standard concepts of fluorescence spectroscopy have been incorporated into a diffusion-based picture that is sometimes referred to as photon migration. The novelty of this work lies in the successful quantitative recovery of fluorescence lifetimes, absolute fluorescence quantum yields, fluorophore concentrations, emission spectra, and both scattering and absorption coefficients at the emission wavelength from a tissue-like medium. All of these parameters are sensitive to the fluorophore local environment and hence are indicators of the tissue's physiological state. One application demonstrating the capabilities of frequency-domain lifetime spectroscopy in tissue-like media is a study of the binding of ethidium bromide to bovine leukocytes in fresh milk. Ethidium bromide is a fluorescent dye that is commonly used to label DNA, and hence visualize chromosomes in cells. The lifetime of

  15. Effect of pharmacologically induced retinal degeneration on retinal autofluorescence lifetimes in mice.

    Science.gov (United States)

    Dysli, Chantal; Dysli, Muriel; Zinkernagel, Martin S; Enzmann, Volker

    2016-12-01

    Fluorescence lifetime imaging ophthalmoscopy (FLIO) was used to investigate retinal autofluorescence lifetimes in mouse models of pharmacologically induced retinal degeneration over time. Sodium iodate (NaIO 3 , 35 mg/kg intravenously) was used to induce retinal pigment epithelium (RPE) degeneration with subsequent loss of photoreceptors (PR) whereas N-methyl-N-nitrosourea (MNU, 45 mg/kg intraperitoneally) was employed for degeneration of the photoreceptor cell layer alone. All mice were measured at day 3, 7, 14, and 28 after the respective injection of NaIO 3 , MNU or NaCl (control). Fluorescence lifetime imaging was performed using a fluorescence lifetime imaging ophthalmoscope (Heidelberg Engineering, Heidelberg, Germany). Fluorescence was excited at 473 nm and fluorescence lifetimes were measured in a short and a long spectral channel (498-560 nm and 560-720 nm). Corresponding optical coherence tomography (OCT) images were consecutively acquired and histology was performed at the end of the experiments. Segmentation of OCT images and histology verified the cell type-specific degeneration process over time. Retinal autofluorescence lifetimes increased from day 3 to day 28 in mice after NaIO 3 treatment. Finally, at day 28, fluorescence lifetimes were prolonged by 8% in the short and 61% in the long spectral channel compared to control animals (p = 0.21 and p = 0.004, respectively). In mice after MNU treatment, the mean retinal autofluorescence lifetimes were already decreased at day 3 and retinal lifetimes were finally shortened by 27% in the short and 51% in the long spectral channel at day 28 (p = 0.0028). In conclusion, degeneration of the RPE with subsequent photoreceptor degeneration by NaIO 3 lead to longer mean fluorescence lifetimes of the retina compared to control mice, whereas during specific degeneration of the photoreceptor layer induced by MNU shorter lifetimes were measured. Therefore, short retinal fluorescence lifetimes may originate

  16. Mining the bulk positron lifetime

    International Nuclear Information System (INIS)

    Aourag, H.; Guittom, A.

    2009-01-01

    We introduce a new approach to investigate the bulk positron lifetimes of new systems based on data-mining techniques. Through data mining of bulk positron lifetimes, we demonstrate the ability to predict the positron lifetimes of new semiconductors on the basis of available semiconductor data already studied. Informatics techniques have been applied to bulk positron lifetimes for different tetrahedrally bounded semiconductors in order to discover computational design rules. (copyright 2009 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  17. Energy Savings Lifetimes and Persistence

    Energy Technology Data Exchange (ETDEWEB)

    Hoffman, Ian M. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Schiller, Steven R. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Todd, Annika [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Billingsley, Megan A. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Goldman, Charles A. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Schwartz, Lisa C. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

    2016-02-01

    This technical brief explains the concepts of energy savings lifetimes and savings persistence and discusses how program administrators use these factors to calculate savings for efficiency measures, programs and portfolios. Savings lifetime is the length of time that one or more energy efficiency measures or activities save energy, and savings persistence is the change in savings throughout the functional life of a given efficiency measure or activity. Savings lifetimes are essential for assessing the lifecycle benefits and cost effectiveness of efficiency activities and for forecasting loads in resource planning. The brief also provides estimates of savings lifetimes derived from a national collection of costs and savings for electric efficiency programs and portfolios.

  18. Positron lifetimes in deformed copper

    International Nuclear Information System (INIS)

    Hinode, Kenji; Tanigawa, Shoichiro; Doyama, Masao

    1976-01-01

    Positron lifetime measurements were performed for Cu samples with different densities of lattice defects. The lifetime spectra were successfully resolved into two components with the help of the well established analysis program. Obtained results were quite consistent with those expected from the trapping model. The positron trapping mechanism from free to trapped states and the initial condition of the model were especially checked. Deduced values obtained for tau sub(c) (lifetime of free positrons) and tau sub(t) (lifetime of trapped positrons) were 122+-5 psec and 176+-5 psec, respectively. (auth.)

  19. Lifetime of Mechanical Equipment

    Energy Technology Data Exchange (ETDEWEB)

    Leland, K.

    1999-07-01

    The gas plant at Kaarstoe was built as part of the Statpipe gas transport system and went on stream in 1985. In 1993 another line was routed from the Sleipner field to carry condensate, and the plant was extended accordingly. Today heavy additional supply- and export lines are under construction, and the plant is extended more than ever. The main role of the factory is to separate the raw gas into commercial products and to pump or ship it to the markets. The site covers a large number of well-known mechanical equipment. This presentation deals with piping, mechanical and structural disciplines. The lifetime of mechanical equipment is often difficult to predict as it depends on many factors, and the subject is complex. Mechanical equipment has been kept in-house, which provides detailed knowledge of the stages from a new to a 14 years old plant. The production regularity has always been very high, as required. The standard of the equipment is well kept, support systems are efficient, and human improvisation is extremely valuable.

  20. Accurate Rapid Lifetime Determination on Time-Gated FLIM Microscopy with Optical Sectioning.

    Science.gov (United States)

    Silva, Susana F; Domingues, José Paulo; Morgado, António Miguel

    2018-01-01

    Time-gated fluorescence lifetime imaging microscopy (FLIM) is a powerful technique to assess the biochemistry of cells and tissues. When applied to living thick samples, it is hampered by the lack of optical sectioning and the need of acquiring many images for an accurate measurement of fluorescence lifetimes. Here, we report on the use of processing techniques to overcome these limitations, minimizing the acquisition time, while providing optical sectioning. We evaluated the application of the HiLo and the rapid lifetime determination (RLD) techniques for accurate measurement of fluorescence lifetimes with optical sectioning. HiLo provides optical sectioning by combining the high-frequency content from a standard image, obtained with uniform illumination, with the low-frequency content of a second image, acquired using structured illumination. Our results show that HiLo produces optical sectioning on thick samples without degrading the accuracy of the measured lifetimes. We also show that instrument response function (IRF) deconvolution can be applied with the RLD technique on HiLo images, improving greatly the accuracy of the measured lifetimes. These results open the possibility of using the RLD technique with pulsed diode laser sources to determine accurately fluorescence lifetimes in the subnanosecond range on thick multilayer samples, providing that offline processing is allowed.

  1. Fluorescence spectroscopy

    DEFF Research Database (Denmark)

    Bagatolli, Luis

    2016-01-01

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

  2. Silica nanodisks as platforms for fluorescence lifetime-based ...

    Indian Academy of Sciences (India)

    Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400 076, India ... The nanoconjugates are found to sense the pH of the medium, through systematic ..... form pH sensing with a good level of sensitivity, with.

  3. Lifetime Improvement by Battery Scheduling

    NARCIS (Netherlands)

    Jongerden, M.R.; Schmitt, Jens B.; Haverkort, Boudewijn R.H.M.

    The use of mobile devices is often limited by the lifetime of their batteries. For devices that have multiple batteries or that have the option to connect an extra battery, battery scheduling, thereby exploiting the recovery properties of the batteries, can help to extend the system lifetime. Due to

  4. Lifetime improvement by battery scheduling

    NARCIS (Netherlands)

    Jongerden, M.R.; Haverkort, Boudewijn R.H.M.

    The use of mobile devices is often limited by the lifetime of its battery. For devices that have multiple batteries or that have the option to connect an extra battery, battery scheduling, thereby exploiting the recovery properties of the batteries, can help to extend the system lifetime. Due to the

  5. Fluorescent nanoparticles for intracellular sensing: A review

    International Nuclear Information System (INIS)

    Ruedas-Rama, Maria J.; Walters, Jamie D.; Orte, Angel; Hall, Elizabeth A.H.

    2012-01-01

    Highlights: ► Analytical applications of fluorescent nanoparticles (NPs) in intracellular sensing. ► Critical review on performance of QDots, metal NPs, silica NPs, and polymer NPs. ► Highlighted potential of fluorescence lifetime imaging microscopy (FLIM). - Abstract: Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy.

  6. Fluorescent nanoparticles for intracellular sensing: A review

    Energy Technology Data Exchange (ETDEWEB)

    Ruedas-Rama, Maria J., E-mail: mjruedas@ugr.esmailto [Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Campus Cartuja, 18071, Granada (Spain); Walters, Jamie D. [Department of Chemical Engineering and Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, UK CB2 1QT (United Kingdom); Orte, Angel [Department of Physical Chemistry, Faculty of Pharmacy, University of Granada, Campus Cartuja, 18071, Granada (Spain); Hall, Elizabeth A.H., E-mail: lisa.hall@biotech.cam.ac.uk [Department of Chemical Engineering and Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QT (United Kingdom)

    2012-11-02

    Highlights: Black-Right-Pointing-Pointer Analytical applications of fluorescent nanoparticles (NPs) in intracellular sensing. Black-Right-Pointing-Pointer Critical review on performance of QDots, metal NPs, silica NPs, and polymer NPs. Black-Right-Pointing-Pointer Highlighted potential of fluorescence lifetime imaging microscopy (FLIM). - Abstract: Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy.

  7. High speed fluorescence imaging with compressed ultrafast photography

    Science.gov (United States)

    Thompson, J. V.; Mason, J. D.; Beier, H. T.; Bixler, J. N.

    2017-02-01

    Fluorescent lifetime imaging is an optical technique that facilitates imaging molecular interactions and cellular functions. Because the excited lifetime of a fluorophore is sensitive to its local microenvironment,1, 2 measurement of fluorescent lifetimes can be used to accurately detect regional changes in temperature, pH, and ion concentration. However, typical state of the art fluorescent lifetime methods are severely limited when it comes to acquisition time (on the order of seconds to minutes) and video rate imaging. Here we show that compressed ultrafast photography (CUP) can be used in conjunction with fluorescent lifetime imaging to overcome these acquisition rate limitations. Frame rates up to one hundred billion frames per second have been demonstrated with compressed ultrafast photography using a streak camera.3 These rates are achieved by encoding time in the spatial direction with a pseudo-random binary pattern. The time domain information is then reconstructed using a compressed sensing algorithm, resulting in a cube of data (x,y,t) for each readout image. Thus, application of compressed ultrafast photography will allow us to acquire an entire fluorescent lifetime image with a single laser pulse. Using a streak camera with a high-speed CMOS camera, acquisition rates of 100 frames per second can be achieved, which will significantly enhance our ability to quantitatively measure complex biological events with high spatial and temporal resolution. In particular, we will demonstrate the ability of this technique to do single-shot fluorescent lifetime imaging of cells and microspheres.

  8. Platelet Count and Plateletcrit

    African Journals Online (AJOL)

    strated that neonates with late onset sepsis (bacteremia after 3 days of age) had a dramatic increase in MPV and. PDW18. We hypothesize that as the MPV and PDW increase and platelet count and PCT decrease in sick children, intui- tively, the ratio of MPV to PCT; MPV to Platelet count,. PDW to PCT, PDW to platelet ...

  9. EcoCount

    Directory of Open Access Journals (Sweden)

    Phillip P. Allen

    2014-05-01

    Full Text Available Techniques that analyze biological remains from sediment sequences for environmental reconstructions are well established and widely used. Yet, identifying, counting, and recording biological evidence such as pollen grains remain a highly skilled, demanding, and time-consuming task. Standard procedure requires the classification and recording of between 300 and 500 pollen grains from each representative sample. Recording the data from a pollen count requires significant effort and focused resources from the palynologist. However, when an adaptation to the recording procedure is utilized, efficiency and time economy improve. We describe EcoCount, which represents a development in environmental data recording procedure. EcoCount is a voice activated fully customizable digital count sheet that allows the investigator to continuously interact with a field of view during the data recording. Continuous viewing allows the palynologist the opportunity to remain engaged with the essential task, identification, for longer, making pollen counting more efficient and economical. EcoCount is a versatile software package that can be used to record a variety of environmental evidence and can be installed onto different computer platforms, making the adoption by users and laboratories simple and inexpensive. The user-friendly format of EcoCount allows any novice to be competent and functional in a very short time.

  10. Clean Hands Count

    Medline Plus

    Full Text Available ... starting stop Loading... Watch Queue Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos with ... ads? Get YouTube Red. Working... Not now Try it free Find out why Close Clean Hands Count ...

  11. Counting It Twice.

    Science.gov (United States)

    Schattschneider, Doris

    1991-01-01

    Provided are examples from many domains of mathematics that illustrate the Fubini Principle in its discrete version: the value of a summation over a rectangular array is independent of the order of summation. Included are: counting using partitions as in proof by pictures, combinatorial arguments, indirect counting as in the inclusion-exclusion…

  12. Lifetime costs of cerebral palsy

    DEFF Research Database (Denmark)

    Kruse, Marie; Michelsen, Susan Ishøy; Flachs, Esben Meulengracht

    2009-01-01

    This study quantified the lifetime costs of cerebral palsy (CP) in a register-based setting. It was the first study outside the US to assess the lifetime costs of CP. The lifetime costs attributable to CP were divided into three categories: health care costs, productivity costs, and social costs....... social care costs and productivity costs associated with CP point to a potential gain from labour market interventions that benefit individuals with CP.......This study quantified the lifetime costs of cerebral palsy (CP) in a register-based setting. It was the first study outside the US to assess the lifetime costs of CP. The lifetime costs attributable to CP were divided into three categories: health care costs, productivity costs, and social costs...... in 2000. The prevalence of CP in eastern Denmark was approximately 1.7 per 1000. Information on productivity and the use of health care was retrieved from registers. The lifetime cost of CP was about euro860 000 for men and about euro800 000 for women. The largest component was social care costs...

  13. Wide-field time-correlated single photon counting (TCSPC) microscopy with time resolution below the frame exposure time

    Energy Technology Data Exchange (ETDEWEB)

    Hirvonen, Liisa M. [Department of Physics, King' s College London, Strand, London WC2R 2LS (United Kingdom); Petrášek, Zdeněk [Max Planck Institute of Biochemistry, Department of Cellular and Molecular Biophysics, Am Klopferspitz 18, D-82152 Martinsried (Germany); Suhling, Klaus, E-mail: klaus.suhling@kcl.ac.uk [Department of Physics, King' s College London, Strand, London WC2R 2LS (United Kingdom)

    2015-07-01

    Fast frame rate CMOS cameras in combination with photon counting intensifiers can be used for fluorescence imaging with single photon sensitivity at kHz frame rates. We show here how the phosphor decay of the image intensifier can be exploited for accurate timing of photon arrival well below the camera exposure time. This is achieved by taking ratios of the intensity of the photon events in two subsequent frames, and effectively allows wide-field TCSPC. This technique was used for measuring decays of ruthenium compound Ru(dpp) with lifetimes as low as 1 μs with 18.5 μs frame exposure time, including in living HeLa cells, using around 0.1 μW excitation power. We speculate that by using an image intensifier with a faster phosphor decay to match a higher camera frame rate, photon arrival time measurements on the nanosecond time scale could well be possible.

  14. Polar plot representation of time-resolved fluorescence.

    Science.gov (United States)

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

    2014-01-01

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

  15. Electricite de France: Lifetime Project

    International Nuclear Information System (INIS)

    Combes, Jean-Pierre

    1991-01-01

    Electricite de France produces almost 80% of its electricity by means of standardized PWR nuclear power stations. Starting in 1986, therefore, a project known as the 'Lifetime Project' was developed, whose aim was initially to ensure that the lifetime defined at design stage (40 years in general) could be attained without major difficulty (follow up of the aging process). It then became apparent that it would be useful to know just how far it would be technically and economically possible to go. As a result, the project is now working towards increasing the lifetime of power stations. (author)

  16. Differential white cell count by centrifugal microfluidics.

    Energy Technology Data Exchange (ETDEWEB)

    Sommer, Gregory Jon; Tentori, Augusto M.; Schaff, Ulrich Y.

    2010-07-01

    We present a method for counting white blood cells that is uniquely compatible with centrifugation based microfluidics. Blood is deposited on top of one or more layers of density media within a microfluidic disk. Spinning the disk causes the cell populations within whole blood to settle through the media, reaching an equilibrium based on the density of each cell type. Separation and fluorescence measurement of cell types stained with a DNA dye is demonstrated using this technique. The integrated signal from bands of fluorescent microspheres is shown to be proportional to their initial concentration in suspension. Among the current generation of medical diagnostics are devices based on the principle of centrifuging a CD sized disk functionalized with microfluidics. These portable 'lab on a disk' devices are capable of conducting multiple assays directly from a blood sample, embodied by platforms developed by Gyros, Samsung, and Abaxis. [1,2] However, no centrifugal platform to date includes a differential white blood cell count, which is an important metric complimentary to diagnostic assays. Measuring the differential white blood cell count (the relative fraction of granulocytes, lymphocytes, and monocytes) is a standard medical diagnostic technique useful for identifying sepsis, leukemia, AIDS, radiation exposure, and a host of other conditions that affect the immune system. Several methods exist for measuring the relative white blood cell count including flow cytometry, electrical impedance, and visual identification from a stained drop of blood under a microscope. However, none of these methods is easily incorporated into a centrifugal microfluidic diagnostic platform.

  17. Characterization of porcine eyes based on autofluorescence lifetime imaging

    Science.gov (United States)

    Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Morgado, António Miguel; König, Karsten

    2015-03-01

    Multiphoton microscopy is a non-invasive imaging technique with ideal characteristics for biological applications. In this study, we propose to characterize three major structures of the porcine eye, the cornea, crystalline lens, and retina using two-photon excitation fluorescence lifetime imaging microscopy (2PE-FLIM). Samples were imaged using a laser-scanning microscope, consisting of a broadband sub-15 femtosecond (fs) near-infrared laser. Signal detection was performed using a 16-channel photomultiplier tube (PMT) detector (PML-16PMT). Therefore, spectral analysis of the fluorescence lifetime data was possible. To ensure a correct spectral analysis of the autofluorescence lifetime data, the spectra of the individual endogenous fluorophores were acquired with the 16-channel PMT and with a spectrometer. All experiments were performed within 12h of the porcine eye enucleation. We were able to image the cornea, crystalline lens, and retina at multiple depths. Discrimination of each structure based on their autofluorescence intensity and lifetimes was possible. Furthermore, discrimination between different layers of the same structure was also possible. To the best of our knowledge, this was the first time that 2PE-FLIM was used for porcine lens imaging and layer discrimination. With this study we further demonstrated the feasibility of 2PE-FLIM to image and differentiate three of the main components of the eye and its potential as an ophthalmologic technique.

  18. Measurement of Charm Meson Lifetimes

    International Nuclear Information System (INIS)

    Bonvicini, G.; Cinabro, D.; Greene, R.; Perera, L.P.; Zhou, G.J.; Chan, S.; Eigen, G.; Lipeles, E.; Schmidtler, M.; Shapiro, A.; Sun, W.M.; Urheim, J.; Weinstein, A.J.; Wuerthwein, F.; Jaffe, D.E.; Masek, G.; Paar, H.P.; Potter, E.M.; Prell, S.; Sharma, V.; Asner, D.M.; Eppich, A.; Gronberg, J.; Hill, T.S.; Korte, C.M.; Lange, D.J.; Morrison, R.J.; Nelson, H.N.; Nelson, T.K.; Roberts, D.; Tajima, H.; Behrens, B.H.; Ford, W.T.; Gritsan, A.; Krieg, H.; Roy, J.; Smith, J.G.; Alexander, J.P.; Baker, R.; Bebek, C.; Berger, B.E.; Berkelman, K.; Boisvert, V.; Cassel, D.G.; Crowcroft, D.S.; Dickson, M.; Dombrowski, S. von; Drell, P.S.; Dumas, D.J.; Ecklund, K.M.; Ehrlich, R.; Foland, A.D.; Gaidarev, P.; Gibbons, L.; Gittelman, B.; Gray, S.W.; Hartill, D.L.; Heltsley, B.K.; Henderson, S.; Hopman, P.I.; Katayama, N.; Kreinick, D.L.; Lee, T.; Liu, Y.; Meyer, T.O.; Mistry, N.B.; Ng, C.R.; Nordberg, E.; Ogg, M.; Patterson, J.R.; Peterson, D.; Riley, D.; Soffer, A.; Thayer, J.G.; Thies, P.G.; Valant-Spaight, B.; Warburton, A.; Ward, C.; Athanas, M.; Avery, P.; Jones, C.D.; Lohner, M.; Prescott, C.; Rubiera, A.I.; Yelton, J.; Zheng, J.; Brandenburg, G.; Briere, R.A.; Ershov, A.; Gao, Y.S.; Kim, D.Y.; Wilson, R.; Browder, T.E.; Li, Y.; Rodriguez, J.L.; Yamamoto, H.; Bergfeld, T.; Eisenstein, B.I.; Ernst, J.; Gladding, G.E.; Gollin, G.D

    1999-01-01

    We report measurements of the D 0 , D + , and D + s meson lifetimes using 3.7 fb -1 of e + e - annihilation data collected near the Υ(4S) resonance with the CLEO detector. The measured lifetimes of the D 0 , D + , and D + s mesons are 408.5±4.1 +3.5 -3.4 fs , 1033.6±22.1 +9.9 -12.7 fs , and 486.3±15.0 +4.9 -5.1 fs . The precision of these lifetimes are comparable to those of the best previous measurements, and the systematic errors are very different. In a single experiment we find that the ratio of the D + s and D 0 lifetimes is 1.19±0.04 . copyright 1999 The American Physical Society

  19. Deconvolution of Positrons' Lifetime spectra

    International Nuclear Information System (INIS)

    Calderin Hidalgo, L.; Ortega Villafuerte, Y.

    1996-01-01

    In this paper, we explain the iterative method previously develop for the deconvolution of Doppler broadening spectra using the mathematical optimization theory. Also, we start the adaptation and application of this method to the deconvolution of positrons' lifetime annihilation spectra

  20. The fluorescence behaviour of methyl and phenyl salicylate

    Science.gov (United States)

    Ford, D.; Thistlethwaite, P. J.; Woolfe, G. J.

    1980-01-01

    Fluorcsccnce lifetimes tor the 450 nm emission of methyl and phenyl salicylate in various solvents have been measured. Qucnching studics on the 340 nm fluorescence of these molecules point to the existence of three distinct ground state conformers.

  1. Clean Hands Count

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    Medline Plus

    Full Text Available ... Clean Hands Count Centers for Disease Control and Prevention (CDC) Loading... Unsubscribe from Centers for Disease Control and Prevention (CDC)? Cancel Unsubscribe Working... Subscribe Subscribed Unsubscribe 66K ...

  3. Housing Inventory Count

    Data.gov (United States)

    Department of Housing and Urban Development — This report displays the data communities reported to HUD about the nature of their dedicated homeless inventory, referred to as their Housing Inventory Count (HIC)....

  4. Scintillation counting apparatus

    International Nuclear Information System (INIS)

    Noakes, J.E.

    1978-01-01

    Apparatus is described for the accurate measurement of radiation by means of scintillation counters and in particular for the liquid scintillation counting of both soft beta radiation and gamma radiation. Full constructional and operating details are given. (UK)

  5. Allegheny County Traffic Counts

    Data.gov (United States)

    Allegheny County / City of Pittsburgh / Western PA Regional Data Center — Traffic sensors at over 1,200 locations in Allegheny County collect vehicle counts for the Pennsylvania Department of Transportation. Data included in the Health...

  6. Counting Knights and Knaves

    Science.gov (United States)

    Levin,Oscar; Roberts, Gerri M.

    2013-01-01

    To understand better some of the classic knights and knaves puzzles, we count them. Doing so reveals a surprising connection between puzzles and solutions, and highlights some beautiful combinatorial identities.

  7. Clean Hands Count

    Medline Plus

    Full Text Available ... why Close Clean Hands Count Centers for Disease Control and Prevention (CDC) Loading... Unsubscribe from Centers for Disease Control and Prevention (CDC)? Cancel Unsubscribe Working... Subscribe Subscribed ...

  8. Clean Hands Count

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  9. The Kruskal Count

    OpenAIRE

    Lagarias, Jeffrey C.; Rains, Eric; Vanderbei, Robert J.

    2001-01-01

    The Kruskal Count is a card trick invented by Martin J. Kruskal in which a magician "guesses" a card selected by a subject according to a certain counting procedure. With high probability the magician can correctly "guess" the card. The success of the trick is based on a mathematical principle related to coupling methods for Markov chains. This paper analyzes in detail two simplified variants of the trick and estimates the probability of success. The model predictions are compared with simula...

  10. Increasing the collection efficiency of time-correlated single-photon counting with single-photon avalanche diodes using immersion lenses.

    Science.gov (United States)

    Pichette, Charles; Giudice, Andrea; Thibault, Simon; Bérubé-Lauzière, Yves

    2016-11-20

    Single-photon avalanche diodes (SPADs) achieving high timing resolution (≈20-50  ps) developed for time-correlated single-photon counting (TCSPC) generally have very small photosensitive areas (25-100 μm in diameter). This limits the achievable photon counting rate and signal-to-noise ratio and may lead to long counting times. This is detrimental in applications requiring several measurements, such as fluorescence lifetime imaging (FLIM) microscopy, which requires scanning, and time-domain diffuse optical tomography (TD-DOT). We show in this work that the use of an immersion lens directly affixed onto the photosensitive area of the SPAD helps alleviate this problem by allowing more light to be concentrated onto the detector. Following careful optical design and simulations, our experimental results show that it is actually possible to achieve the predicted theoretical increase in the photon counting rate (we achieve a factor of ≈4 here). This work is of high relevance in high timing resolution TCSPC with small photosensitive area detectors and should find widespread interest in FLIM and TD-DOT with SPADs.

  11. Occupational risk and lifetime exposure

    International Nuclear Information System (INIS)

    Lapp, R.E.

    1991-01-01

    Any lowering of annual radiation limits for occupational exposure should be based on industry experience with lifetime doses and not on a worst case career exposure of 47 years. Two decades of experience show a lifetime accumulation of less than 1.5 rem for workers with measurable exposure. This is 5% of the normal lifetime exposure of Americans to natural and medical radiation. Any epidemiology of the US nuclear power workforce's two decade long exposure would have to focus on excess leukemia. Application of the Hiroshima and Nagasaki cancer mortality shows that too few leukemias would be expressed to permit a feasible epidemiology. Ionizing radiation appears to be a mild carcinogen as compared to physical and chemical agents presented in the occupational environment. A realistic factor in determining any change in occupational exposure limits for ionizing radiation should take into account the past performance of the licensee and potential health effects applicable to the workplace. Specifically, the lifetime exposure data for workers at nuclear power plants and naval shipyards should be considered. The nuclear industry and the US Navy have detailed data on the annual exposure of workers with a combined collective exposure approaching 1 million worker-rem. The lifetime dose for naval personnel and shipyard workers averages 1.1 rem J 1990. Shipyard workers have an annual dose of 0.28 rem per work-year and a mean exposure time of 4.4 years. The data apply to workers with measurable dose

  12. Confocal fluorescence techniques in industrial application

    Science.gov (United States)

    Eggeling, Christian; Gall, Karsten; Palo, Kaupo; Kask, Peet; Brand, Leif

    2003-06-01

    The FCS+plus family of evaluation tools for confocal fluorescence spectroscopy, which was developed during recent years, offers a comprehensive view to a series of fluorescence properties. Originating in fluorescence correlation spectroscopy (FCS) and using similar experimental equipment, a system of signal processing methods such as fluorescence intensity distribution analysis (FIDA) was created to analyze in detail the fluctuation behavior of fluorescent particles within a small area of detection. Giving simultaneous access to molecular parameters like concentration, translational and rotational diffusion, molecular brightness, and multicolor coincidence, this portfolio was enhanced by more traditional techniques of fluorescence lifetime as well as time-resolved anisotropy determination. The cornerstones of the FCS+plus methodology will be shortly described. The inhibition of a phosphatase enzyme activity gives a comprehensive industrial application that demonstrates FCS+plus' versatility and its potential for pharmaceutical drug discovery.

  13. Low White Blood Cell Count

    Science.gov (United States)

    Symptoms Low white blood cell count By Mayo Clinic Staff A low white blood cell count (leukopenia) is a decrease ... of white blood cell (neutrophil). The definition of low white blood cell count varies from one medical ...

  14. Photon counting and fluctuation of molecular movement

    International Nuclear Information System (INIS)

    Inohara, Koichi

    1978-01-01

    The direct measurement of the fluctuation of molecular motions, which provides with useful information on the molecular movement, was conducted by introducing photon counting method. The utilization of photon counting makes it possible to treat the molecular system consisting of a small number of molecules like a radioisotope in the detection of a small number of atoms, which are significant in biological systems. This method is based on counting the number of photons of the definite polarization emitted in a definite time interval from the fluorescent molecules excited by pulsed light, which are bound to the marked large molecules found in a definite spatial region. Using the probability of finding a number of molecules oriented in a definite direction in the definite spatial region, the probability of counting a number of photons in a definite time interval can be calculated. Thus the measurable count rate of photons can be related with the fluctuation of molecular movement. The measurement was carried out under the condition, in which the probability of the simultaneous arrival of more than two photons at a detector is less than 1/100. As the experimental results, the resolving power of photon-counting apparatus, the frequency distribution of the number of photons of some definite polarization counted for 1 nanosecond are shown. In the solution, the variance of the number of molecules of 500 on the average is 1200, which was estimated from the experimental data by assuming normal distribution. This departure from the Poisson distribution means that a certain correlation does exist in molecular movement. In solid solution, no significant deviation was observed. The correlation existing in molecular movement can be expressed in terms of the fluctuation of the number of molecules. (Nakai, Y.)

  15. Plasmonic enhancement of ultraviolet fluorescence

    Science.gov (United States)

    Jiao, Xiaojin

    Plasmonics relates to the interaction between electromagnetic radiation and conduction electrons at metallic interfaces or in metallic nanostructures. Surface plasmons are collective electron oscillations at a metal surface, which can be manipulated by shape, texture and material composition. Plasmonic applications cover a broad spectrum from visible to near infrared, including biosensing, nanolithography, spectroscopy, optoelectronics, photovoltaics and so on. However, there remains a gap in this activity in the ultraviolet (UV, research. Motivating factors in the study of UV Plasmonics are the direct access to biomolecular resonances and native fluorescence, resonant Raman scattering interactions, and the potential for exerting control over photochemical reactions. This dissertation aims to fill in the gap of Plasmonics in the UV with efforts of design, fabrication and characterization of aluminium (Al) and magnesium (Mg) nanostructures for the application of label-free bimolecular detection via native UV fluorescence. The first contribution of this dissertation addresses the design of Al nanostructures in the context of UV fluorescence enhancement. A design method that combines analytical analysis with numerical simulation has been developed. Performance of three canonical plasmonic structures---the dipole antenna, bullseye nanoaperture and nanoaperture array---has been compared. The optimal geometrical parameters have been determined. A novel design of a compound bullseye structure has been proposed and numerically analyzed for the purpose of compensating for the large Stokes shift typical of UV fluorescence. Second, UV lifetime modification of diffusing molecules by Al nanoapertures has been experimentally demonstrated for the first time. Lifetime reductions of ~3.5x have been observed for the high quantum yield (QY) laser dye p-terphenyl in a 60 nm diameter aperture with 50 nm undercut. Furthermore, quantum-yield-dependence of lifetime reduction has been

  16. Alpha scintillation radon counting

    International Nuclear Information System (INIS)

    Lucas, H.F. Jr.

    1977-01-01

    Radon counting chambers which utilize the alpha-scintillation properties of silver activated zinc sulfide are simple to construct, have a high efficiency, and, with proper design, may be relatively insensitive to variations in the pressure or purity of the counter filling. Chambers which were constructed from glass, metal, or plastic in a wide variety of shapes and sizes were evaluated for the accuracy and the precision of the radon counting. The principles affecting the alpha-scintillation radon counting chamber design and an analytic system suitable for a large scale study of the 222 Rn and 226 Ra content of either air or other environmental samples are described. Particular note is taken of those factors which affect the accuracy and the precision of the method for monitoring radioactivity around uranium mines

  17. Principles of correlation counting

    International Nuclear Information System (INIS)

    Mueller, J.W.

    1975-01-01

    A review is given of the various applications which have been made of correlation techniques in the field of nuclear physics, in particular for absolute counting. Whereas in most cases the usual coincidence method will be preferable for its simplicity, correlation counting may be the only possible approach in such cases where the two radiations of the cascade cannot be well separated or when there is a longliving intermediate state. The measurement of half-lives and of count rates of spurious pulses is also briefly discussed. The various experimental situations lead to different ways the correlation method is best applied (covariance technique with one or with two detectors, application of correlation functions, etc.). Formulae are given for some simple model cases, neglecting dead-time corrections

  18. Interpretation of galaxy counts

    International Nuclear Information System (INIS)

    Tinsely, B.M.

    1980-01-01

    New models are presented for the interpretation of recent counts of galaxies to 24th magnitude, and predictions are shown to 28th magnitude for future comparison with data from the Space Telescope. The results supersede earlier, more schematic models by the author. Tyson and Jarvis found in their counts a ''local'' density enhancement at 17th magnitude, on comparison with the earlier models; the excess is no longer significant when a more realistic mixture of galaxy colors is used. Bruzual and Kron's conclusion that Kron's counts show evidence for evolution at faint magnitudes is confirmed, and it is predicted that some 23d magnitude galaxies have redshifts greater than unity. These may include spheroidal systems, elliptical galaxies, and the bulges of early-type spirals and S0's, seen during their primeval rapid star formation

  19. Lifetime results from heavy quark systems

    International Nuclear Information System (INIS)

    Papadimitriou, V.

    1997-11-01

    We present the latest measurements of weakly decaying b-hadrons from experiments at e + e - and p anti p colliders. These measurements include the average lifetime of b-hadrons, lifetimes of the B - , B 0 and B 0 s mesons, the average lifetime of b-baryons and lifetimes of the Λ b and Ξ b baryons

  20. Computerized radioautographic grain counting

    International Nuclear Information System (INIS)

    McKanna, J.A.; Casagrande, V.A.

    1985-01-01

    In recent years, radiolabeling techniques have become fundamental assays in physiology and biochemistry experiments. They also have assumed increasingly important roles in morphologic studies. Characteristically, radioautographic analysis of structure has been qualitative rather than quantitative, however, microcomputers have opened the door to several methods for quantifying grain counts and density. The overall goal of this chapter is to describe grain counting using the Bioquant, an image analysis package based originally on the Apple II+, and now available for several popular microcomputers. The authors discuss their image analysis procedures by applying them to a study of development in the central nervous system

  1. Rainflow counting revisited

    Energy Technology Data Exchange (ETDEWEB)

    Soeker, H [Deutsches Windenergie-Institut (Germany)

    1996-09-01

    As state of the art method the rainflow counting technique is presently applied everywhere in fatigue analysis. However, the author feels that the potential of the technique is not fully recognized in wind energy industries as it is used, most of the times, as a mere data reduction technique disregarding some of the inherent information of the rainflow counting results. The ideas described in the following aim at exploitation of this information and making it available for use in the design and verification process. (au)

  2. Fluorescent Nanodiamonds in Biomedical Applications.

    Science.gov (United States)

    Mitura, Katarzyna Anna; Włodarczyk, Elżbieta

    2018-04-18

    Nanoparticles have an extended surface and a large surface area, which is the ratio of the size of the surfacearea to the volume. A functionalized surface can give rise to more modifications and therefore allows this nanomaterial to have new properties. Fluorescent molecules contain fluorophore, which is capable of being excited via the absorption of light energy at a specific wavelength and subsequently emitting radiation energy of a longer wavelength. A chemically modified surface of nanodiamond (ND; by carboxylation) demonstrated biocompatibility with DNA, cytochrome C, and antigens. In turn, fluorescent nanodiamonds (FNDs) belong to a group of new nanomaterials. Their surface can be modified by joining functional groups such as carboxyl, hydroxyl, or amino, after which they can be employed as a fluorescence agent. Their fluorescent properties result from defects in the crystal lattice. FNDs reach dimensions of 4-100 nm, have attributes such as photostability, long fluorescence lifetimes (10 ns), and fluorescence emission between 600 and 700 nm. They are also nontoxic, chemically inert, biocompatible, and environmentally harmless. The main purpose of this article was to present the medical applications of various types of modified NDs.

  3. A time-domain fluorescence diffusion optical tomography system for breast tumor diagnosis

    Science.gov (United States)

    Zhang, Wei; Gao, Feng; Wu, LinHui; Ma, Wenjuan; Yang, Fang; Zhou, Zhongxing; Zhang, Limin; Zhao, Huijuan

    2011-02-01

    A prototype time-domain fluorescence diffusion optical tomography (FDOT) system using near-infrared light is presented. The system employs two pulsed light sources, 32 source fibers and 32 detection channels, working separately for acquiring the temporal distribution of the photon flux on the tissue surface. The light sources are provided by low power picosecond pulsed diode lasers at wavelengths of 780 nm and 830 nm, and a 1×32-fiber-optic-switch sequentially directs light sources to the object surface through 32 source fibers. The light signals re-emitted from the object are collected by 32 detection fibers connected to four 8×1 fiber-optic-switch and then routed to four time-resolved measuring channels, each of which consists of a collimator, a filter wheel, a photomultiplier tube (PMT) photon-counting head and a time-correlated single photon counting (TCSPC) channel. The performance and efficacy of the designed multi-channel PMT-TCSPC system are assessed by reconstructing the fluorescent yield and lifetime images of a solid phantom.

  4. Lifetime value in business process

    Directory of Open Access Journals (Sweden)

    Martin Souček

    2011-01-01

    Full Text Available The paper focuses on lifetime value assessment and its implementation and application in business processes. The lifetime value is closely connected to customer relationship management. The paper presents results of three consecutive researches devoted to issues of customer relationship management. The first two from 2008 and 2010 were conducted as quantitative ones; the one from 2009 had qualitative nature. The respondents were representatives of particular companies. The means for data collection was provided by ReLa system. We will focus on individual attributes of lifetime value of a customer, and relate them to approaches of authors mentioned in introduction. Based on the qualitative research data, the paper focuses on individual customer lifetime value parameters. These parameters include: the cost to the customer relationship acquisition and maintenance, profit generated from a particular customer, customer awareness value, the level of preparedness to adopt new products, the value of references and customer loyalty level. For each of these parameters, the paper provides specific recommendations. Moreover, it is possible to learn about the nature of these parameter assessments in the Czech environment.

  5. Lifetime of heavy flavour particles

    International Nuclear Information System (INIS)

    Lueth, V.

    1985-10-01

    Recent measurements of the lifetime of the tau leptons and charm and beauty hadrons are reviewed and their significance for the couplings of the charged weak current, flavour mixing, and models relating quarks to hadron decay are discussed. 70 refs., 17 figs., 5 tabs

  6. Clean Hands Count

    Medline Plus

    Full Text Available ... Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos with zero ads? Get YouTube Red. ... 824 views 1:36 Wash 'Em - Hand Hygiene Music Video - Duration: 5:46. Jefferson Health 409,492 ...

  7. Clean Hands Count

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    Full Text Available ... Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos with zero ads? Get YouTube Red. ... 786 views 1:36 Wash 'Em - Hand Hygiene Music Video - Duration: 5:46. Jefferson Health 413,702 ...

  8. Clean Hands Count

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    Full Text Available ... Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos with zero ads? Get YouTube Red. ... 414 views 3:10 Wash 'Em - Hand Hygiene Music Video - Duration: 5:46. Thomas Jefferson University & Jefferson ...

  9. Clean Hands Count

    Medline Plus

    Full Text Available ... Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos with zero ads? Get YouTube Red. ... 869 views 1:36 Wash 'Em - Hand Hygiene Music Video - Duration: 5:46. Jefferson Health 410,052 ...

  10. Clean Hands Count

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  11. Clean Hands Count

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  12. Clean Hands Count

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  13. Clean Hands Count

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  14. Clean Hands Count

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  15. Clean Hands Count

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  16. Clean Hands Count

    Medline Plus

    Full Text Available ... Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos with zero ads? Get YouTube Red. ... 585 views 3:10 Wash 'Em - Hand Hygiene Music Video - Duration: 5:46. Jefferson Health 413,097 ...

  17. Detection and counting systems

    International Nuclear Information System (INIS)

    Abreu, M.A.N. de

    1976-01-01

    Detection devices based on gaseous ionization are analysed, such as: electroscopes ionization chambers, proportional counters and Geiger-Mueller counters. Scintillation methods are also commented. A revision of the basic concepts in electronics is done and the main equipment for counting is detailed. In the study of gama spectrometry, scintillation and semiconductor detectors are analysed [pt

  18. Clean Hands Count

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  19. Clean Hands Count

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    Full Text Available ... Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos with zero ads? Get YouTube Red. ... 285 views 1:36 Wash 'Em - Hand Hygiene Music Video - Duration: 5:46. Jefferson Health 410,052 ...

  20. Clean Hands Count

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    Full Text Available ... Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos with zero ads? Get YouTube Red. ... 033 views 1:36 Wash 'Em - Hand Hygiene Music Video - Duration: 5:46. Jefferson Health 410,052 ...

  1. Reticulocyte Count Test

    Science.gov (United States)

    ... htm. (2004 Summer). Immature Reticulocyte Fraction(IRF). The Pathology Center Newsletter v9(1). [On-line information]. Available ... Company, Philadelphia, PA [18th Edition]. Levin, M. (2007 March 8, Updated). Reticulocyte Count. MedlinePlus Medical Encyclopedia [On- ...

  2. Clean Hands Count

    Medline Plus

    Full Text Available ... is starting stop Loading... Watch Queue Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos ... empower patients to play a role in their care by asking or reminding healthcare providers to clean ...

  3. Radiation intensity counting system

    International Nuclear Information System (INIS)

    Peterson, R.J.

    1982-01-01

    A method is described of excluding the natural dead time of the radiation detector (or eg Geiger-Mueller counter) in a ratemeter counting circuit, thus eliminating the need for dead time corrections. Using a pulse generator an artificial dead time is introduced which is longer than the natural dead time of the detector. (U.K.)

  4. Clean Hands Count

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    Full Text Available ... Queue __count__/__total__ It’s YouTube. Uninterrupted. Loading... Want music and videos with zero ads? Get YouTube Red. ... 043 views 1:36 Wash 'Em - Hand Hygiene Music Video - Duration: 5:46. Jefferson Health 411,292 ...

  5. Calorie count - fast food

    Science.gov (United States)

    ... GO About MedlinePlus Site Map FAQs Customer Support Health Topics Drugs & Supplements Videos & Tools Español You Are Here: Home → Medical Encyclopedia → Calorie count - fast food URL of this page: //medlineplus.gov/ency/patientinstructions/ ...

  6. Fast counting electronics for neutron coincidence counting

    International Nuclear Information System (INIS)

    Swansen, J.E.

    1987-01-01

    This patent describes a high speed circuit for accurate neutron coincidence counting comprising: neutron detecting means for providing an above-threshold signal upon neutron detection; amplifying means inputted by the neutron detecting means for providing a pulse output having a pulse width of about 0.5 microseconds upon the input of each above threshold signal; digital processing means inputted by the pulse output of the amplifying means for generating a pulse responsive to each input pulse from the amplifying means and having a pulse width of about 50 nanoseconds effective for processing an expected neutron event rate of about 1 Mpps: pulse stretching means inputted by the digital processing means for producing a pulse having a pulse width of several milliseconds for each pulse received form the digital processing means; visual indicating means inputted by the pulse stretching means for producing a visual output for each pulse received from the digital processing means; and derandomizing means effective to receive the 50 ns neutron event pulses from the digital processing means for storage at a rate up to the neutron event rate of 1 Mpps and having first counter means for storing the input neutron event pulses

  7. Heritability of lifetime ecstasy use.

    Science.gov (United States)

    Verweij, Karin J H; Treur, Jorien L; Vreeker, Annabel; Brunt, Tibor M; Willemsen, Gonneke; Boomsma, Dorret I; Vink, Jacqueline M

    2017-09-01

    Ecstasy is a widely used psychoactive drug that users often take because they experience positive effects such as increased euphoria, sociability, elevated mood, and heightened sensations. Ecstasy use is not harmless and several immediate and long term side effects have been identified. Lifetime ecstasy use is likely to be partly influenced by genetic factors, but no twin study has determined the heritability. Here, we apply a classical twin design to a large sample of twins and siblings to estimate the heritability of lifetime ecstasy use. The sample comprised 8500 twins and siblings aged between 18 and 45 years from 5402 families registered at the Netherlands Twin Registry. In 2013-2014 participants filled out a questionnaire including a question whether they had ever used ecstasy. We used the classical twin design to partition the individual differences in liability to ecstasy use into that due to genetic, shared environmental, and residual components. Overall, 10.4% of the sample had used ecstasy during their lifetime, with a somewhat higher prevalence in males than females. Twin modelling indicated that individual differences in liability to lifetime ecstasy use are for 74% due to genetic differences between individuals, whereas shared environmental and residual factors explain a small proportion of its liability (5% and 21%, respectively). Although heritability estimates appeared to be higher for females than males, this difference was not significant. Lifetime ecstasy use is a highly heritable trait, which indicates that some people are genetically more vulnerable to start using ecstasy than others. Copyright © 2017. Published by Elsevier B.V.

  8. Study on the fluorescence characteristics of carbon dots

    Science.gov (United States)

    Mao, Xiao-Jiao; Zheng, Hu-Zhi; Long, Yi-Juan; Du, Juan; Hao, Jian-Yu; Wang, Ling-Ling; Zhou, Dong-Bo

    2010-02-01

    Herein, we prepared water-soluble fluorescent carbon dots with diameter about 1.5 nm from cheap commercial lampblack. These fluorescent carbon nanoparticles are stable toward photobleaching and stable in water for more than half a year without fluorescence decrease. In order to improve its fluorescence properties, we passivated these nanoparticles with bisamino-terminated polyethylene glycol (PEG 1500N). Therefore, both fluorescence quantum yield and lifetime increased after this progress. In addition, the passivated carbon dots were more inert to solvent than the bare one and showed different responses to pH change.

  9. Riboflavin enhanced fluorescence of highly reduced graphene oxide

    Science.gov (United States)

    Iliut, Maria; Gabudean, Ana-Maria; Leordean, Cosmin; Simon, Timea; Teodorescu, Cristian-Mihail; Astilean, Simion

    2013-10-01

    The improvement of graphene derivates' fluorescence properties is a challenging topic and very few ways were reported up to now. In this Letter we propose an easy method to enhance the fluorescence of highly reduced graphene oxide (rGO) through non-covalent binding to a molecular fluorophore, namely the riboflavin (Rb). While the fluorescence of Rb is quenched, the Rb - decorated rGO exhibits strong blue fluorescence and significantly increased fluorescence lifetime, as compared to its pristine form. The data reported here represent a promising start towards tailoring the optical properties of rGOs, having utmost importance in optical applications.

  10. Dual fluorescence of single LH2 antenna nanorings

    International Nuclear Information System (INIS)

    Freiberg, A.; Raetsep, M.; Timpmann, K.; Trinkunas, G.

    2004-01-01

    A dual nature of fluorescence from LH2 pigment-protein complexes, which is a part of the light harvesting system of purple bacteria, is confirmed by fluorescence-lifetime dependence on recording wavelength and spectrally selective spectroscopy. An analysis based on the Holstein molecular crystal model, modified by allowing diagonal disorder, suggests coexistence of large- and small-radius self-trapped excitons, which serve as the origin of the dual fluorescence

  11. Emerging biomedical applications of time-resolved fluorescence spectroscopy

    Science.gov (United States)

    Lakowicz, Joseph R.; Szmacinski, Henryk; Koen, Peter A.

    1994-07-01

    Time-resolved fluorescence spectroscopy is presently regarded as a research tool in biochemistry, biophysics, and chemical physics. Advances in laser technology, the development of long-wavelength probes, and the use of lifetime-based methods are resulting in the rapid migration of time-resolved fluorescence to the clinical chemistry lab, to the patient's bedside, to flow cytometers, to the doctor's office, and even to home health care. Additionally, time-resolved imaging is now a reality in fluorescence microscopy, and will provide chemical imaging of a variety of intracellular analytes and/or cellular phenomena. In this overview paper we attempt to describe some of the opportunities available using chemical sensing based on fluorescence lifetimes, and to predict those applications of lifetime-based sensing which are most likely in the near future.

  12. Lifetime of a black hole

    International Nuclear Information System (INIS)

    Carlitz, R.D.; Willey, R.S.

    1987-01-01

    We study the constraints placed by quantum mechanics upon the lifetime of a black hole. In the context of a moving-mirror analog model for the Hawking radiation process, we conclude that the period of Hawking radiation must be followed by a much longer period during which the remnant mass (of order m/sub P/) may be radiated away. We are able to place a lower bound on the time required for this radiation process, which translates into a lower bound for the lifetime of the black hole. Particles which are emitted during the decay of the remnant, like the particles which comprise the Hawking flux, may be uncorrelated with each other. But each particle emitted from the decaying remnant is correlated with one particle emitted as Hawking radiation. The state which results after the remnant has evaporated is one which locally appears to be thermal, but which on a much larger scale is marked by extensive correlations

  13. Luminosity lifetime in the Tevatron

    International Nuclear Information System (INIS)

    Jackson, G.; Finley, D.; Johnson, R.P.; Kerns, Q.; McCarthy, J.; Siemann, R.; Zhang, P.

    1988-01-01

    Since the inauguration of colliding proton-antiproton operations in 1987, the Tevatron has exhibited luminosity lifetimes shorter than expected. During a typical colliding beam storage period, called a store, luminosity is calculated periodically by measuring the charge and emittances of each bunch. The growth of the transverse bunch emittances is the dominant cause of luminosity deterioration. Throughout, this period, the position spectrum of the bunches exhibited betatron signals larger than expected from Schottky noise. A model assuming externally driven betatron oscillations explains both the betatron signals and the emittance growth. A program is underway to improve the Tevatron luminosity lifetime. The abort kickers have been identified as sources of emittance growth, and some quadrupole power supplies are further candidates. Because the horizontal dispersion through the RF cavities is nonzero, RF phase noise has been investigated. Noise in the main dipole regulation circuit has also been studied. 13 refs., 4 figs

  14. Angular distributions as lifetime probes

    Energy Technology Data Exchange (ETDEWEB)

    Dror, Jeff Asaf; Grossman, Yuval [Department of Physics, LEPP, Cornell University,Ithaca, NY 14853 (United States)

    2014-06-27

    If new TeV scale particles are discovered, it will be important to determine their width. There is, however, a problematic region, where the width is too small to be determined directly, and too large to generate a secondary vertex. For a collection of colored, spin polarized particles, hadronization depolarizes the particles prior to their decay. The amount of depolarization can be used to probe the lifetime in the problematic region. In this paper we apply this method to a realistic scenario of a top-like particle that can be produced at the LHC. We study how depolarization affects the angular distributions of the decay products and derive an equation for the distributions that is sensitive to the lifetime.

  15. Fluorescence microscopy.

    Science.gov (United States)

    Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D

    2014-10-01

    Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

  16. Maintenance engineering of lifetime management programs

    International Nuclear Information System (INIS)

    Hervia Ruperez, F.

    1997-01-01

    The complexity of nuclear power plants obliges to stablish the adecuated management of its lifetime. This article describes the methodologies and the improvement the evaluation of lifetime programs and specially in Garona and Vandellos II Nuclear Power Plants. (Author)

  17. Review of charm and beauty lifetimes

    International Nuclear Information System (INIS)

    Cheung, Harry W. K.

    1999-01-01

    A review of the latest experimental results on charm and beauty particle lifetimes is presented together with a brief summary of measurement methods used for beauty particle lifetime measurements. There have been significant updates to the D s + /D 0 , B + /B d 0 and Λ b 0 /B d 0 lifetime ratios which have some theoretical implications. However more precise measurements are still needed before one can make conclusive statements about the theory used to calculate the particle lifetimes

  18. CalCOFI Egg Counts

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Fish egg counts and standardized counts for eggs captured in CalCOFI icthyoplankton nets (primarily vertical [Calvet or Pairovet], oblique [bongo or ring nets], and...

  19. On luminescence lifetimes in quartz

    International Nuclear Information System (INIS)

    Chithambo, M.L.; Galloway, R.B.

    2000-01-01

    In this paper we present results of investigations concerning the time dependence of luminescence emission relative to the time of stimulation in quartz. Measurements of time-resolved spectra were performed on a new versatile pulsed light emitting diode system using 525 nm stimulation, an 11 μs duration pulse, a repetition rate of 11 kHz and a 64 μs dynamic range. Effects on luminescence lifetime resulting from sample treatments such as optical stimulation, irradiation, and preheating, are reported

  20. Lifetime measurement in 144Gd

    International Nuclear Information System (INIS)

    Jensen, H.J.; Gast, W.; Georgiev, A.; Jaeger, H.M.; Lieder, R.M.; Utzelmann, S.; Gierlik, M.; Morek, T.; Przestrzelska, K.; Rzaca-Urban, T.; Dewald, A.; Kuehn, R.; Meier, C.; Ender, C.; Haertlein, T.

    1996-01-01

    The lifetime measurements of excited states in 144 Gd were carried out using the Koeln RDM-plunger together with the 2 x 3 CLUSTER detector setup in Heidelberg. The nucleus was populated in the 100 Mo( 48 Ti,4n) 144 Gd reaction at a beam energy of 205 MeV giving a recoil velocity of v/c = 2.6 %. Three and higher fold γ-ray coincidences were measured at 12 target-stopper distances ranged from 0 to 400 μm. Both the dipole and quadrupole bands in 144 Gd have been observed. The analysis is in progress

  1. Lifetime of superheated steam components

    International Nuclear Information System (INIS)

    Stoklossa, K.H.; Oude-Hengel, H.H.; Kraechter, H.J.

    1974-01-01

    The current evaluation schemes in use for judging the lifetime expectations of superheated steam components are compared with each other. The influence of pressure and temperature fluctuations, the differences in the strength of the wall, and the spread band of constant-strainrates are critically investigated. The distribution of these contributory effects are demonstrated in the hight of numerous measuring results. As an important supplement to these evaluation schemes a newly developed technique is introduced which is designed to calculate failure probabilities. (orig./RW) [de

  2. The mass-lifetime relation

    Science.gov (United States)

    LoPresto, Michael C.

    2018-05-01

    In a recent "AstroNote," I described a simple exercise on the mass-luminosity relation for main sequence stars as an example of exposing students in a general education science course of lower mathematical level to the use of quantitative skills such as collecting and analyzing data. Here I present another attempt at a meaningful experience for such students that again involves both the gathering and analysis of numerical data and comparison with accepted result, this time on the relationship of the mass and lifetimes of main sequence stars. This experiment can stand alone or be used as an extension of the previous mass-luminosity relationship experiment.

  3. The puzzle of neutron lifetime

    International Nuclear Information System (INIS)

    Paul, Stephan

    2009-01-01

    In this paper we review the role of the neutron lifetime and discuss the present status of measurements. In view of the large discrepancy observed by the two most precise individual measurements so far we describe the different techniques and point out the principle strengths and weaknesses. In particular we discuss the estimation of systematic uncertainties and its correlation to the statistical ones. In order to solve the present puzzle, many new experiments are either ongoing or being proposed. An overview on their possible contribution to this field will be given.

  4. Personality, IQ, and Lifetime Earnings

    DEFF Research Database (Denmark)

    Gensowski, Miriam

    2018-01-01

    This paper estimates the effects of personality traits and IQ on lifetime earnings of the men and women of the Terman study, a high-IQ U.S. sample. Age-by-age earnings profiles allow a study of when personality traits affect earnings most, and for whom the effects are strongest. I document...... a concave life-cycle pattern in the payoffs to personality traits, with the largest effects between the ages of 40 and 60. An interaction of traits with education reveals that personality matters most for highly educated men. The largest effects are found for Conscientiousness, Extraversion...

  5. Lifetime measurement in 136Pm

    International Nuclear Information System (INIS)

    Toney, D.; Zhong, Q.; De Angelis, G.

    2005-01-01

    The aim of the present work is to investigate the electromagnetic transition probabilities in the doublet bands of 136 Pm. These two bands have been observed up to Iπ = (21 + ). Contrary to the case of 134 Pr, the B(M1)/B(E2) ratios take similar values within the error bars in 136 Pm. This is a strong indication that there is considerable difference between the two nuclei. However, a lifetime measurement in 136 Pm is needed to shed light on the scale and the origin of the difference

  6. Nanodiamond arrays on glass for quantification and fluorescence characterisation.

    Science.gov (United States)

    Heffernan, Ashleigh H; Greentree, Andrew D; Gibson, Brant C

    2017-08-23

    Quantifying the variation in emission properties of fluorescent nanodiamonds is important for developing their wide-ranging applicability. Directed self-assembly techniques show promise for positioning nanodiamonds precisely enabling such quantification. Here we show an approach for depositing nanodiamonds in pre-determined arrays which are used to gather statistical information about fluorescent lifetimes. The arrays were created via a layer of photoresist patterned with grids of apertures using electron beam lithography and then drop-cast with nanodiamonds. Electron microscopy revealed a 90% average deposition yield across 3,376 populated array sites, with an average of 20 nanodiamonds per site. Confocal microscopy, optimised for nitrogen vacancy fluorescence collection, revealed a broad distribution of fluorescent lifetimes in agreement with literature. This method for statistically quantifying fluorescent nanoparticles provides a step towards fabrication of hybrid photonic devices for applications from quantum cryptography to sensing.

  7. Counting statistics in radioactivity measurements

    International Nuclear Information System (INIS)

    Martin, J.

    1975-01-01

    The application of statistical methods to radioactivity measurement problems is analyzed in several chapters devoted successively to: the statistical nature of radioactivity counts; the application to radioactive counting of two theoretical probability distributions, Poisson's distribution law and the Laplace-Gauss law; true counting laws; corrections related to the nature of the apparatus; statistical techniques in gamma spectrometry [fr

  8. Do your syringes count?

    International Nuclear Information System (INIS)

    Brewster, K.

    2002-01-01

    Full text: This study was designed to investigate anecdotal evidence that residual Sestamibi (MIBI) activity vaned in certain situations. For rest studies different brands of syringes were tested to see if the residuals varied. The period of time MIBI doses remained in the syringe between dispensing and injection was also considered as a possible source of increased residual counts. Stress Mibi syringe residual activities were measured to assess if the method of stress test affected residual activity. MIBI was reconstituted using 13 Gbq of Technetium in 3mls of normal saline then boiled for 10 minutes. Doses were dispensed according to department protocol and injected via cannula. Residual syringes were collected for three syringe types. In each case the barrel and plunger were measured separately. As the syringe is flushed during the exercise stress test and not the pharmacological stress test the chosen method was recorded. No relationship was demonstrated between the time MIBI remained in a syringe prior to injection and residual activity. Residual activity was not affected by method of stress test used. Actual injected activity can be calculated if the amount of activity remaining in the syringe post injection is known. Imaging time can be adjusted for residual activity to optimise count statistics. Preliminary results in this study indicate there is no difference in residual activity between syringe brands.Copyright (2002) The Australian and New Zealand Society of Nuclear Medicine Inc

  9. Enhancement of uranyl fluorescence using trimesic acid: Ligand sensitization and co-fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Maji, S. [Chemistry Group, Materials Chemistry Division, Indira Gandhi Centre for Atomic Research, Kalpakkam 603102 (India); Viswanathan, K.S., E-mail: vish@igcar.gov.in [Chemistry Group, Materials Chemistry Division, Indira Gandhi Centre for Atomic Research, Kalpakkam 603102 (India)

    2011-09-15

    Trimesic acid (TMA) was shown to sensitize and enhance uranyl fluorescence in aqueous medium, with the enhancement being a maximum at pH 5.0. Fluorescence spectra and lifetime data together suggest that TMA complexes with uranyl (UO{sub 2}{sup 2+}). The fluorescence of UO{sub 2}{sup 2+} in its acid complex is further enhanced by more than two orders of magnitude following the addition of Y{sup 3+}; a process referred to as co-fluorescence, leading to the possibility of detecting uranium at sub ng/mL level. The present study demonstrates, for the first time, fluorescence enhancement of the uranyl species due to co-fluorescence. - Highlights: > Trimesic acid was shown to sensitize and enhance the fluorescence of uranium in aqueous medium. > This ligand also exhibited co-fluorescence of uranium with Y{sup 3+}. > To the best of our knowledge this is the first report of co-fluorescence in uranium. > The enhancement of uranium fluorescence, resulted in detection limits in the ng/mL regime.

  10. Uncertainties in the proton lifetime

    International Nuclear Information System (INIS)

    Ellis, J.; Nanopoulos, D.V.; Rudaz, S.; Gaillard, M.K.

    1980-04-01

    We discuss the masses of the leptoquark bosons m(x) and the proton lifetime in Grand Unified Theories based principally on SU(5). It is emphasized that estimates of m(x) based on the QCD coupling and the fine structure constant are probably more reliable than those using the experimental value of sin 2 theta(w). Uncertainties in the QCD Λ parameter and the correct value of α are discussed. We estimate higher order effects on the evolution of coupling constants in a momentum space renormalization scheme. It is shown that increasing the number of generations of fermions beyond the minimal three increases m(X) by almost a factor of 2 per generation. Additional uncertainties exist for each generation of technifermions that may exist. We discuss and discount the possibility that proton decay could be 'Cabibbo-rotated' away, and a speculation that Lorentz invariance may be violated in proton decay at a detectable level. We estimate that in the absence of any substantial new physics beyond that in the minimal SU(5) model the proton lifetimes is 8 x 10 30+-2 years

  11. Fluorescent nanoparticles for intracellular sensing: a review.

    Science.gov (United States)

    Ruedas-Rama, Maria J; Walters, Jamie D; Orte, Angel; Hall, Elizabeth A H

    2012-11-02

    Fluorescent nanoparticles (NPs), including semiconductor NPs (Quantum Dots), metal NPs, silica NPs, polymer NPs, etc., have been a major focus of research and development during the past decade. The fluorescent nanoparticles show unique chemical and optical properties, such as brighter fluorescence, higher photostability and higher biocompatibility, compared to classical fluorescent organic dyes. Moreover, the nanoparticles can also act as multivalent scaffolds for the realization of supramolecular assemblies, since their high surface to volume ratio allow distinct spatial domains to be functionalized, which can provide a versatile synthetic platform for the implementation of different sensing schemes. Their excellent properties make them one of the most useful tools that chemistry has supplied to biomedical research, enabling the intracellular monitoring of many different species for medical and biological purposes. In this review, we focus on the developments and analytical applications of fluorescent nanoparticles in chemical and biological sensing within the intracellular environment. The review also points out the great potential of fluorescent NPs for fluorescence lifetime imaging microscopy (FLIM). Finally, we also give an overview of the current methods for delivering of fluorescent NPs into cells, where critically examine the benefits and liabilities of each strategy. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. Modal Logics with Counting

    Science.gov (United States)

    Areces, Carlos; Hoffmann, Guillaume; Denis, Alexandre

    We present a modal language that includes explicit operators to count the number of elements that a model might include in the extension of a formula, and we discuss how this logic has been previously investigated under different guises. We show that the language is related to graded modalities and to hybrid logics. We illustrate a possible application of the language to the treatment of plural objects and queries in natural language. We investigate the expressive power of this logic via bisimulations, discuss the complexity of its satisfiability problem, define a new reasoning task that retrieves the cardinality bound of the extension of a given input formula, and provide an algorithm to solve it.

  13. Digital coincidence counting

    International Nuclear Information System (INIS)

    Buckman, S.M.; Ius, D.

    1996-01-01

    This paper reports on the development of a digital coincidence-counting system which comprises a custom-built data acquisition card and associated PC software. The system has been designed to digitise the pulse-trains from two radiation detectors at a rate of 20 MSamples/s with 12-bit resolution. Through hardware compression of the data, the system can continuously record both individual pulse-shapes and the time intervals between pulses. Software-based circuits are used to process the stored pulse trains. These circuits are constructed simply by linking together icons representing various components such as coincidence mixers, time delays, single-channel analysers, deadtimes and scalers. This system enables a pair of pulse trains to be processed repeatedly using any number of different methods. Some preliminary results are presented in order to demonstrate the versatility and efficiency of this new method. (orig.)

  14. Digital coincidence counting

    Science.gov (United States)

    Buckman, S. M.; Ius, D.

    1996-02-01

    This paper reports on the development of a digital coincidence-counting system which comprises a custom-built data acquisition card and associated PC software. The system has been designed to digitise the pulse-trains from two radiation detectors at a rate of 20 MSamples/s with 12-bit resolution. Through hardware compression of the data, the system can continuously record both individual pulse-shapes and the time intervals between pulses. Software-based circuits are used to process the stored pulse trains. These circuits are constructed simply by linking together icons representing various components such as coincidence mixers, time delays, single-channel analysers, deadtimes and scalers. This system enables a pair of pulse trains to be processed repeatedly using any number of different methods. Some preliminary results are presented in order to demonstrate the versatility and efficiency of this new method.

  15. Steady state and time-resolved fluorescence spectroscopy of quinine sulfate dication bound to sodium dodecylsulfate micelles: Fluorescent complex formation

    Energy Technology Data Exchange (ETDEWEB)

    Joshi, Sunita; Pant, Debi D., E-mail: ddpant@pilani.bits-pilani.ac.in

    2014-01-15

    Interaction of quinine sulfate dication (QSD) with anionic, sodium dodecylsulphate (SDS) surfactant has been studied at different premicellar, micellar and postmicellar concentrations in aqueous phase using steady state, time-resolved fluorescence and fluorescence anisotropy techniques. At premicellar concentrations of SDS, the decrease in absorbance, appearance of an extra fluorescence band at lower wavelengths and tri-exponential decay behavior of fluorescence, are attributed to complex formation between QSD molecules and surfactant monomers. At postmicellar concentrations the red shift in fluorescence spectrum, increase in quantum yield and increase in fluorescence lifetimes are attributed to incorporation of solute molecules to micelles. At lower concentrations of SDS, a large shift in fluorescence is observed on excitation at the red edge of absorption spectrum and this is explained in terms of distribution of ion pairs of different energies in the ground state and the observed fluorescence lifetime behavior corroborates with this model. The temporal fluorescence anisotropy decay of QSD in SDS micelles allowed determination of restriction on the motion of the fluorophore. All the different techniques used in this study reveal that the photophysics of QSD is very sensitive to the microenvironments of SDS micelles and QSD molecules reside at the water-micelle interface. -- Highlights: • Probe molecule is very sensitive to microenvironment of micelles. • Highly fluorescent ion-pair formation has been observed. • Modulated photophysics of probe molecule in micellar solutions has been observed. • Probe molecules strongly bind with micelles and reside at probe–micelle interface.

  16. Combination of confocal principle and aperture stop separation improves suppression of crystalline lens fluorescence in an eye model

    OpenAIRE

    Klemm, Matthias; Blum, Johannes; Link, Dietmar; Hammer, Martin; Haueisen, Jens; Schweitzer, Dietrich

    2016-01-01

    Fluorescence lifetime imaging ophthalmoscopy (FLIO) is a new technique to detect changes in the human retina. The autofluorescence decay over time, generated by endogenous fluorophores, is measured in vivo. The strong autofluorescence of the crystalline lens, however, superimposes the intensity decay of the retina fluorescence, as the confocal principle is not able to suppress it sufficiently. Thus, the crystalline lens autofluorescence causes artifacts in the retinal fluorescence lifetimes d...

  17. Counting statistics in low level radioactivity measurements fluctuating counting efficiency

    International Nuclear Information System (INIS)

    Pazdur, M.F.

    1976-01-01

    A divergence between the probability distribution of the number of nuclear disintegrations and the number of observed counts, caused by counting efficiency fluctuation, is discussed. The negative binominal distribution is proposed to describe the probability distribution of the number of counts, instead of Poisson distribution, which is assumed to hold for the number of nuclear disintegrations only. From actual measurements the r.m.s. amplitude of counting efficiency fluctuation is estimated. Some consequences of counting efficiency fluctuation are investigated and the corresponding formulae are derived: (1) for detection limit as a function of the number of partial measurements and the relative amplitude of counting efficiency fluctuation, and (2) for optimum allocation of the number of partial measurements between sample and background. (author)

  18. Personality, IQ, and Lifetime Earnings

    DEFF Research Database (Denmark)

    Gensowski, Miriam

    2014-01-01

    Talented individuals are seen as drivers of long-term growth, but how do they realize their full potential? In this paper, I show that lifetime earnings of high-IQ men and women are substantially influenced by their personality traits, in addition to intelligence and education. Personality traits......, as identified in a factor model, significantly affect earnings, but not for young workers. The effects are furthermore heterogeneous by educational attainment. For women, personality traits do not affect family earnings in the same way as own earnings. Personality and IQ also influence earnings indirectly...... through education, which has sizeable positive rates of return for men in this sample. Women’s returns to education past a bachelor’s degree are lowered through worse marriage prospects, which offset gains to education in terms of own earnings. The causal effect of education is identified through matching...

  19. Measurement of the BS lifetime

    International Nuclear Information System (INIS)

    Siccama, I.

    1996-01-01

    This thesis presents a measurement of the B s lifetime using 3 million hadronic Z decays collected by the DELPHI detector at LEP from 1991 to 1994. Decays of B s mesons are tagged by the reconstruction of a D s - →φπ - or D s - →K *0 K - decay (including the charge conjugated states of these decay modes). The decay time is obtained by reconstructing both the B s momentum and the B s flight distance. The combined result for the D s -lepton and D s -hadron samples is: τ(B s )=1.54±0.31±0.15 ps where the first error is statistical and the second is systematic. (orig./HSI)

  20. Estimation of atomic interaction parameters by photon counting

    DEFF Research Database (Denmark)

    Kiilerich, Alexander Holm; Mølmer, Klaus

    2014-01-01

    Detection of radiation signals is at the heart of precision metrology and sensing. In this article we show how the fluctuations in photon counting signals can be exploited to optimally extract information about the physical parameters that govern the dynamics of the emitter. For a simple two......-level emitter subject to photon counting, we show that the Fisher information and the Cram\\'er- Rao sensitivity bound based on the full detection record can be evaluated from the waiting time distribution in the fluorescence signal which can, in turn, be calculated for both perfect and imperfect detectors...

  1. Advanced Photon Counting Imaging Detectors with 100ps Timing for Astronomical and Space Sensing Applications

    Science.gov (United States)

    Siegmund, O.; Vallerga, J.; Welsh, B.; Rabin, M.; Bloch, J.

    In recent years EAG has implemented a variety of high-resolution, large format, photon-counting MCP detectors in space instrumentation for satellite FUSE, GALEX, IMAGE, SOHO, HST-COS, rocket, and shuttle payloads. Our scheme of choice has been delay line readouts encoding photon event position centroids, by determination of the difference in arrival time of the event charge at the two ends of a distributed resistive-capacitive (RC) delay line. Our most commonly used delay line configuration is the cross delay line (XDL). In its simplest form the delay-line encoding electronics consists of a fast amplifier for each end of the delay line, followed by time-to-digital converters (TDC's). We have achieved resolutions of Pulsar with a telescope as small as 1m. Although microchannel plate delay line detectors meet many of the imaging and timing demands of various applications, they have limitations. The relatively high gain (107) reduces lifetime and local counting rate, and the fixed delay (10's of ns) makes multiple simultaneous event recording problematic. To overcome these limitations we have begun development of cross strip readout anodes for microchannel plate detectors. The cross strip (XS) anode is a coarse (~0.5 mm) multi-layer metal and ceramic pattern of crossed fingers on an alumina substrate. The charge cloud is matched to the anode period so that it is collected on several neighboring fingers to ensure an accurate event charge centroid can be determined. Each finger of the anode is connected to a low noise charge sensitive amplifier and followed by subsequent A/D conversion of individual strip charge values and a hardware centroid determination of better than 1/100 of a strip are possible. Recently we have commissioned a full 32 x 32 mm XS open face laboratory detector and demonstrated excellent resolution (Los Alamos National Laboratory, NASA and NSF we are developing high rate (>107 Hz) XS encoding electronics that will encode temporally simultaneous

  2. Lifetime of Organic Photovoltaics: Status and Predictions

    DEFF Research Database (Denmark)

    Gevorgyan, Suren; Madsen, Morten Vesterager; Roth, Bérenger

    2016-01-01

    The results of a meta-analysis conducted on organic photovoltaics (OPV) lifetime data reported in the literature is presented through the compilation of an extensive OPV lifetime database based on a large number of articles, followed by analysis of the large body of data. We fully reveal the prog......The results of a meta-analysis conducted on organic photovoltaics (OPV) lifetime data reported in the literature is presented through the compilation of an extensive OPV lifetime database based on a large number of articles, followed by analysis of the large body of data. We fully reveal...... the progress of reported OPV lifetimes. Furthermore, a generic lifetime marker has been defi ned, which helps with gauging and comparing the performance of different architectures and materials from the perspective of device stability. Based on the analysis, conclusions are drawn on the bottlenecks...

  3. Let's Make Data Count

    Science.gov (United States)

    Budden, A. E.; Abrams, S.; Chodacki, J.; Cruse, P.; Fenner, M.; Jones, M. B.; Lowenberg, D.; Rueda, L.; Vieglais, D.

    2017-12-01

    The impact of research has traditionally been measured by citations to journal publications and used extensively for evaluation and assessment in academia, but this process misses the impact and reach of data and software as first-class scientific products. For traditional publications, Article-Level Metrics (ALM) capture the multitude of ways in which research is disseminated and used, such as references and citations within social media and other journal articles. Here we present on the extension of usage and citation metrics collection to include other artifacts of research, namely datasets. The Make Data Count (MDC) project will enable measuring the impact of research data in a manner similar to what is currently done with publications. Data-level metrics (DLM) are a multidimensional suite of indicators measuring the broad reach and use of data as legitimate research outputs. By making data metrics openly available for reuse in a number of different ways, the MDC project represents an important first step on the path towards the full integration of data metrics into the research data management ecosystem. By assuring researchers that their contributions to scholarly progress represented by data corpora are acknowledged, data level metrics provide a foundation for streamlining the advancement of knowledge by actively promoting desirable best practices regarding research data management, publication, and sharing.

  4. LAWRENCE RADIATION LABORATORY COUNTING HANDBOOK

    Energy Technology Data Exchange (ETDEWEB)

    Group, Nuclear Instrumentation

    1966-10-01

    The Counting Handbook is a compilation of operational techniques and performance specifications on counting equipment in use at the Lawrence Radiation Laboratory, Berkeley. Counting notes have been written from the viewpoint of the user rather than that of the designer or maintenance man. The only maintenance instructions that have been included are those that can easily be performed by the experimenter to assure that the equipment is operating properly.

  5. The Susquehanna plant lifetime excellence program

    International Nuclear Information System (INIS)

    McNamara, R.W.

    1988-01-01

    This paper discusses how the Susquehanna plant lifetime excellence program (SPLEX) blends many of the objectives of a new managing for excellence program with plant life extension objectives to achieve excellence in the lifetime operation and availability of the two-unit Susquehanna steam electric station. Investments in lifetime excellence improvements will provide near-term, as well as plant life extension, benefits. A high-quality lifetime experience record, together with extensive, periodic technical assessments and cost-benefit analyses, will provide conclusive justification for future extensions of the unit operating licenses

  6. SUMS Counts-Related Projects

    Data.gov (United States)

    Social Security Administration — Staging Instance for all SUMs Counts related projects including: Redeterminations/Limited Issue, Continuing Disability Resolution, CDR Performance Measures, Initial...

  7. Fluorescence spectroscopy and confocal microscopy of the mycotoxin citrinin in condensed phase and hydrogel films.

    Science.gov (United States)

    Lauer, Milena H; Gehlen, Marcelo H; de Jesus, Karen; Berlinck, Roberto G S

    2014-05-01

    The emission spectra, quantum yields and fluorescence lifetimes of citrinin in organic solvents and hydrogel films have been determined. Citrinin shows complex fluorescence decays due to the presence of two tautomers in solution and interconversion from excited-state double proton transfer (ESDPT) process. The fluorescence decay times associated with the two tautomers have values near 1 and 5 ns depending on the medium. In hydrogel films of agarose and alginate, fluorescence imaging showed that citrinin is not homogeneously dispersed and highly emissive micrometer spots may be formed. Fluorescence spectrum and decay analysis are used to recognize the presence of citrinin in hydrogel films using confocal fluorescence microscopy and spectroscopy.

  8. Compton suppression gamma-counting: The effect of count rate

    Science.gov (United States)

    Millard, H.T.

    1984-01-01

    Past research has shown that anti-coincidence shielded Ge(Li) spectrometers enhanced the signal-to-background ratios for gamma-photopeaks, which are situated on high Compton backgrounds. Ordinarily, an anti- or non-coincidence spectrum (A) and a coincidence spectrum (C) are collected simultaneously with these systems. To be useful in neutron activation analysis (NAA), the fractions of the photopeak counts routed to the two spectra must be constant from sample to sample to variations must be corrected quantitatively. Most Compton suppression counting has been done at low count rate, but in NAA applications, count rates may be much higher. To operate over the wider dynamic range, the effect of count rate on the ratio of the photopeak counts in the two spectra (A/C) was studied. It was found that as the count rate increases, A/C decreases for gammas not coincident with other gammas from the same decay. For gammas coincident with other gammas, A/C increases to a maximum and then decreases. These results suggest that calibration curves are required to correct photopeak areas so quantitative data can be obtained at higher count rates. ?? 1984.

  9. Lifetimes for some excited states of sodium; Vidas medias de niveles excitados del atomo del sodio

    Energy Technology Data Exchange (ETDEWEB)

    Thomas, P; Campos, J

    1979-07-01

    The lifetimes of some s,p and d levels of sodium have been measured by the delayed coincidence method, using a single-photon counting technique. The results are compared with the calculated values of the present work, and with other results. The lifetimes of the ns, np, and levels up to n10; of the nf levels up to n-9;and of the ng, nh,n1 and nk levels up to n-8, have been calculated and the transition probabilities of lines with origin in these levels are given. (Author) 38 refs.

  10. Double-gated spectral snapshots for biomolecular fluorescence

    International Nuclear Information System (INIS)

    Nakamura, Ryosuke; Hamada, Norio; Ichida, Hideki; Tokunaga, Fumio; Kanematsu, Yasuo

    2007-01-01

    A versatile method to take femtosecond spectral snapshots of fluorescence has been developed based on a double gating technique in the combination of an optical Kerr gate and an image intensifier as an electrically driven gate set in front of a charge-coupled device detector. The application of a conventional optical-Kerr-gate method is limited to molecules with the short fluorescence lifetime up to a few hundred picoseconds, because long-lifetime fluorescence itself behaves as a source of the background signal due to insufficiency of the extinction ratio of polarizers employed for the Kerr gate. By using the image intensifier with the gate time of 200 ps, we have successfully suppressed the background signal and overcome the application limit of optical-Kerr-gate method. The system performance has been demonstrated by measuring time-resolved fluorescence spectra for laser dye solution and the riboflavin solution as a typical sample of biomolecule

  11. Arraycount, an algorithm for automatic cell counting in microwell arrays

    OpenAIRE

    Kachouie, Nezamoddin N.; Kang, Lifeng; Khademhosseini, Ali

    2009-01-01

    Microscale technologies have emerged as a powerful tool for studying and manipulating biological systems and miniaturizing experiments. However, the lack of software complementing these techniques has made it difficult to apply them for many high-throughput experiments. This work establishes Arraycount, an approach to automatically count cells in microwell arrays. The procedure consists of fluorescent microscope imaging of cells that are seeded in microwells of a microarray system and then an...

  12. Fusion-component lifetime analysis

    International Nuclear Information System (INIS)

    Mattas, R.F.

    1982-09-01

    A one-dimensional computer code has been developed to examine the lifetime of first-wall and impurity-control components. The code incorporates the operating and design parameters, the material characteristics, and the appropriate failure criteria for the individual components. The major emphasis of the modeling effort has been to calculate the temperature-stress-strain-radiation effects history of a component so that the synergystic effects between sputtering erosion, swelling, creep, fatigue, and crack growth can be examined. The general forms of the property equations are the same for all materials in order to provide the greatest flexibility for materials selection in the code. The individual coefficients within the equations are different for each material. The code is capable of determining the behavior of a plate, composed of either a single or dual material structure, that is either totally constrained or constrained from bending but not from expansion. The code has been utilized to analyze the first walls for FED/INTOR and DEMO and to analyze the limiter for FED/INTOR

  13. Aspects of silicon bulk lifetimes

    Science.gov (United States)

    Landsberg, P. T.

    1985-01-01

    The best lifetimes attained for bulk crytalline silicon as a function of doping concentrations are analyzed. It is assumed that the dopants which set the Fermi level do not contribute to the recombination traffic which is due to the unknown defect. This defect is assumed to have two charge states: neutral and negative, the neutral defect concentration is frozen-in at some temperature T sub f. The higher doping concentrations should include the band-band Auger effect by using a generalization of the Shockley-Read-Hall (SRH) mechanism. The generalization of the SRH mechanism is discussed. This formulation gives a straightforward procedure for incorporating both band-band and band-trap Auger effects in the SRH procedure. Two related questions arise in this context: (1) it may sometimes be useful to write the steady-state occupation probability of the traps implied by SRH procedure in a form which approximates to the Fermi-Dirac distribution; and (2) the effect on the SRH mechanism of spreading N sub t levels at one energy uniformly over a range of energies is discussed.

  14. Track counting in radon dosimetry

    International Nuclear Information System (INIS)

    Fesenbeck, Ingo; Koehler, Bernd; Reichert, Klaus-Martin

    2013-01-01

    The newly developed, computer-controlled track counting system is capable of imaging and analyzing the entire area of nuclear track detectors. The high optical resolution allows a new analysis approach for the process of automated counting using digital image processing technologies. This way, higher exposed detectors can be evaluated reliably by an automated process as well. (orig.)

  15. Baselines for Lifetime of Organic Solar Cells

    DEFF Research Database (Denmark)

    Gevorgyan, Suren; Espinosa Martinez, Nieves; Ciammaruchi, Laura

    2016-01-01

    The process of accurately gauging lifetime improvements in organic photovoltaics (OPVs) or other similar emerging technologies, such as perovskites solar cells is still a major challenge. The presented work is part of a larger effort of developing a worldwide database of lifetimes that can help...

  16. Maximizing System Lifetime by Battery Scheduling

    NARCIS (Netherlands)

    Jongerden, M.R.; Haverkort, Boudewijn R.H.M.; Bohnenkamp, H.C.; Katoen, Joost P.

    2009-01-01

    The use of mobile devices is limited by the battery lifetime. Some devices have the option to connect an extra battery, or to use smart battery-packs with multiple cells to extend the lifetime. In these cases, scheduling the batteries over the load to exploit recovery properties usually extends the

  17. Galaxy number counts: Pt. 2

    International Nuclear Information System (INIS)

    Metcalfe, N.; Shanks, T.; Fong, R.; Jones, L.R.

    1991-01-01

    Using the Prime Focus CCD Camera at the Isaac Newton Telescope we have determined the form of the B and R galaxy number-magnitude count relations in 12 independent fields for 21 m ccd m and 19 m ccd m 5. The average galaxy count relations lie in the middle of the wide range previously encompassed by photographic data. The field-to-field variation of the counts is small enough to define the faint (B m 5) galaxy count to ±10 per cent and this variation is consistent with that expected from galaxy clustering considerations. Our new data confirm that the B, and also the R, galaxy counts show evidence for strong galaxy luminosity evolution, and that the majority of the evolving galaxies are of moderately blue colour. (author)

  18. Hanford whole body counting manual

    International Nuclear Information System (INIS)

    Palmer, H.E.; Brim, C.P.; Rieksts, G.A.; Rhoads, M.C.

    1987-05-01

    This document, a reprint of the Whole Body Counting Manual, was compiled to train personnel, document operation procedures, and outline quality assurance procedures. The current manual contains information on: the location, availability, and scope of services of Hanford's whole body counting facilities; the administrative aspect of the whole body counting operation; Hanford's whole body counting facilities; the step-by-step procedure involved in the different types of in vivo measurements; the detectors, preamplifiers and amplifiers, and spectroscopy equipment; the quality assurance aspect of equipment calibration and recordkeeping; data processing, record storage, results verification, report preparation, count summaries, and unit cost accounting; and the topics of minimum detectable amount and measurement accuracy and precision. 12 refs., 13 tabs

  19. Evidence of shallow positron traps in ion-implanted InP observed by maximum entropy reconstruction of positron lifetime distribution: a test of MELT

    International Nuclear Information System (INIS)

    Chen, Z.Q.; Wang, S.J.

    1999-01-01

    A newly developed maximum entropy method, which was realized by the computer program MELT introduced by Shukla et al., was used to analyze positron lifetime spectra measured in semiconductors. Several simulation studies were done to test the performance of this algorithm. Reliable reconstruction of positron lifetime distributions can be extracted at relatively lower counts, which shows the applicability and superiority of this method. Two positron lifetime spectra measured in ion-implanted p-InP(Zn) at 140 and 280 K, respectively were analyzed by this program. The lifetime distribution differed greatly for the two temperatures, giving direct evidence of the existence of shallow positron traps at low temperature

  20. Measuring upconversion nanoparticles photoluminescence lifetime with FastFLIM and phasor plots

    Science.gov (United States)

    Sun, Yuansheng; Lee, Hsien-Ming; Qiu, Hailin; Liao, Shih-Chu Jeff; Coskun, Ulas; Barbieri, Beniamino

    2018-02-01

    Photon upconversion is a nonlinear process in which the sequential of absorption of two or more photons leads to the anti-stoke emission. Different than the conventional multiphoton excitation process, upconversion can be efficiently performed at low excitation densities. Recent developments in lanthanide-doped upconversion nanoparticles (UCNPs) have led to a diversity of applications, including detecting and sensing of biomolecules, imaging of live cells, tissues and animals, cancer diagnostic and therapy, etc. Measuring the upconversion lifetime provides a new dimension of its imaging and opens a new window for its applications. Due to the long metastable intermediate excited state, UCNP typically has a long excited state lifetime ranging from sub-microseconds to milliseconds. Here, we present a novel development using the FastFLIM technique to measure UCNP lifetime by laser scanning confocal microscopy. FastFLIM is capable of measuring lifetime from 100 ps to 100 ms and features the high data collection efficiency (up to 140-million counts per second). Other than the traditional nonlinear least-square fitting analysis, the raw data acquired by FastFLIM can be directly processed by the model-free phasor plots approach for instant and unbiased lifetime results, providing the ideal routine for the UCNP photoluminescence lifetime microscopy imaging.

  1. Autofluorescence lifetime imaging during transoral robotic surgery: a clinical validation study of tumor detection (Conference Presentation)

    Science.gov (United States)

    Lagarto, João. L.; Phipps, Jennifer E.; Unger, Jakob; Faller, Leta M.; Gorpas, Dimitris; Ma, Dinglong M.; Bec, Julien; Moore, Michael G.; Bewley, Arnaud F.; Yankelevich, Diego R.; Sorger, Jonathan M.; Farwell, Gregory D.; Marcu, Laura

    2017-02-01

    Autofluorescence lifetime spectroscopy is a promising non-invasive label-free tool for characterization of biological tissues and shows potential to report structural and biochemical alterations in tissue owing to pathological transformations. In particular, when combined with fiber-optic based instruments, autofluorescence lifetime measurements can enhance intraoperative diagnosis and provide guidance in surgical procedures. We investigate the potential of a fiber-optic based multi-spectral time-resolved fluorescence spectroscopy instrument to characterize the autofluorescence fingerprint associated with histologic, morphologic and metabolic changes in tissue that can provide real-time contrast between healthy and tumor regions in vivo and guide clinicians during resection of diseased areas during transoral robotic surgery. To provide immediate feedback to the surgeons, we employ tracking of an aiming beam that co-registers our point measurements with the robot camera images and allows visualization of the surgical area augmented with autofluorescence lifetime data in the surgeon's console in real-time. For each patient, autofluorescence lifetime measurements were acquired from normal, diseased and surgically altered tissue, both in vivo (pre- and post-resection) and ex vivo. Initial results indicate tumor and normal regions can be distinguished based on changes in lifetime parameters measured in vivo, when the tumor is located superficially. In particular, results show that autofluorescence lifetime of tumor is shorter than that of normal tissue (p robot assisted cancer removal interventions.

  2. In vivo counting of uranium

    International Nuclear Information System (INIS)

    Palmer, H.E.

    1985-03-01

    A state-of-the-art radiation detector system consisting of six individually mounted intrinsic germanium planar detectors, each 20 cm 2 by 13 mm thick, mounted together such that the angle of the whole system can be changed to match the slope of the chest of the person being counted, is described. The sensitivity of the system for counting uranium and plutonium in vivo and the precedures used in calibrating the system are also described. Some results of counts done on uranium mill workers are presented. 15 figs., 2 tabs

  3. Nanostructure induced changes in lifetime and enhanced second-harmonic response of organic-plasmonic hybrids

    Energy Technology Data Exchange (ETDEWEB)

    Leißner, Till [NanoSYD, Mads Clausen Institute, University of Southern Denmark, Alsion 2, 6400 Sønderborg (Denmark); Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230 Odense (Denmark); Kostiučenko, Oksana; Rubahn, Horst-Günter; Fiutowski, Jacek, E-mail: fiutowski@mci.sdu.dk [NanoSYD, Mads Clausen Institute, University of Southern Denmark, Alsion 2, 6400 Sønderborg (Denmark); Brewer, Jonathan R. [Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230 Odense (Denmark)

    2015-12-21

    In this letter we show that the optical response of organic nanofibers, grown from functionalized para-quaterphenylene molecules, can be controlled by forming organic-plasmonic hybrid systems. The interaction between nanofibers and supporting regular arrays of nanostructures leads to a strongly enhanced second harmonic response. At the same time, the fluorescence lifetime of the nanofibers is reduced from 0.32 ns for unstructured gold films to 0.22 ns for gold nanosquare arrays, demonstrating efficient organic–plasmonic interaction. To study the origin of these effects, we applied two-photon laser scanning microscopy and fluorescence lifetime imaging microscopy. These findings provide an effective approach for plasmon-enhanced second-harmonic generation at the nanoscale, which is attractive for nanophotonic circuitry.

  4. Fluorescein Derivatives in Intravital Fluorescence Imaging

    Directory of Open Access Journals (Sweden)

    Michael S. Roberts

    2013-08-01

    Full Text Available Intravital fluorescence microscopy enables the direct imaging of fluorophores in vivo and advanced techniques such as fluorescence lifetime imaging (FLIM enable the simultaneous detection of multiple fluorophores. Consequently, it is now possible to record distribution and metabolism of a chemical in vivo and to optimise the delivery of fluorophores in vivo. Recent clinical applications with fluorescein and other intravital fluorescent stains have occurred in neurosurgery, dermatology [including photodynamic therapy (PDT] and endomicroscopy. Potential uses have been identified in periodontal disease, skin graft and cancer surgery. Animal studies have demonstrated that diseased tissue can be specifically stained with fluorophore conjugates. This review focuses on the fluorescein derived fluorophores in common clinical use and provides examples of novel applications from studies in tissue samples.

  5. Nuclear Power Plant Lifetime Management Study (I)

    Energy Technology Data Exchange (ETDEWEB)

    Hong, Sung Yull; Jeong, Ill Seok; Jang, Chang Heui; Song, Taek Ho; Song, Woo Young [Korea Electric Power Research Institute, Taejon (Korea, Republic of); Jin, Tae Eun [Korea Power Engineering Company Consulting and Architecture Engineers, (Korea, Republic of); Kim, Woo Chul [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of)

    1997-12-31

    As the operation-year of nuclear power plant increases and finding sites for new nuclear power plant becomes harder, a comprehensive and systematic nuclear plant lifetime management(PLIM) program including life extension has to be established for stable and safe supply of electricity. A feasibility study was conducted to systematically evaluate technical, economic and regulatory aspect of plant lifetime managements and plant life extension for Kori-1 nuclear power plant. For technical evaluation of nuclear power plant, 13 major components were selected for lifetime evaluation by screening system. structure, and components(SSCs) of the plant. It was found that except reactor pressure vessel, which needs detailed integrity analysis, and low pressure turbine, which is scheduled to be replaced, 11 out of 13 major components have sufficient service life, for more than 40 years. Because domestic rules and regulations related to license renewal has not yet been written, review on the regulatory aspect of life extensions was conducted using US NRC rules and regulations. A cooperative effort with nuclear regulatory body is needed for early completion of license renewal rules and regulations. For economic evaluation of plant lifetime extension, a computer program was developed and used. It was found that 10 to 20 year of extension operation of Kori-1 nuclear power plant was proved. Based on the results, next phase of plant lifetime management program for detailed lifetime evaluation and presenting detailed implementation schedule for plant refurbishment for lifetime extension should be followed. (author). 74 refs., figs.

  6. Nuclear Power Plant Lifetime Management Study (I)

    Energy Technology Data Exchange (ETDEWEB)

    Hong, Sung Yull; Jeong, Ill Seok; Jang, Chang Heui; Song, Taek Ho; Song, Woo Young [Korea Electric Power Research Institute, Taejon (Korea, Republic of); Jin, Tae Eun [Korea Power Engineering Company Consulting and Architecture Engineers, (Korea, Republic of); Kim, Woo Chul [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of)

    1996-12-31

    As the operation-year of nuclear power plant increases and finding sites for new nuclear power plant becomes harder, a comprehensive and systematic nuclear plant lifetime management(PLIM) program including life extension has to be established for stable and safe supply of electricity. A feasibility study was conducted to systematically evaluate technical, economic and regulatory aspect of plant lifetime managements and plant life extension for Kori-1 nuclear power plant. For technical evaluation of nuclear power plant, 13 major components were selected for lifetime evaluation by screening system. structure, and components(SSCs) of the plant. It was found that except reactor pressure vessel, which needs detailed integrity analysis, and low pressure turbine, which is scheduled to be replaced, 11 out of 13 major components have sufficient service life, for more than 40 years. Because domestic rules and regulations related to license renewal has not yet been written, review on the regulatory aspect of life extensions was conducted using US NRC rules and regulations. A cooperative effort with nuclear regulatory body is needed for early completion of license renewal rules and regulations. For economic evaluation of plant lifetime extension, a computer program was developed and used. It was found that 10 to 20 year of extension operation of Kori-1 nuclear power plant was proved. Based on the results, next phase of plant lifetime management program for detailed lifetime evaluation and presenting detailed implementation schedule for plant refurbishment for lifetime extension should be followed. (author). 74 refs., figs.

  7. Lifetime measurement of ATF damping ring

    International Nuclear Information System (INIS)

    Okugi, T.; Hayano, H.; Kubo, K.; Naito, T.; Terunuma, N.; Urakawa, J.; Zimmermann, F.

    1998-06-01

    The purpose of the ATF damping ring is the development of technologies for producing a low emittance beam required in future linear colliders such as JLC. The lifetime of the damping ring is very short (typically a few minutes). It is limited by elastic beam-gas scattering along with a small dynamic aperture, and by single intra-beam scattering (Touschek effect). The Touschek lifetime strongly depends upon the charge density of the beam, especially, the size of the vertical emittance. In this paper, the authors report the results of beam lifetime measurements in the ATF damping ring and the estimation of the vertical emittance from these measurements

  8. Lifetime of B hadrons from CDF

    International Nuclear Information System (INIS)

    Miao, Ting.

    1996-08-01

    A review of the lifetimes of B hadrons measured by the CDF collaboration at Fermilab is presented. The data corresponds to 110 pb -1 of p anti p collisions at √s = 1.8 TeV. The inclusive B hadron lifetime is measured using a high statistics sample of B → J/ΨΧ decays. Species specific lifetimes of the B + , B 0 , B 0 s , and Λ 0 b are determined using both fully reconstructed decays and partially reconstructed decays consisting of a lepton associated with a charm hadron

  9. Models for Battery Reliability and Lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Smith, K.; Wood, E.; Santhanagopalan, S.; Kim, G. H.; Neubauer, J.; Pesaran, A.

    2014-03-01

    Models describing battery degradation physics are needed to more accurately understand how battery usage and next-generation battery designs can be optimized for performance and lifetime. Such lifetime models may also reduce the cost of battery aging experiments and shorten the time required to validate battery lifetime. Models for chemical degradation and mechanical stress are reviewed. Experimental analysis of aging data from a commercial iron-phosphate lithium-ion (Li-ion) cell elucidates the relative importance of several mechanical stress-induced degradation mechanisms.

  10. Statistical Models and Methods for Lifetime Data

    CERN Document Server

    Lawless, Jerald F

    2011-01-01

    Praise for the First Edition"An indispensable addition to any serious collection on lifetime data analysis and . . . a valuable contribution to the statistical literature. Highly recommended . . ."-Choice"This is an important book, which will appeal to statisticians working on survival analysis problems."-Biometrics"A thorough, unified treatment of statistical models and methods used in the analysis of lifetime data . . . this is a highly competent and agreeable statistical textbook."-Statistics in MedicineThe statistical analysis of lifetime or response time data is a key tool in engineering,

  11. Study on lifetime of C stripping foils

    International Nuclear Information System (INIS)

    Zhang Hongbin; Lu Ziwei; Zhao Yongtao; Li Zhankui; Xu Hushan; Xiao Guoqing; Wang Yuyu; Zhang Ling; Li Longcai; Fang Yan

    2007-01-01

    The carbon stripping foils can be prepared with the AC and DC arc discharge methods, or even sandwiched with AC-DC alternative layers. The lifetime of the carbon stripping foils of 19 μg/cm 2 prepared with different methods and/or structures was measured. The factors affecting the bombarding lifetime of the carbon stripping foils, especially the method of the foil preparation and the structure of the carbon stripping foils, were discussed. It is observed that the foils prepared with the DC arc discharge method have a longer bombarding lifetime than those prepared with the AC arc discharge method. (authors)

  12. Photoluminescence decay lifetime measurements of hemicyanine derivatives of different alkyl chain lengths

    International Nuclear Information System (INIS)

    Shim, Taekyu; Lee, Myounghee; Kim, Sungho; Sung, Jaeho; Rhee, Bum Ku; Kim, Doseok; Kim, Hyunsung; Yoon, Kyung Byung

    2004-01-01

    The fluorescence upconversion setup for the detection of photoluminescence (PL) decay lifetime with subpicosecond time resolution was constructed, and the photoluminescence phenomena of several hemicyanine dyes with alkyl chains of different chain lengths tethered to the N atom of the pyridine moiety (HC-n, n=6, 15, 22) in methanol were investigated. The average decay lifetimes of the solutions determined from the measured data by multi-order exponential decay curve fitting were ∼27 ps at the PL peak wavelength. It was found that the PL decay properties did not depend on the alkyl chain length in the molecule, implying that the twist of the alkylpyridinium ring of the molecule is not possible as a nonfluorescing relaxation pathway. The time-dependent PL spectra constructed from the PL lifetime data showed the dynamic Stokes shift of ∼1000 cm -1

  13. Lifetimes of partial charge transfer exciplexes of 9-cyanophenanthrene and 9-cyanoanthracene

    OpenAIRE

    Dolotova, Elena; Dogadkin, Denis; Soboleva, Irina; Kuzmin, Michael; Nicolet, Olivier; Vauthey, Eric

    2003-01-01

    The fluorescence decays of several exciplexes with partial charge transfer have been investigated in solvents of various polarity. The measured lifetimes are found to be in reasonable agreement with the activation enthalpy and entropy of exciplex decay obtained earlier from the temperature dependence of the exciplex emission quantum yields. For exciplexes with 9-cyanophenanthrene substantial contribution of the higher local excited state into the exciplex electronic structure is found and bor...

  14. Origins of fluorescence in evolved bacteriophytochromes.

    Science.gov (United States)

    Bhattacharya, Shyamosree; Auldridge, Michele E; Lehtivuori, Heli; Ihalainen, Janne A; Forest, Katrina T

    2014-11-14

    Use of fluorescent proteins to study in vivo processes in mammals requires near-infrared (NIR) biomarkers that exploit the ability of light in this range to penetrate tissue. Bacteriophytochromes (BphPs) are photoreceptors that couple absorbance of NIR light to photoisomerization, protein conformational changes, and signal transduction. BphPs have been engineered to form NIR fluorophores, including IFP1.4, Wi-Phy, and the iRFP series, initially by replacement of Asp-207 by His. This position was suggestive because its main chain carbonyl is within hydrogen-bonding distance to pyrrole ring nitrogens of the biliverdin chromophore, thus potentially functioning as a crucial transient proton sink during photoconversion. To explain the origin of fluorescence in these phytofluors, we solved the crystal structures of IFP1.4 and a comparison non-fluorescent monomeric phytochrome DrCBDmon. Met-186 and Val-288 in IFP1.4 are responsible for the formation of a tightly packed hydrophobic hub around the biliverdin D ring. Met-186 is also largely responsible for the blue-shifted IFP1.4 excitation maximum relative to the parent BphP. The structure of IFP1.4 revealed decreased structural heterogeneity and a contraction of two surface regions as direct consequences of side chain substitutions. Unexpectedly, IFP1.4 with Asp-207 reinstalled (IFPrev) has a higher fluorescence quantum yield (∼9%) than most NIR phytofluors published to date. In agreement, fluorescence lifetime measurements confirm the exceptionally long excited state lifetimes, up to 815 ps, in IFP1.4 and IFPrev. Our research helps delineate the origin of fluorescence in engineered BphPs and will facilitate the wide-spread adoption of phytofluors as biomarkers. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  15. Proton detection in the neutron lifetime experiment PENeLOPE

    Energy Technology Data Exchange (ETDEWEB)

    Tietze, Christian [Technische Universitaet Muenchen, Physik Department E18 (Germany); Collaboration: PENeLOPE-Collaboration

    2015-07-01

    Although neutron lifetime plays an important role in the Standard Model of particle physics, τ{sub n} is not very precisely know and often discussed. The official PDG mean value has been lowered during the last years by more than 6σ to the new value of 880.3 ± 1.1 s. The new precision experiment PENeLOPE, which is currently developed at Technische Universitaet Muenchen, will help to clear this up. Ultra-cold neutrons are lossless stored in a magneto-gravitational trap, formed by superconducting coils. The combined determination of τ{sub n} by counting the surviving neutrons after each storage cycle on one side and in-situ detection of the decay protons on the other side together with a very good handle on systematic errors leads to an unprecedented precision of the neutron lifetime value of 0.1s. This contribution will give an overview of the challenges concerning proton detection under the exceptional requirements of this experiment. The developed concept of using avalanche photodiodes for direct proton detection will be presented as well as results from first measurements with a prototype detector read out by particular developed electronics.

  16. Use phase signals to promote lifetime extension for Windows PCs.

    Science.gov (United States)

    Hickey, Stewart; Fitzpatrick, Colin; O'Connell, Maurice; Johnson, Michael

    2009-04-01

    This paper proposes a signaling methodology for personal computers. Signaling may be viewed as an ecodesign strategy that can positively influence the consumer to consumer (C2C) market process. A number of parameters are identified that can provide the basis for signal implementation. These include operating time, operating temperature, operating voltage, power cycle counts, hard disk drive (HDD) self-monitoring, and reporting technology (SMART) attributes and operating system (OS) event information. All these parameters are currently attainable or derivable via embedded technologies in modern desktop systems. A case study detailing a technical implementation of how the development of signals can be achieved in personal computers that incorporate Microsoft Windows operating systems is presented. Collation of lifetime temperature data from a system processor is demonstrated as a possible means of characterizing a usage profile for a desktop system. In addition, event log data is utilized for devising signals indicative of OS quality. The provision of lifetime usage data in the form of intuitive signals indicative of both hardware and software quality can in conjunction with consumer education facilitate an optimal remarketing strategy for used systems. This implementation requires no additional hardware.

  17. Complete Blood Count (For Parents)

    Science.gov (United States)

    ... Kids Deal With Injections and Blood Tests Blood Culture Anemia Blood Test: Basic Metabolic Panel (BMP) Blood Test: Hemoglobin Basic Blood Chemistry Tests Word! Complete Blood Count (CBC) Medical Tests and Procedures ( ...

  18. Make My Trip Count 2015

    Data.gov (United States)

    Allegheny County / City of Pittsburgh / Western PA Regional Data Center — The Make My Trip Count (MMTC) commuter survey, conducted in September and October 2015 by GBA, the Pittsburgh 2030 District, and 10 other regional transportation...

  19. Counting Triangles to Sum Squares

    Science.gov (United States)

    DeMaio, Joe

    2012-01-01

    Counting complete subgraphs of three vertices in complete graphs, yields combinatorial arguments for identities for sums of squares of integers, odd integers, even integers and sums of the triangular numbers.

  20. Counting Word Frequencies with Python

    Directory of Open Access Journals (Sweden)

    William J. Turkel

    2012-07-01

    Full Text Available Your list is now clean enough that you can begin analyzing its contents in meaningful ways. Counting the frequency of specific words in the list can provide illustrative data. Python has an easy way to count frequencies, but it requires the use of a new type of variable: the dictionary. Before you begin working with a dictionary, consider the processes used to calculate frequencies in a list.

  1. Liquid scintillation counting of chlorophyll

    International Nuclear Information System (INIS)

    Fric, F.; Horickova, B.; Haspel-Horvatovic, E.

    1975-01-01

    A precise and reproducible method of liquid scintillation counting was worked out for measuring the radioactivity of 14 C-labelled chlorophyll a and chlorophyll b solutions without previous bleaching. The spurious count rate caused by luminescence of the scintillant-chlorophyll system is eliminated by using a suitable scintillant and by measuring the radioactivity at 4 to 8 0 C after an appropriate time of dark adaptation. Bleaching of the chlorophyll solutions is necessary only for measuring of very low radioactivity. (author)

  2. Long term optical stability of fluorescent solar concentrator plates

    NARCIS (Netherlands)

    Slooff, L.H.; Bakker, N.J.; Sommeling, P.M.; Büchtemann, A.; Wedel, A.; Sark, W.G.J.H.M. van

    2014-01-01

    Fluorescent solar concentrators offer an alternative approach for low-cost photovoltaic energy conversion. For successful application, not only the power conversion efficiency and cost are important, but also lifetime or stability of the devices. As today’s concentrator is made of polymer sheets

  3. Long-term optical stability of fluorescent solar concentrator plates

    NARCIS (Netherlands)

    Slooff, Lenneke H.; Bakker, Nicolaas J.; Sommeling, Paul M.; Büchtemann, Andreas; Wedel, Armin; Van Sark, Wilfried G J H M

    2014-01-01

    Fluorescent solar concentrators offer an alternative approach for low-cost photovoltaic energy conversion. For successful application, not only the power conversion efficiency and cost are important, but also lifetime or stability of the devices. As today's concentrator is made of polymer sheets

  4. Prospects for a new cold neutron beam measurement of the neutron lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Dewey, M., E-mail: mdewey@nist.go [National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States); Coakley, K., E-mail: kevin.coakley@nist.go [National Institute of Standards and Technology, Boulder, CO 80305 (United States); Gilliam, D., E-mail: david.gilliam@nist.go [National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States); Greene, G., E-mail: greenegl@ornl.go [Department of Physics, University of Tennessee, Knoxville, TN 37996 (United States); Physics Division, Oak Ridge National Lab, Building 6010, Oak Ridge, TN 37831 (United States); Laptev, A., E-mail: alaptev@nist.go [Los Alamos National Laboratory, Los Alamos, NM 87545 (United States); Nico, J., E-mail: jnico@nist.go [National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States); Snow, W., E-mail: wsnow@indiana.ed [Indiana University/IUCF, Bloomington, IN 47408 (United States); Wietfeldt, F., E-mail: few@tulane.ed [Tulane University, New Orleans, LA 70118 (United States); Yue, A., E-mail: ayue@nist.go [Department of Physics, University of Tennessee, Knoxville, TN 37996 (United States)

    2009-12-11

    In the most accurate cold neutron beam determination of the neutron lifetime based on the absolute counting of decay protons, the largest uncertainty was attributed to the absolute determination of the capture flux of the cold neutron beam. Currently an experimental effort is underway at the National Institute of Standards and Technology (NIST) that will significantly reduce this contribution to the uncertainty in the lifetime determination. The next largest source of uncertainty is the determination of the absolute count rate of decay protons, which contributes to the experimental uncertainty approximately at the 1 s level. Experience with the recent neutron radiative decay experiment, which used the neutron lifetime apparatus, has provided valuable insights into ways to reduce other uncertainties. In addition, the cold neutron fluence rate at NIST is presently 1.5 times greater than in the 2003 measurement, and there is the prospect for a significantly higher rate with the new guide hall expansion. This paper discusses an approach for achieving a determination of the neutron lifetime with an accuracy of approximately 1 s.

  5. Wide-field fluorescent microscopy and fluorescent imaging flow cytometry on a cell-phone.

    Science.gov (United States)

    Zhu, Hongying; Ozcan, Aydogan

    2013-04-11

    Fluorescent microscopy and flow cytometry are widely used tools in biomedical research and clinical diagnosis. However these devices are in general relatively bulky and costly, making them less effective in the resource limited settings. To potentially address these limitations, we have recently demonstrated the integration of wide-field fluorescent microscopy and imaging flow cytometry tools on cell-phones using compact, light-weight, and cost-effective opto-fluidic attachments. In our flow cytometry design, fluorescently labeled cells are flushed through a microfluidic channel that is positioned above the existing cell-phone camera unit. Battery powered light-emitting diodes (LEDs) are butt-coupled to the side of this microfluidic chip, which effectively acts as a multi-mode slab waveguide, where the excitation light is guided to uniformly excite the fluorescent targets. The cell-phone camera records a time lapse movie of the fluorescent cells flowing through the microfluidic channel, where the digital frames of this movie are processed to count the number of the labeled cells within the target solution of interest. Using a similar opto-fluidic design, we can also image these fluorescently labeled cells in static mode by e.g. sandwiching the fluorescent particles between two glass slides and capturing their fluorescent images using the cell-phone camera, which can achieve a spatial resolution of e.g. - 10 μm over a very large field-of-view of - 81 mm(2). This cell-phone based fluorescent imaging flow cytometry and microscopy platform might be useful especially in resource limited settings, for e.g. counting of CD4+ T cells toward monitoring of HIV+ patients or for detection of water-borne parasites in drinking water.

  6. RDM lifetime measurements in 187Tl

    International Nuclear Information System (INIS)

    Chamoli, S.K.; Joshi, P.; Kumar, A.; Govil, I.M.; Mukherjee, G.; Singh, R.P.; Muralithar, S.; Bhowmik, R.K.

    2003-01-01

    The present work is an attempt to study the shape changes in 187 Tl through a measurement of electromagnetic transition probabilities of the high spin states. The Doppler shifted recoil distance technique was used to measure the lifetimes

  7. Improved b lifetime measurement from MAC

    International Nuclear Information System (INIS)

    Ford, W.T.

    1984-03-01

    Two recent publications, from the MAC and Mark II collaborations, have reported the somewhat surprising result that the lifetime of particles made up of b quarks is in the 1 to 2 picosecond range, or somewhat longer than the lifetimes of charm particles. Although the charm decays are favored transitions while those of b particles depend upon off-diagonal elements of the weak flavor mixing matrix, the smallness of the b decay rates in face of the large available phase space indicates that the off-diagonal elements are indeed very small. The possibility for complete determination of the mixing matrix was brought significantly nearer by the availability of the lifetime information; what is needed now is to reduce the uncertainty of the measurements, which was about 33% for both experiments. We describe here an extension of the b lifetime study with the MAC detector, incorporating some new data and improvements in the analysis. 12 references

  8. Lifetime measurements of excited Co I levels

    CERN Document Server

    Klotz, W D; Gobel, L H

    1977-01-01

    In the region of 3500 AA the lifetimes of eight excited Cobalt I levels have been measured by means of the zero field level crossing method. The measured lifetimes belong to the odd configurations 3d/sup 7/4s4p and 3d/sup 8/4p and are of the accuracy of about 5%. The hyperfine structure of levels with I not=J has to be taken into account in evaluating lifetimes from level crossing data, because the nuclear spin of the natural isotope /sup 59/Co is I=7/2. Therefore the influence of the line profile of the exciting resonance lines on the lifetimes has been investigated. The results are compared with those of other authors. Furthermore absolute oscillator strengths were calculated with known branching ratios and a new absolute scale has been established. (23 refs).

  9. Quantum lifetime in electron storage rings

    International Nuclear Information System (INIS)

    Chao, A.W.

    1977-02-01

    One of the mechanisms which contribute to beam lifetime in electron storage rings is the quantum emission of energetic photons causing particles to be lost from the rf bucket. This quantum lifetime is among other things important in defining the required aperture in a storage ring. An approximate expression of quantum lifetime, predicted by a one-dimensional model which takes into account only the betatron motion, has been used in most storage ring designs. If the beam is aperture-limited at a position with nonzero dispersion, both the betatron and synchrotron motions have to be included and a two-dimensional model must be used. An exact expression of quantum lifetime for the one-dimensional case and an approximate expression for the two-dimensional case are given

  10. Quantum lifetime in electron storage rings

    International Nuclear Information System (INIS)

    Chao, A.W.

    1977-01-01

    One of the mechanisms which contributes to beam lifetime in electron storage rings is the quantum emission of energetic photons causing particles to be lost from the rf bucket. This quantum lifetime is among other things important in defining the required aperture in a storage ring. An approximate expression of quantum lifetime, predicted by a one-dimensional model which takes into account only the betatron motion, has been used in most storage ring designs. If the beam is aperture-limited at a position with nonzero dispersion, both the betatron and synchrotron motions have to be included, and a two-dimensional model must be used. An exact expression of quantum lifetime for the one-dimensional case and an approximate expression for the two-dimensional case are given

  11. Lifetime measurements of hadrons containing heavy quarks

    International Nuclear Information System (INIS)

    Forden, G.E.

    1985-01-01

    Recent lifetime measurements of heavy particles at PETRA and PEP are reviewed. A comparison of the methods used is given. The world averages for the lifetimes of the D 0 and D +- mesons are found to be (tau/dub D/ 0 ) - 3.97 +/- 0.3 x 10 -13 sec and (tau/dub D +-/) = 8.6 +/- 0.7 x 10 -13 sec. This difference in lifetimes is discussed in light of recent information about exclusive decays. The world average for the lifetime of bottom hadrons is determined to be (tau/sub b/) = 11.0 +/- 1.5 x 10 -13 sec and new estimates for the b quark mixing elements, absolute value V/sub bu/ and absolute value V/sub bc/, are given

  12. Lifetime measurement in {sup 195}Po

    Energy Technology Data Exchange (ETDEWEB)

    Grahn, T.; Page, R.D. [University of Liverpool, Department of Physics, Oliver Lodge Laboratory, Liverpool (United Kingdom); Dewald, A.; Jolie, J.; Melon, B.; Pissulla, T. [Universitaet zu Koeln, Institut fuer Kernphysik, Koeln (Germany); Greenlees, P.T.; Jakobsson, U.; Jones, P.; Julin, R.; Juutinen, S.; Ketelhut, S.; Leino, M.; Nyman, M.; Peura, P.; Rahkila, P.; Saren, J.; Scholey, C.; Sorri, J.; Uusitalo, J. [University of Jyvaeskylae, Department of Physics, P.O. Box 35, Jyvaeskylae (Finland); Kroell, T.; Kruecken, R.; Maierbeck, P. [TU Muenchen, Physik-Department E12, Garching (Germany)

    2009-03-15

    The lifetime of the 17/2{sup +} yrast state in {sup 195}Po has been measured using the recoil distance Doppler-shift technique to be {tau}=43(11) ps. The lifetime was extracted from the singles {gamma}-ray spectra obtained by using the recoil-decay tagging method. The present work provides more information of the coupling schemes, shapes and configuration mixing in neutron-deficient odd-mass Po nuclei. (orig.)

  13. The measurement of subnanosecond nuclear lifetimes

    International Nuclear Information System (INIS)

    White, D.C.S.

    1974-01-01

    This research dealt with the measurement of subnanosecond nuclear lifetimes using the pulsed beam delayed-coincidence technique. Measurements were performed on isotopes in the f7/2 shell and specifically the isotopes of titanium and vanadium. Experimental investigations were also pursued in 59 Ni and 65 Zn. Several new lifetimes were determined and confirmation was obtained for some previous values which were measured with different techniques. More information was also obtained on certain levels where previous results are in disagreement. (author)

  14. Masses and lifetimes of B hadrons

    International Nuclear Information System (INIS)

    Kkkroll, I.J.

    1996-02-01

    The latest measurements of the masses and lifetimes of weakly decaying B hadrons from experiments at e + e - and p bar p colliders are presented. These measurements include the lifetimes of the bar B o , bar B o s , B - and b baryons, as well as searches for the B c meson. The observation of B*, p-wave B mesons (B**), and excited b baryons using inclusive and exclusive B hadron reconstruction are discussed. Results on b quark flavour tagging are given

  15. Λc photoproduction and lifetime measurement

    International Nuclear Information System (INIS)

    Amendolia, S.R.; Bagliesi, G.; Batignani, G.; Bertolucci, E.; Bettoni, D.; Bizetti, A.; Bosisio, L.; Bottigli, U.; Bradaschia, C.; Dell'Orso, M.; Fidecaro, F.; Foa, L.; Focardi, E.; Giannetti, P.; Giorgi, M.A.; Marrocchesi, P.S.; Menzione, A.; Raso, G.; Ristori, L.; Scribano, A.; Stefanini, A.; Tenchini, R.; Tonelli, G.; Triggiani, G.; Beck, G.A.; Bologna, G.; D'Ettorre Piazzoli, B.; Picchi, P.; Budinich, M.; Liello, F.; Milotti, E.; Rolandi, L.; Carter, J.; Green, M.G.; Landon, M.P.J.; March, P.V.; Sacks, L.; Sanjari, A.H.; Strong, J.A.; Ciocci, M.A.; Enorini, M.; Fabbri, F.L.; Laurelli, P.; Mannocchi, G.; Simonelli, L.; Spillantini, P.; Zallo, A.

    1987-01-01

    A measurement of the lifetime of the Λ c baryon photoproduced coherently of a germanium-silicon target is presented. A signal of Λ c → ΔΚ * → pKππ 0 has been observed and the two different decay diagrams for this process are compared. A sample of 9 Λ c decays give a lifetime of 1.1(+0.8-0.4)10 13 s. (orig.)

  16. Measurement of the Omega0(c) lifetime

    International Nuclear Information System (INIS)

    Iori, M.

    2007-01-01

    The authors report a precise measurement of the (Omega) c 0 lifetime. The data were taken by the SELEX (E781) experiment using 600 GeV/c Σ - , π - and p beams. The measurement has been made using 83 ± 19 reconstructed (Omega) c 0 in the (Omega) - π - π + π + and (Omega) - π + decay modes. The lifetime of the (Omega) c 0 is measured to be 65 ± 13(stat) ± 9(sys) fs

  17. The total lifetime costs of smoking

    DEFF Research Database (Denmark)

    Rasmussen, S.R.; Prescott, E.; Sørensen, T.I.A.

    2004-01-01

    Net costs of smoking in a lifetime perspective and, hence, the economic interests in antismoking policies have been questioned. It has been proposed that the health-related costs of smoking are balanced by smaller expenditure due to shorter life expectancy.......Net costs of smoking in a lifetime perspective and, hence, the economic interests in antismoking policies have been questioned. It has been proposed that the health-related costs of smoking are balanced by smaller expenditure due to shorter life expectancy....

  18. Positron lifetime studies on thorium oxide powders

    International Nuclear Information System (INIS)

    Upadhyaya, D.D.; Muraleedharan, R.V.; Sharma, B.D.

    1982-01-01

    Positron lifetime spectra have been studied for ThO 2 powders, calcined at different temperatures and having different particle sizes. Three lifetime components could be resolved, the longest component being of low intensity. An observed strong dependence on the particle size of the annihilation process and the variation of positronium diffusion constant is explained on the basis of defect density variations in these powders. (author)

  19. Standardization of 241Am by digital coincidence counting, liquid scintillation counting and defined solid angle counting

    International Nuclear Information System (INIS)

    Balpardo, C.; Capoulat, M.E.; Rodrigues, D.; Arenillas, P.

    2010-01-01

    The nuclide 241 Am decays by alpha emission to 237 Np. Most of the decays (84.6%) populate the excited level of 237 Np with energy of 59.54 keV. Digital coincidence counting was applied to standardize a solution of 241 Am by alpha-gamma coincidence counting with efficiency extrapolation. Electronic discrimination was implemented with a pressurized proportional counter and the results were compared with two other independent techniques: Liquid scintillation counting using the logical sum of double coincidences in a TDCR array and defined solid angle counting taking into account activity inhomogeneity in the active deposit. The results show consistency between the three methods within a limit of a 0.3%. An ampoule of this solution will be sent to the International Reference System (SIR) during 2009. Uncertainties were analysed and compared in detail for the three applied methods.

  20. Measurements of heavy quark and lepton lifetimes

    International Nuclear Information System (INIS)

    Jaros, J.A.

    1985-02-01

    The PEP/PETRA energy range has proved to be well-suited for the study of the lifetimes of hadrons containing the b and c quarks and the tau lepton for several reasons. First, these states comprise a large fraction of the total interaction rate in e + e - annihilation and can be cleanly identified. Second, the storage rings have operated at high luminosity and so produced these exotic states copiously. And finally, thanks to the interplay of the Fermi coupling strength, the quark and lepton masses, and the beam energy, the expected decay lengths are in the 1/2 mm range and so are comparatively easy to measure. This pleasant coincidence of cleanly identified and abundant signal with potentially large effects has made possible the first measurements of two fundamental weak couplings, tau → nu/sub tau/W and b → cW. These measurements have provided a sharp test of the standard model and allowed, for the first time, the full determination of the magnitudes of the quark mixing matrix. This paper reviews the lifetime studies made at PEP during the past year. It begins with a brief review of the three detectors, DELCO, MAC and MARK II, which have reported lifetime measurements. Next it discusses two new measurements of the tau lifetime, and briefly reviews a measurement of the D 0 lifetime. Finally, it turns to measurements of the B lifetime, which are discussed in some detail. 18 references, 14 figures, 1 table

  1. Lifetimes of charm and beauty hadrons

    International Nuclear Information System (INIS)

    Bellini, G.; Dornan, P.J.

    1997-01-01

    Major breakthroughs have been achieved in the determination of the lifetimes of charm and beauty hadrons. Much larger data samples than previously have become available and new experimental devices and techniques have been developed and employed. The lifetimes of all weakly decaying singly charmed hadrons have been measured, some with an accuracy of a few percent. The difference in the shortest lifetime - τ(Ω c ) - and the longest one - τ(D + ) - is given by a factor of close to ten. The experimental status of beauty lifetimes, while less complete, has still reached a new level of quality and is now better than 5% for the commoner states. New theoretical tools, based mainly on heavy quark expansions, have been developed; they incorporate as well as transcend earlier phenomenological descriptions. The observed pattern in the charm lifetime ratios is reproduced in a semi-quantitative manner as well as could be expected; as far as the beauty lifetime ratios are concerned some problems may well be emerging. The maturity level achieved in the measurements bodes quite well for future challenges where reliable and efficient tracking of the decay vertices will be crucial. (orig.)

  2. Kinetics of bacterial fluorescence staining with 3,3'-diethylthiacyanine.

    Science.gov (United States)

    Thomas, Marlon S; Nuñez, Vicente; Upadhyayula, Srigokul; Zielins, Elizabeth R; Bao, Duoduo; Vasquez, Jacob M; Bahmani, Baharak; Vullev, Valentine I

    2010-06-15

    For more than a century, colorimetric and fluorescence staining have been the foundation of a broad range of key bioanalytical techniques. The dynamics of such staining processes, however, still remains largely unexplored. We investigated the kinetics of fluorescence staining of two gram-negative and two gram-positive species with 3,3'-diethylthiacyanine (THIA) iodide. An increase in the THIA fluorescence quantum yield, induced by the bacterial dye uptake, was the principal reason for the observed emission enhancement. The fluorescence quantum yield of THIA depended on the media viscosity and not on the media polarity, which suggested that the microenvironment of the dye molecules taken up by the cells was restrictive. The kinetics of fluorescence staining did not manifest a statistically significant dependence neither on the dye concentration, nor on the cell count. In the presence of surfactant additives, however, the fluorescence-enhancement kinetic patterns manifested species specificity with statistically significant discernibility.

  3. Hanford whole body counting manual

    International Nuclear Information System (INIS)

    Palmer, H.E.; Rieksts, G.A.; Lynch, T.P.

    1990-06-01

    This document describes the Hanford Whole Body Counting Program as it is administered by Pacific Northwest Laboratory (PNL) in support of the US Department of Energy--Richland Operations Office (DOE-RL) and its Hanford contractors. Program services include providing in vivo measurements of internally deposited radioactivity in Hanford employees (or visitors). Specific chapters of this manual deal with the following subjects: program operational charter, authority, administration, and practices, including interpreting applicable DOE Orders, regulations, and guidance into criteria for in vivo measurement frequency, etc., for the plant-wide whole body counting services; state-of-the-art facilities and equipment used to provide the best in vivo measurement results possible for the approximately 11,000 measurements made annually; procedures for performing the various in vivo measurements at the Whole Body Counter (WBC) and related facilities including whole body counts; operation and maintenance of counting equipment, quality assurance provisions of the program, WBC data processing functions, statistical aspects of in vivo measurements, and whole body counting records and associated guidance documents. 16 refs., 48 figs., 22 tabs

  4. Effects of lifetime ingestion of 90Sr in beagle dogs

    International Nuclear Information System (INIS)

    Book, S.A.; Spangler, W.L.; Swartz, L.A.

    1982-01-01

    To investigate the effects of lifetime 90 Sr ingestion, fifteen beagle dogs were fed the equivalent of 1.3, 4.0, and 12.0 μCi 90 Sr. Exposures prior to weaning were made by maternal administration of one of the same 90 Sr levels, begining at 21 days of gestation. Median survival was 12.5 years for the 1.3 μCi/day group, 6.5 years for the 4 μCi/day group, and 5.2 years of the 12 μCi/day group, compared to unirradiated control values of 14.5 to 15 years. The normal life span of more than half of the 1.3 μCi/day group is remarkable, considering they ingested 5900 to 7500 μCi 90 Sr during their lifetimes. One of seven 1.3 μCi/day dogs died of mycloproliferative syndrome (MPS), while one of four 4 μCi/day and one of four 12 μCi/day dogs died from MPS. In addition, another 12 μCi/day dog died of osteosarcoma and two others in the same group showed skeletal changes related to 90 Sr exposure. Other deaths were as could be expected in normal canine populations. Lifetime skeletal doses, determined by periodic whole-body counting, were 1990-3750, 1880-9230, and 6360-14,680 rad for the 1.3, 4, and 12 μCi/day groups, respectively. Comparison of these values with those from the large 90 Sr toxicity study at Davis in which 90 Sr feeding ended at 18 months of age indicates similar average skeletal doses from the two types of feeding regimens. The similarity of the skeletal doses may relate to the inability of 90 Sr to be incorporated to any great extent into a mature skeleton that no longer has the high calcium turnover associated with early life. As a result, deaths from lifetime 90 Sr ingestion appeared no earlier than when 90 Sr ingestion ended in early adulthood

  5. Temporal trends in sperm count

    DEFF Research Database (Denmark)

    Levine, Hagai; Jørgensen, Niels; Martino-Andrade, Anderson

    2017-01-01

    a predefined protocol 7518 abstracts were screened and 2510 full articles reporting primary data on SC were reviewed. A total of 244 estimates of SC and TSC from 185 studies of 42 935 men who provided semen samples in 1973-2011 were extracted for meta-regression analysis, as well as information on years.......006, respectively). WIDER IMPLICATIONS: This comprehensive meta-regression analysis reports a significant decline in sperm counts (as measured by SC and TSC) between 1973 and 2011, driven by a 50-60% decline among men unselected by fertility from North America, Europe, Australia and New Zealand. Because......BACKGROUND: Reported declines in sperm counts remain controversial today and recent trends are unknown. A definitive meta-analysis is critical given the predictive value of sperm count for fertility, morbidity and mortality. OBJECTIVE AND RATIONALE: To provide a systematic review and meta-regression...

  6. Reviews in fluorescence 2010

    CERN Document Server

    Geddes, Chris D

    2011-01-01

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

  7. Multiwavelength FLIM: new concept for fluorescence diagnosis

    Science.gov (United States)

    Rück, Angelika; Lorenz, S.; Hauser, Carmen; Mosch, S.; Kalinina, S.

    2012-03-01

    Fluorescence guided tumor resection is very well accepted in the case of bladder cancer and brain tumor, respectively. However, false positive results are one of the major problems, which will make the discrimination between tumor tissue and inflammation difficult. In contrast fluorescence lifetime imaging (FLIM) and especially spectral resolved FLIM (SLIM) can significantly improve the analysis. The fluorescence decay of a fluorophore in many cases does not show a simple monoexponential profile. A very complex situation arises, when more than one compound has to be analyzed. This could be the case when endogenous fluorophores of living cells and tissues have to be discriminated to identify oxidative metabolic changes. Other examples are PDT, when different photosensitizer metabolites are observed simultaneously. In those cases a considerable improvement could be achieved when time-resolved and spectral-resolved techniques are simultaneously incorporated. Within this presentation the principles of spectral and time-resolved fluorescence imaging will be discussed. Successful applications as autofluorescence and 5-ALA induced porphyrin fluorescence will be described in more detail.

  8. Photon-counting image sensors

    CERN Document Server

    Teranishi, Nobukazu; Theuwissen, Albert; Stoppa, David; Charbon, Edoardo

    2017-01-01

    The field of photon-counting image sensors is advancing rapidly with the development of various solid-state image sensor technologies including single photon avalanche detectors (SPADs) and deep-sub-electron read noise CMOS image sensor pixels. This foundational platform technology will enable opportunities for new imaging modalities and instrumentation for science and industry, as well as new consumer applications. Papers discussing various photon-counting image sensor technologies and selected new applications are presented in this all-invited Special Issue.

  9. ConA-based glucose sensing using the long-lifetime azadioxatriangulenium fluorophore

    Science.gov (United States)

    Cummins, Brian; Simpson, Jonathan; Gryczynski, Zygmunt; Sørensen, Thomas Just; Laursen, Bo W.; Graham, Duncan; Birch, David; Coté, Gerard

    2014-02-01

    Fluorescent glucose sensing technologies have been identified as possible alternatives to current continuous glucose monitoring approaches. We have recently introduced a new, smart fluorescent ligand to overcome the traditional problems of ConA-based glucose sensors. For this assay to be translated into a continuous glucose monitoring device where both components are free in solution, the molecular weight of the smart fluorescent ligand must be increased. We have identified ovalbumin as a naturally-occurring glycoprotein that could serve as the core-component of a 2nd generation smart fluorescent ligand. It has a single asparagine residue that is capable of displaying an N-linked glycan and a similar isoelectric point to ConA. Thus, binding between ConA and ovalbumin can potentially be monovalent and sugar specific. This work is the preliminary implementation of fluorescently-labeled ovalbumin in the ConA-based assay. We conjugate the red-emitting, long-lifetime azadioxatriangulenium (ADOTA+) dye to ovalbumin, as ADOTA have many advantageous properties to track the equilibrium binding of the assay. The ADOTA-labeled ovalbumin is paired with Alexa Fluor 647-labeled ConA to create a Förster Resonance Energy Transfer (FRET) assay that is glucose dependent. The assay responds across the physiologically relevant glucose range (0-500 mg/dL) with increasing intensity from the ADOTA-ovalbumin, showing that the strategy may allow for the translation of the smart fluorescent ligand concept into a continuous glucose monitoring device.

  10. Fluorescent intensifying screens: contribution of secondary X-rays

    International Nuclear Information System (INIS)

    Barroso, R.C.; Goncalves, O.D.; Eichler, J.; Lopes, R.T.; Cardoso, S.C.

    1996-01-01

    The counting rate and angular distribution of secondary X-rays produced by fluorescent intensifying screens are studied. A source of 241 Am - gamma radiation of 59.54 keV - is used. Fluorescent intensifying screens reduce the radiation dose in radiology since they produce visible light which increases the efficiency of the film. In addition, secondary X-rays arise due to the photoelectric effect, elastic (Rayleigh) and inelastic (Compton) scattering

  11. Plant lifetime management at Jose Cabrera NPP

    International Nuclear Information System (INIS)

    Martin, Jorge; Garcia, Piedad

    1998-01-01

    This paper presents the results obtained during the development and implementation of the Jose Cabrera NPP Lifetime Management Program according to the methodology applied in the Plant. The implementation of the Lifetime Management Program began in 1995 with the elaboration of the annual revision document 'Lifetime Management Plan', which describes the level of development of the Lifetime Management activities, the results that have been obtained during the implementation of the Program, and the schedule of the upcoming activities. The drawing up of a weighted list of 135 important components and the elaboration of 17 dossiers integrating the ageing mechanisms analysis and its corresponding evaluation, control and mitigation methods, were the result of the activities completed during 1996. A group of 62 component/degradation phenomena pairs with a high degradation risk classification has been considered within the scope of the activity 'Assessment of Maintenance Practices. Improvement Proposal', performed by the plant during 1997 and the first term of 1998 in parallel with other Lifetime Management related activities. The results obtained within this activity have revealed for the components included in the scope of the assessment that the associated degradation phenomena are practically covered by the current maintenance, inspection and testing practices. Recommendations and improvements of the maintenance practices have been particularly proposed from a technical, supporting, proceeding and documentary point of view, and currently an analysis is being made in relation to the feasibility of implementing them at the Jose Cabrera NPP. (author)

  12. Positron lifetime studies of electron irradiated copper

    International Nuclear Information System (INIS)

    Hadnagy, T.D.

    1976-01-01

    Single-crystal copper was irradiated with 4.5-MeV electrons producing simple Frenkel defects as well as a significant concentration of divacancies. Mean positron lifetime characteristics, which are sensitive to the presence of vacancies and multivacancies in copper, was monitored after isochronal anneals between 80 and 800 0 K to determine the relative change of characteristic mean lifetimes and their associated intensities. Also a study of the dependence of the mean positron lifetime on the total electron fluence was made and compared with existing theories relating these lifetimes to vacancy or multivacancy concentrations. Numerical data from curve fitting procedures using a conventional trapping model for defect-induced changes in positron lifetimes indicate that upon irradiation with 4.5-MeV electrons at 80 0 K, about 8 percent of the defects produced are divacancy units. Divacancy units appear to be several times more effective in trapping positrons than are monovacancies. Further, the experimental data suggest that the stage III annealing processes in electron-irradiated copper most probably involve the motion and removal of both monovacancies and divacancies. A conglomerate (multivacancy) unit appears to exist as a stable entity even after annealing procedures are carried out at temperatures slightly above the stage III region. Such a stable unit could serve as a nucleation center for the appearance of voids

  13. Fluorescence energy transfer on erythrocyte membranes

    International Nuclear Information System (INIS)

    Fuchs, H.M.; Hof, M.; Lawaczeck, R.

    1995-08-01

    Stationary and time-dependent fluorescence have been measured for a donor/acceptor (DA) pair bound to membrane proteins of bovine erythrocyte ghosts. The donor N-(p-(2-benzoxazolyl)phenyl)-maleimid (BMI) and the acceptor fluram bind to SH- and NH 2 -residues, respectively. The fluorescence spectra and the time-dependent emission are consistent with a radiationless fluorescence energy transfer (RET). The density of RET-effective acceptor binding sites c=0.072 nm -2 was calculated on the basis of the two-dimensional Foerster-kinetic. Band3 protein is the only membrane spanning protein with accessible SH-groups, and therefore only effective binding sites on the band3 protein are counted for the RET measurements performed. (author). 23 refs, 4 figs, 2 tabs

  14. Liquid scintillation, counting, and compositions

    International Nuclear Information System (INIS)

    Sena, E.A.; Tolbert, B.M.; Sutula, C.L.

    1975-01-01

    The emissions of radioactive isotopes in both aqueous and organic samples can be measured by liquid scintillation counting in micellar systems. The micellar systems are made up of scintillation solvent, scintillation solute and a mixture of surfactants, preferably at least one of which is relatively oil-soluble water-insoluble and another which is relatively water-soluble oil-insoluble

  15. Phase space quark counting rule

    International Nuclear Information System (INIS)

    Wei-gin, C.; Lo, S.

    1980-01-01

    A simple quark counting rule based on phase space consideration suggested before is used to fit all 39 recent experimental data points on inclusive reactions. Parameter free relations are found to agree with experiments. Excellent detail fits are obtained for 11 inclusive reactions

  16. Counting a Culture of Mealworms

    Science.gov (United States)

    Ashbrook, Peggy

    2007-01-01

    Math is not the only topic that will be discussed when young children are asked to care for and count "mealworms," a type of insect larvae (just as caterpillars are the babies of butterflies, these larvae are babies of beetles). The following activity can take place over two months as the beetles undergo metamorphosis from larvae to adults. As the…

  17. Counting problems for number rings

    NARCIS (Netherlands)

    Brakenhoff, Johannes Franciscus

    2009-01-01

    In this thesis we look at three counting problems connected to orders in number fields. First we study the probability that for a random polynomial f in Z[X] the ring Z[X]/f is the maximal order in Q[X]/f. Connected to this is the probability that a random polynomial has a squarefree

  18. On Counting the Rational Numbers

    Science.gov (United States)

    Almada, Carlos

    2010-01-01

    In this study, we show how to construct a function from the set N of natural numbers that explicitly counts the set Q[superscript +] of all positive rational numbers using a very intuitive approach. The function has the appeal of Cantor's function and it has the advantage that any high school student can understand the main idea at a glance…

  19. Logistic regression for dichotomized counts.

    Science.gov (United States)

    Preisser, John S; Das, Kalyan; Benecha, Habtamu; Stamm, John W

    2016-12-01

    Sometimes there is interest in a dichotomized outcome indicating whether a count variable is positive or zero. Under this scenario, the application of ordinary logistic regression may result in efficiency loss, which is quantifiable under an assumed model for the counts. In such situations, a shared-parameter hurdle model is investigated for more efficient estimation of regression parameters relating to overall effects of covariates on the dichotomous outcome, while handling count data with many zeroes. One model part provides a logistic regression containing marginal log odds ratio effects of primary interest, while an ancillary model part describes the mean count of a Poisson or negative binomial process in terms of nuisance regression parameters. Asymptotic efficiency of the logistic model parameter estimators of the two-part models is evaluated with respect to ordinary logistic regression. Simulations are used to assess the properties of the models with respect to power and Type I error, the latter investigated under both misspecified and correctly specified models. The methods are applied to data from a randomized clinical trial of three toothpaste formulations to prevent incident dental caries in a large population of Scottish schoolchildren. © The Author(s) 2014.

  20. Vote Counting as Mathematical Proof

    DEFF Research Database (Denmark)

    Schürmann, Carsten; Pattinson, Dirk

    2015-01-01

    then consists of a sequence (or tree) of rule applications and provides an independently checkable certificate of the validity of the result. This reduces the need to trust, or otherwise verify, the correctness of the vote counting software once the certificate has been validated. Using a rule...

  1. Counting SET-free sets

    OpenAIRE

    Harman, Nate

    2016-01-01

    We consider the following counting problem related to the card game SET: How many $k$-element SET-free sets are there in an $n$-dimensional SET deck? Through a series of algebraic reformulations and reinterpretations, we show the answer to this question satisfies two polynomiality conditions.

  2. Improved lifetime of microchannel-plate PMTs

    Energy Technology Data Exchange (ETDEWEB)

    Lehmann, A., E-mail: lehmann@physik.uni-erlangen.de [Physikalisches Institut IV, Friedrich Alexander-University of Erlangen-Nuremberg, Erlangen (Germany); Britting, A.; Eyrich, W.; Uhlig, F. [Physikalisches Institut IV, Friedrich Alexander-University of Erlangen-Nuremberg, Erlangen (Germany); Dzhygadlo, R.; Gerhardt, A.; Götzen, K.; Höhler, R.; Kalicy, G.; Kumawat, H.; Lehmann, D.; Lewandowski, B.; Patsyuk, M.; Peters, K.; Schepers, G.; Schmitt, L.; Schwarz, C.; Schwiening, J.; Traxler, M.; Zühlsdorf, M. [GSI Helmholtzzentrum für Schwerionenforschung GmbH, Darmstadt (Germany); and others

    2014-12-01

    The charged particle identification at the PANDA experiment will be mainly performed with DIRC detectors. Because of their advantageous properties the preferred photon sensors are MCP-PMTs. However, until recently these devices showed serious aging problems which resulted in a diminishing quantum efficiency (QE) of the photo cathode. By applying innovative countermeasures against the aging causes, the manufacturers recently succeeded in drastically improving the lifetime of MCP-PMTs. Especially the application of an ALD coating technique to seal the material of the micro-channels proves very powerful and results in a lifetime of ≈6C/cm{sup 2} integrated anode charge without a substantial QE degradation for the latest PHOTONIS XP85112. This paper will present a comparative measurement of the lifetime of several older and recent MCP-PMTs demonstrating this progress.

  3. Measurement of the $\\tau$ lepton lifetime

    CERN Document Server

    Buskulic, Damir; De Bonis, I; Décamp, D; Ghez, P; Goy, C; Lees, J P; Lucotte, A; Minard, M N; Odier, P; Pietrzyk, B; Ariztizabal, F; Chmeissani, M; Crespo, J M; Efthymiopoulos, I; Fernández, E; Fernández-Bosman, M; Gaitan, V; Garrido, L; Martínez, M; Orteu, S; Pacheco, A; Padilla, C; Palla, Fabrizio; Pascual, A; Perlas, J A; Sánchez, F; Teubert, F; Colaleo, A; Creanza, D; De Palma, M; Farilla, A; Gelao, G; Girone, M; Iaselli, Giuseppe; Maggi, G; Maggi, M; Marinelli, N; Natali, S; Nuzzo, S; Ranieri, A; Raso, G; Romano, F; Ruggieri, F; Selvaggi, G; Silvestris, L; Tempesta, P; Zito, G; Huang, X; Lin, J; Ouyang, Q; Wang, T; Xie, Y; Xu, R; Xue, S; Zhang, J; Zhang, L; Zhao, W; Bonvicini, G; Cattaneo, M; Comas, P; Coyle, P; Drevermann, H; Engelhardt, A; Forty, Roger W; Frank, M; Hagelberg, R; Harvey, J; Jacobsen, R; Janot, P; Jost, B; Kneringer, E; Knobloch, J; Lehraus, Ivan; Markou, C; Martin, E B; Mato, P; Minten, Adolf G; Miquel, R; Oest, T; Palazzi, P; Pater, J R; Pusztaszeri, J F; Ranjard, F; Rensing, P E; Rolandi, Luigi; Schlatter, W D; Schmelling, M; Schneider, O; Tejessy, W; Tomalin, I R; Venturi, A; Wachsmuth, H W; Wiedenmann, W; Wildish, T; Witzeling, W; Wotschack, J; Ajaltouni, Ziad J; Bardadin-Otwinowska, Maria; Barrès, A; Boyer, C; Falvard, A; Gay, P; Guicheney, C; Henrard, P; Jousset, J; Michel, B; Monteil, S; Pallin, D; Perret, P; Podlyski, F; Proriol, J; Rossignol, J M; Saadi, F; Fearnley, Tom; Hansen, J B; Hansen, J D; Hansen, J R; Hansen, P H; Nilsson, B S; Kyriakis, A; Simopoulou, Errietta; Siotis, I; Vayaki, Anna; Zachariadou, K; Blondel, A; Bonneaud, G R; Brient, J C; Bourdon, P; Passalacqua, L; Rougé, A; Rumpf, M; Tanaka, R; Valassi, Andrea; Verderi, M; Videau, H L; Candlin, D J; Parsons, M I; Focardi, E; Parrini, G; Corden, M; Delfino, M C; Georgiopoulos, C H; Jaffe, D E; Antonelli, A; Bencivenni, G; Bologna, G; Bossi, F; Campana, P; Capon, G; Chiarella, V; Felici, G; Laurelli, P; Mannocchi, G; Murtas, F; Murtas, G P; Pepé-Altarelli, M; Dorris, S J; Halley, A W; ten Have, I; Knowles, I G; Lynch, J G; Morton, W T; O'Shea, V; Raine, C; Reeves, P; Scarr, J M; Smith, K; Smith, M G; Thompson, A S; Thomson, F; Thorn, S; Turnbull, R M; Becker, U; Braun, O; Geweniger, C; Graefe, G; Hanke, P; Hepp, V; Kluge, E E; Putzer, A; Rensch, B; Schmidt, M; Sommer, J; Stenzel, H; Tittel, K; Werner, S; Wunsch, M; Beuselinck, R; Binnie, David M; Cameron, W; Colling, D J; Dornan, Peter J; Konstantinidis, N P; Moneta, L; Moutoussi, A; Nash, J; San Martin, G; Sedgbeer, J K; Stacey, A M; Dissertori, G; Girtler, P; Kuhn, D; Rudolph, G; Bowdery, C K; Brodbeck, T J; Colrain, P; Crawford, G; Finch, A J; Foster, F; Hughes, G; Sloan, Terence; Whelan, E P; Williams, M I; Galla, A; Greene, A M; Kleinknecht, K; Quast, G; Raab, J; Renk, B; Sander, H G; Wanke, R; Van Gemmeren, P; Zeitnitz, C; Aubert, Jean-Jacques; Bencheikh, A M; Benchouk, C; Bonissent, A; Bujosa, G; Calvet, D; Carr, J; Diaconu, C A; Etienne, F; Thulasidas, M; Nicod, D; Payre, P; Rousseau, D; Talby, M; Abt, I; Assmann, R W; Bauer, C; Blum, Walter; Brown, D; Dietl, H; Dydak, Friedrich; Ganis, G; Gotzhein, C; Jakobs, K; Kroha, H; Lütjens, G; Lutz, Gerhard; Männer, W; Moser, H G; Richter, R H; Rosado-Schlosser, A; Schael, S; Settles, Ronald; Seywerd, H C J; Saint-Denis, R; Wolf, G; Alemany, R; Boucrot, J; Callot, O; Cordier, A; Courault, F; Davier, M; Duflot, L; Grivaz, J F; Heusse, P; Jacquet, M; Kim, D W; Le Diberder, F R; Lefrançois, J; Lutz, A M; Musolino, G; Nikolic, I A; Park, H J; Park, I C; Schune, M H; Simion, S; Veillet, J J; Videau, I; Abbaneo, D; Azzurri, P; Bagliesi, G; Batignani, G; Bettarini, S; Bozzi, C; Calderini, G; Carpinelli, M; Ciocci, M A; Ciulli, V; Dell'Orso, R; Fantechi, R; Ferrante, I; Fidecaro, F; Foà, L; Forti, F; Giassi, A; Giorgi, M A; Gregorio, A; Ligabue, F; Lusiani, A; Marrocchesi, P S; Messineo, A; Rizzo, G; Sanguinetti, G; Sciabà, A; Spagnolo, P; Steinberger, Jack; Tenchini, Roberto; Tonelli, G; Triggiani, G; Vannini, C; Verdini, P G; Walsh, J; Betteridge, A P; Blair, G A; Bryant, L M; Cerutti, F; Gao, Y; Green, M G; Johnson, D L; Medcalf, T; Mir, L M; Perrodo, P; Strong, J A; Bertin, V; Botterill, David R; Clifft, R W; Edgecock, T R; Haywood, S; Edwards, M; Maley, P; Norton, P R; Thompson, J C; Bloch-Devaux, B; Colas, P; Emery, S; Kozanecki, Witold; Lançon, E; Lemaire, M C; Locci, E; Marx, B; Pérez, P; Rander, J; Renardy, J F; Roussarie, A; Schuller, J P; Schwindling, J; Trabelsi, A; Vallage, B; Johnson, R P; Kim, H Y; Litke, A M; McNeil, M A; Taylor, G; Beddall, A; Booth, C N; Boswell, R; Cartwright, S L; Combley, F; Dawson, I; Köksal, A; Letho, M; Newton, W M; Rankin, C; Thompson, L F; Böhrer, A; Brandt, S; Cowan, G D; Feigl, E; Grupen, Claus; Lutters, G; Minguet-Rodríguez, J A; Rivera, F; Saraiva, P; Smolik, L; Stephan, F; Apollonio, M; Bosisio, L; Della Marina, R; Giannini, G; Gobbo, B; Ragusa, F; Rothberg, J E; Wasserbaech, S R; Armstrong, S R; Bellantoni, L; Elmer, P; Feng, Z; Ferguson, D P S; Gao, Y S; González, S; Grahl, J; Harton, J L; Hayes, O J; Hu, H; McNamara, P A; Nachtman, J M; Orejudos, W; Pan, Y B; Saadi, Y; Schmitt, M; Scott, I J; Sharma, V; Turk, J; Walsh, A M; Wu Sau Lan; Wu, X; Yamartino, J M; Zheng, M; Zobernig, G

    1996-01-01

    The mean lifetime of the \\tau lepton is measured in a sample of 25700 \\tau pairs collected in 1992 with the ALEPH detector at LEP. A new analysis of the 1-1 topology events is introduced. In this analysis, the dependence of the impact parameter sum distribution on the daughter track momenta is taken into account, yielding improved precision compared to other impact parameter sum methods. Three other analyses of the one- and three-prong \\tau decays are updated with increased statistics. The measured lifetime is 293.5 \\pm 3.1 \\pm 1.7 \\fs. Including previous (1989--1991) ALEPH measurements, the combined \\tau lifetime is 293.7 \\pm 2.7 \\pm 1.6 \\fs.

  4. Lifetime for the Ti X spectrum

    International Nuclear Information System (INIS)

    Singh, Jagjit; Jha, A K S; Mohan, M

    2010-01-01

    We present configuration interaction calculations for the lifetime of 294 fine-structure levels of the Ti X spectrum in the LSJ coupling scheme. The calculations include all the major correlation effects. The relativistic effects are included by adding the mass correction term, Darwin term and spin-orbit interaction term to the non-relativistic Hamiltonian in the Breit-Pauli approximation. The calculated lifetime values are in very close agreement with other available experimental and theoretical results. We have predicted new lifetime results for levels belonging to 3p 2 3d, 3s 2 4p, 3s3p4s, 3s3p4p and various other configurations of Ti X, where no other theoretical and experimental results are available.

  5. Repeatability of differential goat bulk milk culture and associations with somatic cell count, total bacterial count, and standard plate count

    OpenAIRE

    Koop, G.; Dik, N.; Nielen, M.; Lipman, L.J.A.

    2010-01-01

    The aims of this study were to assess how different bacterial groups in bulk milk are related to bulk milk somatic cell count (SCC), bulk milk total bacterial count (TBC), and bulk milk standard plate count (SPC) and to measure the repeatability of bulk milk culturing. On 53 Dutch dairy goat farms, 3 bulk milk samples were collected at intervals of 2 wk. The samples were cultured for SPC, coliform count, and staphylococcal count and for the presence of Staphylococcus aureus. Furthermore, SCC ...

  6. Reviews in fluorescence 2008

    CERN Document Server

    Geddes, Chris D

    2010-01-01

    This volume serves as a comprehensive collection of current trends and emerging hot topics in the field of fluorescence spectroscopy. It summarizes the year's progress in fluorescence and its applications as well as includes authoritative analytical reviews.

  7. Fluorescent optical position sensor

    Science.gov (United States)

    Weiss, Jonathan D.

    2005-11-15

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

  8. Vibrational lifetimes of protein amide modes

    International Nuclear Information System (INIS)

    Peterson, K.A.; Rella, C.A.

    1995-01-01

    Measurement of the lifetimes of vibrational modes in proteins has been achieved with a single frequency infrared pump-probe technique using the Stanford Picosecond Free-electron Laser, These are the first direct measurements of vibrational dynamics in the polyamide structure of proteins. In this study, modes associated with the protein backbone are investigated. Results for the amide I band, which consists mainly of the stretching motion of the carbonyl unit of the amide linkage, show that relaxation from the first vibrational excited level (v=1) to the vibrational ground state (v=0) occurs within 1.5 picoseconds with apparent first order kinetics. Comparison of lifetimes for myoglobin and azurin, which have differing secondary structures, show a small but significant difference. The lifetime for the amide I band of myoglobin is 300 femtoseconds shorter than for azurin. Further measurements are in progress on other backbone vibrational modes and on the temperature dependence of the lifetimes. Comparison of vibrational dynamics for proteins with differing secondary structure and for different vibrational modes within a protein will lead to a greater understanding of energy transfer and dissipation in biological systems. In addition, these results have relevance to tissue ablation studies which have been conducted with pulsed infrared lasers. Vibrational lifetimes are necessary for calculating the rate at which the energy from absorbed infrared photons is converted to equilibrium thermal energy within the irradiated volume. The very fast vibrational lifetimes measured here indicate that mechanisms which involve direct vibrational up-pumping of the amide modes with consecutive laser pulses, leading to bond breakage or weakening, are not valid

  9. Safe biodegradable fluorescent particles

    Science.gov (United States)

    Martin, Sue I [Berkeley, CA; Fergenson, David P [Alamo, CA; Srivastava, Abneesh [Santa Clara, CA; Bogan, Michael J [Dublin, CA; Riot, Vincent J [Oakland, CA; Frank, Matthias [Oakland, CA

    2010-08-24

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

  10. Permeability log using new lifetime measurements

    International Nuclear Information System (INIS)

    Dowling, D.J.; Boyd, J.F.; Fuchs, J.A.

    1975-01-01

    Comparative measurements of thermal neutron decay time are obtained for a formation after irradiation with a pulsed neutron source. Chloride ions in formation fluids are concentrated by the electrosmosis effect using charged poles on a well logging sonde. The formation is irradiated with fast neutrons and a first comparative measure of the thermal neutron decay time or neutron lifetime is taken. The chloride ions are then dispersed by acoustic pumping with a magnetostrictive transducer. The formation is then again irradiated with fast neutrons and a comparative measure of neutron lifetime is taken. The comparison is a function of the variation in chloride concentration between the two measurements which is related to formation permeability

  11. An approach for longer lifetime MCFCs

    Energy Technology Data Exchange (ETDEWEB)

    Matsumoto, Masaru; Tatsumi, Masahiko; Hayano, Takuro [MCFC Research Association, Tokyo (Japan)] [and others

    1996-12-31

    For entering into commercialization of MCFC power plants in the beginning of the 21st century, we will devote to research for increasing lifetime as long as 40,000 hours with cell performance decay rate of 0.25 %/1000hrs as the target in FY 1999. This paper will discuss on our approach for longer lifetime MCFCs through electrolyte-loss management and NiO precipitation management as well as micro-structural control of electrodes and matrix plates. Cell voltage decay rate will be estimated by simulation through series of experiments on accelerated conditions.

  12. Lifetimes of some b-flavored hadrons

    International Nuclear Information System (INIS)

    Stone, S.

    2014-06-01

    Recent measurements of lifetimes of some b-flavored hadrons are presented and interpreted in the context of theoretical models, especially the Heavy Quark Expansion. Decay widths and decay width differences in the B s 0 - B-bar s 0 system are discussed from the studies of decays into the final states J/ψK + K - , J/ψπ + π - , D s + D s - , K + K - and D s ± π ± . Lifetime measurements of the baryons Λ b 0 , Ξ b - , Ξ b 0 , and Ω b - are also shown. (author)

  13. Positron lifetime in vanadium oxide bronzes

    International Nuclear Information System (INIS)

    Dryzek, J.; Dryzek, E.

    2003-01-01

    The positron lifetime (PL) and Doppler broadening (DB) of annihilation line measurements have been performed in vanadium oxide bronzes M x V 2 O 5 . The dependence of these annihilation characteristics on the kind and concentration of the metal M donor has been observed. In the PL spectrum only one lifetime component has been detected in all studied bronzes. The results indicate the positron localization in the structural tunnels present in the crystalline lattice of the vanadium oxide bronzes. (copyright 2003 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  14. Quantum system lifetimes and measurement perturbations

    International Nuclear Information System (INIS)

    Najakov, E.

    1977-05-01

    The recently proposed description of quantum system decay in terms of repeated measurement perturbations is modified. The possibility of retarded reductions to a unique quantum state, due to ineffective localization of the decay products at initial time measurements, is simply taken into account. The exponential decay law is verified again. A modified equation giving the observed lifetime in terms of unperturbed quantum decay law, measurement frequency and reduction law is derived. It predicts deviations of the observed lifetime from the umperturbed one, together with a dependence on experimental procedures. The influence of different model unperturbed decay laws and reduction laws on this effect is studied

  15. Extension of the nuclear power plant lifetime

    International Nuclear Information System (INIS)

    Keramsi, Alain

    2011-01-01

    After a presentation of the French nuclear context (history of the reactor fleet, choice of reactor type, PWR operation principle, competitiveness, environmental performance), this Power Point presentation addresses the context and challenges of the operation lifetime (average fleet age in different countries, examples of extensions, case of the United States, what is at stake with lifetime extension, decennial visits, EDF strategy), discusses the EDF's safety objectives (definition of the three main safety functions, impact of the operation duration and of the coexistence of two generations for the safety functions), discusses how to manage the ageing phenomenon for replaceable and non-replaceable components

  16. Lifetime, money and cost-benefit analysis

    International Nuclear Information System (INIS)

    Bengtsson, G.

    1984-01-01

    The paper describes briefly many methods for explicit or implicit valuation of the loss of lifetime expectancy due to radiation exposures or other hazards. The health gain from investment in protection is compared with the health gain from a general increase in wealth. It is concluded that in many instances lifetime is valued at 1 to 10 times the gross national product produced in this time. This seems to be reasonable for rich countries whereas it may be questionable for poorer countries. Here, any investment that raises the level of living of the poorer segment of the population may have a greater effect on life expectancy. (author)

  17. Magnon lifetimes in terbium at low temperatures

    International Nuclear Information System (INIS)

    Bjerrum Moeller, H.; Mackintosh, A.R.

    1979-01-01

    The lifetimes of magnons propagating in the c-direction of Tb at 4.2 K have been measured by inelastic neutron scattering. In contrast to the behaviour at higher temperatures, where magnon-magnon scattering predominates, the broadening of the magnons increases towards the boundary of the single Brillouin zone, both in the acoustic and optical branches. This suggests that the scattering of the magnons by conduction electrons is important, and the observed lifetimes are consistent with a recent estimate of the magnitude of this effect. The acoustic magnons of very long wavelength behave anomalously, presumably due to dipolar interactions

  18. Measurement of the Bs0 lifetime

    Science.gov (United States)

    Buskulic, D.; de Bonis, I.; Decamp, D.; Ghez, P.; Goy, C.; Lees, J.-P.; Minard, M.-N.; Odier, P.; Pietrzyk, B.; Ariztizabal, F.; Comas, P.; Crespo, J. M.; Efthymiopoulos, I.; Fernandez, E.; Fernandez-Bosman, M.; Gaitan, V.; Garrido, Ll.; Martinez, M.; Mattison, T.; Orteu, S.; Pacheco, A.; Padilla, C.; Pascual, A.; Creanza, D.; de Palma, M.; Farilla, A.; Iaselli, G.; Maggi, G.; Marinelli, N.; Natali, S.; Nuzzo, S.; Ranieri, A.; Raso, G.; Romano, F.; Ruggieri, F.; Selvaggi, G.; Silvestris, L.; Tempesta, P.; Zito, G.; Chai, Y.; Hu, H.; Huang, D.; Huang, X.; Lin, J.; Wang, T.; Xie, Y.; Xu, D.; Xu, R.; Zhang, J.; Zhang, L.; Zhao, W.; Bonvicini, G.; Boudreau, J.; Casper, D.; Drevermann, H.; Forty, R. W.; Ganis, G.; Gay, C.; Girone, M.; Hagelberg, R.; Harvey, J.; Hilgart, J.; Jacobsen, R.; Jost, B.; Knobloch, J.; Lehraus, I.; Maggi, M.; Markou, C.; Mato, P.; Meinhard, H.; Minten, A.; Miquel, R.; Moser, H.-G.; Palazzi, P.; Pater, J. R.; Perlas, J. A.; Perrodo, P.; Pusztaszeri, J.-F.; Ranjard, F.; Rolandi, L.; Rothberg, J.; Ruan, T.; Saich, M.; Schlatter, D.; Schmelling, M.; Sefkow, F.; Tejessy, W.; Tomalin, I. R.; Veenhof, R.; Wachsmuth, H.; Wasserbaech, S.; Wiedenmann, W.; Wildish, T.; Witzeling, W.; Wotschack, J.; Ajaltouni, Z.; Bardadin-Otwinowska, M.; Barres, A.; Boyer, C.; Falvard, A.; Gay, P.; Guicheney, C.; Henrard, P.; Jousset, J.; Michel, B.; Montret, J.-C.; Pallin, D.; Perret, P.; Podlyski, F.; Proriol, J.; Saadi, F.; Fearnley, T.; Hansen, J. B.; Hansen, J. D.; Hansen, J. R.; Hansen, P. H.; Johnson, S. D.; Møllerud, R.; Nilsson, B. S.; Kyriakis, A.; Simopoulou, E.; Siotis, I.; Vayaki, A.; Zachariadou, K.; Badier, J.; Blondel, A.; Bonneaud, G.; Brient, J. C.; Bourdon, P.; Fouque, G.; Passalacqua, L.; Rougé, A.; Rumpf, M.; Tanaka, R.; Verderi, M.; Videau, H.; Candlin, D. J.; Parsons, M. I.; Veitch, E.; Focardi, E.; Moneta, L.; Parrini, G.; Corden, M.; Delfino, M.; Georgiopoulos, C.; Jaffe, D. E.; Levinthal, D.; Antonelli, A.; Bencivenni, G.; Bologna, G.; Bossi, F.; Campana, P.; Capon, G.; Cerutti, F.; Chiarella, V.; Felici, G.; Laurelli, P.; Mannocchi, G.; Murtas, F.; Murtas, G. P.; Pepe-Altarelli, M.; Salomone, S.; Colrain, P.; Ten Have, I.; Knowles, I. G.; Lynch, J. G.; Maitland, W.; Morton, W. T.; Raine, C.; Reeves, P.; Scarr, J. M.; Smith, K.; Smith, M. G.; Thompson, A. S.; Thorn, S.; Turnbull, R. M.; Brandl, B.; Braun, O.; Geweniger, C.; Graefe, G.; Hanke, P.; Hepp, V.; Karger, C.; Kluge, E. E.; Maumary, Y.; Putzer, A.; Rensch, B.; Stahl, A.; Tittel, K.; Wunsch, M.; Beuselinck, R.; Binnie, D. M.; Cameron, W.; Cattaneo, M.; Colling, D. J.; Dornan, P. J.; Hassard, J. F.; Lieske, N. M.; Moutoussi, A.; Nash, J.; Patton, S.; Payne, D. G.; Phillips, M. J.; San Martin, G.; Sedgbeer, J. K.; Wright, A. G.; Girtler, P.; Kuhn, D.; Rudolph, G.; Vogl, R.; Bowdery, C. K.; Brodbeck, T. J.; Finch, A. J.; Foster, F.; Hughes, G.; Jackson, D.; Keemer, N. R.; Nuttall, M.; Patel, A.; Sloan, T.; Snow, S. W.; Whelan, E. P.; Galla, A.; Greene, A. M.; Kleinknecht, K.; Raab, J.; Renk, B.; Sander, H.-G.; Schmidt, H.; Walther, S. M.; Wanke, R.; Wolf, B.; Bencheikh, A. M.; Benchouk, C.; Bonissent, A.; Calvet, D.; Carr, J.; Coyle, P.; Diaconu, C.; Drinkard, J.; Etienne, F.; Nicod, D.; Payre, P.; Ross, L.; Rousseau, D.; Schwemling, P.; Talby, M.; Adlung, S.; Assmann, R.; Bauer, C.; Blum, W.; Brown, D.; Cattaneo, P.; Dehning, B.; Dietl, H.; Dydak, F.; Frank, M.; Halley, A. W.; Jakobs, K.; Lauber, J.; Lütjens, G.; Lutz, G.; Männer, W.; Richter, R.; Schröder, J.; Schwarz, A. S.; Settles, R.; Seywerd, H.; Stierlin, U.; Stiegler, U.; Denis, R. St.; Wolf, G.; Alemany, R.; Boucrot, J.; Callot, O.; Cordier, A.; Davier, M.; Duflot, L.; Grivaz, J.-F.; Heusse, Ph.; Janot, P.; Kimfn 19, D. W.; Le Diberder, F.; Lefrançois, J.; Lutz, A.-M.; Musolino, G.; Schune, M.-H.; Veillet, J.-J.; Videau, I.; Abbaneo, D.; Bagliesi, G.; Batignani, G.; Bottigli, U.; Bozzi, C.; Calderini, G.; Carpinelli, M.; Ciocci, M. A.; Ciulli, V.; Dell'Orso, R.; Ferrante, I.; Fidecaro, F.; Foà, L.; Forti, F.; Giassi, A.; Giorgi, M. A.; Gregorio, A.; Ligabue, F.; Lusiani, A.; Mannelli, E. B.; Marrocchesi, P. S.; Messineo, A.; Palla, F.; Rizzo, G.; Sanguinetti, G.; Spagnolo, P.; Steinberger, J.; Tenchini, R.; Tonelli, G.; Triggiani, G.; Valassi, A.; Vannini, C.; Venturi, A.; Verdini, P. G.; Walsh, J.; Betteridge, A. P.; Gao, Y.; Green, M. G.; Johnson, D. L.; March, P. V.; Medcalf, T.; Mir, Ll. M.; Quazi, I. S.; Strong, J. A.; Bertin, V.; Botterill, D. R.; Clifft, R. W.; Edgecock, T. R.; Haywood, S.; Edwards, M.; Norton, P. R.; Thompson, J. C.; Bloch-Devaux, B.; Colas, P.; Duarte, H.; Emery, S.; Kozanecki, W.; Lançon, E.; Lemaire, M. C.; Locci, E.; Marx, B.; Perez, P.; Rander, J.; Renardy, J.-F.; Rosowsky, A.; Roussarie, A.; Schuller, J.-P.; Schwindling, J.; Si Mohand, D.; Vallage, B.; Johnson, R. P.; Litke, A. M.; Taylor, G.; Wear, J.; Babbage, W.; Booth, C. N.; Buttar, C.; Cartwright, S.; Combley, F.; Dawson, I.; Thompson, L. F.; Barberio, E.; Böhrer, A.; Brandt, S.; Cowan, G.; Grupen, C.; Lutters, G.; Rivera, F.; Schäfer, U.; Smolik, L.; Bosisio, L.; Della Marina, R.; Giannini, G.; Gobbo, B.; Pitis, L.; Ragusa, F.; Bellantoni, L.; Chen, W.; Conway, J. S.; Feng, Z.; Ferguson, D. P. S.; Gao, Y. S.; Grahl, J.; Harton, J. L.; Hayes, O. J.; Nachtman, J. M.; Pan, Y. B.; Saadi, Y.; Schmitt, M.; Scott, I.; Sharma, V.; Shi, Z. H.; Turk, J. D.; Walsh, A. M.; Weber, F. V.; Lan Wu, Sau; Wu, X.; Zheng, M.; Zobernig, G.; Aleph Collaboration

    1994-02-01

    The lifetime of the Bs0 has been measured in a data sample of 8890000 hadronic events recorded with the ALEPH detector at LEP. After background subtraction 30.8 ± 6.9 events are attributed to the semileptonic decay of the Bs0 to a Ds- and an opposite-sign lepton. A maximum-likelihood fit to the distribution of the proper times of these events yields a Bs0 lifetime of τBs = 1.92 -0.35+0.45 ± 0.04 ps.

  19. Prompt Neutron Lifetime for the NBSR Reactor

    Energy Technology Data Exchange (ETDEWEB)

    Hanson, A.L.; Diamond, D.

    2012-06-24

    In preparation for the proposed conversion of the National Institute of Standards and Technology (NIST) research reactor (NBSR) from high-enriched uranium (HEU) to low-enriched uranium (LEU) fuel, certain point kinetics parameters must be calculated. We report here values of the prompt neutron lifetime that have been calculated using three independent methods. All three sets of calculations demonstrate that the prompt neutron lifetime is shorter for the LEU fuel when compared to the HEU fuel and longer for the equilibrium end-of-cycle (EOC) condition when compared to the equilibrium startup (SU) condition for both the HEU and LEU fuels.

  20. Optimization of fluorescent proteins

    NARCIS (Netherlands)

    Bindels, D.S.; Goedhart, J.; Hink, M.A.; van Weeren, L.; Joosen, L.; Gadella (jr.), T.W.J.; Engelborghs, Y.; Visser, A.J.W.G.

    2014-01-01

    Nowadays, fluorescent protein (FP) variants have been engineered to fluoresce in all different colors; to display photoswitchable, or photochromic, behavior; or to show yet other beneficial properties that enable or enhance a still growing set of new fluorescence spectroscopy and microcopy

  1. Smart optical probes for near-infrared fluorescence imaging of Alzheimer's disease pathology

    International Nuclear Information System (INIS)

    Raymond, Scott B.; Bacskai, Brian J.; Skoch, Jesse; Hills, Ivory D.; Swager, Timothy M.; Nesterov, Evgueni E.

    2008-01-01

    Near-infrared fluorescent probes for amyloid-beta (Aβ) are an exciting option for molecular imaging in Alzheimer's disease research and may translate to clinical diagnostics. However, Aβ-targeted optical probes often suffer from poor specificity and slow clearance from the brain. We are designing smart optical probes that emit characteristic fluorescence signal only when bound to Aβ. We synthesized a family of dyes and tested Aβ-binding sensitivity with fluorescence spectroscopy and tissue-staining. Select compounds exhibited Aβ-dependent changes in fluorescence quantum yield, lifetime, and emission spectra that may be imaged microscopically or in vivo using new lifetime and spectral fluorescence imaging techniques. Smart optical probes that turn on when bound to Aβ will improve amyloid detection and may enable quantitative molecular imaging in vivo. (orig.)

  2. PMP, a novel solute for liquid and plastic scintillation counting

    International Nuclear Information System (INIS)

    Gusten, Hans

    1983-01-01

    The excellent fluorescence properties of PMP ( 11-phenyl-3-mesityl-2-pyrazoline) such as long wavelength emission of over 400 nm, and high fluorescence quantum yield with a short decay time together with a solubility of more than one Mol/L in toluene make this compound a promising solute for scintillation counting. The Stokes' shift of PMP of over 10,000 cm -1 is twice as large as that of the commonly used PPO. Due to this unusually large Stokes' shift PMP can be used as a primary solute without requiring a secondary solute as wavelength shifter. A comparison of the scintillation properties of PMP and PPO in toluene reveals that the counting efficiency for 14 C is better for PMP while the 3 H efficiency is equally good. Due to the large Stokes' shift, PMP is about 50 percent less sensitive to color quenching than PPO. Compared to the solute combinations PPO/secondary solutes, the scintillation counting efficiency of PMP for 14 C in toluene or xylene is the same, while the absolute 3 H efficiency of PPO/secondary solutes in cocktails with emulsifiers is about 10 percent higher. The PMP scintillation efficiency for 14 C as well as 3 H in chemical quenching by urine is more or less the same as for PPO/dimethyl-POPOP. PMP is more sensitive to quenching by halogenated solvents. In the dioxane-based scintillation, this sensitivity to chemical quenching by CHCl 3 vanishes and the counting efficiencies for 14 C and 3 H are as good as for PPO/dimethyl-POPOP or PPO/bis-MSB. Due to the large Stokes' shift, the self-absorption of the scintillation light by PMP is lower than in conventional scintillators. This offers good possibilities in very large-volume applications of liquid as well as plastic scintillators

  3. Two-photon excited fluorescence microscopy application for ex vivo investigation of ocular fundus samples

    Science.gov (United States)

    Peters, Sven; Hammer, Martin; Schweitzer, Dietrich

    2011-07-01

    Two-photon excited fluorescence (TPEF) imaging of ocular tissue has recently become a promising tool in ophthalmology for diagnostic and research purposes. The feasibility and the advantages of TPEF imaging, namely deeper tissue penetration and improved high-resolution imaging of microstructures, have been demonstrated lately using human ocular samples. The autofluorescence properties of endogenous fluorophores in ocular fundus tissue are well known from spectrophotometric analysis. But fluorophores, especially when it comes to fluorescence lifetime, typically display a dependence of their fluorescence properties on local environmental parameters. Hence, a more detailed investigation of ocular fundus autofluorescence ideally in vivo is of utmost interest. The aim of this study is to determine space-resolved the stationary and time-resolved fluorescence properties of endogenous fluorophores in ex vivo porcine ocular fundus samples by means of two-photon excited fluorescence spectrum and lifetime imaging microscopy (FSIM/FLIM). By our first results, we characterized the autofluorescence of individual anatomical structures of porcine retina samples excited at 760 nm. The fluorescence properties of almost all investigated retinal layers are relatively homogenous. But as previously unknown, ganglion cell bodies show a significantly shorter fluorescence lifetime compared to the adjacent mueller cells. Since all retinal layers exhibit bi-exponential autofluorescence decays, we were able to achieve a more precise characterization of fluorescence properties of endogenous fluorophores compared to a present in vivo FLIM approach by confocal scanning laser ophthalmoscope (cSLO).

  4. Lifetime of MCP-PMTs and other performance features

    Science.gov (United States)

    Lehmann, A.; Böhm, M.; Eyrich, W.; Miehling, D.; Pfaffinger, M.; Stelter, S.; Uhlig, F.; Ali, A.; Belias, A.; Dzhygadlo, R.; Gerhardt, A.; Götzen, K.; Kalicy, G.; Krebs, M.; Lehmann, D.; Nerling, F.; Patsyuk, M.; Peters, K.; Schepers, G.; Schmitt, L.; Schwarz, C.; Schwiening, J.; Traxler, M.; Düren, M.; Etzelmüller, E.; Föhl, K.; Hayrapetyan, A.; Kreutzfeld, K.; Merle, O.; Rieke, J.; Schmidt, M.; Wasem, T.; Achenbach, P.; Cardinali, M.; Hoek, M.; Lauth, W.; Schlimme, S.; Sfienti, C.; Thiel, M.

    2018-02-01

    The bar PANDA experiment at FAIR will use DIRC detectors for the separation of hadrons. The compactness of the bar PANDA detector requires the image planes of these detectors to be placed inside the magnetic field of the solenoid. Due to this and other boundary conditions MCP-PMTs were identified as the only suitable photon sensors. Until recently the major obstacle for an application of MCP-PMTs in high rate experiments like bar PANDA were serious aging problems which led to damage at the photo-cathode and a fast declining quantum efficiency as the integrated anode charge (IAC) increased. With new countermeasures against the aging, in particular due to the application of an atomic layer deposition (ALD) technique to coat the MCP pores, the lifetime of MCP-PMTs has meanwhile increased by a factor >50 which is fully sufficient for bar PANDA. The recent results of our long-term lifetime measurements are discussed. New 2-inch MCP-PMT prototypes from Hamamatsu show an encouraging behavior. However, the currently best performing MCP-PMT is a 2-inch PHOTONIS tube with two ALD-layers which reaches an IAC of >16 C/cm2 without any visible sign of aging. In the second part of these proceedings a new data acquisition system of the PADIWA/TRB type is presented which allows a quasi-parallel measurement of many MCP-PMT performance parameters. Especially unwanted effects like dark-count rate, crosstalk, ion after-pulsing, and recoil electrons can be studied in more detail than ever before. Exemplary results for these parameters are shown. The discussed DAQ system will be used for the comprehensive data quality checks of the MCP-PMTs being built into the DIRCs.

  5. Cerenkov counting and Cerenkov-scintillation counting with high refractive index organic liquids using a liquid scintillation counter

    Energy Technology Data Exchange (ETDEWEB)

    Wiebe, L I; Helus, F; Maier-Borst, W [Deutsches Krebsforschungszentrum, Heidelberg (Germany, F.R.). Inst. fuer Nuklearmedizin

    1978-06-01

    /sup 18/F and /sup 14/C radioactivity was measured in methyl salicylate (MS), a high refractive index hybrid Cherenkov-scintillation generating medium, using a liquid scintillation counter. At concentrations of up to 21.4%, in MS, dimethyl sulfoxide (DMSO) quenched /sup 14/C fluorescence, and with a 10-fold excess of DMSO over MS, /sup 18/F count rates were reduced below that for DMSO alone, probably as a result of concentration-independent self-quenching due to 'dark-complex' formation. DMSO in lower concentrations did not reduce the counting efficiency of /sup 18/F in MS. Nitrobenzene was a concentration-dependent quencher for both /sup 14/C and /sup 18/F in MS. Chlorobenzene (CB) and DMSO were both found to be weak Cherenkov generators with /sup 18/F. Counting efficiencies for /sup 18/F in MS, CB, and DMSO were 50.3, 7.8 and 4.3% respectively in the coincidence counting mode, and 58.1, 13.0 and 6.8% in the singles mode. /sup 14/C efficiencies were 14.4 and 22.3% for coincidence and singles respectively, and 15.3 and 42.0% using a modern counter designed for coincidence and single photon counting. The high /sup 14/C and /sup 18/F counting efficiency in MS are discussed with respect to excitation mechanism, on the basis of quench and channels ratios changes observed. It is proposed that MS functions as an efficient Cherenkov-scintillation generator for high-energy beta emitters such as /sup 18/F, and as a low-efficiency scintillator for weak beta emitting radionuclides such as /sup 14/C.

  6. Cerenkov counting and Cerenkov-scintillation counting with high refractive index organic liquids using a liquid scintillation counter

    International Nuclear Information System (INIS)

    Wiebe, L.I.; Helus, F.; Maier-Borst, W.

    1978-01-01

    18 F and 14 C radioactivity was measured in methyl salicylate (MS), a high refractive index hybrid Cherenkov-scintillation generating medium, using a liquid scintillation counter. At concentrations of up to 21.4%, in MS, dimethyl sulfoxide (DMSO) quenched 14 C fluorescence, and with a 10-fold excess of DMSO over MS, 18 F count rates were reduced below that for DMSO alone, probably as a result of concentration-independent self-quenching due to 'dark-complex' formation. DMSO in lower concentrations did not reduce the counting efficiency of 18 F in MS. Nitrobenzene was a concentration-dependent quencher for both 14 C and 18 F in MS. Chlorobenzene (CB) and DMSO were both found to be weak Cherenkov generators with 18 F. Counting efficiencies for 18 F in MS, CB, and DMSO were 50.3, 7.8 and 4.3% respectively in the coincidence counting mode, and 58.1, 13.0 and 6.8% in the singles mode. 14 C efficiencies were 14.4 and 22.3% for coincidence and singles respectively, and 15.3 and 42.0% using a modern counter designed for coincidence and single photon counting. The high 14 C and 18 F counting efficiency in MS are discussed with respect to excitation mechanism, on the basis of quench and channels ratios changes observed. It is proposed that MS functions as an efficient Cherenkov-scintillation generator for high-energy beta emitters such as 18 F, and as a low-efficiency scintillator for weak beta emitting radionuclides such as 14 C. (author)

  7. Lifetime Reliability Assessment of Concrete Slab Bridges

    DEFF Research Database (Denmark)

    Thoft-Christensen, Palle

    A procedure for lifetime assesment of the reliability of short concrete slab bridges is presented in the paper. Corrosion of the reinforcement is the deterioration mechanism used for estimating the reliability profiles for such bridges. The importance of using sensitivity measures is stressed....... Finally the produce is illustrated on 6 existing UK bridges....

  8. SIRTF thermal design modifications to increase lifetime

    Science.gov (United States)

    Petrick, S. W.

    1993-01-01

    An effort was made to increase the predicted lifetime of the SIRTF dewar by lowering the exterior shell temperature, increasing the radiated energy from the vapor cooled shields and reconfiguring the vapor cooled shields. The lifetime increases can be used to increase the scientific return from the mission and as a trade-off against mass and cost. This paper describes the configurations studied, the steady state thermal model used, the analytical methods and the results of the analysis. Much of the heat input to the outside dewar shell is radiative heat transfer from the solar panel. To lower the shell temperature, radiative cooled shields were placed between the solar panel and the dewar shell and between the bus and the dewar shell. Analysis showed that placing a radiator on the outer vapor cooled shield had a significant effect on lifetime. Lengthening the distance between the outer shell and the point where the vapor cooled shields are attached to the support straps also improved lifetime.

  9. Updated measurement of the $\\tau$ lepton lifetime

    CERN Document Server

    Barate, R; Décamp, D; Ghez, P; Goy, C; Lees, J P; Lucotte, A; Minard, M N; Nief, J Y; Pietrzyk, B; Casado, M P; Chmeissani, M; Comas, P; Crespo, J M; Delfino, M C; Fernández, E; Fernández-Bosman, M; Garrido, L; Juste, A; Martínez, M; Merino, G; Miquel, R; Mir, L M; Padilla, C; Park, I C; Pascual, A; Perlas, J A; Riu, I; Sánchez, F; Teubert, F; Colaleo, A; Creanza, D; De Palma, M; Gelao, G; Iaselli, Giuseppe; Maggi, G; Maggi, M; Marinelli, N; Nuzzo, S; Ranieri, A; Raso, G; Ruggieri, F; Selvaggi, G; Silvestris, L; Tempesta, P; Tricomi, A; Zito, G; Huang, X; Lin, J; Ouyang, Q; Wang, T; Xie, Y; Xu, R; Xue, S; Zhang, J; Zhang, L; Zhao, W; Abbaneo, D; Alemany, R; Becker, U; Bazarko, A O; Bright-Thomas, P G; Cattaneo, M; Cerutti, F; Dissertori, G; Drevermann, H; Forty, Roger W; Frank, M; Hagelberg, R; Hansen, J B; Harvey, J; Janot, P; Jost, B; Kneringer, E; Knobloch, J; Lehraus, Ivan; Mato, P; Minten, Adolf G; Moneta, L; Pacheco, A; Pusztaszeri, J F; Ranjard, F; Rizzo, G; Rolandi, Luigi; Rousseau, D; Schlatter, W D; Schmitt, M; Schneider, O; Tejessy, W; Tomalin, I R; Wachsmuth, H W; Wagner, A; Ajaltouni, Ziad J; Barrès, A; Boyer, C; Falvard, A; Ferdi, C; Gay, P; Guicheney, C; Henrard, P; Jousset, J; Michel, B; Monteil, S; Montret, J C; Pallin, D; Perret, P; Podlyski, F; Proriol, J; Rosnet, P; Rossignol, J M; Fearnley, Tom; Hansen, J D; Hansen, J R; Hansen, P H; Nilsson, B S; Rensch, B; Wäänänen, A; Daskalakis, G; Kyriakis, A; Markou, C; Simopoulou, Errietta; Siotis, I; Vayaki, Anna; Blondel, A; Bonneaud, G R; Brient, J C; Bourdon, P; Rougé, A; Rumpf, M; Valassi, Andrea; Verderi, M; Videau, H L; Candlin, D J; Parsons, M I; Focardi, E; Parrini, G; Zachariadou, K; Corden, M; Georgiopoulos, C H; Jaffe, D E; Antonelli, A; Bencivenni, G; Bologna, G; Bossi, F; Campana, P; Capon, G; Casper, David William; Chiarella, V; Felici, G; Laurelli, P; Mannocchi, G; Murtas, F; Murtas, G P; Passalacqua, L; Pepé-Altarelli, M; Curtis, L; Dorris, S J; Halley, A W; Knowles, I G; Lynch, J G; O'Shea, V; Raine, C; Scarr, J M; Smith, K; Teixeira-Dias, P; Thompson, A S; Thomson, E; Thomson, F; Turnbull, R M; Buchmüller, O L; Dhamotharan, S; Geweniger, C; Graefe, G; Hanke, P; Hansper, G; Hepp, V; Kluge, E E; Putzer, A; Sommer, J; Tittel, K; Werner, S; Wunsch, M; Beuselinck, R; Binnie, David M; Cameron, W; Dornan, Peter J; Girone, M; Goodsir, S M; Martin, E B; Moutoussi, A; Nash, J; Sedgbeer, J K; Spagnolo, P; Stacey, A M; Williams, M D; Ghete, V M; Girtler, P; Kuhn, D; Rudolph, G; Betteridge, A P; Bowdery, C K; Colrain, P; Crawford, G; Finch, A J; Foster, F; Hughes, G; Jones, R W L; Sloan, Terence; Williams, M I; Galla, A; Giehl, I; Greene, A M; Hoffmann, C; Jakobs, K; Kleinknecht, K; Quast, G; Renk, B; Rohne, E; Sander, H G; Van Gemmeren, P; Zeitnitz, C; Aubert, Jean-Jacques; Benchouk, C; Bonissent, A; Bujosa, G; Carr, J; Coyle, P; Diaconu, C A; Etienne, F; Konstantinidis, N P; Leroy, O; Motsch, F; Payre, P; Talby, M; Sadouki, A; Thulasidas, M; Trabelsi, K; Aleppo, M; Antonelli, M; Ragusa, F; Berlich, R; Blum, Walter; Büscher, V; Dietl, H; Ganis, G; Gotzhein, C; Kroha, H; Lütjens, G; Lutz, Gerhard; Männer, W; Moser, H G; Richter, R H; Rosado-Schlosser, A; Schael, S; Settles, Ronald; Seywerd, H C J; Saint-Denis, R; Stenzel, H; Wiedenmann, W; Wolf, G; Boucrot, J; Callot, O; Chen, S; Choi, Y; Cordier, A; Davier, M; Duflot, L; Grivaz, J F; Heusse, P; Höcker, A; Jacholkowska, A; Jacquet, M; Kim, D W; Le Diberder, F R; Lefrançois, J; Lutz, A M; Nikolic, I A; Schune, M H; Simion, S; Tournefier, E; Veillet, J J; Videau, I; Zerwas, D; Azzurri, P; Bagliesi, G; Batignani, G; Bettarini, S; Bozzi, C; Calderini, G; Carpinelli, M; Ciocci, M A; Ciulli, V; Dell'Orso, R; Fantechi, R; Ferrante, I; Foà, L; Forti, F; Giassi, A; Giorgi, M A; Gregorio, A; Ligabue, F; Lusiani, A; Marrocchesi, P S; Messineo, A; Palla, Fabrizio; Sanguinetti, G; Sciabà, A; Steinberger, Jack; Tenchini, Roberto; Tonelli, G; Vannini, C; Venturi, A; Verdini, P G; Blair, G A; Bryant, L M; Chambers, J T; Gao, Y; Green, M G; Medcalf, T; Perrodo, P; Strong, J A; Von Wimmersperg-Töller, J H; Botterill, David R; Clifft, R W; Edgecock, T R; Haywood, S; Norton, P R; Thompson, J C; Wright, A E; Bloch-Devaux, B; Colas, P; Emery, S; Kozanecki, Witold; Lançon, E; Lemaire, M C; Locci, E; Pérez, P; Rander, J; Renardy, J F; Roussarie, A; Schuller, J P; Schwindling, J; Trabelsi, A; Vallage, B; Black, S N; Dann, J H; Johnson, R P; Kim, H Y; Litke, A M; McNeil, M A; Taylor, G; Booth, C N; Boswell, R; Brew, C A J; Cartwright, S L; Combley, F; Kelly, M S; Lehto, M H; Newton, W M; Reeve, J; Thompson, L F; Böhrer, A; Brandt, S; Cowan, G D; Grupen, Claus; Lutters, G; Saraiva, P; Smolik, L; Stephan, F; Apollonio, M; Bosisio, L; Della Marina, R; Giannini, G; Gobbo, B; Musolino, G; Pütz, J; Rothberg, J E; Wasserbaech, S R; Armstrong, S R; Charles, E; Elmer, P; Ferguson, D P S; González, S; Greening, T C; Hayes, O J; Hu, H; Jin, S; McNamara, P A; Nachtman, J M; Nielsen, J; Orejudos, W; Pan, Y B; Saadi, Y; Scott, I J; Walsh, J; Wu Sau Lan; Wu, X; Yamartino, J M; Zobernig, G

    1997-01-01

    A new measurement of the mean lifetime of the tau lepton is presented. Three different analysis methods are applied to a sample of 90000 tau pairs, collected in 1993 and 1994 with the ALEPH detector at LEP. The average of this measurement and those previously published by ALEPH is tau_tau = 290.1 +- 1.5 +- 1.1 fs.

  10. Lifetime measurements of the rare earths

    International Nuclear Information System (INIS)

    Stahnke, H.J.

    1981-01-01

    The lifetime of excited energy levels of Praseodymium, Neodymium, Gadolinium, Holmium and Erbium are measured. The measurements were done on atomic beams excited by laser radiation. The experimental results allow an interpretation of the electronic structure of the rare earths. (BEF)

  11. Materials Education: Opportunities over a Lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Iver E.; Schwartz, Lyle H.; Faber, Katherine T.; Cargill III, G. Slade; Houston, Betsy

    2003-10-28

    A report, in the form of abbreviated notes, of the 17th Biennial Conference on National Materials Policy ''Materials Education: Opportunities over a Lifetime'' held May 20-21, 2002 in College Park, MD, sponsored by the Federation of Materials Societies and the University Materials Council.

  12. Smoking expands expected lifetime with musculoskeletal disease

    DEFF Research Database (Denmark)

    Brønnum-Hansen, Henrik; Juel, Knud

    2003-01-01

    By indirect estimation of mortality from smoking and life table methods we estimated expected lifetime without musculoskeletal diseases among never smokers, ex-smokers, and smokers. We found that although life expectancy of a heavy smoker is 7 years shorter than that of a never smoker, heavy...

  13. Disc Golf: Teaching a Lifetime Activity

    Science.gov (United States)

    Eastham, Susan L.

    2015-01-01

    Disc golf is a lifetime activity that can be enjoyed by students of varying skill levels and abilities. Disc golf follows the principles of ball golf but is generally easier for students to play and enjoy success. The object of disc golf is similar to ball golf and involves throwing a disc from the teeing area to the target in as few throws as…

  14. Lifetime and spin measurements in 40Ar

    International Nuclear Information System (INIS)

    Southon, J.

    1976-01-01

    Lifetimes of levels in 40 Ar populated by the 40 Ar(p,p') reaction have been measured using the Doppler shift attenuation method with a p-γ coincidence technique. A solid argon target was used. The lifetimes determined were (in psec.): 1461 keV level, 1.95 +- 0.15; 2121 keV, >25; 2524 keV, 0.53 +- 0.06; 2893 keV, 4.4 [+2.6,-1.3]; 3208 keV, 0.27. A comprehensive set of branching ratios was also derived and the spins and parities of the 3208 and 4481 keV states were determined to be 2 + and 1 +- respectively. Some of these results suggest that 2 particle -2 hole and 4 particle - 4 hole components are strongly mixed in the low-lying positive parity states in a manner similar to the 2 particle and 4 particle - 2 hole mixing that occurs in 42 Ca. An additional lifetime measurement for the recently discovered high spin state at 3464 keV was carried out using direct electronic timing. The level was excited by the 37 Cl(α,p) reaction and was found to have a lifetime of 1.00 +- 0.03 nsec, which taken together with other evidence indicates that its spin and parity are 6 + . The E2 transition strengths of the 40 Ar 6 + - 4 + - 2 + - 0 + cascade can be simply interpreted in terms of a weak coupling model. (author)

  15. A measurement of the Ds lifetime

    International Nuclear Information System (INIS)

    Braunschweig, W.; Gerhards, R.; Kirschfink, F.J.; Martyn, H.U.; Rosskamp, P.; Kolanoski, A.; Balkwill, C.; Bowler, M.G.; Burrows, P.N.; Cashmore, R.J.; Dauncey, P.; Heath, G.P.; Mellor, D.J.; Ratoff, P.; Tomalin, I.; Yelton, J.M.; Baranko, G.; Caldwell, A.; Cherney, M.; Izen, J.M.; Muller, D.; Ritz, S.; Storm, D.; Takashima, M.; Wicklung, E.; Wu Saulan; Zobernig, G.

    1987-01-01

    The lifetime of the D S meson has been measured using the TASSO detector at PETRA and found to be (5.7 (+3.6-2.6)±0.9) x 10 -13 s. The method used was to reconstruct fully the decay vertex of the channel D s → φπ ± , φ → K + K - . (orig.)

  16. Updated measurement of the τ lepton lifetime

    Science.gov (United States)

    ALEPH Collaboration; Barate, R.; Buskulic, D.; Decamp, D.; Ghez, P.; Goy, C.; Lees, J.-P.; Lucotte, A.; Minard, M.-N.; Nief, J.-Y.; Pietrzyk, B.; Casado, M. P.; Chmeissani, M.; Comas, P.; Crespo, J. M.; Delfino, M.; Fernandez, E.; Fernandez-Bosman, M.; Garrido, Ll.; Juste, A.; Martinez, M.; Merino, G.; Miquel, R.; Mir, Ll. M.; Padilla, C.; Park, I. C.; Pascual, A.; Perlas, J. A.; Riu, I.; Sanchez, F.; Teubert, F.; Colaleo, A.; Creanza, D.; de Palma, M.; Gelao, G.; Iaselli, G.; Maggi, G.; Maggi, M.; Marinelli, N.; Nuzzo, S.; Ranieri, A.; Raso, G.; Ruggieri, F.; Selvaggi, G.; Silvestris, L.; Tempesta, P.; Tricomi, A.; Zito, G.; Huang, X.; Lin, J.; Ouyang, Q.; Wang, T.; Xie, Y.; Xu, R.; Xue, S.; Zhang, J.; Zhang, L.; Zhao, W.; Abbaneo, D.; Alemany, R.; Becker, U.; Bazarko, A. O.; Bright-Thomas, P.; Cattaneo, M.; Cerutti, F.; Dissertori, G.; Drevermann, H.; Forty, R. W.; Frank, M.; Hagelberg, R.; Hansen, J. B.; Harvey, J.; Janot, P.; Jost, B.; Kneringer, E.; Knobloch, J.; Lehraus, I.; Mato, P.; Minten, A.; Moneta, L.; Pacheco, A.; Pusztaszeri, J.-F.; Ranjard, F.; Rizzo, G.; Rolandi, L.; Rousseau, D.; Schlatter, D.; Schmitt, M.; Schneider, O.; Tejessy, W.; Tomalin, I. R.; Wachsmuth, H.; Wagner, A.; Ajaltouni, Z.; Barrès, A.; Boyer, C.; Falvard, A.; Ferdi, C.; Gay, P.; Guicheney, C.; Henrard, P.; Jousset, J.; Michel, B.; Monteil, S.; Montret, J.-C.; Pallin, D.; Perret, P.; Podlyski, F.; Proriol, J.; Rosnet, P.; Rossignol, J.-M.; Fearnley, T.; Hansen, J. D.; Hansen, J. R.; Hansen, P. H.; Nilsson, B. S.; Rensch, B.; Wäänänen, A.; Daskalakis, G.; Kyriakis, A.; Markou, C.; Simopoulou, E.; Siotis, I.; Vayaki, A.; Blondel, A.; Bonneaud, G.; Brient, J. C.; Bourdon, P.; Rougé, A.; Rumpf, M.; Valassi, A.; Verderi, M.; Videau, H.; Candlin, D. J.; Parsons, M. I.; Focardi, E.; Parrini, G.; Zachariadou, K.; Corden, M.; Georgiopoulos, C.; Jaffe, D. E.; Antonelli, A.; Bencivenni, G.; Bologna, G.; Bossi, F.; Campana, P.; Capon, G.; Casper, D.; Chiarella, V.; Felici, G.; Laurelli, P.; Mannocchi, G.; Murtas, F.; Murtas, G. P.; Passalacqua, L.; Pepe-Altarelli, M.; Curtis, L.; Dorris, S. J.; Halley, A. W.; Knowles, I. G.; Lynch, J. G.; O'Shea, V.; Raine, C.; Scarr, J. M.; Smith, K.; Teixeira-Dias, P.; Thompson, A. S.; Thomson, E.; Thomson, F.; Turnbull, R. M.; Buchmüller, O.; Dhamotharan, S.; Geweniger, C.; Graefe, G.; Hanke, P.; Hansper, G.; Hepp, V.; Kluge, E. E.; Putzer, A.; Sommer, J.; Tittel, K.; Werner, S.; Wunsch, M.; Beuselinck, R.; Binnie, D. M.; Cameron, W.; Dornan, P. J.; Girone, M.; Goodsir, S.; Martin, E. B.; Moutoussi, A.; Nash, J.; Sedgbeer, J. K.; Spagnolo, P.; Stacey, A. M.; Williams, M. D.; Ghete, V. M.; Girtler, P.; Kuhn, D.; Rudolph, G.; Betteridge, A. P.; Bowdery, C. K.; Colrain, P.; Crawford, G.; Finch, A. J.; Foster, F.; Hughes, G.; Jones, R. W. L.; Sloan, T.; Williams, M. I.; Galla, A.; Giehl, I.; Greene, A. M.; Hoffmann, C.; Jakobs, K.; Kleinknecht, K.; Quast, G.; Renk, B.; Rohne, E.; Sander, H.-G.; van Gemmeren, P.; Zeitnitz, C.; Aubert, J. J.; Benchouk, C.; Bonissent, A.; Bujosa, G.; Carr, J.; Coyle, P.; Diaconu, C.; Etienne, F.; Konstantinidis, N.; Leroy, O.; Motsch, F.; Payre, P.; Talby, M.; Sadouki, A.; Thulasidas, M.; Trabelsi, K.; Aleppo, M.; Antonelli, M.; Ragusa, F.; Berlich, R.; Blum, W.; Büscher, V.; Dietl, H.; Ganis, G.; Gotzhein, C.; Kroha, H.; Lütjens, G.; Lutz, G.; Männer, W.; Moser, H.-G.; Richter, R.; Rosado-Schlosser, A.; Schael, S.; Settles, R.; Seywerd, H.; St. Denis, R.; Stenzel, H.; Wiedenmann, W.; Wolf, G.; Boucrot, J.; Callot, O.; Chen, S.; Choi, Y.; Cordier, A.; Davier, M.; Duflot, L.; Grivaz, J.-F.; Heusse, Ph.; Höcker, A.; Jacholkowska, A.; Jacquet, M.; Kim, D. W.; Le Diberder, F.; Lefrançois, J.; Lutz, A.-M.; Nikolic, I.; Schune, M.-H.; Simion, S.; Tournefier, E.; Veillet, J.-J.; Videau, I.; Zerwas, D.; Azzurri, P.; Bagliesi, G.; Batignani, G.; Bettarini, S.; Bozzi, C.; Calderini, G.; Carpinelli, M.; Ciocci, M. A.; Ciulli, V.; dell'Orso, R.; Fantechi, R.; Ferrante, I.; Foà, L.; Forti, F.; Giassi, A.; Giorgi, M. A.; Gregorio, A.; Ligabue, F.; Lusiani, A.; Marrocchesi, P. S.; Messineo, A.; Palla, F.; Sanguinetti, G.; Sciabà, A.; Steinberger, J.; Tenchini, R.; Tonelli, G.; Vannini, C.; Venturi, A.; Verdini, P. G.; Blair, G. A.; Bryant, L. M.; Chambers, J. T.; Gao, Y.; Green, M. G.; Medcalf, T.; Perrodo, P.; Strong, J. A.; von Wimmersperg-Toeller, J. H.; Botterill, D. R.; Clifft, R. W.; Edgecock, T. R.; Haywood, S.; Norton, P. R.; Thompson, J. C.; Wright, A. E.; Bloch-Devaux, B.; Colas, P.; Emery, S.; Kozanecki, W.; Lançon, E.; Lemaire, M. C.; Locci, E.; Perez, P.; Rander, J.; Renardy, J.-F.; Roussarie, A.; Schuller, J.-P.; Schwindling, J.; Trabelsi, A.; Vallage, B.; Black, S. N.; Dann, J. H.; Johnson, R. P.; Kim, H. Y.; Litke, A. M.; McNeil, M. A.; Taylor, G.; Booth, C. N.; Boswell, R.; Brew, C. A. J.; Cartwright, S.; Combley, F.; Kelly, M. S.; Lehto, M.; Newton, W. M.; Reeve, J.; Thompson, L. F.; Böhrer, A.; Brandt, S.; Cowan, G.; Grupen, C.; Lutters, G.; Saraiva, P.; Smolik, L.; Stephan, F.; Apollonio, M.; Bosisio, L.; della Marina, R.; Giannini, G.; Gobbo, B.; Musolino, G.; Putz, J.; Rothberg, J.; Wasserbaech, S.; Armstrong, S. R.; Charles, E.; Elmer, P.; Ferguson, D. P. S.; González, S.; Greening, T. C.; Hayes, O. J.; Hu, H.; Jin, S.; McNamara, P. A., III; Nachtman, J. M.; Nielsen, J.; Orejudos, W.; Pan, Y. B.; Saadi, Y.; Scott, I. J.; Walsh, J.; Wu, Sau Lan; Wu, X.; Yamartino, J. M.; Zobernig, G.

    1997-11-01

    A new measurement of the mean lifetime of the τ lepton is presented. Three different analysis methods are applied to a sample of 90 000 τ pairs, collected in 1993 and 1994 with the ALEPH detector at LEP. The average of this measurement and those previously published by ALEPH is ττ=290.1+/-1.5+/-1.1 fs.

  17. Cosmological constraints on the neutron lifetime

    Energy Technology Data Exchange (ETDEWEB)

    Salvati, L.; Pagano, L.; Melchiorri, A. [Physics Department, Università di Roma ' ' La Sapienza' ' , Piazzale Aldo Moro 2, 00185, Rome (Italy); Consiglio, R., E-mail: laura.salvati@roma1.infn.it, E-mail: luca.pagano@roma1.infn.it, E-mail: rconsiglio@na.infn.it, E-mail: alessandro.melchiorri@roma1.infn.it [Physics Department, Università di Napoli ' ' Federico II' ' , Complesso Universitario Monte S. Angelo, Via Cintia, I-80126 Napoli (Italy)

    2016-03-01

    We derive new constraints on the neutron lifetime based on the recent Planck 2015 observations of temperature and polarization anisotropies of the CMB. Under the assumption of standard Big Bang Nucleosynthesis, we show that Planck data constrains the neutron lifetime to τ{sub n} = (907±69) [s] at 68% c.l.. Moreover, by including the direct measurements of primordial Helium abundance of Aver et al. (2015) and Izotov et al. (2014), we show that cosmological data provide the stringent constraints τ{sub n} = (875±19) [s] and τ{sub n} = (921±11) [s] respectively. The latter appears to be in tension with neutron lifetime value quoted by the Particle Data Group (τ{sub n} = (880.3±1.1) [s]). Future CMB surveys as COrE+, in combination with a weak lensing survey as EUCLID, could constrain the neutron lifetime up to a ∼ 6 [s] precision.

  18. Increasing the lifetime of fuel cell catalysts

    NARCIS (Netherlands)

    Latsuzbaia, R.

    2015-01-01

    In this thesis, I discuss a novel idea of fuel cell catalyst regeneration to increase lifetime of the PEM fuel cell electrode/catalyst operation and, therefore, reduce the catalyst costs. As many of the catalyst degradation mechanisms are difficult to avoid, the regeneration is alternative option to

  19. Lifetime oriented maintenance planning in the Netherlands

    NARCIS (Netherlands)

    Straub, A.

    2003-01-01

    In this paper we set up a framework for lifetime oriented maintenance planning as an outcome and input for strategic housing stock management. The maintenance planning holds maintenance activities and costs in the longer term. We consider the maintenance planning as a tool to calculate and implement

  20. Lifetime tests for MAC vertex chamber

    International Nuclear Information System (INIS)

    Nelson, H.N.

    1986-07-01

    A vertex chamber for MAC was proposed to increase precision in the measurement of the B hadron and tau lepton lifetimes. Thin-walled aluminized mylar drift tubes were used for detector elements. A study of radiation hardness was conducted under the conditions of the proposed design using different gases and different operating conditions