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Sample records for fluorescence depolarization measurements

  1. Fluorescence depolarization measurements under shock compression

    NASA Astrophysics Data System (ADS)

    Wang, Jue; Banishev, Alexandr; Bassett, Will P.; Dlott, Dana D.

    2017-01-01

    Measurements of the time-dependent fluorescence depolarization of emissive probe molecules enable real-time observations of molecular rotations in shocked materials. In shocked solids, molecular rotations occur as a result of shear deformations. An apparatus is described to measure time-dependent fluorescence depolarization of shocked materials using laser-driven flyer plates and either a picosecond or a nanosecond probe laser. The emission was separated into parallel and perpendicular channels and imaged onto a streak camera. Time-dependent fluorescence depolarization of rhodamine 6G (R6G) dye dissolved in poly-methyl methacrylate (PMMA) was measured with a 16 ns duration impact at 1 km s-1. A partial depolarization of the dye emission was observed to occur during a 150 ns period after the shock.

  2. Reexamination of depolarization in lidar measurements.

    PubMed

    Gimmestad, Gary G

    2008-07-20

    Almost all of the depolarization papers in the lidar literature employ a physically inappropriate notation and they use a definition of the depolarization ratio that is not linear in the quantity of interest. This depolarization lidar legacy is misleading and confusing. In particular, subscripts meaning parallel and perpendicular do not apply to atmospheric parameters, such as the volume backscatter coefficient, because (for linear polarization) the two components of the backscattered light are polarized in the transmitted sense and completely unpolarized; the unpolarized component is not "perpendicular." An analysis of lidar depolarization measurements with a particle scattering matrix recently provided in the literature yields algorithms for retrieving the depolarization parameter from either linear or circular depolarization lidar measurements. The analysis, notation, and definitions recommended here harmonize lidar depolarization analysis with radiative transfer theory, particle scattering theory, and standard polarization measurement techniques.

  3. Identifiability analysis of rotational diffusion tensor and electronic transition moments measured in time-resolved fluorescence depolarization experiment.

    PubMed

    Szubiakowski, Jacek P

    2014-06-14

    The subject of this paper is studies of the deterministic identifiability of molecular parameters, such as rotational diffusion tensor components and orientation of electronic transition moments, resulting from the time-resolved fluorescence anisotropy experiment. In the most general case considered, a pair of perpendicularly polarized emissions enables the unique determination of all the rotational diffusion tensor's principal components. The influence of the tensor's symmetry and the associated degeneration of its eigenvalues on the identifiability of the electronic transitions moments is systematically investigated. The analysis reveals that independently of the rotational diffusion tensor's symmetry, the transition moments involved in photoselection and emission processes cannot be uniquely identified without a priori information about their mutual orientation or their orientation with respect to the principal axes of the tensor. Moreover, it is shown that increasing the symmetry of the rotational diffusion tensor deteriorates the degree of the transition moments identifiability. To obtain these results analytically, a novel approach to solve bilinear system of equations for Markov parameters is applied. The effect of the additional information, obtained from fluorescence measurements for different molecular mobilities, to improve the identifiability at various levels of analysis is shown. The effectiveness and reliability of the target analysis method for experimental determination of the molecular parameters is also discussed.

  4. Identifiability analysis of rotational diffusion tensor and electronic transition moments measured in time-resolved fluorescence depolarization experiment

    SciTech Connect

    Szubiakowski, Jacek P.

    2014-06-14

    The subject of this paper is studies of the deterministic identifiability of molecular parameters, such as rotational diffusion tensor components and orientation of electronic transition moments, resulting from the time-resolved fluorescence anisotropy experiment. In the most general case considered, a pair of perpendicularly polarized emissions enables the unique determination of all the rotational diffusion tensor's principal components. The influence of the tensor's symmetry and the associated degeneration of its eigenvalues on the identifiability of the electronic transitions moments is systematically investigated. The analysis reveals that independently of the rotational diffusion tensor's symmetry, the transition moments involved in photoselection and emission processes cannot be uniquely identified without a priori information about their mutual orientation or their orientation with respect to the principal axes of the tensor. Moreover, it is shown that increasing the symmetry of the rotational diffusion tensor deteriorates the degree of the transition moments identifiability. To obtain these results analytically, a novel approach to solve bilinear system of equations for Markov parameters is applied. The effect of the additional information, obtained from fluorescence measurements for different molecular mobilities, to improve the identifiability at various levels of analysis is shown. The effectiveness and reliability of the target analysis method for experimental determination of the molecular parameters is also discussed.

  5. Improved calibration method for depolarization lidar measurement.

    PubMed

    Liu, Bo; Wang, Zhien

    2013-06-17

    An improved calibration method for lidar depolarization measurement is described. With this method the system constants including the electronic gain ratio of the parallel and perpendicular channels, the optical reflectance and transmission parameters of the polarizing beam splitter, and the linear polarization ratio of the emitting laser beam can be determined conveniently by using lidar measurements with a half-wave plate oriented at selected angles.

  6. Effect of multiple scattering on depolarization measurements with spaceborne lidars.

    PubMed

    Reichardt, Susanne; Reichardt, Jens

    2003-06-20

    An analytical model based on the integration of the scattering-angle and light-path manifold has been developed to quantify the effect of multiple scattering on cirrus measurements obtained with elastic polarization lidars from space. Light scattering by molecules and by a horizontally homogeneous cloud is taken into account. Lidar parameter, including laser beam divergence, can be freely chosen. Up to 3 orders of scattering are calculated. Furthermore, an inversion technique for the retrieval of cloud extinction profiles from measurements with elastic-backscatter lidars is proposed that explicitly takes multiple scattering into account. It is found that for typical lidar system parameters such as those of the Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observations (CALIPSO) instrument multiple scattering does not significantly affect depolarization-ratio measurements in cirrus clouds with small to moderate optical depths. For all simulated clouds, the absolute value of the difference between measured and single-scattering volume depolarization ratio is < 0.006. The particle depolarization ratio can be calculated from the measured volume depolarization ratio and the retrieved backscatter ratio without degradation of accuracy; thus characterization of the various cirrus categories in terms of the particle depolarization ratio and retrieval of cloud microphysical properties is feasible from space. The results of this study apply to polar stratospheric clouds as well.

  7. Depolarization of Surface-Enhanced Fluorescence: An Approach to Fluorescence Polarization Assays

    PubMed Central

    Szmacinski, Henryk; Lakowicz, Joseph R.

    2009-01-01

    Localized surface plasmons of metallic particles of sub-wavelength sizes strongly modify the spectral properties of nearby fluorophores. The enhanced radiative decay rate leads to high fluorescence efficiencies and decreased fluorescence lifetimes. In this report we show that metal-enhanced fluorescence generated by the presence of the silver islands on the glass substrate displays high depolarization. Intensities, lifetimes, and emission anisotropies of several fluorophore protein conjugates have been studied in the absence and presence of metallic nanostructures. Despite highly decreased lifetimes of about 10-fold and immobilization of conjugates on the solid substrate, the observed emission anisotropies for all fluorophores on the metal-enhanced substrate decreased 300–500% compared to that in solution. This observation implies a new generation of fluorescence polarization immunoassays with broad applications because of no restrictions to the lifetime of the probe and the size of labeled biomolecules. The changes in polarization are due to binding that occur on the bioactive surface localized near the metal particles. PMID:18627176

  8. Developing Ocean Subsurface Data Record from CALIPSO Depolarization Ratio Measurements

    NASA Astrophysics Data System (ADS)

    Hu, Y.; Rodier, S. D.; Zhai, P.; Josset, D. B.; Omar, A. H.

    2012-12-01

    CALIOP, the dual wavelength, polarization sensitive lidar flying aboard the CALIPSO satellite, has been operating since June 2006 and is expected to continue for several more years. CALIOP's depolarization ratio is one of the best calibrated measurements made by the A-Train sensors. Over the life of the CALIPSO mission, the stability of the CALIOP depolarization ratio calibration has remained within 1%. CALIOP's depolarization ratio measurements can be used for studying changes in the backscatter of ocean subsurface particulates. The ocean surface/subsurface depolarization ratio measurements from CALIOP together with collocated A-train instruments can be used for estimating the subsurface particulate backscatter coefficient (bbp) and the cross polarization component of the column integrated ocean subsurface backscatter signal. To derive the cross component of inte¬grated ocean subsurface backscatter signal from the depolarization ratio of CALIOP integrated ocean surface/subsurface range bins, we need theoretical estimates of the ocean surface back¬scatter cross section, which can be derived accurately from CloudSat ocean surface backscatter measurements or from AMSR-E wind speeds. Using the CALIOP cross component of the integrated ocean subsurface backscatter together with diffuse attenuation coefficient derived from MODIS, bbp can be estimated. Preliminary CALIOP data analysis shows that in the Southern Oceans, ocean subsurface backscatter has increased by about 5% since the beginning of the CALIPSO mission in June 2006. This study will: (1) introduce the CALIPSO ocean subsurface backscatter data products; (2) assess the uncertainties in the data products and comparing with in situ optics measurements, radiative transfer modeling and aircraft lidar measurements; and (3) summarize the global and regional (e.g., Gulf of Mexico region) statistics and temporal variations of the ocean subsurface backscatter from CALIPSO measurements.

  9. Fluorescence imaging of heat-stress induced mitochondrial long-term depolarization in breast cancer cells.

    PubMed

    Dressler, Cathrin; Beuthan, Juergen; Mueller, Gerhard; Zabarylo, Urszula; Minet, Olaf

    2006-09-01

    Various thermotherapies are based on the induction of lethal heat in target tissues. Spatial and temporal instabilities of elevated temperatures induced in therapy targets require optimized treatment protocols and reliable temperature control methods during thermotherapies. Heat-stress induced effects on mitochondrial transmembrane potentials were analyzed in breast cancer cells, species MX1, using the potential sensor JC-1 (Molecular Probes, Invitrogen, Germany). Potential dependant labeling of heat-stressed cells was imaged and evaluated by fluorescence microscopy and compared with control cells. JC-1 stains mitochondria in cells with high mitochondrial potentials by forming orange-red fluorescent J-aggregates while in cells with depolarized or damaged mitochondria the sensor dye exists as green fluorescent monomers. In MX1 cells orange-red and green fluorescence intensities were correlated with each other after various heat-stress treatments and states of mitochondrial membrane potentials were deduced from the image data. With increasing stress temperatures the intensity of red fluorescent J-aggregates decreased while the green fluorescence intensity of JC-1 monomers increased. This heat-stress response happened in a nonlinear manner with increasing temperatures resulting in a nonlinear increase of red/green fluorescence ratios. These data indicated that mitochondria in MX1 cells were increasingly depolarized in response to increasing ambient temperatures.

  10. A Critique of Asphaltene Fluorescence Decay and Depolarization-Based Claims about Molecular Weight and Molecular Architecture

    SciTech Connect

    Strausz,O.; Safarik, I.; Lown, E.; Morales-Izquierdo, A.

    2008-01-01

    Relying on experimental and theoretical data available from the literature, it is shown that the conclusions derived from measurements of fluorescence decay and depolarization kinetic times as reported in a series of papers over the past decade are egregiously wrong. To start with, the decay time measurements were done with inappropriate instrumentation which resulted in misleading results. Misinterpretation of the results led to the mistaken conclusion that bichromophoric type molecules are absent from petroleum asphaltene and therefore the architecture of the asphaltene molecule features a single condensed cyclic core spiked with some alkyl chains, in spite of irrefutable chemical evidence to the contrary. It was further concluded that if the asphaltene core is a single condensed ring, then the fluorescence depolarization with rotational correlation time method is applicable for the molecular weight determination of asphaltene. This is definitely not so, since, regardless of any other considerations, asphaltene is a mixture of a plethora of different, unknown components, with unknown concentrations along with innumerable different, unknown and some known chromophores portraying widely different absorption coefficients, fluorescence quantum yields, and kinetic decay times. Consequently, asphaltene fluorescence is a highly complex function of the above attributes and as such it is a totally unsuitable property for its molecular weight determination. The injection of an incorrect, single condensed ring core architecture for asphaltene has caused some confusion in asphaltene chemistry that has now hopefully been settled.

  11. Depolarization after resonance energy transfer (DARET): a sensitive fluorescence-based assay for botulinum neurotoxin protease activity.

    PubMed

    Gilmore, Marcella A; Williams, Dudley; Okawa, Yumiko; Holguin, Bret; James, Nicholas G; Ross, Justin A; Roger Aoki, K; Jameson, David M; Steward, Lance E

    2011-06-01

    The DARET (depolarization after resonance energy transfer) assay is a coupled Förster resonance energy transfer (FRET)-fluorescence polarization assay for botulinum neurotoxin type A or E (BoNT/A or BoNT/E) proteolytic activity that relies on a fully recombinant substrate. The substrate consists of blue fluorescent protein (BFP) and green fluorescent protein (GFP) flanking SNAP-25 (synaptosome-associated protein of 25 kDa) residues 134-206. In this assay, the substrate is excited with polarized light at 387 nm, which primarily excites the BFP, whereas emission from the GFP is monitored at 509 nm. Energy transfer from the BFP to the GFP in the intact substrate results in a substantial depolarization of the GFP emission. The energy transfer is eliminated when the fluorescent domains separate on cleavage by the endopeptidase, and emission from the directly excited GFP product fragment is then highly polarized, resulting in an overall increase in polarization. This increase in polarization can be monitored to assay the proteolytic activity of BoNT/A and BoNT/E in real time. It allows determination of the turnover rate of the substrate and the kinetic constants (V(max) and k(cat)) based on the concentration of cleaved substrate determined directly from the measurements using the additivity properties of polarization. The assay is amenable to high-throughput applications.

  12. Local mobility of polymer chain grafted onto polyethylene monitored by fluorescence depolarization

    NASA Astrophysics Data System (ADS)

    Tsuneda, Satoshi; Endo, Toshihiro; Saito, Kyoichi; Sugita, Kazuyuki; Horie, Kazuyuki; Yamashita, Takashi; Sugo, Takanobu

    1997-08-01

    The fluorescence depolarization method was used for investigating the local mobility of polymer chains grafted onto a porous polyethylene membrane. The real value of the rotational diffusion coefficient of a dansyl probe attached to the grafted polymer chain was obtained by using a correction method which eliminated the effect of multiple scattering on fluorescence anisotropy. The rotational mobility of the dansyl probe attached to the grafted polymer chain was sensitive to both degree of grafting and solvent polarity, which indicated that the conformation of the grafted polymer chain and the pore size of the base membrane strongly governed the dynamic parameters of the grafted polymer chain.

  13. Depolarization of the intrinsic and extrinsic fluorescence of pepsinogen and pepsin

    PubMed Central

    Teale, F. W. J.; Badley, R. A.

    1970-01-01

    1. The effects on the intrinsic tryptophan emission anisotropy of pepsin and pepsinogen solutions produced by (a) changes in temperature, (b) increases in viscosity with added glycerol at constant temperature and (c) decreases in lifetime through collisional quenching by potassium iodide were measured at several excitation wavelengths. The rotational-relaxation times calculated from results provided by method (b) approximate to the theoretical values for the two proteins, on taking hydration and shape factors into account, on the basis of random orientation of the tryptophan groups within the macromolecules. Differences between the results provided by methods (b) and (c) are attributable to inter-tryptophan resonance-energy-transfer depolarization, and the anomalous values recorded in method (a) can be attributed to the temperature-dependence of the limiting anisotropies. 2. Two different monomeric conjugates of pepsin, each containing one extrinsic fluorescent group per macromolecule, gave widely different relaxation times. This difference may arise from a specific orientation of the emission dipole in the enzyme. In active-site-labelled pepsin (1-dimethylaminonaphthalene-5-sulphonylphenylalanine–pepsin) this orientation would be approximately parallel to the symmetry axis of the equivalent ellipsoid, whereas in the other conjugate (1-dimethylaminonaphthalene-5-sulphonyl-pepsin) the orientation may be roughly normal to this direction, or some independent rotation of parts of the protein molecule is possible. PMID:4907957

  14. Suppression of Rayleigh scattering noise in sodium laser guide stars by hyperfine depolarization of fluorescence.

    PubMed

    Guillet de Chatellus, Hugues; Moldovan, Ioana; Fesquet, Vincent; Pique, Jean-Paul

    2006-11-27

    We propose what we believe is a novel method for enabling the complete suppression of noise due to Rayleigh scattering in sodium laser guide star systems by means of selective discrimination between Rayleigh and fluorescence signals based on polarization properties. We show that, contrary to the nearly 100% polarized Rayleigh scattering, fluorescence from the D(2) sodium line is strongly depolarized under excitation by a modeless laser. This offers the possibility of completely cancelling the effects of the Rayleigh scattering background while preserving the fluorescence signal to about 40% of its maximal value, leading to an improvement of the signal-to-noise ratio by several orders of magnitude. Both theoretical and experimental data confirm this new proposal.

  15. Bipolar and fixable probe targeting mitochondria to trace local depolarization via two-photon fluorescence lifetime imaging.

    PubMed

    Wang, Benlei; Zhang, Xinfu; Wang, Chao; Chen, Lingcheng; Xiao, Yi; Pang, Yi

    2015-08-21

    Polarization/depolarization levels of different single mitochondria in a cell are inhomogeneous, and always varying. Because depolarization is an indicator of mitochondrial dysfunction, tracing local depolarization is highly desirable. The existing fluorescent probes, however, are not well suited for this task, although they are applicable to assess the average polarization extents of whole cells. A multifunctional and bipolar probe MITFPS is thus developed, which includes a positively charged hydrophilic group and an environment sensitive fluorophore. In the probe design, the hydrophilic anchoring unit is chemically immobilized on a membrane protein, while the lipophilic fluorophore can be inserted deep into the phospholipid layer. The probe exhibits a sensitive response to the local variation in polarization by changing its fluorescence lifetime. MITFPS's applicability is confirmed by real-time in situ imaging of the complete process of an uncoupler-induced depolarization under a two-photon fluorescence lifetime microscope. The imaging result reveals that one mitochondrion could have quite different polarization than the other, even though they are in the same cell.

  16. Aerosol Properties over Southeastern China from Multi-Wavelength Raman and Depolarization Lidar Measurements

    NASA Astrophysics Data System (ADS)

    Heese, Birgit; Althausen, Dietrich; Baars, Holger; Bohlmann, Stephanie; Deng, Ruru

    2016-06-01

    A dataset of particle optical properties of highly polluted urban aerosol over the Pearl River Delta, Guangzhou, China is presented. The data were derived from multi-wavelengths Raman and depolarization lidar PollyXT and AERONET sun photometer measurements. The measurement campaign was conducted from Nov 2011 to June 2012. High aerosol optical depth was observed in the polluted atmosphere over this megacity, with a mean value of 0.54 ± 0.33 and a peak value of even 1.9. For the particle characterization the lidar ratio and the linear particle depolarization ratio, both at 532 nm, were used. The mean values of these properties are 48.0 sr ± 10.7 sr for the lidar ratio and 4%+-4% for the particle depolarization ratio, which means most depolarization measurements stayed below 10%. So far, most of these results indicate urban pollution particles mixed with particles arisen from biomass and industrial burning.

  17. Three-Signal Method for Accurate Measurements of Depolarization Ratio with Lidar

    NASA Technical Reports Server (NTRS)

    Reichardt, Jens; Baumgart, Rudolf; McGee, Thomsa J.

    2003-01-01

    A method is presented that permits the determination of atmospheric depolarization-ratio profiles from three elastic-backscatter lidar signals with different sensitivity to the state of polarization of the backscattered light. The three-signal method is insensitive to experimental errors and does not require calibration of the measurement, which could cause large systematic uncertainties of the results, as is the case in the lidar technique conventionally used for the observation of depolarization ratios.

  18. Light depolarization measurements in malaria: A new job for an old friend.

    PubMed

    Rebelo, Maria; Tempera, Carolina; Bispo, Claudia; Andrade, Claudia; Gardner, Rui; Shapiro, Howard M; Hänscheid, Thomas

    2015-05-01

    The use of flow cytometry in malaria research has increased over the last decade. Most approaches use nucleic acid stains to detect parasite DNA and RNA and require complex multi-color, multi-parameter analysis to reliably detect infected red blood cells (iRBCs). We recently described a novel and simpler approach to parasite detection based on flow cytometric measurement of scattered light depolarization caused by hemozoin (Hz), a pigment formed by parasite digestion of hemoglobin in iRBCs. Depolarization measurement by flow cytometry was described in 1987; however, patent issues restricted its use to a single manufacturer's hematology analyzers until 2009. Although we recently demonstrated that depolarization measurement of Hz, easily implemented on a bench top flow cytometer (Cyflow), provided useful information for malaria work, doubts regarding its application and utility remain in both the flow cytometry and malaria communities, at least in part because instrument manufacturers do not offer the option of measuring depolarized scatter. Under such circumstances, providing other researchers with guidance as to how to do this seemed to offer the most expeditious way to resolve the issue. We accordingly examined how several commercially available flow cytometers (CyFlow SL, MoFLo, Attune and Accuri C6) could be modified to detect depolarization due to the presence of free Hz on solution, or of Hz in leukocytes or erythrocytes from rodent or human blood. All were readily adapted, with substantially equivalent results obtained with lasers emitting over a wide wavelength range. Other instruments now available may also be modifiable for Hz measurement. Cytometric detection of Hz using depolarization is useful to study different aspects of malaria. Adding additional parameters, such as DNA content and base composition and RNA content, can demonstrably provide improved accuracy and sensitivity of parasite detection and characterization, allowing malaria researchers and

  19. A New Technique for Measurements of Cloud Properties Using Lidar Depolarization

    NASA Astrophysics Data System (ADS)

    McCullough, E. M.; Perro, C. W.; Sica, R. J.; Duck, T.; Walker, K. A.; Drummond, J. R.

    2013-12-01

    The radiative behaviour of clouds is dependent on cloud particle phase. Water droplets can exist in temperatures well below 0° C for extended periods. Lidar depolarization ratio measurements allow liquid and solid states to be differentiated in individual clouds at high spatial-temporal resolution. The 2012 and 2013 Canadian Arctic ACE Validation Campaigns in Eureka, Nunavut, Canada (80°N, 86°W) provided an opportunity to make nearly continuous depolarization measurements using the CANDAC Rayleigh-Mie-Raman Lidar (CRL) in the troposphere. Depolarization ratio measurements using two techniques are presented here. The CRL transmits linearly polarized 532 nm laser pulses. The depolarizing effect of the cloud particles on the backscattered 532 nm photons is expressed as the linear depolarization ratio. A typical lidar uses two detection channels to determine the ratio: a 'Parallel' channel and a 'Perpendicular' channel. Backscattered photons may all remain polarized parallel to the transmitted beam (depol ratio = 0), or be depolarized, up to half parallel and half perpendicular (depol ratio = 1; light unpolarized). Thus, even before taking receiver optics into account, maximum possible count rates differ by a factor of two between the channels. In the CRL, optics further suppress perpendicular photons, reducing perpendicular rates to an order of magnitude smaller than the parallel rates, making the conventional retrieval technique challenging. To use the conventional technique, perpendicular signals require considerable co-adding to meet acceptable signal-to-noise thresholds, i.e. greater than 30 minutes even in ice clouds where signal rates are highest. Clear-sky retrievals are impossible. The CRL has been experimenting with an alternate approach: using the parallel channel in conjunction with the familiar polarization-insensitive Rayleigh Elastic channel. Sufficient parallel and Rayleigh elastic photons are recorded that depolarization ratio calculations with the

  20. Arctic polar stratospheric cloud measurements by means of a four wavelength depolarization lidar

    NASA Technical Reports Server (NTRS)

    Stefanutti, L.; Castagnoli, F.; Delguasta, M.; Flesia, C.; Godin, S.; Kolenda, J.; Kneipp, H.; Kyro, Esko; Matthey, R.; Morandi, M.

    1994-01-01

    A four wavelength depolarization backscattering lidar has been operated during the European Arctic Stratospheric Ozone Experiment (EASOE) in Sodankyl, in the Finnish Arctic. The lidar performed measurements during the months of December 1991, January, February and March 1992. The Finnish Meteorological Institute during the same period launched regularly three Radiosondes per day, and three Ozone sondes per week. Both Mt. Pinatubo aerosols and Polar Stratospheric Clouds were measured. The use of four wavelengths, respectively at 355 nm, 532 nm , 750 nm, and 850 nm permits an inversion of the lidar data to determine aerosol particle size. The depolarization technique permits the identification of Polar Stratospheric Clouds. Frequent correlation between Ozone minima and peaks in the Mt. Pinatubo aerosol maxima were detected. Measurements were carried out both within and outside the Polar Vortex.

  1. Measurement of the depolarization ratio of Rayleigh scattering at absorption bands

    NASA Astrophysics Data System (ADS)

    Anglister, J.; Steinberg, I. Z.

    1981-01-01

    Measurements of the depolarization ratio ρv of light scattered by the pigments lycopene and β-carotene at the red part of their absorption bands yielded values which are very close to the theoretical value 1/3 of a fully anisotropic molecular polarizability, i.e., that due to an electric dipole moment. Measurements of ρv at the blue edge of the visible absorption band of pinacyanol chloride yielded a value of 0.75 at 472.2 nm, which is the maximum value that a depolarization ratio can assume, and is attained if the average molecular polarizability is zero. This is possible only if the diagonalized polarizability tensor has at least one negative element to counterbalance the positive ones. A negative refractive index at the blue edge of the absorption band is thus experimentally demonstrated.

  2. Detection of mitochondrial depolarization/recovery during ischaemia–reperfusion using spectral properties of confocally recorded TMRM fluorescence

    PubMed Central

    Venable, Paul W; Taylor, Tyson G; Sciuto, Katie J; Zhao, Jerry; Shibayama, Junko; Warren, Mark; Spitzer, Kenneth W; Zaitsev, Alexey V

    2013-01-01

    Timing and pattern of mitochondrial potential (ΔΨm) depolarization during no-flow ischaemia–reperfusion (I–R) remain controversial, at least in part due to difficulties in interpreting the changes in the fluorescence of ΔΨm-sensitive dyes such as TMRM. The objective of this study was to develop a new approach for interpreting confocal TMRM signals during I–R based on spatial periodicity of mitochondrial packaging in ventricular cardiomyocytes. TMRM fluorescence (FTMRM) was recorded from Langendorff-perfused rabbit hearts immobilized with blebbistatin using either a confocal microscope or an optical mapping system. The hearts were studied under normal conditions, during mitochondrial uncoupling using the protonophore FCCP, and during I–R. Confocal images of FTMRM were subjected to spatial Fourier transform which revealed distinct peaks at a spatial frequency of ∼2 μm−1. The area under the peak (MPA) progressively decreased upon application of increasing concentrations of FCCP (0.3–20 μm), becoming undetectable at 5–20 μm FCCP. During ischaemia, a dramatic decrease in MPA, reaching the low/undetectable level comparable to that induced by 5–20 μm FCCP, was observed between 27 and 69 min of ischaemia. Upon reperfusion, a heterogeneous MPA recovery was observed, but not a de novo MPA decrease. Both confocal and wide-field imaging registered a consistent decrease in spatially averaged FTMRM in the presence of 5 μm FCCP, but no consistent change in this parameter during I–R. We conclude that MPA derived from confocal images provides a sensitive and specific indicator of significant mitochondrial depolarization or recovery during I–R. In contrast, spatially averaged FTMRM is not a reliable indicator of ΔΨm changes during I–R. PMID:23529126

  3. Single ice crystal measurements during nucleation experiments with the depolarization detector IODE

    NASA Astrophysics Data System (ADS)

    Nicolet, M.; Stetzer, O.; Lüönd, F.; Möhler, O.; Lohmann, U.

    2010-01-01

    In order to determine the efficiency of different aerosol particles to nucleate ice, an Ice Optical DEpolarization detector (IODE) was developed to distinguish between water droplets and ice crystals in ice nucleation chambers. A laser beam polarized linearly (power: 50 mW, wavelength: 407 nm) is directed through the chamber. The scattered light intensity from particles is measured at a scattering angle of Θ=175° in both polarization components (parallel and perpendicular). The ratio between the perpendicular intensity over the total one yields the depolarization ratio δ. Single particle detection is possible, using a peak detection algorithm. For high particle concentrations, a real-time signal averaging method can also be run simultaneously. The IODE detector was used in connection with the Zurich ice nucleation chamber during the ICIS 2007 workshop where ice nucleation experiments were performed with several aerosol types. In presence of ice crystals, a depolarization ratio could be measured on a particle-by-particle basis. Mean values of δ ranged from 0.24 to 0.37 and agree well with theoretical calculations.

  4. Single ice crystal measurements during nucleation experiments with the depolarization detector IODE

    NASA Astrophysics Data System (ADS)

    Nicolet, M.; Stetzer, O.; Lohmann, U.; Möhler, O.

    2008-12-01

    In order to determine the efficiency of aerosol particles of several types to nucleate ice, an Ice Optical DEpolarization detector (IODE) was developed to distinguish between water droplets and ice crystals in ice nucleation chambers. A laser beam polarized linearly (power: 50 mW, wavelength: 407 nm) is directed through the chamber. The scattered light intensity from particles is measured at a scattering angle of Θ=175° in both polarization components (parallel and perpendicular). The ratio between the perpendicular intensity over the total one gives the depolarization ratio δ. Single particle detection is possible, using a peak detection algorithm. For high particle concentrations, a real-time signal averaging method can also be run simultaneously. The IODE detector was used in connection with the Zurich ice nucleation chamber during the ICIS 2007 workshop where ice nucleation experiments were performed with several aerosol types. In presence of ice crystals, peaks were detected in both channels, generating depolarization signals. Mean values of δ ranged from 0.24 to 0.37.

  5. Femtosecond fluorescence depolarization study of photosynthetic antenna proteins: observation of ultrafast energy transfer in trimeric C-phycocyanin and allophycocyanin

    NASA Astrophysics Data System (ADS)

    Xie, Sunney; Du, Mei; Mets, Laurens; Fleming, Graham R.

    1992-04-01

    C-phycocyanin (CPC) and Allophycocyanin (APC) are pigment-protein complexes isolated from antenna systems in cyanobacteria. The crystal structure of CPC has recently been solved and APC has a similar structure. CPC and APC have a trimeric structure, monomeric subunits are composed of an (alpha) and (beta) polypeptide chain, each has a tetrapyrrole chromophore chemically bound to position 84. In CPC and APC trimers, the (alpha) 84 and (beta) 84 chromophores in adjacent monomers are in close proximity, forming relatively strong coupled pairs. Calculation of pairwise energy transfer rates using Foerster theory has suggested an extremely fast transfer (> 1 ps-1) between the (alpha) 84 and (beta) 84 pair in CPC. A femtosecond fluorescence up-conversion apparatus was constructed which achieves subhundred femtosecond time resolution. This allows experimental observation of the fast energy transfer process between the (alpha) 84 and (beta) 84 pair in both CPC and APC. There was also a wavelength dependence of the fluorescence depolarization kinetics which is inconsistent with Foerster inductive resonance energy transfer theory.

  6. Impacts of depolarization calibration methods on cloud phase interpretation at Eureka during 2013 and 2014 CRL lidar measurement campaigns

    NASA Astrophysics Data System (ADS)

    McCullough, E. M.; Perro, C. W.; Gamage, S. M.; Hopper, J.; Sica, R. J.; Duck, T.; Walker, K. A.; Drummond, J. R.

    2014-12-01

    The radiative behaviour of clouds is dependent on cloud particle phase. Water droplets can exist in temperatures well below 0° C for extended periods. Lidar depolarization measurements allow liquid and solid states to be differentiated in individual clouds at high spatial-temporal resolution. The 2012, 2013 and 2014 Canadian Arctic ACE Validation Campaigns in Eureka, Nunavut, Canada (80°N, 86°W) provided an opportunity to make extensive depolarization measurements using the CANDAC Rayleigh-Mie-Raman Lidar (CRL) in the troposphere.To date, most calibration methods in the literature are applicable to lidars which do not have non-ideal polarizing optics upstream of the polarizing analyzers in the receiver. We demonstrate a more complete matrix algebra calibration of the CRL to take the extra optics from six upstream lidar channels into account.Differences in depolarization parameter from 2013 and 2014 measurements show the advantage of the more extensive calibration for this lidar compared to the simpler traditional approach. The largest differences are found for depolarization parameter values around d = 0.50 (corresponding to δ = 0.33). Depolarization ratio values of δ = 0.2 to 0.3 are generally taken to be the cutoff between interpretations of ice (higher δ) or water (lower δ), and many CRL measurements lie in this particularly diagnostic range. An uncertainty analysis becomes important when extending the depolarization parameters to interpretation of the clouds in the atmosphere above Eureka.

  7. Fluorescence depolarization and contact angle investigation of dynamic and static interfacial tension of liquid crystal display materials.

    PubMed

    Quintella, Cristina M; Lima, Angelo M V; Gonçalves, Cristiane C; Watanabe, Yuji N; Mammana, Alaide P; Schreiner, Marcos A; Pepe, Iuri; Pizzo, Angela A

    2003-06-01

    Interfacial interactions control two processes empirically known to be critical for molecular anchoring in twisted nematic liquid crystal displays technology (TN-LCDs): surface treatment and filling procedure. Static and dynamical interfacial tensions (Gamma(SL)) between liquids and several substrates with similar roughness were observed respectively by contact angle (theta(c)) of sessile drops and by fluorescence depolarization of thin liquid films flowing at high velocity. Gamma(SL) decreased when glass was coated with tin dioxide and increased with polyvinyl alcohol (PVA) deposition. Drops were circular for all substrates except rubbed PVA, where they flowed spontaneously along the rubbing direction, reaching an oblong form that had theta(c) parallel and perpendicular to the rubbing direction respectively greater and smaller than theta(c) for non-rubbed PVA. This is attributed to polar group alignment generating an asymmetric Gamma(SL) distribution with nanometric preferential direction, inducing a capillary-like flow. Polarization and anisotropy maps for high-velocity flow parallel to the PVA rubbing direction showed an increase in the net alignment of molecular domains and a widening of the region where it occurred. This is attributed to preferential anchoring in the downstream direction, instead of in several directions, as for non-rubbed PVA. This explains why filling direction is crucial for TN-LCDs homogeneous behavior.

  8. Observation of wall wettability under imposed flow by fluorescence depolarization: dependence on surface oxygen content and degree of polymer branching.

    PubMed

    Quintella, Cristina M; Musse, Ana Paula S; Castro, Martha T P O; Gonçalves, Cristiane C; Watanabe, Yuji N

    2005-01-01

    This work was concerned with the dependence of the interfacial tension (Gamma(SL)) on surface degree of oxygen content and on polymer branching degree. The static Gamma(SL) was evaluated by contact angle (theta;(c)) and the dynamic Gamma(SL) by fluorescence depolarization of molecular probes seeded in induced flows of monoethylene glycol. The latter results were interpreted using statistical covariant analysis. Two different systems of flowing films were studied: free films flowing on the surfaces on which they impinge and films flowing inside 1-mm-thick microflow cells. The solid surfaces were polyethylene of low density, medium density, high density, and linear with low density, polypropylene, vinyl acetate co-polymer with oxygen content of 15% and 28%, borosilicate, and tin dioxide. Increase in oxygen content of the surface decreased both the static and the dynamic Gamma(SL), which demonstrated that the presence of oxygen atoms hindered wetting. Only the dynamical Gamma(SL) was sensitive to polymer branching, and it increased as branching degree decreased. This was attributed to the higher hydrogen-atom density at the surface, which favored temporary intermolecular bonds between the surface and the flowing liquid.

  9. Concept Design of a Multiwavelength Aerosol Lidar System With Mitigated Diattenuation Effects and Depolarization-Measurement Capability

    NASA Astrophysics Data System (ADS)

    Comerón, Adolfo; Sicard, Michaël; Vidal, Eric; Barragán, Rubén; Muñoz, Constantino; Rodríguez, Alejandro; Tiana-Alsina, Jordi; Rocadenbosch, Francesc; García-Vizcaíno, David

    2016-06-01

    It is known that the retrieval of aerosol extinction and backscatter coefficients from lidar data acquired through so-called total-power channels - intended to measure the backscattered power irrespective of the polarization - can be adversely affected by varying depolarization effects produced by the aerosol under measurement. This effect can be particularly noticeable in advanced multiwavelength systems, where different wavelengths are separated using a system of dichroic beam splitters, because in general the reflection and transmission coefficients of the beam splitters will be different for fields with polarization parallel or perpendicular to the incidence plane. Here we propose a setup for multiwavelength aerosol lidars alleviating diattenuation effects due to changing depolarization conditions while allowing measure linear depolarization.

  10. Backscatter and depolarization measurements of aerosolized biological simulants using a chamber lidar system

    NASA Astrophysics Data System (ADS)

    Brown, David M.; Thrush, Evan P.; Thomas, Michael E.; Santarpia, Josh; Quizon, Jason; Carter, Christopher C.

    2010-04-01

    To ensure agent optical cross sections are well understood from the UV to the LWIR, volume integrated measurements of aerosolized agent material at a few key wavelengths is required to validate existing simulations. Ultimately these simulations will be used to assess the detection performance of various classes of lidar technology spanning the entire range of the optical spectrum. The present work demonstrates an optical measurement architecture based on lidar allowing the measurement of backscatter and depolarization ratio from biological aerosols released in a refereed, 1-m cubic chamber. During 2009, various upgrades have been made to the chamber LIDAR system, which operates at 1.064 μm with sub nanosecond pulses at a 120 Hz repetition rate. The first build of the system demonstrated a sensitivity of aerosolized Bacillus atrophaeus (BG) on the order of 5×105 ppl with 1 GHz InGaAs detectors. To increase the sensitivity and reduce noise, the InGaAs detectors were replaced with larger-area silicon avalanche photodiodes for the second build of the system. In addition, computer controlled step variable neutral density filters are now incorporated to facilitate calibrating the system for absolute back-scatter measurements. Calibrated hard target measurements will be combined with data from the ground truth instruments for cross-section determination of the material aerosolized in the chamber. Measured results are compared to theoretical simulations of cross-sections.

  11. Studying Photosynthesis by Measuring Fluorescence

    ERIC Educational Resources Information Center

    Sanchez, Jose Francisco; Quiles, Maria Jose

    2006-01-01

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

  12. Structure of the pyridine-chloranil complex in solution: a surprise from depolarized hyper-Rayleigh scattering measurements.

    PubMed

    Pandey, Ravindra; Mukhopadhyay, S; Ramasesha, S; Das, Puspendu K

    2011-12-01

    In this article, we report the structure of a 1:1 charge transfer complex between pyridine (PYR) and chloranil (CHL) in solution (CHCl(3)) from the measurement of hyperpolarizability (β(HRS)) and linear and circular depolarization ratios, D and D', respectively, by the hyper-Rayleigh scattering technique and state-of-the-art quantum chemical calculations. Using linearly (electric field vector along X) and circularly polarized incident light, respectively, we have measured two macroscopic depolarization ratios D = I(X,X)(2ω)/I(X,Z)(2ω) and D' = I(X,C)(2ω)/I(Z,C)(2ω) in the laboratory fixed XYZ frame by detecting the second harmonic (SH) scattered light in a polarization resolved fashion. The stabilization energy and the optical gap calculated through the MP2/cc-pVDZ method using Gaussian09 were not significantly different to distinguish between the cofacial and T-shape structures. Only when the experimentally obtained β(HRS) and the depolarization ratios, D and D', were matched with the theoretically computed values from single and double configuration interaction (SDCI) calculations performed using the ZINDO-SCRF technique, we concluded that the room temperature equilibrium structure of the complex is cofacial. This is in sharp contrast to an earlier theoretical prediction of the T-shape structure of the complex.

  13. Dynamic changes in cortical NADH fluorescence in rat focal ischemia: evaluation of the effects of hypothermia on propagation of peri-infarct depolarization by temporal and spatial analysis.

    PubMed

    Sasaki, Toshihiro; Takeda, Yoshimasa; Taninishi, Hideki; Arai, Minako; Shiraishi, Kensuke; Morita, Kiyoshi

    2009-01-02

    Suppression of peri-infarct depolarizations (PIDs) is one of the major mechanisms of hypothermic protection against transient focal cerebral ischemia. Previous studies have shown the lack of hypothermic protection against permanent focal ischemia. We hypothesized the lack of hypothermic protection was due to the poor efficacy in suppression of PIDs. To examine the hypothesis, we elucidated the effects of hypothermia on the manner of propagation of PIDs with temporal and spatial resolutions using NADH (reduced nicotinamide adenine dinucleotide) fluorescence images by illuminating the parietal-temporal cortex with ultraviolet light. Spontaneously hypertensive rats (n=14) were subjected to permanent focal ischemia by occlusion of the middle cerebral and left common carotid arteries. 2-h hypothermia (30 degrees C) was initiated before ischemia. Although hypothermia delayed the appearance of PIDs, it did not suppress their appearance. Furthermore, 54% of the PIDs enlarged the high-intensity area of NADH fluorescence in the hypothermia group, similar to the normothermia group (53%). The high-intensity area of NADH fluorescence widened by each PID was larger in the hypothermia group than in the normothermia group. These findings suggest that PIDs even in hypothermia are one of the major factors causing growth of infarction, emphasizing the importance of therapy that targets suppression of PIDs even during hypothermia.

  14. Dynamics of helical worm-like chains. VIII. Higher-order subspace approximations to dielectric and magnetic relaxation and fluorescence depolarization for flexible chains

    NASA Astrophysics Data System (ADS)

    Yamakawa, Hiromi; Yoshizaki, Takenao; Fujii, Motoharu

    1986-04-01

    Following the general scheme developed in the preceding paper (paper VII), dielectric and magnetic relaxation and fluorescence depolarization for flexible chain polymers in dilute solution are reinvestigated on the basis of the discrete helical worm-like chain in the higher-order subspace approximation. A comparison of theory with experiment is made with respect to the dielectric correlation time τD, the spin-lattice relaxation time T1, the spin-spin relaxation time T2, the nuclear Overhauser enhancement (NOE), the fluorescence emission anisotropy r(t), the average fluorescence anisotropy r¯, and the fluorescence correlation time τF. It is found that there is agreement between the diameters of the chains determined from these dynamic properties and those from chemical structures, better than in the previous crude subspace approximation, indicating that the theory is remarkably improved in the present approximation. The magnetic correlation time τM is in general not an observable, and therefore an empirical equation to be used for its determination from the observed T1 is constructed. It is then found that there is good correlation between the dynamic chain stiffness τX/τ0X and the static chain stiffness λ-1, where τ0X is the correlation time of the isolated subbody (monomer unit) with X=D, M, and F; τX/τ0X is a monotonically increasing function of λ-1 nearly independent of X as far as perpendicular dipoles are concerned. An explanation of this result is given. However, the dependence of τX on temperature cannot be explained very satisfactorily.

  15. Ice crystal habits from cloud chamber studies obtained by in-line holographic microscopy related to depolarization measurements.

    PubMed

    Amsler, Peter; Stetzer, Olaf; Schnaiter, Martin; Hesse, Evelyn; Benz, Stefan; Moehler, Ottmar; Lohmann, Ulrike

    2009-10-20

    We investigate hydrometeor habits at the AIDA chamber with a newly developed in-line holographic microscope HOLographic Imager for Microscopic Objects (HOLIMO). Sizes and habits of ice crystals and droplets in a mixed-phase cloud experiment are related to relative humidity with respect to ice (RH(ice)), temperature (T), and experiment time. This experiment is initiated with supercooled water drops. As a result, ice crystals within a maximum particle diameter size range of 2 to 118 microm (average size of 19 microm) are detected and 63% of them reveal regular habits. The observed particle habits match those predicted for a given RH(ice) and T. Two different growth modes emerge from this cloud. The first one appears during water injection and reveals mainly optical particle sizes in the range of 5 to 250 microm. The second mode grows to sizes of 5 to 63 microm, just after the particles of the first one fall out. It is found that an increasing aspect ratio chi of maximum length over thickness from 2 to 20 as obtained by HOLIMO corresponds to a decreasing linear depolarization ratio from 0.1 to 0.04, as independently obtained by depolarization measurements.

  16. Depolarization ratio and attenuated backscatter for nine cloud types: analyses based on collocated CALIPSO lidar and MODIS measurements.

    PubMed

    Cho, Hyoun-Myoung; Yang, Ping; Kattawar, George W; Nasiri, Shaima L; Hu, Yongxiang; Minnis, Patrick; Trepte, Charles; Winker, David

    2008-03-17

    This paper reports on the relationship between lidar backscatter and the corresponding depolarization ratio for nine types of cloud systems. The data used in this study are the lidar returns measured by the Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) aboard the Cloud- Aerosol Lidar and Infrared Pathfinder Satellite Observations (CALIPSO) satellite and the collocated cloud products derived from the observations made by the Moderate Resolution Imaging Spectroradiometer (MODIS) aboard Aqua satellite. Specifically, the operational MODIS cloud optical thickness and cloud-top pressure products are used to classify cloud types on the basis of the International Satellite Cloud Climatology Project (ISCCP) cloud classification scheme. While the CALIPSO observations provide information for up to 10 cloud layers, in the present study only the uppermost clouds are considered. The layer-averaged attenuated backscatter (gamma') and layer-averaged depolarization ratio (delta) from the CALIPSO measurements show both water- and ice-phase features for global cirrus, cirrostratus, and deep convective cloud classes. Furthermore, we screen both the MODIS and CALIPSO data to eliminate cases in which CALIPSO detected two- or multi-layered clouds. It is shown that low gamma' values corresponding to uppermost thin clouds are largely eliminated in the CALIPSO delta-gamma' relationship for single-layered clouds. For mid-latitude and polar regions corresponding, respectively, to latitude belts 30 degrees -60 degrees and 60 degrees -90 degrees in both the hemispheres, a mixture of water and ice is also observed in the case of the altostratus class. MODIS cloud phase flags are also used to screen ice clouds. The resultant water clouds flagged by the MODIS algorithm show only water phase feature in the delta-gamma' relation observed by CALIOP; however, in the case of the ice clouds flagged by the MODIS algorithm, the co-existence of ice- and water-phase clouds is still observed in

  17. Containerless Atomic-Fluorescence Property Measurements

    NASA Technical Reports Server (NTRS)

    Nordine, P.; Schiffman, R.; Walker, C.

    1987-01-01

    Report describes studies conducted to establish and verify use of laser-induced fluorescence in monitoring and controlling high-temperature containerless processes. Specimens levitated by gas jets or electromagnetic fields and heated by laser beams or electromagnetic induction while being irradiated and detected by fluorescence technique. Makes quantitative and qualitative comparisons among three new methods of temperature measurement; all rely on laser-induced fluorescence. One method gas-density thermometry with seed gas. Other two methods involve measurements of velocities of evaporating atoms or of population ratios of different electronic states.

  18. Calculations of Light Scattering Measurements Predicting Sensitivity of Depolarization to Shape Changes of Spores and Bacteria

    DTIC Science & Technology

    2008-04-01

    either spores or bacteria. 44 LITERATURE CITED 1. Grund, C.J; Brilliant, N.; Bjork, C.; Craig, T. Eyesafe, Multi-function Coherent Doppler Lidar for...an aerosol containing biological particles. This is of interest both for lidar backscatter and for point measurements where samples of the aerosol are... laser light. Each of the added spheres was tangent to the center sphere but did not overlap it. The scattering calculated for the cluster of seven

  19. Rotational diffusion of receptors for epidermal growth factor measured by time-resolved phosphorescence depolarization

    NASA Astrophysics Data System (ADS)

    Zidovetzki, Raphael; Johnson, David A.; Arndt-Jovin, Donna J.; Jovin, Thomas M.

    1991-06-01

    The cell surface receptor for epidermal growth factor (EGFR) is one of the most studied integral membrane proteins. The receptor is widely distributed in cells and tissues of mammalian and avian tissues and plays an important role in growth control. Binding of the epidermal growth factor (EGF) to EGFR initiates a complex biological response, which includes self-phosphorylation of the receptor due to an intrinsic tyrosine kinase activity, phosphorylation of other membrane proteins, increased intake of metabolites, and increased proliferation. Complete amino acid sequence of EGFR revealed a high degree of homology with viral oncogenes and allowed tentative identification of an external hormone binding domain, a transmembrane domain, and a cytoplasmic domain that includes tyrosine kinase activity. EGF binding induces rapid aggregation of EGFR, a process which was also observed on other receptor systems. These and other observations led to a hypothesis that microaggregation of EGFR is a necessary prerequisite for the biological response of EGF. A direct approach to study the processes of oligomerization of cell membrane proteins is to measure their mobility under various conditions. The lateral mobility of the EGFR was studied on mouse 3T3 fibroblasts and on A431 cells. However, an examination of the equations for the lateral and rotational diffusion in membranes shows that only rotational diffusion is strongly dependent on the size of the diffusing entity. A method of measuring protein rotational diffusion by time-resolved phosphorescence has proved to be very useful in the analysis of both in vivo and in vitro systems. The authors apply this method to study the mobility of EGFR on living A431 cells and membrane preparations.

  20. Measurement of Rydberg positronium fluorescence lifetimes

    NASA Astrophysics Data System (ADS)

    Deller, A.; Alonso, A. M.; Cooper, B. S.; Hogan, S. D.; Cassidy, D. B.

    2016-06-01

    We report measurements of the fluorescence lifetimes of positronium (Ps) atoms with principal quantum numbers n =10 -19 . Ps atoms in Rydberg-Stark states were produced via a two-color two-step 1 3S→2 3P→n 3S/n measured time-of-flight distributions were used to determine the mean lifetimes of the Rydberg levels, yielding values ranging from 3 μ s to 26 μ s . Our data are in accord with the expected radiative lifetimes of Rydberg-Stark states of Ps.

  1. Laser-excited fluorescence for measuring atmospheric pollution

    NASA Technical Reports Server (NTRS)

    Menzies, R. T.

    1975-01-01

    System measures amount of given pollutant at specific location. Infrared laser aimed at location has wavelength that will cause molecules of pollutant to fluoresce. Detector separates fluorescence from other radiation and measures its intensity to indicate concentration of pollutant.

  2. Plant stress detection by remote measurement of fluorescence

    USGS Publications Warehouse

    McFarlane, J. C.; Watson, Robert D.; Theisen, Arnold F.; Jackson, R. D.; Ehrler, W. L.; Pinter, P. J.; Idso, S. B.; Reginato, R. J.

    1980-01-01

    Chlorophyll fluorescence of mature lemon trees was measured with a Fraunhofer line discriminator (FLD). An increase in fluorescence was correlated with plant water stress as measured by stomatal resistance and twig water potential.

  3. Fluorescence emission spectral shift measurements of membrane potential in single cells.

    PubMed

    Kao, W Y; Davis, C E; Kim, Y I; Beach, J M

    2001-08-01

    Previous measurements of transmembrane potential using the electrochromic probe di-8-ANEPPS have used the excitation spectral shift response by alternating excitation between two wavelengths centered at voltage-sensitive portions of the excitation spectrum and recording at a single wavelength near the peak of the emission spectrum. Recently, the emission spectral shift associated with the change in transmembrane potential has been used for continuous membrane potential monitoring. To characterize this form of the electrochromic response from di-8-ANEPPS, we have obtained fluorescence signals from single cells in response to step changes in transmembrane potentials set with a patch electrode, using single wavelength excitation near the peak of the dye absorption spectrum. Fluorescence changes at two wavelengths near voltage-sensitive portions of the emission spectrum and shifts in the complete emission spectrum were determined for emission from plasma membrane and internal membrane. We found that the fluorescence ratio from either dual-wavelength recordings, or from opposite sides of the emission spectrum, varied linearly with the amplitude of the transmembrane potential step between -80 and +60 mV. Voltage dependence of difference spectra exhibit a crossover point near the peak of the emission spectra with approximately equal gain and loss of fluorescence intensity on each side of the spectrum and equal response amplitude for depolarization and hyperpolarization. These results are consistent with an electrochromic mechanism of action and demonstrate how the emission spectral shift response can be used to measure the transmembrane potential in single cells.

  4. Oligomeric state of human erythrocyte band 3 measured by fluorescence resonance energy homotransfer.

    PubMed Central

    Blackman, S M; Piston, D W; Beth, A H

    1998-01-01

    The oligomeric state of the erythrocyte anion exchange protein, band 3, has been assayed by resonance energy homotransfer. Homotransfer between oligomeric subunits, labeled with eosin-5-maleimide at Lys430 in the transmembrane domain, has been demonstrated by steady-state and time-resolved fluorescence spectroscopy, and is readily observed by its depolarization of the eosin fluorescence. Polarized fluorescence measurements of HPLC-purified band 3 oligomers indicate that eosin homotransfer increases progressively with increasing species size. This shows that homotransfer also occurs between labeled band 3 dimers as well as within the dimers, making fluorescence anisotropy measurements sensitive to band 3 self-association. Treatment of ghost membranes with either Zn2+ or melittin, agents that cluster band 3, significantly decreases the anisotropy as a result of the increased homotransfer within the band 3 clusters. By comparison with the anisotropy of species of known oligomeric state, the anisotropy of erythrocyte ghost membranes at 37 degrees C is consistent with dimeric and/or tetrameric band 3, and does not require postulation of a fraction of large clusters. Proteolytic removal of the cytoplasmic domain of band 3, which significantly increases the rotational mobility of the transmembrane domain, does not affect its oligomeric state, as reported by eosin homotransfer. These results support a model in which interaction with the membrane skeleton restricts the mobility of band 3 without significantly altering its self-association state. PMID:9675213

  5. Shape-induced Gravitational Sorting of Saharan Dust During Transatlantic Voyage: Evidence from CALIOP Lidar Depolarization Measurements

    NASA Technical Reports Server (NTRS)

    Yang, Weidong; Marshak, Alexander; Kostinski, Alexander B.; Varnai, Tamas

    2013-01-01

    Motivated by the physical picture of shape-dependent air resistance and, consequently, shape-induced differential sedimentation of dust particles, we searched for and found evidence of dust particle asphericity affecting the evolution and distribution of dust-scattered light depolarization ratio (delta). Specifically, we examined a large data set of Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) observations of Saharan dust from June to August 2007. Observing along a typical transatlantic dust track, we find that (1) median delta is uniformly distributed between 2 and 5?km altitudes as the elevated dust leaves the west coast of Africa, thereby indicating uniformly random mixing of particle shapes with height; (2) vertical homogeneity of median delta breaks down during the westward transport: between 2 and 5?km delta increases with altitude and this increase becomes more pronounced with westward progress; (3) delta tends to increase at higher altitude (greater than 4?km) and decrease at lower altitude (less than 4?km) during the westward transport. All these features are captured qualitatively by a minimal model (two shapes only), suggesting that shape-induced differential settling and consequent sorting indeed contribute significantly to the observed temporal evolution and vertical stratification of dust properties. By implicating particle shape as a likely cause of gravitational sorting, these results will affect the estimates of radiative transfer through Saharan dust layers.

  6. Apparatus for eliminating background interference in fluorescence measurements

    DOEpatents

    Martin, J.C.; Jett, J.H.

    1986-03-04

    The disclosure is directed to an apparatus for eliminating background interference during fluorescence measurements in a multiple laser flow cytometer. A biological particle stained with fluorescent dyes is excited by a laser. A fluorescence detector detects the fluorescence. The particle scatters light and a gate signal is generated and delayed until the biological particle reaches the next laser. The delayed signal turns on this next laser, which excites a different stained component of the same biological particle. 8 figs.

  7. Apparatus for eliminating background interference in fluorescence measurements

    DOEpatents

    Martin, J.C.; Jett, J.H.

    1984-01-06

    The disclosure is directed to an apparatus for eliminating background interference during fluorescence measurements in a multiple laser flow cytometer. A biological particle stained with fluorescent dyes is excited by a laser. A fluorescence detector detects the fluorescence. The particle scatters light and a gate signal is generated and delayed until the biological particle reaches the next laser. The delayed signal turns on this next laser which excites a different stained component of the same biological particle.

  8. Apparatus for eliminating background interference in fluorescence measurements

    DOEpatents

    Martin, John C.; Jett, James H.

    1986-01-01

    The disclosure is directed to an apparatus for eliminating background interference during fluorescence measurements in a multiple laser flow cytometer. A biological particle stained with fluorescent dyes is excited by a laser. A fluorescence detector detects the fluorescence. The particle scatters light and a gate signal is generated and delayed until the biological particle reaches the next laser. The delayed signal turns on this next laser, which excites a different stained component of the same biological particle.

  9. Depolarization in polarizing supermirrors

    NASA Astrophysics Data System (ADS)

    Klauser, Christine; Bigault, Thierry; Böni, Peter; Courtois, Pierre; Devishvili, Anton; Rebrova, Nataliya; Schneider, Michael; Soldner, Torsten

    2016-12-01

    We present data on depolarizing effects in polarizing mirrors. At typical magnetizing field strengths used in polarizing devices, depolarizations rise up to the percent level in the specular region and are shown to be successfully suppressed to 10-4 when increasing the magnetizing field. We show evidence linking a part of this depolarization to lateral correlation of the magnetization fluctuations in the ferromagnetic layers. Effects of the supermirror factor (m), wavelength and incidence angle are studied. The findings are applied to a crossed supermirror geometry and we report a neutron beam polarization of 99.97(1)% for a beam of wavelength λ = 5.3 Å, Δλ/λ = 0.1 (FWHM).

  10. Time-resolved fluorescence decay measurements for flowing particles

    DOEpatents

    Deka, Chiranjit; Steinkamp, John A.

    1999-01-01

    Time-resolved fluorescence decay measurements for flowing particles. An apparatus and method for the measurement and analysis of fluorescence for individual cells and particles in flow are described, wherein the rapid measurement capabilities of flow cytometry and the robust measurement and analysis procedures of time-domain fluorescence lifetime spectroscopy are combined. A pulse-modulated cw laser is employed for excitation of the particles. The characteristics and the repetition rate of the excitation pulses can be readily adjusted to accommodate for fluorescence decays having a wide range of lifetimes.

  11. Time-resolved fluorescence decay measurements for flowing particles

    DOEpatents

    Deka, C.; Steinkamp, J.A.

    1999-06-01

    Time-resolved fluorescence decay measurements are disclosed for flowing particles. An apparatus and method for the measurement and analysis of fluorescence for individual cells and particles in flow are described, wherein the rapid measurement capabilities of flow cytometry and the robust measurement and analysis procedures of time-domain fluorescence lifetime spectroscopy are combined. A pulse-modulated CW laser is employed for excitation of the particles. The characteristics and the repetition rate of the excitation pulses can be readily adjusted to accommodate for fluorescence decays having a wide range of lifetimes. 12 figs.

  12. Depolarizing differential Mueller matrices.

    PubMed

    Ortega-Quijano, Noé; Arce-Diego, José Luis

    2011-07-01

    The evolution of a polarized beam can be described by the differential formulation of Mueller calculus. The nondepolarizing differential Mueller matrices are well known. However, they only account for 7 out of the 16 independent parameters that are necessary to model a general anisotropic depolarizing medium. In this work we present the nine differential Mueller matrices for general depolarizing media, highlighting the physical implications of each of them. Group theory is applied to establish the relationship between the differential matrix and the set of transformation generators in the Minkowski space, of which Lorentz generators constitute a particular subgroup.

  13. Photon correlation system for fluorescence lifetime measurements

    NASA Astrophysics Data System (ADS)

    Morgan, C. G.; Murray, J. G.; Mitchell, A. C.

    1995-07-01

    The construction and testing of a dual-channel photon correlator is reported for the frequency domain imaging of fluorescence lifetimes using photon-counting detection. A light source modulated at radio frequency excites fluorescence, which is detected using an imaging single-photon detector. After discrimination, single-photon events are processed in parallel by the correlation circuit, the purpose of which is to allow both the mean phase delay and the demodulation of fluorescence to be calculated relative to a reference signal derived from the modulated excitation source. Outputs from the correlator are integrated in a computer, resulting in accumulation of images which have been statistically filtered by sine and cosine transforms, and which can be manipulated within the computer to generate a resultant image where contrast depends on fluorescence lifetime rather than fluorescence intensity.

  14. Suitability of fluorescence measurements to quantify sulfate-reducing bacteria.

    PubMed

    Barton, Larry L; Carpenter, Claire M

    2013-06-01

    Fluorescence activity has been used to identify Desulfovibrio and has been termed the 'desulfoviridin test'. This fluorescence is attributed to the prosthetic group of bisulfite reductase, a key enzyme in dissimilatory sulfate reduction. We have pursued the use of fluorescence measurements to quantify sulfate-reducing bacteria. Cells of D. desulfuricans and D. gigas were treated with NaOH and produced two fluorescence spectra: one with maximum fluorescence with an excitation at 395 nm and an emission at 605 nm and another with an excitation at 320 nm and emission at 360 nm. Using the fluorescence with excitation at 395 nm and emission at 605 nm, we explored a series of parameters to measure Desulfovibrio in pure cultures and environmental samples. Fluorescence measurements are reliable provided the cells are treated with 1.75 N NaOH and the chromophore released from the cells is not exposed to strong light intensity, and is not exposed to temperatures greater than 20 °C, and measurements are done within a few minutes of extraction. Bleaching of fluorescence was attributed to metal ions in solution which was not observed until metal concentrations reached 1.5mM. We propose that D. desulfuricans is appropriate as the reference organism for measurement of sulfate-reducing bacteria by fluorescence and by using fluorescence intensity, 10(5) cells/ml can be readily detected in environmental samples.

  15. Fluorescent Protein Based FRET Pairs with Improved Dynamic Range for Fluorescence Lifetime Measurements

    PubMed Central

    George Abraham, Bobin; Sarkisyan, Karen S.; Mishin, Alexander S.; Santala, Ville; Tkachenko, Nikolai V.; Karp, Matti

    2015-01-01

    Fluorescence Resonance Energy Transfer (FRET) using fluorescent protein variants is widely used to study biochemical processes in living cells. FRET detection by fluorescence lifetime measurements is the most direct and robust method to measure FRET. The traditional cyan-yellow fluorescent protein based FRET pairs are getting replaced by green-red fluorescent protein variants. The green-red pair enables excitation at a longer wavelength which reduces cellular autofluorescence and phototoxicity while monitoring FRET. Despite the advances in FRET based sensors, the low FRET efficiency and dynamic range still complicates their use in cell biology and high throughput screening. In this paper, we utilized the higher lifetime of NowGFP and screened red fluorescent protein variants to develop FRET pairs with high dynamic range and FRET efficiency. The FRET variations were analyzed by proteolytic activity and detected by steady-state and time-resolved measurements. Based on the results, NowGFP-tdTomato and NowGFP-mRuby2 have shown high potentials as FRET pairs with large fluorescence lifetime dynamic range. The in vitro measurements revealed that the NowGFP-tdTomato has the highest Förster radius for any fluorescent protein based FRET pairs yet used in biological studies. The developed FRET pairs will be useful for designing FRET based sensors and studies employing Fluorescence Lifetime Imaging Microscopy (FLIM). PMID:26237400

  16. Models of fluorescence and photosynthesis for interpreting measurements of solar-induced chlorophyll fluorescence

    PubMed Central

    van der Tol, C; Berry, J A; Campbell, P K E; Rascher, U

    2014-01-01

    We have extended a conventional photosynthesis model to simulate field and laboratory measurements of chlorophyll fluorescence at the leaf scale. The fluorescence paramaterization is based on a close nonlinear relationship between the relative light saturation of photosynthesis and nonradiative energy dissipation in plants of different species. This relationship diverged only among examined data sets under stressed (strongly light saturated) conditions, possibly caused by differences in xanthophyll pigment concentrations. The relationship was quantified after analyzing data sets of pulse amplitude modulated measurements of chlorophyll fluorescence and gas exchange of leaves of different species exposed to different levels of light, CO2, temperature, nitrogen fertilization treatments, and drought. We used this relationship in a photosynthesis model. The coupled model enabled us to quantify the relationships between steady state chlorophyll fluorescence yield, electron transport rate, and photosynthesis in leaves under different environmental conditions. Key Points Light saturation of photosynthesis determines quenching of leaf fluorescence We incorporated steady state leaf fluorescence in a photosynthesis model PMID:27398266

  17. One year of urban background fluorescent aerosol measurements

    NASA Astrophysics Data System (ADS)

    Pope, Francis

    2016-04-01

    Online aerosol fluorescence is a popular methodology for detecting bioaerosols in the atmosphere. In recent years there has been considerable effort into refining the technique to be able to distinguish between different bioaerosol classes such as pollen, spores and bacteria. A near continuous record of aerosol fluorescence measurements has been recorded at an urban background observation site in Birmingham, UK for the year 2015. Fluorescence measurements were performed using the Biral aerosol fluorescence spectrometer (AFS) which measures both UV and visible fluorescence resulting from the excitation of aerosol particles at 280 nm. Speciation of the fluorescent particles into different bioaerosol class is possible with the AFS but the lack of particle sizing makes the task difficult compared to other techniques. In addition to the fluorescence measurements, further campaign mode measurements were also generated for size segregated total particle numbers, ozone, nitrogen oxides and other chemical species. These measurements allow for the influence of road traffic on the concentration of fluorescent particle to be determined. This presentation will provide an in depth look into how bioaerosol concentrations and speciation (pollen, spores and bacteria) change throughout the year. These changes will be linked to local and regional meteorology and climate. In particular, the consequences of the unusually warm UK winter upon bioaerosol concentrations will be highlighted.

  18. SIMULTANEOUS MEASUREMENT OF CIRCULAR DICHROISM AND FLUORESCENCE POLARIZATION ANISOTROPY.

    SciTech Connect

    SUTHERLAND,J.C.

    2002-01-19

    Circular dichroism and fluorescence polarization anisotropy are important tools for characterizing biomolecular systems. Both are used extensively in kinetic experiments involving stopped- or continuous flow systems as well as titrations and steady-state spectroscopy. This paper presents the theory for determining circular dichroism and fluorescence polarization anisotropy simultaneously, thus insuring the two parameters are recorded under exactly the same conditions and at exactly the same time in kinetic experiments. The approach to measuring circular dichroism is that used in almost all conventional dichrographs. Two arrangements for measuring fluorescence polarization anisotropy are described. One uses a single fluorescence detector and signal processing with a lock-in amplifier that is similar to the measurement of circular dichroism. The second approach uses classic ''T'' format detection optics, and thus can be used with conventional photon-counting detection electronics. Simple extensions permit the simultaneous measurement of the absorption and excitation intensity corrected fluorescence intensity.

  19. Laser-fluorescence measurement of marine algae

    NASA Technical Reports Server (NTRS)

    Browell, E. V.

    1980-01-01

    Progress in remote sensing of algae by laser-induced fluorescence is subject of comprehensive report. Existing single-wavelength and four-wavelength systems are reviewed, and new expression for power received by airborne sensor is derived. Result differs by as much as factor of 10 from those previously reported. Detailed error analysis evluates factors affecting accuracy of laser-fluorosensor systems.

  20. Intracellular distribution of fluorescent copper and zinc bis(thiosemicarbazonato) complexes measured with fluorescence lifetime spectroscopy.

    PubMed

    Hickey, James L; James, Janine L; Henderson, Clare A; Price, Katherine A; Mot, Alexandra I; Buncic, Gojko; Crouch, Peter J; White, Jonathan M; White, Anthony R; Smith, Trevor A; Donnelly, Paul S

    2015-10-05

    The intracellular distribution of fluorescently labeled copper and zinc bis(thiosemicarbazonato) complexes was investigated in M17 neuroblastoma cells and primary cortical neurons with a view to providing insights into the neuroprotective activity of a copper bis(thiosemicarbazonato) complex known as Cu(II)(atsm). Time-resolved fluorescence measurements allowed the identification of the Cu(II) and Zn(II) complexes as well as the free ligand inside the cells by virtue of the distinct fluorescence lifetime of each species. Confocal fluorescent microscopy of cells treated with the fluorescent copper(II)bis(thiosemicarbazonato) complex revealed significant fluorescence associated with cytoplasmic puncta that were identified to be lysosomes in primary cortical neurons and both lipid droplets and lysosomes in M17 neuroblastoma cells. Fluorescence lifetime imaging microscopy confirmed that the fluorescence signal emanating from the lipid droplets could be attributed to the copper(II) complex but also that some degree of loss of the metal ion led to diffuse cytosolic fluorescence that could be attributed to the metal-free ligand. The accumulation of the copper(II) complex in lipid droplets could be relevant to the neuroprotective activity of Cu(II)(atsm) in models of amyotrophic lateral sclerosis and Parkinson's disease.

  1. Practical aspects of measuring intracellular calcium signals with fluorescent indicators.

    PubMed

    Kao, Joseph P Y; Li, Gong; Auston, Darryl A

    2010-01-01

    The use of fluorescent indicators for monitoring calcium (Ca(2+)) signals and for measuring Ca(2+) concentration ([Ca(2+)]) in living cells is described. The following topics are covered in detail: (1) ratiometric and nonratiometric fluorescent indicators and the principles underlying their use, (2) techniques for loading Ca(2+) indicators and Ca(2+) buffers into living cells, (3) calibration of indicator fluorescence intensity measurements to yield values of intracellular [Ca(2+)], (4) analysis of nonratiometric fluorescence intensity data and caveats relating to their interpretation, (5) techniques for manipulating intracellular and extracellular [Ca(2+)], and (6) the use of fluorescent indicators to monitor Ca(2+) signals in mitochondria. The chapter aims to present these fundamental topics in a manner that is practically useful and intuitively accessible. The origins of key mathematical equations used in the article are outlined in two appendices.

  2. Double-excitation fluorescence spectral imaging: eliminating tissue auto-fluorescence from in vivo PPIX measurements

    NASA Astrophysics Data System (ADS)

    Torosean, Sason; Flynn, Brendan; Samkoe, Kimberley S.; Davis, Scott C.; Gunn, Jason; Axelsson, Johan; Pogue, Brian W.

    2012-02-01

    An ultrasound coupled handheld-probe-based optical fluorescence molecular tomography (FMT) system has been in development for the purpose of quantifying the production of Protoporphyrin IX (PPIX) in aminolevulinic acid treated (ALA), Basal Cell Carcinoma (BCC) in vivo. The design couples fiber-based spectral sampling of PPIX fluorescence emission with a high frequency ultrasound imaging system, allowing regionally localized fluorescence intensities to be quantified [1]. The optical data are obtained by sequential excitation of the tissue with a 633nm laser, at four source locations and five parallel detections at each of the five interspersed detection locations. This method of acquisition permits fluorescence detection for both superficial and deep locations in ultrasound field. The optical boundary data, tissue layers segmented from ultrasound image and diffusion theory are used to estimate the fluorescence in tissue layers. To improve the recovery of the fluorescence signal of PPIX, eliminating tissue autofluorescence is of great importance. Here the approach was to utilize measurements which straddled the steep Qband excitation peak of PPIX, via the integration of an additional laser source, exciting at 637 nm; a wavelength with a 2 fold lower PPIX excitation value than 633nm.The auto-fluorescence spectrum acquired from the 637 nm laser is then used to spectrally decouple the fluorescence data and produce an accurate fluorescence emission signal, because the two wavelengths have very similar auto-fluorescence but substantially different PPIX excitation levels. The accuracy of this method, using a single source detector pair setup, is verified through animal tumor model experiments, and the result is compared to different methods of fluorescence signal recovery.

  3. Optical and microphysical characterization of aerosol layers over South Africa by means of multi-wavelength depolarization and Raman lidar measurements

    NASA Astrophysics Data System (ADS)

    Giannakaki, Elina; van Zyl, Pieter G.; Müller, Detlef; Balis, Dimitris; Komppula, Mika

    2016-07-01

    Optical and microphysical properties of different aerosol types over South Africa measured with a multi-wavelength polarization Raman lidar are presented. This study could assist in bridging existing gaps relating to aerosol properties over South Africa, since limited long-term data of this type are available for this region. The observations were performed under the framework of the EUCAARI campaign in Elandsfontein. The multi-wavelength PollyXT Raman lidar system was used to determine vertical profiles of the aerosol optical properties, i.e. extinction and backscatter coefficients, Ångström exponents, lidar ratio and depolarization ratio. The mean microphysical aerosol properties, i.e. effective radius and single-scattering albedo, were retrieved with an advanced inversion algorithm. Clear differences were observed for the intensive optical properties of atmospheric layers of biomass burning and urban/industrial aerosols. Our results reveal a wide range of optical and microphysical parameters for biomass burning aerosols. This indicates probable mixing of biomass burning aerosols with desert dust particles, as well as the possible continuous influence of urban/industrial aerosol load in the region. The lidar ratio at 355 nm, the lidar ratio at 532 nm, the linear particle depolarization ratio at 355 nm and the extinction-related Ångström exponent from 355 to 532 nm were 52 ± 7 sr, 41 ± 13 sr, 0.9 ± 0.4 % and 2.3 ± 0.5, respectively, for urban/industrial aerosols, while these values were 92 ± 10 sr, 75 ± 14 sr, 3.2 ± 1.3 % and 1.7 ± 0.3, respectively, for biomass burning aerosol layers. Biomass burning particles are larger and slightly less absorbing compared to urban/industrial aerosols. The particle effective radius were found to be 0.10 ± 0.03, 0.17 ± 0.04 and 0.13 ± 0.03 µm for urban/industrial, biomass burning, and mixed aerosols, respectively, while the single-scattering albedo at 532 nm was 0.87 ± 0.06, 0.90 ± 0.06, and 0.88 ± 0.07 (at 532

  4. Optical and microphysical characterization of aerosol layers over South Africa by means of multi-wavelength depolarization and Raman lidar measurements

    NASA Astrophysics Data System (ADS)

    Giannakaki, E.; van Zyl, P. G.; Müller, D.; Balis, D.; Komppula, M.

    2015-12-01

    Optical and microphysical properties of different aerosol types over South Africa measured with a multi-wavelength polarization Raman lidar are presented. This study could assist in bridging existing gaps relating to aerosol properties over South Africa, since limited long-term data of this type is available for this region. The observations were performed under the framework of the EUCAARI campaign in Elandsfontein. The multi-wavelength PollyXT Raman lidar system was used to determine vertical profiles of the aerosol optical properties, i.e. extinction and backscatter coefficients, Ångström exponents, lidar ratio and depolarization ratio. The mean microphysical aerosol proper ties, i.e. effective radius and single scattering, albedo were retrieved with an advanced inversion algorithm. Clear differences were observed for the intensive optical properties of atmospheric layers of biomass burning and urban/industrial aerosols. Our results reveal a wide range of optical and microphysical parameters for biomass burning aerosols. This indicates probable mixing of biomass burning aerosols with desert dust particles, as well as the possible continuous influence of urban/industrial aerosol load in the region. The lidar ratio at 355 nm, the linear particle depolarization ratio at 355 nm and the extinction-related Ångström exponent from 355 to 532 nm were 52 ± 7 sr; 0.9 ± 0.4 % and 2.3 ± 0.5, respectively for urban/industrial aerosols, while these values were 92 ± 10 sr; 3.2 ± 1.3 %; 2.0 ± 0.4 respectively for biomass burning aerosols layers. Biomass burning particles are larger and slightly less absorbing compared to urban/industrial aerosols. The particle effective radius were found to be 0.10 ± 0.03, 0.17 ± 0.04 and 0.13 ± 0.03 μm for urban/industrial, biomass burning, and mixed biomass burning and desert dust aerosols, respectively, while the single scattering albedo at 532 nm were 0.87 ± 0.06, 0.90 ± 0.06, and 0.88 ± 0.07 (at 532 nm), respectively for

  5. Effect of pre-ischaemic conditioning on hypoxic depolarization of dopamine efflux in the rat caudate brain slice measured in real-time with fast cyclic voltammetry.

    PubMed

    Davidson, Colin; Coomber, Ben; Gibson, Claire L; Young, Andrew M J

    2011-10-01

    Fast cyclic voltammetry can be used to measure dopamine release after oxygen and glucose deprivation (OGD) induced anoxic depolarization in vitro. Here we measure dopamine efflux with 1s time resolution, which is appropriate to measure OGD-evoked dopamine efflux accurately. In the present study, we examined whether OGD-evoked dopamine efflux could be used to show pre-ischaemic conditioning in the rat caudate brain slice. Caudate slices were exposed to 0, 2, or 10 min OGD pre-ischaemic conditioning, then 60 min later exposed to a second OGD event of 15 min duration. We measured the OGD-evoked dopamine efflux using fast cyclic voltammetry and in some experiments caudate dopamine and DOPAC tissue levels were measured using HPLC and 20 μm cryostat sections were Nissl stained to indicate neuronal loss. We found that 10 but not 2 min OGD pre-ischaemic conditioning resulted in a longer time to onset of OGD-evoked dopamine efflux on the main OGD event (475 ± 31 and 287 ± 30 s for 10 Vs 0 min pre-ischaemic conditioning respectively). Further, 10 min OGD pre-ischaemic conditioning resulted in less dopamine efflux on the second OGD event (4.23 ± 1.12 and 8.14 ± 0.82 μM for 10 Vs 0 min pre-ischaemic conditioning respectively), despite these slices having similar tissue dopamine content and DOPAC/DA ratio, and the rate of dopamine release was slower in the main OGD event (21 ± 5 and 74 ± 8 nM/s for 10 Vs 0 min pre-ischaemic conditioning respectively). These data suggest that 10 min OGD pre-ischaemic conditioning can evoke tolerance to a second OGD event and that voltammetric recording of OGD-evoked dopamine efflux is a useful model of pre-ischaemic conditioning in neuronal tissue.

  6. System and method for measuring fluorescence of a sample

    DOEpatents

    Riot, Vincent J

    2015-03-24

    The present disclosure provides a system and a method for measuring fluorescence of a sample. The sample may be a polymerase-chain-reaction (PCR) array, a loop-mediated-isothermal amplification array, etc. LEDs are used to excite the sample, and a photodiode is used to collect the sample's fluorescence. An electronic offset signal is used to reduce the effects of background fluorescence and the noises from the measurement system. An integrator integrates the difference between the output of the photodiode and the electronic offset signal over a given period of time. The resulting integral is then converted into digital domain for further processing and storage.

  7. Models of fluorescence and photosynthesis for interpreting measurements of solar-induced chlorophyll fluorescence

    NASA Astrophysics Data System (ADS)

    Tol, C.; Berry, J. A.; Campbell, P. K. E.; Rascher, U.

    2014-12-01

    We have extended a conventional photosynthesis model to simulate field and laboratory measurements of chlorophyll fluorescence at the leaf scale. The fluorescence paramaterization is based on a close nonlinear relationship between the relative light saturation of photosynthesis and nonradiative energy dissipation in plants of different species. This relationship diverged only among examined data sets under stressed (strongly light saturated) conditions, possibly caused by differences in xanthophyll pigment concentrations. The relationship was quantified after analyzing data sets of pulse amplitude modulated measurements of chlorophyll fluorescence and gas exchange of leaves of different species exposed to different levels of light, CO2, temperature, nitrogen fertilization treatments, and drought. We used this relationship in a photosynthesis model. The coupled model enabled us to quantify the relationships between steady state chlorophyll fluorescence yield, electron transport rate, and photosynthesis in leaves under different environmental conditions.

  8. Laser-saturated fluorescence measurements in laminar sooting diffusion flames

    NASA Technical Reports Server (NTRS)

    Wey, Changlie

    1993-01-01

    The hydroxyl radical is known to be one of the most important intermediate species in the combustion processes. The hydroxyl radical has also been considered a dominant oxidizer of soot particles in flames. In this investigation the hydroxyl concentration profiles in sooting diffusion flames were measured by the laser-saturated fluorescence (LSF) method. The temperature distributions in the flames were measured by the two-line LSF technique and by thermocouple. In the sooting region the OH fluorescence was too weak to make accurate temperature measurements. The hydroxyl fluorescence profiles for all four flames presented herein show that the OH fluorescence intensities peaked near the flame front. The OH fluorescence intensity dropped sharply toward the dark region of the flame and continued declining to the sooting region. The OH fluorescence profiles also indicate that the OH fluorescence decreased with increasing height in the flames for all flames investigated. Varying the oxidizer composition resulted in a corresponding variation in the maximum OH concentration and the flame temperature. Furthermore, it appears that the maximum OH concentration for each flame increased with increasing flame temperature.

  9. Fluorescence cross section measurements of biological agent simulants

    SciTech Connect

    Stephens, J.R.

    1996-11-01

    Fluorescence is a powerful technique that has potential uses in detection and characterization of biological aerosols both in the battlefield and in civilian environments. Fluorescence techniques can be used with ultraviolet (UV) light detection and ranging (LIDAR) equipment to detect biological aerosol clouds at a distance, to provide early warning of a biological attack, and to track an potentially noxious cloud. Fluorescence can also be used for detection in a point sensor to monitor biological materials and to distinguish agents from benign aerosols. This work is part of a continuing program by the Army`s Chemical and Biological Defense Command to characterized the optical properties of biological agents. Reported here are ultraviolet fluorescence measurements of Bacillus megaterium and Bacillus Globigii aerosols suspended in an electrodynamic particle trap. Fluorescence spectra of a common atmospheric aerosol, pine pollen, are also presented.

  10. Measurement of the fluorescence quantum yield of bis-MSB

    NASA Astrophysics Data System (ADS)

    Ding, Xue-Feng; Wen, Liang-Jian; Zhou, Xiang; Ding, Ya-Yun; Ye, Xing-Chen; Zhou, Li; Liu, Meng-Chao; Cai, Hao; Cao, Jun

    2015-12-01

    The fluorescence quantum yield of bis-MSB, a widely used liquid scintillator wavelength shifter, was measured to study the photon absorption and re-emission processes in a liquid scintillator. The re-emission process affects the photoelectron yield and distribution, especially in a large liquid scintillator detector, thus must be understood to optimize the liquid scintillator for good energy resolution and to precisely simulate the detector with Monte Carlo. In this study, solutions of different bis-MSB concentration were prepared for absorption and fluorescence emission measurements to cover a broad range of wavelengths. Harmane was used as a standard reference to obtain the absolution fluorescence quantum yield. For the first time we measured the fluorescence quantum yield of bis-MSB up to 430 nm as inputs required by Monte Carlo simulation, which is 0.926±0.053 at λex=350 nm. Supported by National Natural Science Foundation of China (11205183, 11225525, 11390381)

  11. Measurement of Sun Induced Chlorophyll Fluorescence Using Hyperspectral Satellite Imagery

    NASA Astrophysics Data System (ADS)

    Irteza, S. M.; Nichol, J. E.

    2016-06-01

    Solar Induced Chlorophyll Fluorescence (SIF), can be used as an indicator of stress in vegetation. Several scientific approaches have been made and there is considerable evidence that steady state Chlorophyll fluorescence is an accurate indicator of plant stress hence a reliable tool to monitor vegetation health status. Retrieval of Chlorophyll fluorescence provides an insight into photochemical and carbon sequestration processes within vegetation. Detection of Chlorophyll fluorescence has been well understood in the laboratory and field measurement. Fluorescence retrieval methods were applied in and around the atmospheric absorption bands 02B (Red wavelength) approximately 690 nm and 02A (Far red wavelengths) 740 nm. Hyperion satellite images were acquired for the years 2012 to 2015 in different seasons. Atmospheric corrections were applied using the 6S Model. The Fraunhofer Line Discrimanator (FLD) method was applied for retrieval of SIF from the Hyperion images by measuring the signal around the absorption bands in both vegetated and non vegetated land cover types. Absorption values were extracted in all the selected bands and the fluorescence signal was detected. The relationships between NDVI and Fluorescence derived from the satellite images are investigated to understand vegetation response within the absorption bands.

  12. X-ray fluorescence measurements of dissolved gas and cavitation

    NASA Astrophysics Data System (ADS)

    Duke, Daniel J.; Kastengren, Alan L.; Swantek, Andrew B.; Matusik, Katarzyna E.; Powell, Christopher F.

    2016-10-01

    The dynamics of dissolved gas and cavitation are strongly coupled, yet these phenomena are difficult to measure in-situ. Both create voids in the fluid that can be difficult to distinguish. We present an application of X-ray fluorescence in which liquid density and total noncondensible gas concentration (both dissolved and nucleated) are simultaneously measured. The liquid phase is doped with 400 ppm of a bromine tracer, and dissolved air is removed and substituted with krypton. Fluorescent emission at X-ray wavelengths is simultaneously excited from the Br and Kr with a focused monochromatic X-ray beam from a synchrotron source. We measure the flow in a cavitating nozzle 0.5 mm in diameter. From Br fluorescence, total displacement of the liquid is measured. From Kr fluorescence, the mass fraction of both dissolved and nucleated gas is measured. Volumetric displacement of liquid due to both cavitation and gas precipitation can be separated through estimation of the local equilibrium dissolved mass fraction. The uncertainty in the line of sight projected densities of the liquid and gas phases is 4-6 %. The high fluorescence yields and energies of Br and Kr allow small mass fractions of gas to be measured, down to 10-5, with an uncertainty of 8 %. These quantitative measurements complement existing optical diagnostic techniques and provide new insight into the diffusion of gas into cavitation bubbles, which can increase their internal density, pressure and lifetimes by orders of magnitude.

  13. X-ray fluorescence measurements of dissolved gas and cavitation

    SciTech Connect

    Duke, Daniel J.; Kastengren, Alan L.; Swantek, Andrew B.; Matusik, Katarzyna E.; Powell, Christopher F.

    2016-09-28

    The dynamics of dissolved gas and cavitation are strongly coupled, yet these phenomena are difficult to measure in-situ. Both create voids in the fluid that can be difficult to distinguish. In this paper, we present an application of X-ray fluorescence in which liquid density and total noncondensible gas concentration (both dissolved and nucleated) are simultaneously measured. The liquid phase is doped with 400 ppm of a bromine tracer, and dissolved air is removed and substituted with krypton. Fluorescent emission at X-ray wavelengths is simultaneously excited from the Br and Kr with a focused monochromatic X-ray beam from a synchrotron source. We measure the flow in a cavitating nozzle 0.5 mm in diameter. From Br fluorescence, total displacement of the liquid is measured. From Kr fluorescence, the mass fraction of both dissolved and nucleated gas is measured. Volumetric displacement of liquid due to both cavitation and gas precipitation can be separated through estimation of the local equilibrium dissolved mass fraction. The uncertainty in the line of sight projected densities of the liquid and gas phases is 4–6 %. The high fluorescence yields and energies of Br and Kr allow small mass fractions of gas to be measured, down to 10-5, with an uncertainty of 8 %. Finally, these quantitative measurements complement existing optical diagnostic techniques and provide new insight into the diffusion of gas into cavitation bubbles, which can increase their internal density, pressure and lifetimes by orders of magnitude.

  14. Collisional depolarization of OH(A) with Ar: Experiment and theory.

    PubMed

    Brouard, M; Bryant, A; Chang, Y-P; Cireasa, R; Eyles, C J; Green, A M; Marinakis, S; Aoiz, F J; Kłos, J

    2009-01-28

    Zeeman quantum beat spectroscopy has been used to measure the 300 K rate constants for the angular momentum depolarization of OH(A (2)Sigma(+)) in the presence of Ar. We show that the beat amplitude at short times, in the absence of collisions, is well described by previously developed line strength theory for (1+1) laser induced fluorescence. The subsequent pressure dependent decay of the beat amplitude is used to extract depolarization rate constants and estimates of collisional depolarization cross sections. Depolarization accompanies both inelastic collisions, giving rise to rotational energy transfer, and elastic collisions, which change m(j) but conserve j. Previous experimental studies, as well as classical theory, suggest that elastic scattering contributes around 20% to the observed total depolarization rate at low j. Simulation of the experimental beat amplitudes, using theoretical calculations presented in the preceding paper, reveals that depolarization of OH(A) by Ar has a rate constant comparable to, if not larger than, that for energy transfer. This is consistent with a significant tilting or realignment of j(') away from j on collision. The experimental data are used to provide a detailed test of quantum mechanical and quasiclassical trajectory scattering calculations performed on a recently developed ab initio potential energy surface of Kłos et al. [J. Chem. Phys. 129, 054301 (2008)]. The calculations and simulations account well for the observed cross sections at high N, but underestimate the experimental results by between 10% and 20% at low N, possibly due to remaining inaccuracies in the potential energy surface or perhaps to limitations in the dynamical approximations made, particularly the freezing of the OH(A) bond.

  15. Molecular anisotropy effects in carbon K-edge scattering: depolarized diffuse scattering and optical anisotropy

    SciTech Connect

    Stone, Kevin H.

    2014-07-14

    Some polymer properties, such as conductivity, are very sensitive to short- and intermediate-range orientational and positional ordering of anisotropic molecular functional groups, and yet means to characterize orientational order in disordered systems are very limited. We demonstrate that resonant scattering at the carbon K-edge is uniquely sensitive to short-range orientation correlations in polymers through depolarized scattering at high momentum transfers, using atactic polystyrene as a well-characterized test system. Depolarized scattering is found to coexist with unpolarized fluorescence, and to exhibit pronounced anisotropy. We also quantify the spatially averaged optical anisotropy from low-angle reflectivity measurements, finding anisotropy consistent with prior visible, x-ray absorption, and theoretical studies. The average anisotropy is much smaller than that in the depolarized scattering and the two have different character. Both measurements exhibit clear spectral signatures from the phenyl rings and the polyethylene-like backbone. Discussion focuses on analysis considerations and prospects for using this depolarized scattering for studies of disorder in soft condensed matter.

  16. Fluorescent-Antibody Measurement Of Cancer-Cell Urokinase

    NASA Technical Reports Server (NTRS)

    Morrison, Dennis R.

    1993-01-01

    Combination of laboratory techniques provides measurements of amounts of urokinase in and between normal and cancer cells. Includes use of fluorescent antibodies specific against different forms of urokinase-type plasminogen activator, (uPA), fluorescence microscopy, quantitative analysis of images of sections of tumor tissue, and flow cytometry of different uPA's and deoxyribonucleic acid (DNA) found in suspended-tumor-cell preparations. Measurements provide statistical method for indicating or predicting metastatic potentials of some invasive tumors. Assessments of metastatic potentials based on such measurements used in determining appropriate follow-up procedures after surgical removal of tumors.

  17. Breast cancer: in vitro measurements of native fluorescence

    NASA Astrophysics Data System (ADS)

    Lohmann, Wolfgang; Bohle, Rainer M.; Dreyer, Thomas; Haas, Sabine; Wallenfels, Heike; Schwemmle, Konrad; Schill, Wolf-Bernhard

    1996-12-01

    Unfixed, HE stained cryosections of breast tissue obtained from 67 patients during surgery were illuminated with 395 - 440 nm and their fluorescence response as well as the 2- dimensional fluorophore distribution were measured. The histological evaluation of the same cryosection, illuminated as usual with a transmitted light obtained from a halogen lamp, revealed 9 patients with healthy tissue, 11 with benign epithelial hyperplasia, 4 with ductal carcinoma in situ, 35 with invasive ductal carcinoma, 7 with invasive lobular carcinoma, and one with invasive tubular carcinoma. A comparison between the fluorescence and the HE images shows that both match very nicely and that the fluorescence images are also characteristic for the different pathological condition of the biopsy sample. Moreover, benign tumors e.g. fibroadenomas, exhibit a fluorescence response different from cancer and healthy tissue.

  18. Measuring and interpreting X-ray fluorescence from planetary surfaces.

    PubMed

    Owens, Alan; Beckhoff, Burkhard; Fraser, George; Kolbe, Michael; Krumrey, Michael; Mantero, Alfonso; Mantler, Michael; Peacock, Anthony; Pia, Maria-Grazia; Pullan, Derek; Schneider, Uwe G; Ulm, Gerhard

    2008-11-15

    As part of a comprehensive study of X-ray emission from planetary surfaces and in particular the planet Mercury, we have measured fluorescent radiation from a number of planetary analog rock samples using monochromatized synchrotron radiation provided by the BESSY II electron storage ring. The experiments were carried out using a purpose built X-ray fluorescence (XRF) spectrometer chamber developed by the Physikalisch-Technische Bundesanstalt, Germany's national metrology institute. The XRF instrumentation is absolutely calibrated and allows for reference-free quantitation of rock sample composition, taking into account secondary photon- and electron-induced enhancement effects. The fluorescence data, in turn, have been used to validate a planetary fluorescence simulation tool based on the GEANT4 transport code. This simulation can be used as a mission analysis tool to predict the time-dependent orbital XRF spectral distributions from planetary surfaces throughout the mapping phase.

  19. Fluorescence lifetime measurements of boronate derivatives to determine glucose concentration

    SciTech Connect

    Gable, J H

    2000-06-01

    A novel investigation into the fluorescence lifetimes of molecules, both established and newly designed, was performed. These molecules are the basis of a continuous, minimally invasive, glucose sensor based on fluorescence lifetime measurements. This sensor, if coupled with an automated insulin delivery device, would effectively create an artificial pancreas allowing for the constant monitoring and control of glucose levels in a person with diabetes. The proposed sensor includes a fluorescent molecule that changes its' fluorescence properties upon binding selectively and reversibly to glucose. One possible sensor molecule is N-methyl-N-(9-methylene anthryl)-2-methylenephenylboronic acid (AB). The fluorescence intensity of AB was shown to change in response to changing glucose concentrations. (James, 1994) James proposed that when glucose binds to AB the fluorescence intensity increases due to an enhancement of the N{yields}B dative bond which prevents photoinduced electron transfer (PET). PET from the amine (N) to the fluorophore (anthracene) quenches the fluorescence. The dative bond between the boron and the amine can prevent PET by involving the lone pair of electrons on the amine in interactions with the boron rather than allowing them to be transferred to the fluorophore. Results of this research show the average fluorescence lifetime of AB also changes with glucose concentration. It is proposed that fluorescence is due to two components: (1) AB with an enhanced N{yields}B interaction, and no PET, and (2) AB with a weak N{yields}B interaction, resulting in fluorescence quenching by PET. Lifetime measurements of AB as a function of both the pH of the solvent and glucose concentration in the solution were made to characterize this two component system and investigate the nature of the N{yields}B bond. Measurements of molecules similar to AB were also performed in order to isolate behavior of specific AB constituents. These molecules are 9-(Methylaminomethyl

  20. Marine fluorescence from high spectrally resolved satellite measurements

    NASA Astrophysics Data System (ADS)

    Wolanin, Aleksandra; Dinter, Tilman; Rozanov, Vladimir; Noël, Stefan; Vountas, Marco; Burrows, John P.; Bracher, Astrid

    2014-05-01

    When chlorophyll molecules absorb light, most of this energy is transformed into chemical energy in a process of photosynthesis. However, a fraction of the energy absorbed is reemitted as fluorescence. As a result of its relationship to photosynthetic e?ciency, information about chlorophyll fluorescence can be used to assess the physiological state of phytoplankton (Falkowski and Kolber,1995). In-situ measurements of chlorophyll fluorescence are widespread in physiological and ecophysiological studies. When retrieved from space, chlorophyll fluorescence can improve our knowledge of global biogeochemical cycles and phytoplankton productivity (Behrenfeld et al., 2009; Huot et al., 2013) by providing high coverage and periodicity. So far, the only satellite retrieval of sun-induced marine fluorescence, Fluorescence Line Height (FLH), was designed for MODIS (Abbott and Letelier, 1999), and later also applied to the similar sensor MERIS (Gower et al., 2004). However, it could so far not be evaluated on global scale. Here, we present a different approach to observe marine chlorophyll fluorescence, based on the Differential Optical Absorption Spectroscopy (DOAS) technique (Perner and Platt, 1979) applied to the hyperspectral data from Scanning Imaging Absorption Spectrometer for Atmospheric Chartography (SCIAMACHY) and Global Ozone Monitoring Experiment-2 (GOME-2). Since fluorescence, as a trans-spectral process, leads to the shift of the wavelength of the radiation, it can be observed in the filling-in of Fraunhofer lines. In our retrieval, we evaluate the filling-in of the Zeeman triplet Fraunhofer line FeI at 684.3 nm, which is located very close to the emission peak of marine fluorescence (~685 nm). In order to conduct the chlorophyll fluorescence retrieval with the DOAS method, we calculated the reference spectra for chlorophyll fluorescence, based on simulations performed with the coupled ocean-atmosphere radiative transfer model SCIATRAN (Rozanov et al., 2014

  1. Mercury mass measurement in fluorescent lamps via neutron activation analysis

    NASA Astrophysics Data System (ADS)

    Viererbl, L.; Vinš, M.; Lahodová, Z.; Fuksa, A.; Kučera, J.; Koleška, M.; Voljanskij, A.

    2015-11-01

    Mercury is an essential component of fluorescent lamps. Not all fluorescent lamps are recycled, resulting in contamination of the environment with toxic mercury, making measurement of the mercury mass used in fluorescent lamps important. Mercury mass measurement of lamps via instrumental neutron activation analysis (NAA) was tested under various conditions in the LVR-15 research reactor. Fluorescent lamps were irradiated in different positions in vertical irradiation channels and a horizontal channel in neutron fields with total fluence rates from 3×108 cm-2 s-1 to 1014 cm-2 s-1. The 202Hg(n,γ)203Hg nuclear reaction was used for mercury mass evaluation. Activities of 203Hg and others induced radionuclides were measured via gamma spectrometry with an HPGe detector at various times after irradiation. Standards containing an Hg2Cl2 compound were used to determine mercury mass. Problems arise from the presence of elements with a large effective cross section in luminescent material (europium, antimony and gadolinium) and glass (boron). The paper describes optimization of the NAA procedure in the LVR-15 research reactor with particular attention to influence of neutron self-absorption in fluorescent lamps.

  2. Characterization of porous media by means of the depolarization metrics

    NASA Astrophysics Data System (ADS)

    Savenkov, S.; Priezzhev, A.; Oberemok, Ye.; Silfsten, P.; Ervasti, T.; Ketolainen, J.; Peiponen, K.-E.

    2012-12-01

    In this paper Mueller polarimetry is applied to study the samples with different porosity compacted from microcrystalline cellulose. We measure the whole Mueller matrices of the samples as a function of the incident angle at a wavelength of 632.8 nm. To quantify separability of the different porous samples based on differences in their Mueller matrix behavior we apply depolarization and anisotropy analysis to measured Mueller matrices by calculating parameters characterizing depolarization (depolarization index, Q(M)-metric, first and second Lorenz indices, Cloude and Lorenz entropy) and anisotropy (values and azimuths of phase and amplitude anisotropy) properties of a sample. The results show that anisotropy parameters are almost completely insensitive to the range of porosity at least at 632.8 nm. Whereas, all depolarization metrics considered are sensitive to the range of porosity. Most sensitive (not worst than 5%) among depolarization metrics are the Lorenz entropy and Q(M)-metric.

  3. Measurements of extrinsic fluorescence in Intralipid and polystyrene microspheres.

    PubMed

    Du Le, Vinh Nguyen; Nie, Zhaojun; Hayward, Joseph E; Farrell, Thomas J; Fang, Qiyin

    2014-08-01

    The fluorescence of Intralipid and polystyrene microspheres with sphere diameter of 1 µm at a representative lipid and microsphere concentration for simulation of mucosal tissue scattering has not been a subject of extensive experimental study. In order to elucidate the quantitative relationship between lipid and microsphere concentration and the respective fluorescent intensity, the extrinsic fluorescence spectra between 360 nm and 650 nm (step size of 5 nm) were measured at different lipid concentrations (from 0.25% to 5%) and different microsphere concentrations (0.00364, 0.0073, 0.0131 spheres per cubic micrometer) using laser excitation at 355 nm with pulse energy of 2.8 µJ. Current findings indicated that Intralipid has a broadband emission between 360 and 650 nm with a primary peak at 500 nm and a secondary peak at 450 nm while polystyrene microspheres have a single peak at 500 nm. In addition, for similar scattering properties the fluorescence of Intralipid solutions is approximately three-fold stronger than that of the microsphere solutions. Furthermore, Intralipid phantoms with lipid concentrations ~2% (simulating the bottom layer of mucosa) produce up to seven times stronger fluorescent emission than phantoms with lipid concentration ~0.25% (simulating the top layer of mucosa). The fluoresence decays of Intralipid and microsphere solutions were also recorded for estimation of fluorescence lifetime.

  4. Velocity measurements by laser resonance fluorescence. [single atom diffusional motion

    NASA Technical Reports Server (NTRS)

    She, C. Y.; Fairbank, W. M., Jr.

    1980-01-01

    The photonburst correlation method was used to detect single atoms in a buffer gas. Real time flow velocity measurements with laser induced resonance fluorescence from single or multiple atoms was demonstrated and this method was investigated as a tool for wind tunnel flow measurement. Investigations show that single atoms and their real time diffusional motion on a buffer gas can be measured by resonance fluorescence. By averaging over many atoms, flow velocities up to 88 m/s were measured in a time of 0.5 sec. It is expected that higher flow speeds can be measured and that the measurement time can be reduced by a factor of 10 or more by careful experimental design. The method is clearly not ready for incorporation in high speed wind tunnels because it is not yet known whether the stray light level will be higher or lower, and it is not known what detection efficiency can be obtained in a wind tunnel situation.

  5. Depolarization of diffusely reflecting man-made objects.

    PubMed

    DeBoo, Brian J; Sasian, Jose M; Chipman, Russell A

    2005-09-10

    The polarization properties of light scattered or diffusely reflected from seven different man-made samples are studied. For each diffusely reflecting sample an in-plane Mueller matrix bidirectional reflectance distribution function is measured at a fixed bistatic angle using a Mueller matrix imaging polarimeter. The measured profile of depolarization index with changing scattering geometry for most samples is well approximated by an inverted Gaussian function. Depolarization is minimum for specular reflection and increases asymptotically in a Gaussian fashion as the angles of incidence and scatter increase. Parameters of the Gaussian profiles fitted to the depolarization data are used to compare samples. The dependence of depolarization on the incident polarization state is compared for each Stokes basis vector: horizontal, vertical, 45 degrees, 135 degrees, and right- and left-circular polarized light. Linear states exhibit similar depolarization profiles that typically differ in value by less than 0.06 (where 1.0 indicates complete depolarization). Circular polarization states are depolarized more than linear states for all samples tested, with the output degree of polarization reduced from that of linear states by as much as 0.15. The depolarization difference between linear and circular states varies significantly between samples.

  6. X-ray fluorescence measurements of dissolved gas and cavitation

    DOE PAGES

    Duke, Daniel J.; Kastengren, Alan L.; Swantek, Andrew B.; ...

    2016-09-28

    The dynamics of dissolved gas and cavitation are strongly coupled, yet these phenomena are difficult to measure in-situ. Both create voids in the fluid that can be difficult to distinguish. In this paper, we present an application of X-ray fluorescence in which liquid density and total noncondensible gas concentration (both dissolved and nucleated) are simultaneously measured. The liquid phase is doped with 400 ppm of a bromine tracer, and dissolved air is removed and substituted with krypton. Fluorescent emission at X-ray wavelengths is simultaneously excited from the Br and Kr with a focused monochromatic X-ray beam from a synchrotron source.more » We measure the flow in a cavitating nozzle 0.5 mm in diameter. From Br fluorescence, total displacement of the liquid is measured. From Kr fluorescence, the mass fraction of both dissolved and nucleated gas is measured. Volumetric displacement of liquid due to both cavitation and gas precipitation can be separated through estimation of the local equilibrium dissolved mass fraction. The uncertainty in the line of sight projected densities of the liquid and gas phases is 4–6 %. The high fluorescence yields and energies of Br and Kr allow small mass fractions of gas to be measured, down to 10-5, with an uncertainty of 8 %. Finally, these quantitative measurements complement existing optical diagnostic techniques and provide new insight into the diffusion of gas into cavitation bubbles, which can increase their internal density, pressure and lifetimes by orders of magnitude.« less

  7. Kr II laser-induced fluorescence for measuring plasma acceleration.

    PubMed

    Hargus, W A; Azarnia, G M; Nakles, M R

    2012-10-01

    We present the application of laser-induced fluorescence of singly ionized krypton as a diagnostic technique for quantifying the electrostatic acceleration within the discharge of a laboratory cross-field plasma accelerator also known as a Hall effect thruster, which has heritage as spacecraft propulsion. The 728.98 nm Kr II transition from the metastable 5d(4)D(7/2) to the 5p(4)P(5/2)(∘) state was used for the measurement of laser-induced fluorescence within the plasma discharge. From these measurements, it is possible to measure velocity as krypton ions are accelerated from near rest to approximately 21 km/s (190 eV). Ion temperature and the ion velocity distributions may also be extracted from the fluorescence data since available hyperfine splitting data allow for the Kr II 5d(4)D(7/2)-5p(4)P(5/2)(∘) transition lineshape to be modeled. From the analysis, the fluorescence lineshape appears to be a reasonable estimate for the relatively broad ion velocity distributions. However, due to an apparent overlap of the ion creation and acceleration regions within the discharge, the distributed velocity distributions increase ion temperature determination uncertainty significantly. Using the most probable ion velocity as a representative, or characteristic, measure of the ion acceleration, overall propellant energy deposition, and effective electric fields may be calculated. With this diagnostic technique, it is possible to nonintrusively characterize the ion acceleration both within the discharge and in the plume.

  8. Kr II laser-induced fluorescence for measuring plasma acceleration

    NASA Astrophysics Data System (ADS)

    Hargus, W. A.; Azarnia, G. M.; Nakles, M. R.

    2012-10-01

    We present the application of laser-induced fluorescence of singly ionized krypton as a diagnostic technique for quantifying the electrostatic acceleration within the discharge of a laboratory cross-field plasma accelerator also known as a Hall effect thruster, which has heritage as spacecraft propulsion. The 728.98 nm Kr II transition from the metastable 5d4D7/2 to the 5p ^4P^circ _{5/2} state was used for the measurement of laser-induced fluorescence within the plasma discharge. From these measurements, it is possible to measure velocity as krypton ions are accelerated from near rest to approximately 21 km/s (190 eV). Ion temperature and the ion velocity distributions may also be extracted from the fluorescence data since available hyperfine splitting data allow for the Kr II 5d4D7/2-5p ^4P^circ _{5/2} transition lineshape to be modeled. From the analysis, the fluorescence lineshape appears to be a reasonable estimate for the relatively broad ion velocity distributions. However, due to an apparent overlap of the ion creation and acceleration regions within the discharge, the distributed velocity distributions increase ion temperature determination uncertainty significantly. Using the most probable ion velocity as a representative, or characteristic, measure of the ion acceleration, overall propellant energy deposition, and effective electric fields may be calculated. With this diagnostic technique, it is possible to nonintrusively characterize the ion acceleration both within the discharge and in the plume.

  9. Photoinhibition of Photosystems I and II Using Chlorophyll Fluorescence Measurements

    ERIC Educational Resources Information Center

    Quiles, Maria Jose

    2005-01-01

    In this study the photoinhibition of photosystems (PS) I and II caused by exposure to high intensity light in oat ("Avena sativa," var Prevision) is measured by the emission of chlorophyll fluorescence in intact leaves adapted to darkness. The maximal quantum yield of PS II was lower in plants grown under high light intensity than in plants grown…

  10. Pulse train fluorescence technique for measuring triplet state dynamics.

    PubMed

    De Boni, Leonardo; Franzen, Paulo L; Gonçalves, Pablo J; Borissevitch, Iouri E; Misoguti, Lino; Mendonça, Cleber R; Zilio, Sergio C

    2011-05-23

    We report on a method to study the dynamics of triplet formation based on the fluorescence signal produced by a pulse train. Basically, the pulse train acts as sequential pump-probe pulses that precisely map the excited-state dynamics in the long time scale. This allows characterizing those processes that affect the population evolution of the first excited singlet state, whose decay gives rise to the fluorescence. The technique was proven to be valuable to measure parameters of triplet formation in organic molecules. Additionally, this single beam technique has the advantages of simplicity, low noise and background-free signal detection.

  11. Developing and Testing a Bayesian Analysis of Fluorescence Lifetime Measurements

    PubMed Central

    Needleman, Daniel J.

    2017-01-01

    FRET measurements can provide dynamic spatial information on length scales smaller than the diffraction limit of light. Several methods exist to measure FRET between fluorophores, including Fluorescence Lifetime Imaging Microscopy (FLIM), which relies on the reduction of fluorescence lifetime when a fluorophore is undergoing FRET. FLIM measurements take the form of histograms of photon arrival times, containing contributions from a mixed population of fluorophores both undergoing and not undergoing FRET, with the measured distribution being a mixture of exponentials of different lifetimes. Here, we present an analysis method based on Bayesian inference that rigorously takes into account several experimental complications. We test the precision and accuracy of our analysis on controlled experimental data and verify that we can faithfully extract model parameters, both in the low-photon and low-fraction regimes. PMID:28060890

  12. Fluorescence quantum yield measurements of fluorescent proteins: a laboratory experiment for a biochemistry or molecular biophysics laboratory course.

    PubMed

    Wall, Kathryn P; Dillon, Rebecca; Knowles, Michelle K

    2015-01-01

    Fluorescent proteins are commonly used in cell biology to assess where proteins are within a cell as a function of time and provide insight into intracellular protein function. However, the usefulness of a fluorescent protein depends directly on the quantum yield. The quantum yield relates the efficiency at which a fluorescent molecule converts absorbed photons into emitted photons and it is necessary to know for assessing what fluorescent protein is the most appropriate for a particular application. In this work, we have designed an upper-level, biochemistry laboratory experiment where students measure the fluorescence quantum yields of fluorescent proteins relative to a standard organic dye. Four fluorescent protein variants, enhanced cyan fluorescent protein (ECFP), enhanced green fluorescent protein (EGFP), mCitrine, and mCherry, were used, however the methods described are useful for the characterization of any fluorescent protein or could be expanded to fluorescent quantum yield measurements of organic dye molecules. The laboratory is designed as a guided inquiry project and takes two, 4 hr laboratory periods. During the first day students design the experiment by selecting the excitation wavelength, choosing the standard, and determining the concentration needed for the quantum yield experiment that takes place in the second laboratory period. Overall, this laboratory provides students with a guided inquiry learning experience and introduces concepts of fluorescence biophysics into a biochemistry laboratory curriculum.

  13. Depolarization of polarized light caused by high altitude clouds. 1: Depolarization of lidar induced by cirrus.

    PubMed

    Sun, Y Y; Li, Z P; Bösenberg, J

    1989-09-01

    A scattering model is described for the investigation of depolarization of polarized light caused by ice clouds. The scattering by a single particle is described by refraction, reflection, and diffraction. The ice cloud is assumed to be a random mixture of hexagonal columns and plates of random orientation and size. Multiple scattering effects are included by means of a Monte Carlo method, where single photon histories are constructed from random samples of the distributions governing the basic scattering parameters. The dependence of depolarization on cloud extinction coefficient, receiver field of view, and mixing ratio of columns to plates are studied. Lidar measurements of depolarization by a high altitude cirrus cloud are presented and discussed within the frame of the present model. Good agreement can be obtained assuming a variation of crystal shape distribution with height.

  14. Elastic depolarization of OH(A) by He and Ar: a comparative study.

    PubMed

    Costen, M L; Livingstone, R; McKendrick, K G; Paterson, G; Brouard, M; Chadwick, H; Chang, Y-P; Eyles, C J; Aoiz, F J; Kłos, J

    2009-12-31

    Two color polarization spectroscopy has been employed to measure the collisional depolarization of OH(A(2)Sigma(+), v = 1) by He and Ar. Complementary experiments using Zeeman quantum beat spectroscopy have also been performed to determine separately the cross sections for rotational energy transfer (RET) out of selected rotational levels of OH(A, v = 0) + Ar, as well as those for elastic depolarization. This has been achieved by dispersing the emission, so as to observe a single fluorescence transition. Elastic depolarization of OH(A) by Ar is found to be significant with that for loss of rotational alignment exceeding that for loss of orientation. In the case of OH(A) + He, the polarization spectroscopy measurements suggest that elastic depolarization plays a relatively minor role in the loss of the polarization signal compared with RET. The experimental data for OH(A) + Ar are compared in detail with the results of quasi-classical trajectory calculations that accommodate the effects of electron spin. These classical calculations are assessed against the results obtained using full close-coupled open shell quantum mechanical scattering methods. Overall the level of agreement between the two experiments, and between experiment and theory, is very reasonable. Surprisingly, at low N the elastic depolarization cross sections for OH(A) + Ar are found to be quite similar in magnitude to those observed for OH(X) + Ar despite the fact that the well depth in the latter system is considerably smaller than that for OH(A)-Ar. However, for OH(A) + Ar the depolarization cross sections are insensitive to N in the range 1-14. It is proposed that this behavior partly reflects the relatively anisotropic nature of the potential energy surface, which exhibits deep wells of different depths at the two linear configurations OH(A)-Ar and Ar-OH(A), and partly the nature of elastic depolarizing collisions, which must occur with a velocity component perpendicular to the plane of rotation

  15. Blind deconvolution estimation of fluorescence measurements through quadratic programming

    NASA Astrophysics Data System (ADS)

    Campos-Delgado, Daniel U.; Gutierrez-Navarro, Omar; Arce-Santana, Edgar R.; Skala, Melissa C.; Walsh, Alex J.; Jo, Javier A.

    2015-07-01

    Time-deconvolution of the instrument response from fluorescence lifetime imaging microscopy (FLIM) data is usually necessary for accurate fluorescence lifetime estimation. In many applications, however, the instrument response is not available. In such cases, a blind deconvolution approach is required. An iterative methodology is proposed to address the blind deconvolution problem departing from a dataset of FLIM measurements. A linear combination of a base conformed by Laguerre functions models the fluorescence impulse response of the sample at each spatial point in our formulation. Our blind deconvolution estimation (BDE) algorithm is formulated as a quadratic approximation problem, where the decision variables are the samples of the instrument response and the scaling coefficients of the basis functions. In the approximation cost function, there is a bilinear dependence on the decision variables. Hence, due to the nonlinear nature of the estimation process, an alternating least-squares scheme iteratively solves the approximation problem. Our proposal searches for the samples of the instrument response with a global perspective, and the scaling coefficients of the basis functions locally at each spatial point. First, the iterative methodology relies on a least-squares solution for the instrument response, and quadratic programming for the scaling coefficients applied just to a subset of the measured fluorescence decays to initially estimate the instrument response to speed up the convergence. After convergence, the final stage computes the fluorescence impulse response at all spatial points. A comprehensive validation stage considers synthetic and experimental FLIM datasets of ex vivo atherosclerotic plaques and human breast cancer cell samples that highlight the advantages of the proposed BDE algorithm under different noise and initial conditions in the iterative scheme and parameters of the proposal.

  16. Measurements of Solar Induced Chlorophyll Fluorescence at 685 nm by Airborne Plant Fluorescence Sensor (APFS)

    NASA Astrophysics Data System (ADS)

    Morgan, F.; Yee, J. H.; Boldt, J.; Cook, W. B.; Corp, L. A.

    2015-12-01

    Solar-induced chlorophyll fluorescence (ChlF) by terrestrial vegetation is linked closely to photosynthetic efficiency that can be exploited to monitor the plant health status and to assess the terrestrial carbon budget from space. The weak, broad continuum ChlF signal can be detected from the fill-in of strong O2 absorption lines or solar Fraunhofer lines in the reflected spectral radiation. The Johns Hopkins University, Applied Physics Laboratory (JHU/APL) Airborne Plant Fluorescence Sensor (APFS) is a triple etalon Fabry-Perot interferometer designed and optimized specifically for the ChlF sensing from an airborne platform using this line fill-in technique. In this paper, we will present the results of APFS ChlF measurements obtained from a NASA Langley King Air during two airborne campaigns (12/12 in 2014 and 5/20 in 2015) over various land, river, and vegetated targets in Virginia during stressed and growth seasons.

  17. Measurement of Nanoparticle Magnetic Hyperthermia Using Fluorescent Microthermal Imaging

    NASA Astrophysics Data System (ADS)

    Zheng, Xiaowan; van Keuren, Edward

    Nanoparticle magnetic hyperthermia uses the application of an AC magnetic field to ferromagnetic nanoparticles to elevate the temperature of cancer cells. The principle of hyperthermia as a true cell-specific therapy is that tumor cells are more sensitive to high temperature, so it is of great importance to control the locality and magnitude of the temperature differences. One technique to measure temperature variations on microscopic length scales is fluorescent microthermal imaging (FMI). Since it is the local temperature that is measured in FMI, effects such as heating due to nearby field coils can be accounted for. A dye, the rare earth chelate europium thenoyltrifluoroacetonate (Eu:TTA), with a strong temperature-dependent fluorescence emission has been incorporated into magnetic nanoparticles dispersed in a polymer films. FMI experiments were carried out on these samples under an applied high frequency magnetic field. Preliminary results show that FMI is a promising technique for characterizing the local generation of heat in nanoparticle magnetic hyperthermia.

  18. Collisional Effects On Laser-Induced Fluorescence Flame Measurements

    NASA Astrophysics Data System (ADS)

    Crosley, David R.

    1981-08-01

    Abstract. Laser-induced fluorescence (LIF) is a method of considerable utility for the measurement of the transient free radicals which are the keys to the chemistry of flames. Collisions experienced by the electronically excited state can alter the magnitude and the spectral form of the fluorescence signals. Recent studies on both quenching and energy transfer collisions, and their influence on LIF measurements, are treated in this review; special emphasis is given to the important and popular OH molecule. Different solutions to the problem of accounting for quenching are considered, and both effects and exploitation of energy transfer within the excited state are discussed. Although further research is needed to better quantify these collisional effects, LIF can currently provide data significant for the understanding of combustion chemistry.

  19. Fluorescence decay time measurement - a new optical sensing scheme

    NASA Astrophysics Data System (ADS)

    Draxler, Sonja; Lippitsch, Max E.

    1994-02-01

    Optical sensors often suffer from poor long-term stability. This drawback can be overcome by using fluorescence decay-time measurement as the sensing principle. In this way calibration- free chemical sensors can be developed. The sensing scheme has been used so far mainly in connection with dynamic quenching, for example in oxygen sensors. We have succeeded in extending it to ground-state indicator-analyte reactions, thus obtaining stable optical sensors for decay-time sensing of various analytes.

  20. Kr II laser-induced fluorescence for measuring plasma acceleration

    SciTech Connect

    Hargus, W. A. Jr.

    2012-10-15

    We present the application of laser-induced fluorescence of singly ionized krypton as a diagnostic technique for quantifying the electrostatic acceleration within the discharge of a laboratory cross-field plasma accelerator also known as a Hall effect thruster, which has heritage as spacecraft propulsion. The 728.98 nm Kr II transition from the metastable 5d{sup 4}D{sub 7/2} to the 5p{sup 4}P{sub 5/2}{sup Ring-Operator} state was used for the measurement of laser-induced fluorescence within the plasma discharge. From these measurements, it is possible to measure velocity as krypton ions are accelerated from near rest to approximately 21 km/s (190 eV). Ion temperature and the ion velocity distributions may also be extracted from the fluorescence data since available hyperfine splitting data allow for the Kr II 5d{sup 4}D{sub 7/2}-5p{sup 4}P{sub 5/2}{sup Ring-Operator} transition lineshape to be modeled. From the analysis, the fluorescence lineshape appears to be a reasonable estimate for the relatively broad ion velocity distributions. However, due to an apparent overlap of the ion creation and acceleration regions within the discharge, the distributed velocity distributions increase ion temperature determination uncertainty significantly. Using the most probable ion velocity as a representative, or characteristic, measure of the ion acceleration, overall propellant energy deposition, and effective electric fields may be calculated. With this diagnostic technique, it is possible to nonintrusively characterize the ion acceleration both within the discharge and in the plume.

  1. Latrunculin A depolarizes starfish oocytes.

    PubMed

    Moccia, F

    2007-12-01

    Depolymerization of the actin cytoskeleton may liberate Ca2+ from InsP3-sensitive stores in some cell types, including starfish oocytes, while inhibiting Ca2+ influx in others. However, no information is available on the modulation of membrane potential (V(m)) by actin. The present study was aimed to ascertain whether the widely employed actin depolymerizing drug, latrunculin A (Lat A), affects V(m) in mature oocytes of the starfish Astropecten aranciacus. Lat A induced a membrane depolarization which was mimicked by cytochalasin D, another popular actin disruptor, and prevented by jasplakinolide, a stabilizer of the actin network. Lat A-elicited depolarization consisted in a positive shift in V(m) which reached the threshold of activation of voltage-gated Ca2+ channels (VGCC), thus triggering an action potential. Lat A-promoted depolarization lacked the action potential in Ca2+-free sea water, while it was abolished upon removal of external Na+. Moreover, membrane depolarization was prevented by pre-injection of BAPTA and heparin, but not ryanodine. These data indicate that Lat A induces a membrane depolarization by releasing Ca2+ from InsP3Rs. The Ca2+ signal in turn activates a Ca2+-dependent Na+ entry, which causes the positive shift in V(m) and stimulates the VGCC.

  2. Translational Diffusion in Phospholipid Monolayers Measured by Fluorescence Microphotolysis

    NASA Astrophysics Data System (ADS)

    Peters, Reiner; Beck, Konrad

    1983-12-01

    A method is described that eliminates surface flow in monolayers at the air-water interface and makes possible diffusion measurements by fluorescence microphotolysis (``photobleaching''). In contrast to previous studies that did not account for surface flow, lipid probe diffusion has been found to be similar in densely packed monolayers and in related bilayers. Furthermore, it seems that lipid diffusion is based on the same molecular mechanism in monolayers, bilayers, and potentially also cell membranes. In monolayers of L-α -dilauroylphosphatidylcholine (Lau2-PtdCho) the translational diffusion coefficient D of the fluorescent lipid probe N-4-nitrobenzo-2-oxa-1,3 diazole egg phosphatidylethanolamine decreased from 110 μ m2/s at a surface pressure Pi =1 mN/m to 15 μ m2/s at Pi =38 mN/m (T = 21-22 degrees C). Data could be fitted by the ``free volume model.'' In monolayers of L-α -dipalmitoylphosphatidylcholine (Pam2-PtdCho) D decreased by >3 orders of magnitude upon increasing Pi at constant temperature, thus indicating a fluid-to-crystalline phase transition. In Lau2-PtdCho/Pam2-PtdCho monolayers phase separation has been visualized in the fluorescence microscope and the effect on D measured. These results suggest that monolayers are a promising model system for studying the molecular mobility of lipids and other cell membrane components.

  3. Cell-free measurements of brightness of fluorescently labeled antibodies

    NASA Astrophysics Data System (ADS)

    Zhou, Haiying; Tourkakis, George; Shi, Dennis; Kim, David M.; Zhang, Hairong; Du, Tommy; Eades, William C.; Berezin, Mikhail Y.

    2017-02-01

    Validation of imaging contrast agents, such as fluorescently labeled imaging antibodies, has been recognized as a critical challenge in clinical and preclinical studies. As the number of applications for imaging antibodies grows, these materials are increasingly being subjected to careful scrutiny. Antibody fluorescent brightness is one of the key parameters that is of critical importance. Direct measurements of the brightness with common spectroscopy methods are challenging, because the fluorescent properties of the imaging antibodies are highly sensitive to the methods of conjugation, degree of labeling, and contamination with free dyes. Traditional methods rely on cell-based assays that lack reproducibility and accuracy. In this manuscript, we present a novel and general approach for measuring the brightness using antibody-avid polystyrene beads and flow cytometry. As compared to a cell-based method, the described technique is rapid, quantitative, and highly reproducible. The proposed method requires less than ten microgram of sample and is applicable for optimizing synthetic conjugation procedures, testing commercial imaging antibodies, and performing high-throughput validation of conjugation procedures.

  4. Cell-free measurements of brightness of fluorescently labeled antibodies

    PubMed Central

    Zhou, Haiying; Tourkakis, George; Shi, Dennis; Kim, David M.; Zhang, Hairong; Du, Tommy; Eades, William C.; Berezin, Mikhail Y.

    2017-01-01

    Validation of imaging contrast agents, such as fluorescently labeled imaging antibodies, has been recognized as a critical challenge in clinical and preclinical studies. As the number of applications for imaging antibodies grows, these materials are increasingly being subjected to careful scrutiny. Antibody fluorescent brightness is one of the key parameters that is of critical importance. Direct measurements of the brightness with common spectroscopy methods are challenging, because the fluorescent properties of the imaging antibodies are highly sensitive to the methods of conjugation, degree of labeling, and contamination with free dyes. Traditional methods rely on cell-based assays that lack reproducibility and accuracy. In this manuscript, we present a novel and general approach for measuring the brightness using antibody-avid polystyrene beads and flow cytometry. As compared to a cell-based method, the described technique is rapid, quantitative, and highly reproducible. The proposed method requires less than ten microgram of sample and is applicable for optimizing synthetic conjugation procedures, testing commercial imaging antibodies, and performing high-throughput validation of conjugation procedures. PMID:28150730

  5. Bloodstain age analysis: toward solid state fluorescent lifetime measurements

    NASA Astrophysics Data System (ADS)

    Guo, Kevin; Zhegalova, Natalia; Achilefu, Samuel; Berezin, Mikhail Y.

    2013-03-01

    One of the most pressing unsolved challenges in forensic science is the determination of time since deposition (TSD) of bloodstains at crime scenes. Despite a number of high profile cases over the past couple hundred years involving controversy over TSD methods, no reliable quantitative method has been established. We present here an approach that has yet to be explored by forensic scientist: measuring the fluorescence lifetime of solid-state blood. Such a method would allow for on-site measurements of bloodstains utilizing the appropriate device, and would allow for rapid results returned in real-time to investigators.

  6. Containerless high temperature property measurements by atomic fluorescence

    NASA Technical Reports Server (NTRS)

    Nordine, P. C.; Schiffman, R. A.

    1982-01-01

    Laser induced fluorescence techniques were developed for the containerless study of high temperature processes, material properties, levitation, and heating techniques for containerless earth-based experimentation. Experiments were performed in which fluorescence of atomic aluminum, mercury, or tungsten were studied. These experiments include measurements of: (1) Al atom evaporation from CW CO2 laser heated and aerodynamically levitated sapphire and alumina spheres, and self-supported sapphire filaments, (2) Al atom reaction with ambient oxygen in the wake of a levitated specimen, (3) Hg atom concentrations in the wake of levitated alumina and sapphire spheres, relative to the ambient Hg atom concentration, (4) Hg atom concentrations in supersonic levitation jets, and (5) metastable, electronically excited W atom concentrations produced by evaporation of an electrically heated tungsten filament.

  7. Influence of the surface hydrophobicity on fluorescence correlation spectroscopy measurements

    NASA Astrophysics Data System (ADS)

    Boutin, Céline; Jaffiol, Rodolphe; Plain, Jérome; Royer, Pascal

    2007-02-01

    Fluorescence correlation spectroscopy (FCS) is a powerful experimental technique used to analyze the diffusion at the single molecule level in solution. FCS is based on the temporal autocorrelation of fluorescent signal generated by dye molecules diffusing through a small confocal volume. These measurements are mostly carried out in a chambered coverglass, close to the glass substrate. In this report, we discuss how the chemical nature of the glass-water interface may interact with the free diffusion of molecules. Our results reveal a strong influence, up to a few μm from the interface, of the surface hydrophobicity degree. This influence is assessed through the relative weight of the two dimension diffusion process observed at the vicinity of the surface.

  8. Compton effect thermally activated depolarization dosimeter

    DOEpatents

    Moran, Paul R.

    1978-01-01

    A dosimetry technique for high-energy gamma radiation or X-radiation employs the Compton effect in conjunction with radiation-induced thermally activated depolarization phenomena. A dielectric material is disposed between two electrodes which are electrically short circuited to produce a dosimeter which is then exposed to the gamma or X radiation. The gamma or X-radiation impinging on the dosimeter interacts with the dielectric material directly or with the metal composing the electrode to produce Compton electrons which are emitted preferentially in the direction in which the radiation was traveling. A portion of these electrons becomes trapped in the dielectric material, consequently inducing a stable electrical polarization in the dielectric material. Subsequent heating of the exposed dosimeter to the point of onset of ionic conductivity with the electrodes still shorted through an ammeter causes the dielectric material to depolarize, and the depolarization signal so emitted can be measured and is proportional to the dose of radiation received by the dosimeter.

  9. Remote chlorophyll fluorescence measurements with the laser-induced fluorescence transient approach.

    PubMed

    Pieruschka, Roland; Klimov, Denis; Berry, Joseph A; Osmond, C Barry; Rascher, Uwe; Kolber, Zbigniew S

    2012-01-01

    The interaction of plants with their environment is very dynamic. Studying the underlying processes is important for understanding and modeling plant response to changing environmental conditions. Photosynthesis varies largely between different plants and at different locations within a canopy of a single plant. Thus, continuous and spatially distributed monitoring is necessary to assess the dynamic response of photosynthesis to the environment. Limited scale of observation with portable instrumentation makes it difficult to examine large numbers of plants under different environmental conditions. We report here on the application of a recently developed technique, laser-induced fluorescence transient (LIFT), for continuous remote measurement of photosynthetic efficiency of selected leaves at a distance of up to 50 m. The ability to make continuous, automatic, and remote measurements of photosynthetic efficiency of leaves with the LIFT provides a new approach for studying the interaction of plants with the environment and may become an important tool in phenotyping photosynthetic properties in field applications.

  10. Containerless high temperature property measurements by atomic fluorescence

    NASA Technical Reports Server (NTRS)

    Schiffman, R. A.; Walker, C. A.

    1984-01-01

    Laser induced fluorescence (LIF) techniques for containerless study of high temperature processes and material properties was studied. Gas jet and electromagnetic levitation and electromagnetic and laser heating techniques are used with LIF in earth-based containerless high temperature experiments. Included are the development of an apparatus and its use in the studies of (1) chemical reactions on Al2O3, molybdenum, tungsten and LaB6 specimens, (2) methods for noncontact specimen temperature measurement, (3) levitation jet properties and (4) radiative lifetime and collisional energy transfer rates for electronically excited atoms.

  11. Containerless high temperature property measurements by atomic fluorescence

    NASA Technical Reports Server (NTRS)

    1983-01-01

    The use of laser induced fluorescence (LIF) techniques for containerless study of high temperature processes and material properties is studied. Gas jet and electromagnetic levitation and electromagnetic and laser heating techniques are used with LIF in Earth-based containerless high temperature experiments. The work to date includes development of an apparatus and its use in studies of chemical reactions on Al2O3, molybdenum, and tungsten specimens, novel methods for noncontact specimen temperature measurement, and levitation jet properties. Brief summaries of these studies are given. The apparatus is described and detailed results for the current reporting period are presented.

  12. Measuring Phagosomal pH by Fluorescence Microscopy.

    PubMed

    Canton, Johnathan; Grinstein, Sergio

    2017-01-01

    Dual wavelength ratiometric imaging has become a powerful tool for the study of pH in intracellular compartments. It allows for the dynamic imaging of live cells while accounting for changes in the focal plane, differential loading of the fluorescent probe, and photobleaching caused by repeated image acquisitions. Ratiometric microscopic imaging has the added advantage over whole population methods of being able to resolve individual cells and even individual organelles. In this chapter we provide a detailed discussion of the basic principles of ratiometric imaging and its application to the measurement of phagosomal pH, including probe selection, the necessary instrumentation, and calibration methods.

  13. Time-resolved fluorescence measurements of actin-phalloidin interactions

    NASA Astrophysics Data System (ADS)

    Helms, Michael K.; French, Todd E.

    2000-03-01

    Compounds that interact with the cytoskeleton affect mobility and division, making them useful for treatment of certain types of cancer. Actin binding drugs such as the phallotoxins (small, bicyclic peptides) bind to and stabilize actin polymers (F-actin) without binding to actin monomers (G-actin). It has been shown that the intensity of fluorescently labeled phallotoxins such as fluorescein- phalloidin and rhodamine-phalloidin increases upon bind F- actin. We used LJL BioSystems' new FLAReTM technology to measure excited state lifetime changes of fluorescein- phalloidin and rhodamine-phalloidin upon binding to F- actin.

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

    NASA Astrophysics Data System (ADS)

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

    2007-10-01

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

  15. Assessment of Vegetation Stress Using Reflectance or Fluorescence Measurements

    NASA Technical Reports Server (NTRS)

    Campbell, P. K. E.; Middleton, E. M.; McMurtrey, J. E.; Corp, L. A.; Chappelle, E. W.

    2007-01-01

    Current methods for large-scale vegetation monitoring rely on multispectral remote sensing, which has serious limitation for the detection of vegetation stress. To contribute to the establishment of a generalized spectral approach for vegetation stress detection, this study compares the ability of high-spectral resolution reflectance (R) and fluorescence (F) foliar measurements to detect vegetation changes associated with common environmental factors affecting plant growth and productivity. To obtain a spectral dataset from a broad range of species and stress conditions, plant material from three experiments was examined, including (i) corn, nitrogen (N) deficiency/excess; (ii) soybean, elevated carbon dioxide, and ozone levels; and (iii) red maple, augmented ultraviolet irradiation. Fluorescence and R spectra (400-800 nm) were measured on the same foliar samples in conjunction with photosynthetic pigments, carbon, and N content For separation of a wide range of treatment levels, hyperspectral (5-10 nm) R indices were superior compared with F or broadband R indices, with the derivative parameters optimal results. For the detection of changes in vegetation physiology, hyperspectral indices can provide a significant improvement over broadband indices. The relationship of treatment levels to R was linear, whereas that to F was curvilinear. Using reflectance measurements, it was not possible to identify the unstressed vegetation condition, which was accomplished in all three experiments using F indices. Large-scale monitoring of vegetation condition and the detection of vegetation stress could be improved by using hyperspectral R and F information, a possible strategy for future remote sensing missions.

  16. Bone lead measured by X-ray fluorescence: epidemiologic methods.

    PubMed Central

    Hu, H; Aro, A; Rotnitzky, A

    1995-01-01

    In vivo X-ray fluorescence (XRF) measurement of bone lead concentration (XRF) has emerged as an important technique for future epidemiological studies of long-term toxicity. Several issues germane to epidemiologic methodology need to be addressed, however. First, sources of variability in measurements of bone lead need to be quantified, including imprecision related to the physical measurement itself and the variability of lead deposition over the two main compartments of bones (cortical vs. trabecular) and within each compartment. Imprecision related to the physical measurement can be estimated for each individual measurement based on the variability of the signal and background. Second, approaches to low-level data need to be debated. We argue for using the minimal detection limit (MDL) to compare instruments and interpret individual measurements; however, with regard to epidemiologic studies, we would abandon the MDL in favor of using all point estimates. In analyses using bone lead as an independent variable, statistical techniques can be used to adjust regression estimates based on estimates of measurement uncertainty and bone lead variability. Third, factors that can be expected to modify the relationship between bone lead and toxicity such as gravida history, endocrinological states, nutrition, and other important influences on bone metabolism, need to be identified and measured in epidemiologic studies. By addressing these issues, investigators will be able to maximize the utility of XRF measurements in environmental epidemiologic studies. Images Figure 2. PMID:7621788

  17. Quantitation of carcinogen bound protein adducts by fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Gan, Liang-Shang; Otteson, Michael S.; Doxtader, Mark M.; Skipper, Paul L.; Dasari, Ramachandra R.; Tannenbaum, Steven R.

    1989-01-01

    A highly significant correlation of aflatoxin B 1 serum albumin adduct level with daily aflatoxin B 1 intake was observed in a molecular epidemiological study of aflatoxin carcinogenesis which used conventional fluorescence spectroscopy methods for adduct quantitation. Synchronous fluorescence spectroscopy and laser induced fluorescence techniques have been employed to quantitate antibenzo[ a]pyrene diol epoxide derived globin peptide adducts. Fast and efficient methods to isolate the peptide adducts as well as eliminate protein fluorescence background are described. A detection limit of several femtomoles has been achieved. Experimental and technical considerations of low temperature synchronous fluorescence spectroscopy and fluorescence line narrowing to improve the detection sensitivities are also presented.

  18. Containerless high temperature property measurements by atomic fluorescence

    NASA Technical Reports Server (NTRS)

    Nordine, Paul C.; Shiffman, Robert A.

    1987-01-01

    Containerless high temperature processing and material property measurements are discussed. Researchers developed methods for non-contact suspension, heating, and property measurement for materials at temperatures up to 3,680K, the melting point of tungsten. New, scientifically interesting results were obtained in Earth-based research. These results and the demonstration of new methods and techniques form a basis for further advances under the low gravity environment of space where containerless conditions are more easily achieved. Containerless high temperature material property investigations that have been completed in this and our earlier projects include measurements of fluorine LaB sub 6 reaction kinetics at 1,000 to 1,500K; optical property measurements on sapphire (Al2O3) at temperatures up to the melting point (2,327K); and vapor pressure measurements for LaB sub 6 at 2,000 to 2,500K, for molybdenum up to 2,890K and for tungsten up to 3,680K. Gas jet levitation which is applicable to any solid material, and electromagnetic levitation of electrical conductors were used to suspend the materials of interest. Non-contact heating and property measurements were achieved by optical techniques, i.e., laser heating, laser induced fluorescence measurements of vapor concentrations, and optical pyrometry for specimen temperatures.

  19. Fluorescent measurements of Zn2+ on a smartphone

    NASA Astrophysics Data System (ADS)

    Hossain, Md. Arafat; Ast, Sandra; Canning, John; Cook, Kevin; Rutledge, Peter J.; Jamalipour, Abbas

    2015-07-01

    Using a smartphone-based portable spectrofluorimeter, measurement of metal ion concentration in water is reported. A UV LED (λex ~ 370 nm), which is powered by the internal source of the smartphone was implemented to function as the excitation source. The emission peak of the UV LED overlaps well with the absorption peak of the Zn2+-responsive molecular probe 6-(1,4,8,11-cyclam-1-yl)ethyl-1,2,3-triazol-4-yl)2-ethyl-naphthalimide fluoro-ionophore (λabs ~ 358 nm). The fluorescence emission of this dye at λem ~ 458 nm is enhanced upon coordination of Zn2+. A customized Android application digitally processes the image from a nano-imprinted polymer diffraction grating and analyses the spectral changes. Zn2+ concentration in water samples were measured with a detection limit of δ ~ 5 μM.

  20. Fluorescence quantum yield measurement in nanoparticle-fluorophore systems by thermal lens spectroscopy

    NASA Astrophysics Data System (ADS)

    Ferreira, M.; Piscitelli, V.

    2016-04-01

    Metallic nanoparticles have been used as a way to tailor the fluorescence properties like quantum yield, but regular fluorescence quantum yield measurements have to counter the reflection and dispersion of a sample for an accurate result. Thermal lens spectroscopy is a good alternative to resolve this problem because doesn't measure the fluorescence intensity but the heat generated by absorption. We studied the changes induced by silver nanoparticles, generated by laser ablation, in the fluorescence peak and quantum yield of Rhodamine B. We fund that the silver nanoparticles lowered the fluorescence peak and quenched the fluorescence of the Rhodamine B and how much is quenched also depends on its concentration.

  1. Polarized fluorescence measurements on ordered photosynthetic antenna complexes

    PubMed Central

    van Amerongen, H.; van Haeringen, B.; van Gurp, M.; van Grondelle, R.

    1991-01-01

    We have used a new and relatively easy approach to study the pigment-organization in chlorosomes from the photosynthetic bacterium Chloroflexus aurantiacus and in B800-850 antenna complexes of the photosynthetic purple bacterium Rhodobacter sphaeroides. These particles were embedded in compressed and uncompressed gels and the polarized fluorescence was determined in a 90° setup. Assuming both a rotational symmetric distribution of the particles in the gel and of the transition dipole moments in the particles, the order parameters and , describing the orientation of the symmetry axis of the particles with respect to the direction of gel expansion can be determined. Moreover, the direction parameters, describing the orientation of the absorption and emission dipole moments with respect to the symmetry axis of the particles can be obtained. The value of is essential for quantitative interpretation of linear dichroism measurements and usually it is estimated from theoretical approaches, which may lead to incorrect results. For the rod-like chlorosomes the value of appears to be the same as predicted by the theoretical approach of Ganago, A. O., M. V. Fok, I. A. Abdourakhmanov, A. A. Solov'ev, and Yu. E. Erokhin (1980. Mol. Biol. [Mosc.]. 14:381-389). The agreement with linear dichroism results, analyzed with this theoretical approach shows that the transition dipole moments are indeed in good approximation distributed in a rotationally symmetric way around the long axis of the chlorosomes. Moreover, it appears those BChl c molecules, which fluoresce, are oriented in the same way with respect to the symmetry axis as the rest of these pigments, with the dipole moments close to parallel to the long axis. The B800-850 complexes appear to orient like discs, whereas the transition dipoles of the BChl a 800- and 850-nm bands are oriented almost perpendicular to the symmetry axis. These findings are in agreement with the minimal model for these complexes

  2. Depolarizing field in ultrathin electrocalorics

    NASA Astrophysics Data System (ADS)

    Glazkova, E.; Chang, C.-M.; Lisenkov, S.; Mani, B. K.; Ponomareva, I.

    2015-08-01

    Ferroelectric thin films are considered to be among the top candidates for room-temperature electrocaloric materials as they exhibit excellent electric properties and allow application of record high electric fields. At the same time, downsizing of ferroelectric electrocalorics brings about an unwanted but unavoidable depolarizing field that could critically alter or even destroy the films' electrocaloric properties. We use an atomistic first-principles-based computational approach that does not rely on the use of Maxwell relations (i) to reveal the critical role of the depolarizing field on the electrocaloric properties of ferroelectric ultrathin films, (ii) to demonstrate the contribution of nanodomains to the electrocaloric effect in such films, and (iii) to revisit the potential limitations of the indirect approach to study electrocaloric effect in nanoscale ferroelectrics.

  3. Localization of subsurface fluorescent lesions using surface spectral measurements

    NASA Astrophysics Data System (ADS)

    Kolste, Kolbein

    Localization of Subsurface Fluorescent Lesions using Surface Spectral Measurements Sponsored by the National Institute of Health, Bethesda, Maryland Kolbein Kolste, Ph.D. Keith Paulsen In neurosurgical tumor resection, maximizing extent of resection plays a major role in the care of cancer patients. To date, ALA is being researched as a technique to guide tumor resection by inducing the accumulation of the endogenous fluorophore PpIX. Most research has focused on the use of blue light excitation of PpIX to visual the tumor. However, due to the high attenuation of blue light by in vivo chromophores, such as oxy- and deoxy-hemoglobin, the source of collected fluorescence emissions is confined to the top layer of cells, and the signal is subject to masking by blood on the surface of the surgical field of view. This issue is particularly a problem at the end of the resection, when the surgeon is evaluating the margin for remaining tumor, but the blue-signal is insensitive to residual tumor that may be located several millimeters beneath the surface. PpIX has an absorption band in the near infrared (NIR), where the absorption due to blood is orders of magnitude lower, enabling the excitation of a fluorophore at depth. In this work, we created a hyperspectral imaging system that attaches to a neurosurgical microscope and is capable of detecting PpIX fluorescence that has been excited at 635 nm. We utilize a dual-waveband technique from the hyperspectral to estimate depth of fluorescence origin and characterize the inherent limitations of the estimated depth. One of the major benefits of this technique is that the estimation is independent of the concentration and size of the fluorophore. This is first demonstrated in phantom studies, where the depths of multiple separate inclusions at various depths are accurately estimated. The technique is verified in animal tumor models and translated into the clinical theater, with pilot data showing the first estimation of depth of

  4. Quantitative Fluorescent Speckle Microscopy (QFSM) to Measure Actin Dynamics

    PubMed Central

    Mendoza, Michelle C.; Besson, Sebastien; Danuser, Gaudenz

    2012-01-01

    Quantitative Fluorescent Speckle Microscopy (QFSM) is a live cell imaging method to analyze the dynamics of macromolecular assemblies with high spatial and temporal resolution. Its greatest successes were in the analysis of actin filament and adhesion dynamics in the context of cell migration and microtubule dynamics in interphase and the meotic/mitotic spindle. Here, we focus on the former application to illustrate the procedures of FSM imaging and the computational image processing that extracts quantitative information from these experiments. QFSM is advantageous over other methods because it measures the movement and turnover kinetics of the actin filament (F-actin) network in living cells across the entire field of view. Experiments begin with microinjection of fluorophore-labeled actin into cells, which generate a low ratio of fluorescently-labeled:endogenous unlabeled actin monomers. Spinning disk confocal or wide-field imaging then visualizes fluorophore clusters (2–8 actin monomers) within the assembled F-actin network as speckles. QFSM software identifies and computationally tracks and utilizes the location, appearance, and disappearance of speckles to derive network flows and maps of the rate of filament assembly and disassembly. PMID:23042526

  5. Rotation of plasma membrane proteins measured by polarized fluorescence depletion

    NASA Astrophysics Data System (ADS)

    Barisas, B. George; Rahman, Noorul A.; Yoshida, Thomas M.; Roess, Deborah A.

    1990-05-01

    We have implemented a new laser microscopic method, polarized fluorescence depletion (PFD), for measuring the rotational dynamics of functional membrane proteins on individual, microscopically selected cells under physiological conditions. This method combines the long lifetimes of triplet-state probes with the sensitivity of fluorescence detection to measure macromolecular rotational correlation times from 10 microsec to > 1 ms. As examples, the rotational correlation time of Fc receptors (FcR) on the surface of 2H3 rat basophilic leukemia cells is 79.9 4.4 microsec at 4°C when labeled with eosin conjugates of IgE. This value is consistent with the known 100 kDa receptor size. When labeled with intact F4 anti-FcR monoclonal antibody, the rotational correlation time for FcER is increased about 2-fold to 170.8 +/- 6.5 microsec, consistent with receptor dimer formation on the plasma membrane and with the ability of this antibody to form FcER dimers on 2H3 cell surfaces. We have also examined the rotational diffusion of the luteinizing hormone receptor on plasma membranes of small ovine luteal cells. Luteinizing hormone receptors (LHR), when occupied by ovine luteinizing hormone (oLH), have a rotational correlation time of 20.5 +/- 0.1 microsec at 4°C. When occupied by human chorionic gonadotropin (hCG), LHR have a rotational correlation time of 46.2 +/- 0.4 microsec suggesting that binding of hCG triggers additional LHR interactions with plasma membrane proteins. Together these studies suggest the utility of PFD measurements in assessing molecular size and molecular association of membrane proteins on individual cells. Relative advantages of time- and frequency-domain implementations of PFD are also discussed.

  6. Analysis of Antimicrobial-Triggered Membrane Depolarization Using Voltage Sensitive Dyes

    PubMed Central

    te Winkel, J. Derk; Gray, Declan A.; Seistrup, Kenneth H.; Hamoen, Leendert W.; Strahl, Henrik

    2016-01-01

    The bacterial cytoplasmic membrane is a major inhibitory target for antimicrobial compounds. Commonly, although not exclusively, these compounds unfold their antimicrobial activity by disrupting the essential barrier function of the cell membrane. As a consequence, membrane permeability assays are central for mode of action studies analysing membrane-targeting antimicrobial compounds. The most frequently used in vivo methods detect changes in membrane permeability by following internalization of normally membrane impermeable and relatively large fluorescent dyes. Unfortunately, these assays are not sensitive to changes in membrane ion permeability which are sufficient to inhibit and kill bacteria by membrane depolarization. In this manuscript, we provide experimental advice how membrane potential, and its changes triggered by membrane-targeting antimicrobials can be accurately assessed in vivo. Optimized protocols are provided for both qualitative and quantitative kinetic measurements of membrane potential. At last, single cell analyses using voltage-sensitive dyes in combination with fluorescence microscopy are introduced and discussed. PMID:27148531

  7. Limitation of fluorescence spectrophotometry in the measurement of naphthenic acids in oil sands process water.

    PubMed

    Lu, Weibing; Ewanchuk, Andrea; Perez-Estrada, Leonidas; Sego, Dave; Ulrich, Ania

    2013-01-01

    Fluorescence spectrophotometry has been proposed as a quick screening technique for the measurement of naphthenic acids (NAs). To evaluate the feasibility of this application, the fluorescence emission spectra of NAs extracted from three oil sands process water sources were compared with that of commercial NAs. The NAs resulting from the bitumen extraction process cannot be differentiated because of the similarity of the fluorescence spectra. Separation of the fluorescent species in NAs using high performance liquid chromatography with fluorescence detector proved unsuccessful. The acidic fraction of NAs is fluorescent but the basic fraction of NAs is not fluorescent, implying that aromatic acids in NAs give rise to the fluorescent signals. The concentrations of NAs in oil sands process water were measured by Fourier transform infrared spectroscopy (FTIR), fluorescence spectrophotometry and ultra high performance liquid chromatography-time of flight/mass spectrometry (UPLC-TOF/MS). Commercial Merichem and Kodak NAs are the best standards to use when measuring NAs concentration with FTIR and fluorescence spectrophotometry. In addition, the NAs concentrations measured by fluorescence spectrophotometry are about 30 times higher than those measured by FTIR and UPLC-TOF/MS. The findings in this study underscore the limitation of fluorescence spectrophotometry in the measurement of NAs.

  8. Measuring Agarwood Formation Ratio Quantitatively by Fluorescence Spectral Imaging Technique.

    PubMed

    Huang, Botao; Nguyen, Duykien; Liu, Tianyi; Jiang, Kaibin; Tan, Jinfen; Liu, Chunxin; Zhao, Jing; Huang, Shaowei

    2015-01-01

    Agarwood is a kind of important and precious traditional Chinese medicine. With the decreasing of natural agarwood, artificial cultivation has become more and more important in recent years. Quantifying the formation of agarwood is an essential work which could provide information for guiding cultivation and controlling quality. But people only can judge the amount of agarwood qualitatively by experience before. Fluorescence multispectral imaging method is presented to measure the agarwood quantitatively in this paper. A spectral cube from 450 nm to 800 nm was captured under the 365 nm excitation sources. The nonagarwood, agarwood, and rotten wood in the same sample were distinguished based on analyzing the spectral cube. Then the area ratio of agarwood to the whole sample was worked out, which is the quantitative information of agarwood area percentage. To our knowledge, this is the first time that the formation of agarwood was quantified accurately and nondestructively.

  9. Measuring Agarwood Formation Ratio Quantitatively by Fluorescence Spectral Imaging Technique

    PubMed Central

    Huang, Botao; Nguyen, Duykien; Jiang, Kaibin; Tan, Jinfen; Liu, Chunxin; Zhao, Jing; Huang, Shaowei

    2015-01-01

    Agarwood is a kind of important and precious traditional Chinese medicine. With the decreasing of natural agarwood, artificial cultivation has become more and more important in recent years. Quantifying the formation of agarwood is an essential work which could provide information for guiding cultivation and controlling quality. But people only can judge the amount of agarwood qualitatively by experience before. Fluorescence multispectral imaging method is presented to measure the agarwood quantitatively in this paper. A spectral cube from 450 nm to 800 nm was captured under the 365 nm excitation sources. The nonagarwood, agarwood, and rotten wood in the same sample were distinguished based on analyzing the spectral cube. Then the area ratio of agarwood to the whole sample was worked out, which is the quantitative information of agarwood area percentage. To our knowledge, this is the first time that the formation of agarwood was quantified accurately and nondestructively. PMID:26089935

  10. Nonspherical nanoparticles characterization by partially depolarized dynamic light scattering

    NASA Astrophysics Data System (ADS)

    Levin, Alexander D.; Shmytkova, Ekaterina A.

    2015-06-01

    The realization of improved depolarized dynamic light scattering method is presented. This technique supports measurement of non-spherical nanoparticals dimensions in liquids. The relations between translational and rotational diffusion coefficients and autocorrelation function of scattered light with polarized and depolarized components in various proportions are derived. Thus measurement of very weak cross-polarized component can be avoided. This improvement permits to reduce measurement time, to improve signal to noise ratio and results precision. The technique was applied for sizing of gold nanorods and multiwalled carbon nanotubes in liquids.

  11. Application of fluorescence lifetime imaging of enhanced green fluorescent protein to intracellular pH measurements.

    PubMed

    Nakabayashi, Takakazu; Wang, Hui-Ping; Kinjo, Masataka; Ohta, Nobuhiro

    2008-06-01

    We have shown that the intracellular pH of a single HeLa cell expressing the enhanced green fluorescent protein (EGFP) can be imaged using the fluorescence lifetime of EGFP, which can be interpreted in terms of the pH-dependent ionic equilibrium of the p-hydroxybenzylidene-imidazolidinone structure of the chromophore of EGFP.

  12. An advanced fluorescence LIDAR system for the acquisition of interleaved active (LIF) and passive (SIF) fluorescence measurements on vegetation

    NASA Astrophysics Data System (ADS)

    Raimondi, Valentina; Palombi, Lorenzo; Di Ninni, Paola

    2015-10-01

    Fluorescence is regarded as a valuable tool to investigate the eco-physiological status of vegetation. Chlorophyll a, which emits a typical fluorescence in the red/far-red region of the e.m. spectrum, plays a key role in the photosynthetic process and its fluorescence is considered an effective proxy of photosynthetic activity of plants. Laser Induced Fluorescence (LIF) has been studied for several decades both at leaf- and canopy-level by means of optical fibers-coupled instrumentation and fluorescence LIDAR systems. On the other hand, Solar-Induced Fluorescence (SIF) has been the object of several scientific studies quite recently, with the aim to investigate the feasibility of measuring the fluorescence of vegetation using passive spectroradiometers in view of global scale monitoring from satellite platforms. This paper presents the main technical features and preliminary tests of a fluorescence LIDAR, recently upgraded to acquire maps of interleaved LIF and SIF measurements at canopy level. In-house developed electronics and software permits the acquisition of interleaved LIF and SIF spectra by switching on/off the laser, the selection of the suitable grating, the setting of the integration time and the synchronization of the Intensified CCD (ICCD) gate opening time. For each pixel of the map, a fluorescence dataset can be acquired containing a LIF spectrum - from 570 nm to 830 nm with a spectral resolution of 0.5 nm - and radiance spectra from 685.53 nm to 690.30 nm with subnanometric spectral resolution containing the molecular oxygen O2-B telluric absorption band. The latter can be exploited for polynomial regression data fit and SIF retrieval.

  13. Critical assessment of fluorescence polarization measurements with a FACS IV cell sorter

    NASA Astrophysics Data System (ADS)

    Muller, Claude P.; Krabichler, Gert

    1988-09-01

    The usefulness and limitations of the Becton-Dickinson fluorescence-activated cell sorter FACS IV for fluorescence polarization measurements were examined. A set of tests to determine the characteristics of the detection geometry, the optical properties of the beam splitter, and the capability to process fluorescence polarization data is presented. Recommendations are provided for correcting instrumental deficiencies.

  14. Characterization of natural and irradiated nails by means of the depolarization metrics

    NASA Astrophysics Data System (ADS)

    Savenkov, Sergey; Priezzhev, Alexander; Oberemok, Yevgen; Sholom, Sergey; Kolomiets, Ivan; Chunikhina, Kateryna

    2016-07-01

    Mueller polarimetry is applied to study the samples of nails: natural (or reference) and irradiated to 2 Gy ionizing radiation dose. We measure the whole Mueller matrices of the samples as a function of the scattering angle at a wavelength of 632.8 nm. We apply depolarization analysis to measured Mueller matrices by calculating the depolarization metrics [depolarization index, Q(M)-metric, first and second Lorenz indices, Cloude and Lorenz entropy] to quantify separability of the different samples of nails under study based on differences in their Mueller matrix. The results show that nail samples strongly depolarize the output light in backscattering, and irradiation in all cases results in increasing of depolarization. Most sensitive among depolarization metrics are the Lorenz entropy and Q(M)-metric.

  15. Acute Ethanol Causes Hepatic Mitochondrial Depolarization in Mice: Role of Ethanol Metabolism

    PubMed Central

    Zhong, Zhi; Ramshesh, Venkat K.; Rehman, Hasibur; Liu, Qinlong; Theruvath, Tom P.; Krishnasamy, Yasodha; Lemasters, John J.

    2014-01-01

    Background/Aims An increase of ethanol metabolism and hepatic mitochondrial respiration occurs in vivo after a single binge of alcohol. Here, our aim was to determine how ethanol intake affects hepatic mitochondrial polarization status in vivo in relation to ethanol metabolism and steatosis. Methods Hepatic mitochondrial polarization, permeability transition (MPT), and reduce pyridine nucleotides, and steatosis in mice were monitored by intravital confocal/multiphoton microscopy of the fluorescence of rhodamine 123 (Rh123), calcein, NAD(P)H, and BODIPY493/503, respectively, after gavage with ethanol (1–6 g/kg). Results Mitochondria depolarized in an all-or-nothing fashion in individual hepatocytes as early as 1 h after alcohol. Depolarization was dose- and time-dependent, peaked after 6 to 12 h and maximally affected 94% of hepatocytes. This mitochondrial depolarization was not due to onset of the MPT. After 24 h, mitochondria of most hepatocytes recovered normal polarization and were indistinguishable from untreated after 7 days. Cell death monitored by propidium iodide staining, histology and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was low throughout. After alcohol, mitochondrial NAD(P)H autofluorescence increased and decreased, respectively, in hepatocytes with polarized and depolarized mitochondria. Ethanol also caused steatosis mainly in hepatocytes with depolarized mitochondria. Depolarization was linked to ethanol metabolism, since deficiency of alcohol dehydrogenase and cytochrome-P450 2E1 (CYP2E1), the major ethanol-metabolizing enzymes, decreased mitochondrial depolarization by ∼70% and ∼20%, respectively. Activation of aldehyde dehydrogenase decreased depolarization, whereas inhibition of aldehyde dehydrogenase enhanced depolarization. Activation of aldehyde dehydrogenase also markedly decreased steatosis. Conclusions Acute ethanol causes reversible hepatic mitochondrial depolarization in vivo that may contribute to

  16. Distinct characteristics of Ca(2+)-induced depolarization of isolated brain and liver mitochondria.

    PubMed

    Vergun, Olga; Reynolds, Ian J

    2005-09-05

    Ca(2+)-induced mitochondrial depolarization was studied in single isolated rat brain and liver mitochondria. Digital imaging techniques and rhodamine 123 were used for mitochondrial membrane potential measurements. Low Ca(2+) concentrations (about 30--100 nM) initiated oscillations of the membrane potential followed by complete depolarization in brain mitochondria. In contrast, liver mitochondria were less sensitive to Ca(2+); 20 microm Ca(2+) was required to depolarize liver mitochondria. Ca(2+) did not initiate oscillatory depolarizations in liver mitochondria, where each individual mitochondrion depolarized abruptly and irreversibly. Adenine nucleotides dramatically reduced the oscillatory depolarization in brain mitochondria and delayed the onset of the depolarization in liver mitochondria. In both type of mitochondria, the stabilizing effect of adenine nucleotides completely abolished by an inhibition of adenine nucleotide translocator function with carboxyatractyloside, but was not sensitive to bongkrekic acid. Inhibitors of mitochondrial permeability transition cyclosporine A and bongkrekic acid also delayed Ca(2+)-depolarization. We hypothesize that the oscillatory depolarization in brain mitochondria is associated with the transient conformational change of the adenine nucleotide translocator from a specific transporter to a non-specific pore, whereas the non-oscillatory depolarization in liver mitochondria is caused by the irreversible opening of the pore.

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

    NASA Astrophysics Data System (ADS)

    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.

  18. Geometry of generalized depolarizing channels

    SciTech Connect

    Burrell, Christian K.

    2009-10-15

    A generalized depolarizing channel acts on an N-dimensional quantum system to compress the 'Bloch ball' in N{sup 2}-1 directions; it has a corresponding compression vector. We investigate the geometry of these compression vectors and prove a conjecture of Dixit and Sudarshan [Phys. Rev. A 78, 032308 (2008)], namely, that when N=2{sup d} (i.e., the system consists of d qubits), and we work in the Pauli basis then the set of all compression vectors forms a simplex. We extend this result by investigating the geometry in other bases; in particular we find precisely when the set of all compression vectors forms a simplex.

  19. A depolarization and attenuation experiment using the CTS satellite. Volume 1: Experiment description

    NASA Technical Reports Server (NTRS)

    Bostian, C. W.; Holt, S. B., Jr.; Kauffman, S. R.; Manus, E. A.; Marshall, R. E.; Stutzman, W. L.; Wiley, P. H.

    1976-01-01

    An experiment for measuring precipitation attenuation and depolarization on the Communications Technology Satellite (CTS) 11.7 GHz downlink is described. Attenuation and depolarization of the signal received from the spacecraft is monitored on a 24 hour basis. Data is correlated with ground weather conditions. Theoretical models for millimeter wave propagation through rain are refined for maximum agreement with observed data. Techniques are developed for predicting and mimimizing the effects of rain scatter and depolarization on future satellite communication systems.

  20. Results of the VPI&SU Comstar experiment. [depolarization and attenuation due to rain

    NASA Technical Reports Server (NTRS)

    Andrews, J. H.; Ozbay, C.; Pratt, T.; Bostian, C. W.; Manus, E. A.; Gaines, J. M.; Marshall, R. E.; Stutzman, W. L.; Wiley, P. H.

    1982-01-01

    This paper summarizes annual and cumulative attenuation data, depolarization data, and associated local rain rate distributions obtained with the Comstar family of 19.04- and 28.56-GHz satellite beacons during the years 1977-1981. It discusses the relationships between attenuation and rain rate and between attenuation and depolarization, compares measured data on the joint distribution of attenuation and depolarization, and examines the limitations that propagation effects will impose on future 20/30-GHz satellite communications systems.

  1. Electron beam fluorescence measurements in the Boeing hypersonic shock tunnel

    NASA Technical Reports Server (NTRS)

    Price, Linwood L.; Williams, W. Dan; Powell, H. M.

    1992-01-01

    The Calspan electron beam fluorescence (EBF) measurement system is described along with the results of measurements made in hypersonic flow. Numerous self-emitting metallic species were identified, many of which may be associated with an aging/erosion process within the B30HST. Because there were only 16 tunnel runs, it was only possible to obtain spectral measurements over a limited range of wavelengths and time sampling periods. Many spectral features of the flow remain uninvestigated. Because flow self-emission is important to all optical diagnostic techniques, it is recommended that additional spectral studies by performed. The three electron beam-excited species that were identified are nitrogen, helium, and nitric oxide. The high metallic radiation background interfered with attempts to obtain the time-wise variation of N2 density and He radiation with the optical fiber/PMT channels. In the case of the N2 density measurements the result of interference was increased uncertainty. Unfortunately, the interference caused the time-wise He measurements to fail completely. It is recommended that the electron beam be modulated to provide discrimination against the background radiation in future N2 density measurements. Careful data reduction produced useful measurements of N2 vibrational temperature, even though the high background from metallic species significantly increased measurement uncertainty. Perhaps the recommended additional spectral studies would reveal N2(+) First Negative System band-pair regions having less background. Detection of the He arrival was easily accomplished with the spectrometer/array detector system. Because of this, it is recommended that this means of detecting He arrival be used in the future. With proper calibrations of the system an He number density could be obtained. Although the flow conditions were out of limits for the run in which the NO spectrum was recorded, the usefulness of the NO spectrum for determination of free

  2. Fluorescence anisotropy of DNA/DAPI complex: torsional dynamics and geometry of the complex.

    PubMed Central

    Barcellona, M L; Gratton, E

    1996-01-01

    Fluorescence depolarization of synthetic polydeoxynucleotide/4'-6-diamidino-2-phenylindole dihydrochloride complexes has been investigated as a function of dye/polymer coverage. At low coverage, fluorescence depolarization is due to local torsional motions of the DNA segment where the dye resides. At relatively high coverage, fluorescence depolarization is dominated by energy transfer to other dye molecules along the DNA. The extent of the observed depolarization due to torsional motion depends on the angle the dye molecule forms with the DNA helical axis. A large torsional motion and a small angle produce the same depolarization as a small torsional motion and a large projection angle. Furthermore, the extent of transfer critically depends on the relative orientation of dye molecules along the DNA. The effect of multiple transfer is examined using a Monte Carlo approach. The measurement of depolarization with transfer, at high coverage, allows determination of the dye orientation about the DNA helical axis. The value of the torsional spring constant is then determined, at very low coverage, for few selected polydeoxynucleotides. Images FIGURE 3 PMID:9172758

  3. Phytoplankton photocompensation from space-based fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Morrison, J. Ruairidh; Goodwin, Deborah S.

    2010-03-01

    Recent satellite-derived observations linked global scale phytoplankton fluorescence variability with iron stress and hinted at photophysiological responses associated with changing light levels. These photocompensation reactions, the sum of photoacclimation and photoadaptation, were examined with climatological data for the Gulf of Maine. Significant seasonal variability was observed in the fluorescence quantum yield that was unrelated to patterns of biomass. Up to 89% of the variability in the fluorescence quantum yield was explained by a physiology-based photocompensation model. Spatial variability in seasonal patterns was associated with differing hydrodynamic regimes. This variability in the quantum yield demonstrates that satellite-based fluorescence is inappropriate for phytoplankton biomass determinations. More importantly, the work presented here provides the modeling foundation for fluorescence-based investigations of temporal and spatial variability in phytoplankton physiology associated with growth irradiance. These space-based physiological observations have the potential to decrease uncertainties in future ocean color derived primary productivity estimates.

  4. In situ measurements of phytoplankton fluorescence using low cost electronics.

    PubMed

    Leeuw, Thomas; Boss, Emmanuel S; Wright, Dana L

    2013-06-19

    Chlorophyll a fluorometry has long been used as a method to study phytoplankton in the ocean. In situ fluorometry is used frequently in oceanography to provide depth-resolved estimates of phytoplankton biomass. However, the high price of commercially manufactured in situ fluorometers has made them unavailable to some individuals and institutions. Presented here is an investigation into building an in situ fluorometer using low cost electronics. The goal was to construct an easily reproducible in situ fluorometer from simple and widely available electronic components. The simplicity and modest cost of the sensor makes it valuable to students and professionals alike. Open source sharing of architecture and software will allow students to reconstruct and customize the sensor on a small budget. Research applications that require numerous in situ fluorometers or expendable fluorometers can also benefit from this study. The sensor costs US$150.00 and can be constructed with little to no previous experience. The sensor uses a blue LED to excite chlorophyll a and measures fluorescence using a silicon photodiode. The sensor is controlled by an Arduino microcontroller that also serves as a data logger.

  5. In situ Measurements of Phytoplankton Fluorescence Using Low Cost Electronics

    PubMed Central

    Leeuw, Thomas; Boss, Emmanuel S.; Wright, Dana L.

    2013-01-01

    Chlorophyll a fluorometry has long been used as a method to study phytoplankton in the ocean. In situ fluorometry is used frequently in oceanography to provide depth-resolved estimates of phytoplankton biomass. However, the high price of commercially manufactured in situ fluorometers has made them unavailable to some individuals and institutions. Presented here is an investigation into building an in situ fluorometer using low cost electronics. The goal was to construct an easily reproducible in situ fluorometer from simple and widely available electronic components. The simplicity and modest cost of the sensor makes it valuable to students and professionals alike. Open source sharing of architecture and software will allow students to reconstruct and customize the sensor on a small budget. Research applications that require numerous in situ fluorometers or expendable fluorometers can also benefit from this study. The sensor costs US$150.00 and can be constructed with little to no previous experience. The sensor uses a blue LED to excite chlorophyll a and measures fluorescence using a silicon photodiode. The sensor is controlled by an Arduino microcontroller that also serves as a data logger. PMID:23783738

  6. Telescope Array UHECR composition measurement via stereoscopic fluorescence observation

    NASA Astrophysics Data System (ADS)

    Stroman, Thomas; Bergman, Douglas; Telescope Array Collaboration

    2016-03-01

    When entering Earth's atmosphere at ultra-high energies, cosmic rays (UHECRs) produce extensive air showers whose longitudinal development is influenced by the incident primary particle's mass. Each longitudinal shower profile reaches its maximum particle count at an atmospheric slant depth Xmax, and the distributions of observed Xmax values can be compared to those predicted by detailed simulations of the air-shower physics and the detector; accurately simulated compositions that most closely resemble that found in nature will produce the best agreement between predicted and observed Xmax distributions. This is the basis of composition measurement at the Telescope Array experiment, the largest and most sensitive UHECR detector in the northern hemisphere. At the perimeter of a large surface-detector array are three fluorescence telescope stations, whose overlapping apertures enable high-precision reconstruction of Xmax from stereoscopic observation of air-shower longitudinal profiles. We present the distribution of Xmax observed during eight years of operation, and from comparisons with several simulated combinations of composition and high-energy hadronic physics, we show that a low primary mass is favored at E >10 18 . 2 eV.

  7. Laser-induced fluorescence, dispersed fluorescence and lifetime measurements of jet-cooled chloro-substituted benzyl radicals

    NASA Astrophysics Data System (ADS)

    Hamatani, Satoshi; Tsuji, Kazuhide; Kawai, Akio; Shibuya, Kazuhiko

    2002-07-01

    We measured the laser-induced fluorescence (LIF) and dispersed fluorescence (DF) spectra of jet-cooled α-, o- and m-chlorobenzyl radicals after they were generated by the 193 nm photolysis of the corresponding parent molecules. The vibronically resolved spectra were obtained to analyze their D1-D0 transitions. The fluorescence lifetimes of α-, o-, m- and p-chlorobenzyls in the zeroth vibrational levels of the D1 states were measured to estimate the oscillator strengths of a series of benzyl derivatives. It was found that the α-substitution is inefficient to break the `accidental forbiddenness' of the D1-D0 transition of benzyl, while the ring-substitution enhances the oscillator strength by 50%.

  8. Fluorescence measurements of the thermal control experiments coatings on LDEF S0069 and A0114

    NASA Technical Reports Server (NTRS)

    Zwiener, J. M.; Mell, R. J.; Peters, P. N.; Gregory, J. C.; Wilkes, D. R.; Miller, E. R.

    1993-01-01

    Fluorescence measurements were made on the thermal control coatings from the Long Duration Experiment Facility (LDEF) S0069, Thermal Control Surfaces Experiment (TCSE); and the A0114, Interaction of Atomic Oxygen with Material Surfaces in Low Earth orbit. Fluorescence was observed in two types of thermal control coatings and is attributed to pigments or binders. In addition, fluorescence measurement on the silver Teflon from the front cover of TCSE led to confirmation of damage (cracking) to the metal layers during application.

  9. Color measurements on prints containing fluorescent whitening agents

    NASA Astrophysics Data System (ADS)

    Andersson, Mattias; Norberg, Ole

    2007-01-01

    Papers with a slightly blue shade are, at least among a majority of observers being perceived as whiter than papers having a more neutral color1. Therefore, practically all commercially available printing papers contain bluish dyes and fluorescent whitening agents (FWA) to give the paper a whiter appearance. Furthermore, in the paper industry, the most frequently used measure for paper whiteness is the CIE-whiteness. The CIE Whiteness formula, does in turn, also favor slightly bluish papers. Excessive examples of high CIE-whiteness values can be observed in the office-paper segment where a high CIE-whiteness value is an important sales argument. As an effect of the FWA, spectrophotometer measurements of optical properties such as paper whiteness are sensitive to the ultraviolet (UV) content of the light source used in the instrument. To address this, the standard spectrophotometers used in the paper industry are equipped with an adjustable filter for calibrating the UV-content of the illumination. In the paper industry, spectrophotometers with d/0 measurement geometry and a light source of type C are used. The graphical arts industry on the other hand, typically measures with spectrophotometers having 45/0 geometry and a light source of type A. Moreover, these instruments have only limited possibilities to adjust the UV-content by the use of different weighting filters. The standard for color measurements in the paper industry governs that measurements should be carried out using D65 standard illumination and the 10 ° standard observer. The corresponding standard for the graphic arts industry specify D50 standard illumination and the 2 ° standard observer. In both cases, the standard illuminants are simulated from the original light source by spectral weighting functions. However, the activation of FWA, which will impact the measured spectral reflectance, depends on the actual UV-content of the illumination used. Therefore, comparisons between measurements on

  10. Improving membrane voltage measurements using FRET with new fluorescent proteins.

    PubMed

    Tsutsui, Hidekazu; Karasawa, Satoshi; Okamura, Yasushi; Miyawaki, Atsushi

    2008-08-01

    We used two new coral fluorescent proteins as fluorescence resonance energy transfer (FRET) donor and acceptor to develop a voltage sensor, named Mermaid, that displays approximately 40% changes in emission ratio per 100 mV, allowing for direct visualization of electrical activities in cultured excitable cells. Notably, Mermaid has fast on-off kinetics at warm (approximately 33 degrees C) temperatures and can report voltage spikes comparable to action potentials.

  11. Determination of biological activity from fluorescence-lifetime measurements in Saccharomyces cerevisiae

    NASA Astrophysics Data System (ADS)

    Rudek, F.; Baselt, T.; Lempe, B.; Taudt, C.; Hartmann, P.

    2015-03-01

    The importance of fluorescence lifetime measurement as an optical analysis tool is growing. Many applications already exist in order to determine the fluorescence lifetime, but the majority of these require the addition of fluorescence-active substances to enable measurements. Every usage of such foreign materials has an associated risk. This paper investigates the use of auto-fluorescing substances in Saccharomyces cerevisiae (Baker's yeast) as a risk free alternative to fluorescence-active substance enabled measurements. The experimental setup uses a nitrogen laser with a pulse length of 350 ps and a wavelength of 337 nm. The excited sample emits light due to fluorescence of NADH/NADPH and collagen. A fast photodiode collects the light at the output of an appropriate high-pass edge-filter at 400 nm. Fluorescence lifetimes can be determined from the decay of the measurement signals, which in turn characterizes the individual materials and their surrounding environment. Information about the quantity of the fluorescence active substances can also be measured based on the received signal intensity. The correlation between the fluorescence lifetime and the metabolic state of Saccharomyces cerevisiae was investigated and is presented here.

  12. Temperature measurement by two-line laser-saturated OH fluorescence in flames.

    PubMed

    Lucht, R P; Laurendeau, N M; Sweeney, D W

    1982-10-15

    A technique is proposed and demonstrated for measuring combustion temperatures using two-line laser-saturated fluorescence. The rotational temperature of OH is determined by saturating two different rotational transitions in the (0,0) band of the A(2)Sigma(+)-X(2)II electronic system and detecting fluorescence emission which originates from the laser-pumped upper rotational levels. Temperature is calculated from the ratio of the fluorescence intensities for the two different excitation-emission pairs. The method is demonstrated by measuring temperature profiles in subatmospheric H(2)/O(2)/Ar flat flames. Temperatures measured by two-line saturated fluorescence are compared with temperatures measured by coated thermocouples and OH absorption and with predictions from an elementary chemical kinetics code. The temperatures measured by the two-line fluorescence technique are accurate to 3-5% and exhibit low random error.

  13. Repetitive Transient Depolarizations of the Inner Mitochondrial Membrane Induced by Proton Pumping

    PubMed Central

    Hattori, Tomohiro; Watanabe, Koichi; Uechi, Yukiko; Yoshioka, Hisashi; Ohta, Yoshihiro

    2005-01-01

    Single mitochondria show the spontaneous fluctuations of ΔΨm. In this study, to examine the mechanism of the fluctuations, we observed ΔΨm in single isolated heart mitochondria using time-resolved fluorescence microscopy. Addition of malate, succinate, or ascorbate plus TMPD to mitochondria induced polarization of the inner membrane followed by repeated cycles of rapid depolarizations and immediate repolarizations. ADP significantly decreased the frequency of the rapid depolarizations, but the ADP effect was counteracted by oligomycin. On the other hand, the rapid depolarizations did not occur when mitochondria were polarized by the efflux of K+ from the matrix. The rapid depolarizations became frequent with the increase in the substrate concentration or pH of the buffer. These results suggest that the rapid depolarizations depend on the net translocation of protons from the matrix. The frequency of the rapid depolarizations was not affected by ROS scavengers, Ca2+, CsA, or BA. In addition, the obvious increase in the permeability of the inner membrane to calcein (MW 623) that was entrapped in the matrix was not observed upon the transient depolarization. The mechanisms of the spontaneous oscillations of ΔΨm are discussed in relation to the matrix pH and the permeability transitions. PMID:15653749

  14. High precision optical fiber fluorescent temperature measurement system and data processing

    NASA Astrophysics Data System (ADS)

    Wang, Yutian; Bo, Xiaoxu; Gui, Feifei

    2010-08-01

    Generally, the theoretical analysis of the fluorescent life time temperature measurement is based on the assumption of the exponential life time characteristic, but in practice, the actual curve of the fluorescence are different from exponential. This is the key-influence on the stability of the high precision fluorescent measurement system. The differences are analyzed base on the theoretical mechanism of fluorescent, and a cutting and normalized method is given to describe the degree of the non-exponent of the actual fluorescent curve defer from the exponential curve. Several kinds of typical fluorescence materials spectrum and its cutting and normalized experiment results verify this theoretical analysis. Some effective measures to improve the non-exponent of the system are taken and are applied to a temperature measurement system based on actual fluorescent curve analysis with resolution 0.1°C, precisions +/-0.2°C, and real-time calibration is carried on. Based the theory base and the actuality of fluorescence optical fiber temperature sensor, two methods about fluorescence decay time constant are proposed. In the mean time, the mathematic model has been formed and analysis, so that the different schemes are selected in different situation.

  15. Frequency domain fluorescence lifetime microwell-plate platform for respirometry measurements

    NASA Astrophysics Data System (ADS)

    Chatni, M. R.; Yale, G.; Van Ryckeghem, A.; Porterfield, D. M.

    2010-04-01

    Traditionally micro-well plate based platforms used in biology utilize fluorescence intensity based methods to measure processes of biological relevance. However, fluorescence intensity measurements suffer from calibration drift due to a variety of factors. Photobleaching and self-quenching of the fluorescent dyes cause the intensity signal to drop over the lifetime of sensor immobilized inside the well. Variation in turbidity of the sample during the course of the measurement affects the measured fluorescence intensity. In comparison, fluorescence lifetime measurements are not significantly affected by these factors because fluorescence lifetime is a physico-chemical property of the fluorescent dye. Reliable and inexpensive frequency domain fluorescence lifetime instrumentation platforms are possible because the greater tolerance for optical alignment, and because they can be performed using inexpensive light sources such as LEDs. In this paper we report the development of a frequency domain fluorescence lifetime well-plate platform utilizing an oxygen sensitive transition-metal ligand complex fluorophore with a lifetime in the microsecond range. The fluorescence lifetime dye is incorporated in a polymer matrix and immobilized on the base of micro-well of a 60 well micro-well plate. Respiration measurements are performed in both aqueous and non-aqueous environment. Respirometry measurements were recorded from single Daphnia magna egg in hard water. Daphnia is an aquatic organism, important in environmental toxicology as a standard bioassay and early warning indicator for water quality monitoring. Also respirometry measurements were recorded from Tribolium castaneum eggs, which are common pests in the processed flour industry. These eggs were subjected to mitochondrial electron transport chain inhibitor such as potassium cyanide (KCN) and its effects on egg respiration were measured in real-time.

  16. Depolarization Diffusion During Weak Suprathreshold Stimulation of Cardiac Tissue

    DTIC Science & Technology

    2007-11-02

    DEPOLARIZATION DIFFUSION DURING WEAK SUPRATHRESHOLD STIMULATION OF CARDIAC TISSUE Vladimir Nikolski, Aleksandre Sambelashvili, and Igor R. Efimov...the depolarized regions. Such an activation pattern appears similar to break activation. The effect of the depolarization diffusion from depolarized...Subtitle Depolarization Diffusion During Weak Suprathreshold Stimulation of Cardiac Tissue Contract Number Grant Number Program Element Number Author(s

  17. Dynamic measurement of the height and volume of migrating cells by a novel fluorescence microscopy technique.

    PubMed

    Bottier, Céline; Gabella, Chiara; Vianay, Benoît; Buscemi, Lara; Sbalzarini, Ivo F; Meister, Jean-Jacques; Verkhovsky, Alexander B

    2011-11-21

    We propose a new technique to measure the volume of adherent migrating cells. The method is based on a negative staining where a fluorescent, non-cell-permeant dye is added to the extracellular medium. The specimen is observed with a conventional fluorescence microscope in a chamber of uniform height. Given that the fluorescence signal depends on the thickness of the emitting layer, the objects excluding the fluorescent dye (i.e., cells) appear dark, and the decrease of the fluorescent signal with respect to the background is expected to give information about the height and the volume of the object. Using a glass microfabricated pattern with steps of defined heights, we show that the drop in fluorescence intensity is indeed proportional to the height of the step and obtain calibration curves relating fluorescence intensity to height. The technique, termed the fluorescence displacement method, is further validated by comparing our measurements with the ones obtained by atomic force microscopy (AFM). We apply our method to measure the real-time volume dynamics of migrating fish epidermal keratocytes subjected to osmotic stress. The fluorescence displacement technique allows fast and precise monitoring of cell height and volume, thus providing a valuable tool for characterizing the three-dimensional behaviour of migrating cells.

  18. Boreal tree pollen sensed by polarization lidar: Depolarizing biogenic chaff

    NASA Astrophysics Data System (ADS)

    Sassen, Kenneth

    2008-09-01

    Polarization (0.694 μm) lidar measurements show that tree pollen can generate strong laser depolarization in the backscatter from the lower atmosphere. Examples are given illustrating that linear depolarization ratios up to 0.3 are measured in plumes of paper birch pollen at the onset of boreal forest green-out. These pollen are ~25 μm in diameter and near-spherical in shape, but with lobes protruding from a surface membrane, which appears to produce the depolarization. Similar lidar findings are frequently observed during the summer at Fairbanks, Alaska, indicating that various types of seasonal pollen releases may be identified by polarization lidar. This scattering behavior is likely a general attribute of pollen and other suspended biogenic debris, which has implications for benefiting human health. This source of laser depolarization should not be confused with the presence of airborne dust or certain pollution particles, but is a natural background aerosol component caused by plant reproduction, as should be recognized in current global polarization lidar aerosol research using the CALIPSO satellite.

  19. Photonic reagents for concentration measurement of flu-orescent proteins with overlapping spectra

    NASA Astrophysics Data System (ADS)

    Goun, Alexei; Bondar, Denys I.; Er, Ali O.; Quine, Zachary; Rabitz, Herschel A.

    2016-05-01

    By exploiting photonic reagents (i.e., coherent control by shaped laser pulses), we employ Optimal Dynamic Discrimination (ODD) as a novel means for quantitatively characterizing mixtures of fluorescent proteins with a large spectral overlap. To illustrate ODD, we simultaneously measured concentrations of in vitro mixtures of Enhanced Blue Fluorescent Protein (EBFP) and Enhanced Cyan Fluorescent Protein (ECFP). Building on this foundational study, the ultimate goal is to exploit the capabilities of ODD for parallel monitoring of genetic and protein circuits by suppressing the spectral cross-talk among multiple fluorescent reporters.

  20. Photonic reagents for concentration measurement of flu-orescent proteins with overlapping spectra

    PubMed Central

    Goun, Alexei; Bondar, Denys I.; Er, Ali O.; Quine, Zachary; Rabitz, Herschel A.

    2016-01-01

    By exploiting photonic reagents (i.e., coherent control by shaped laser pulses), we employ Optimal Dynamic Discrimination (ODD) as a novel means for quantitatively characterizing mixtures of fluorescent proteins with a large spectral overlap. To illustrate ODD, we simultaneously measured concentrations of in vitro mixtures of Enhanced Blue Fluorescent Protein (EBFP) and Enhanced Cyan Fluorescent Protein (ECFP). Building on this foundational study, the ultimate goal is to exploit the capabilities of ODD for parallel monitoring of genetic and protein circuits by suppressing the spectral cross-talk among multiple fluorescent reporters. PMID:27181496

  1. Photonic reagents for concentration measurement of flu-orescent proteins with overlapping spectra.

    PubMed

    Goun, Alexei; Bondar, Denys I; Er, Ali O; Quine, Zachary; Rabitz, Herschel A

    2016-05-16

    By exploiting photonic reagents (i.e., coherent control by shaped laser pulses), we employ Optimal Dynamic Discrimination (ODD) as a novel means for quantitatively characterizing mixtures of fluorescent proteins with a large spectral overlap. To illustrate ODD, we simultaneously measured concentrations of in vitro mixtures of Enhanced Blue Fluorescent Protein (EBFP) and Enhanced Cyan Fluorescent Protein (ECFP). Building on this foundational study, the ultimate goal is to exploit the capabilities of ODD for parallel monitoring of genetic and protein circuits by suppressing the spectral cross-talk among multiple fluorescent reporters.

  2. Investigation of Laser Induced Fluorescence for Concentration Measurements of Diatomic Sulfur.

    DTIC Science & Technology

    1981-12-01

    Once the spectrum was obtained, it was calibrated with N2 laser scatter and a mercury reference run. Fluorescence peaks were identified by com... Freddie , Jr. The Measurement of Quenching Rate Constants Using Fluorescence Emission. MS Thesis. Wright-Patterson Air Force Base, Ohio: Air Force Institute

  3. X-ray Fluorescence Measurements of Turbulent Methane-Oxygen Shear Coaxial Flames

    DTIC Science & Technology

    2015-05-01

    number of positive attributes of x-ray fluorescence physics, particularly insensitivity to chemical bonding (which results in tracers being conserved... rocket engines where, due to the high temperature, it is difficult to obtain quantitative mixing field measurements using conventional optical...temperature flames is attractive due to a number of positive attributes of x-ray fluorescence physics, particularly insensitivity to chemical bonding

  4. Dihydropyridine Receptors as Voltage Sensors for a Depolarization-evoked, IP3R-mediated, Slow Calcium Signal in Skeletal Muscle Cells

    PubMed Central

    Araya, Roberto; Liberona, José L.; Cárdenas, J. César; Riveros, Nora; Estrada, Manuel; Powell, Jeanne A.; Carrasco, M. Angélica; Jaimovich, Enrique

    2003-01-01

    The dihydropyridine receptor (DHPR), normally a voltage-dependent calcium channel, functions in skeletal muscle essentially as a voltage sensor, triggering intracellular calcium release for excitation-contraction coupling. In addition to this fast calcium release, via ryanodine receptor (RYR) channels, depolarization of skeletal myotubes evokes slow calcium waves, unrelated to contraction, that involve the cell nucleus (Jaimovich, E., R. Reyes, J.L. Liberona, and J.A. Powell. 2000. Am. J. Physiol. Cell Physiol. 278:C998–C1010). We tested the hypothesis that DHPR may also be the voltage sensor for these slow calcium signals. In cultures of primary rat myotubes, 10 μM nifedipine (a DHPR inhibitor) completely blocked the slow calcium (fluo-3-fluorescence) transient after 47 mM K+ depolarization and only partially reduced the fast Ca2+ signal. Dysgenic myotubes from the GLT cell line, which do not express the α1 subunit of the DHPR, did not show either type of calcium transient following depolarization. After transfection of the α1 DNA into the GLT cells, K+ depolarization induced slow calcium transients that were similar to those present in normal C2C12 and normal NLT cell lines. Slow calcium transients in transfected cells were blocked by nifedipine as well as by the G protein inhibitor, pertussis toxin, but not by ryanodine, the RYR inhibitor. Since slow Ca2+ transients appear to be mediated by IP3, we measured the increase of IP3 mass after K+ depolarization. The IP3 transient seen in control cells was inhibited by nifedipine and was absent in nontransfected dysgenic cells, but α1-transfected cells recovered the depolarization-induced IP3 transient. In normal myotubes, 10 μM nifedipine, but not ryanodine, inhibited c-jun and c-fos mRNA increase after K+ depolarization. These results suggest a role for DHPR-mediated calcium signals in regulation of early gene expression. A model of excitation-transcription coupling is presented in which both G proteins and IP

  5. Measuring actin dynamics during phagocytosis using photo-switchable fluorescence

    NASA Astrophysics Data System (ADS)

    Kovari, Daniel T.; Curtis, Jennifer E.

    2013-03-01

    Phagocytosis has traditionally been investigated in terms of the relevant biochemical signaling pathways. However, a growing number of studies investigating the physical aspects of phagocytosis have demonstrated that several distinct forces are exerted throughout particle ingestion. We use variations on FRAP (Fluorescence Recovery After Photobleaching) in combination with photo-switchable fluorescent protein to investigate actin dynamics as a phagocyte attempts to engulf its prey. The goal of our actin studies are to determine the recruitment and polymerization rate of actin in the forming phagosome and whether an organized contractile actin ring is present and responsible for phagosome closure, as proposed in the literature. These experiments are ongoing and contribute to our long term effort of developing a physics based model of phagocytosis.

  6. Differential matrix formalism for depolarizing anisotropic media.

    PubMed

    Ossikovski, Razvigor

    2011-06-15

    Azzam's differential matrix formalism [J. Opt. Soc. Am. 68, 1756 (1978)], originally developed for longitudinally inhomogeneous anisotropic nondepolarizing media, is extended to include depolarizing media. The generalization is physically interpreted in terms of means and uncertainties of the elementary optical properties of the medium, as well as of three anisotropy absorption parameters introduced to describe the depolarization. The formalism results in a particularly simple mathematical procedure for the retrieval of the elementary properties of a generally depolarizing anisotropic medium, assumed to be globally homogeneous, from its experimental Mueller matrix. The approach is illustrated on literature data and the conditions of its validity are identified and discussed.

  7. Mechanism of nerve membrane depolarization caused by grayanotoxin I

    PubMed Central

    Narahashi, Toshio; Seyama, Issei

    1974-01-01

    1. The mechanism of depolarization of squid axon membranes caused by grayanotoxin I has been studied by means of internal perfusion and voltage clamp techniques. 2. The depolarization induced by either internal or external application of grayanotoxin I was reversed by decreasing the external sodium concentration from 449 to 1 mm. 3. No depolarization was observed when both external and internal media were devoid of sodium ions, indicating that the depolarization by grayanotoxin I in normal media is due to a specific increase in resting sodium permeability. 4. The resting sodium permeability as measured by voltage clamp was increased to 1·31 × 10-6 cm/sec by internal application of 1 × 10-5 m grayanotoxin I, an increase by a factor of about 90. 5. The apparent dissociation constant of internally applied grayanotoxin I in increasing the resting sodium permeability was estimated to be 4·12 × 10-5 m, and the toxin interacts with the membrane receptor on a one-to-one stoichiometric basis. 6. Tetrodotoxin antagonized the action of grayanotoxin I in increasing the resting sodium permeability in a non-competitive manner. PMID:4455829

  8. Migraine prophylaxis, ischemic depolarizations and stroke outcomes in mice

    PubMed Central

    Eikermann-Haerter, Katharina; Lee, Jeong Hyun; Yalcin, Nilufer; Yu, Esther Sori; Daneshmand, Ali; Wei, Ying; Zheng, Yi; Can, Anil; Sengul, Buse; Ferrari, Michel D.; van den Maagdenberg, Arn M. J. M.; Ayata, Cenk

    2014-01-01

    Background and Purpose Migraine with aura is an established stroke risk factor, and excitatory mechanisms such as spreading depression are implicated in the pathogenesis of both migraine and stroke. Spontaneous spreading depression waves originate within the peri-infarct tissue and exacerbate the metabolic mismatch during focal cerebral ischemia. Genetically enhanced spreading depression susceptibility facilitates anoxic depolarizations and peri-infarct spreading depressions and accelerates infarct growth, suggesting that susceptibility to spreading depression is a critical determinant of vulnerability to ischemic injury. Because chronic treatment with migraine prophylactic drugs suppresses spreading depression susceptibility, we tested whether migraine prophylaxis can also suppress ischemic depolarizations and improve stroke outcome. Methods We measured the cortical susceptibility to spreading depression and ischemic depolarizations, and determined tissue and neurological outcome after middle cerebral artery occlusion in wild type and familial hemiplegic migraine type 1 knock-in mice treated with vehicle, topiramate or lamotrigine daily for 7 weeks or as a single dose shortly before testing. Results Chronic treatment with topiramate or lamotrigine reduces the susceptibility to KCl- or electrical stimulation-induced spreading depressions as well as ischemic depolarizations in both wild-type and familial hemiplegic migraine type 1 mutant mice. Consequently, both tissue and neurological outcomes are improved. Notably, treatment with a single dose of either drug is ineffective. Conclusions These data underscore the importance of hyperexcitability as a mechanism for increased stroke risk in migraineurs, and suggest that migraine prophylaxis may not only prevent migraine attacks but also protect migraineurs against ischemic injury. PMID:25424478

  9. Space-resolved fluorescence spectroscopic measurements with an optical fiber probe

    NASA Astrophysics Data System (ADS)

    Li, Enbang; Qiu, Hialin

    2008-12-01

    By monitoring of the emitted signal from a sample while varying the excitation wavelength, emission wavelength or both of them, fluorescence spectroscopy has become a powerful diagnostic technology. Fluorescence spectrometers can be used to measure and record the fluorescence spectra of a given sample, and have been successfully applied in different areas including biology, biochemistry, chemistry, medicine, environmental science, material science, food industry, and pharmaceutical industry. In order to increase the flexibility and applicability of conventional fluorescence spectrometers, we design an optic fiber probe for conducting the UV/Vis excitation light to a sample under study, and for collecting the fluorescence produced by the sample. Different excitation/emission fiber bundle arrangements have been fabricated and their performances have been evaluated and compared. Fiber adaptors which can be used for different commercial fluorescence spectrometers are also developed. In order to achieve space-resolved fluorescence spectroscopic measurements, we connect the fiber probe to a microscope which is mounted on a 3D traverse stage. Experiments and measurement results using the space-resolved fiber optic fluorescence spectrometer are presented in this paper.

  10. Temperature dependent steady state and picosecond kinetic fluorescence measurements of a photosystem I preparation from spinach

    SciTech Connect

    Mukerji, I.; Sauer, K.

    1988-08-01

    The fluorescence properties of a photosystem I (PSI) preparation from spinach containing approximately 200 chlorophyll (Chl) per reaction center were investigated. The preparation, characterized both spectroscopically and biochemically, contained the peripheral light harvesting antenna associated with PSI. In this study steady state fluorescence measurements were performed as a function of temperature. An emission maximum at 690 nm and a long wavelength shoulder from 710 to 740 nm were observed. The fluorescence yield at 690 nm is temperature independent, while the yield of the long wavelength shoulder increases dramatically with decreasing temperature. Additionally, kinetic measurements using the technique of single photon counting were done at room temperature and 77K. At 295K a four component fit was needed to describe the fluorescence decay; whereas at 77K, an additional 40-50 ps rise component indicative of fluorescence induction was necessary. 28 refs., 13 figs., 1 tab.

  11. Measurements of the Ultraviolet Fluorescence Cross Sections and Spectra of Bacillus Anthracis Simulants

    SciTech Connect

    Stephens, J.R.

    1998-09-01

    Measurements of the ultraviolet autofluorescence spectra and absolute cross sections of the Bacillus anthracis (Ba) simulants Bacillus globigii (Bg), Bacillus megaterium (Bm), Bacillus subtilis (Bs), and Bacillus cereus (Bc) were measured. Fluorescence spectra and cross sections of pine pollen (Pina echinata) were measured for comparison. Both dried vegetative cells and spores separated from the sporulated vegetative material were studied. The spectra were obtained by suspending a small number (<10) of particles in air in our Single Particle Spectroscopy Apparatus (SPSA), illuminating the particles with light from a spectrally filtered arc lamp, and measuring the fluorescence spectra of the particles. The illumination was 280 nm (20 nm FWHM) and the fluorescence spectra was measured between 300 and 450 nm. The fluorescence cross section of vegetative Bg peaks at 320 nm with a maximum cross section of 5 X 10{sup -14} cm{sup 2}/sr-nm-particle while the Bg spore fluorescence peaks at 310 nm with peak fluorescence of 8 X 10{sup -15} cm{sup 2}/sr-nm-particle. Pine pollen particles showed a higher fluorescence peaking at 355 nm with a cross section of 1.7 X 10{sup -13} cm{sup 2}/sr-nm-particle. Integrated cross sections ranged from 3.0 X 10{sup -13} for the Bg spores through 2.25 X 10{sup -12} (cm{sup 2}/sr-particle) for the vegetative cells.

  12. Insect monitoring with fluorescence lidar techniques: field experiments.

    PubMed

    Guan, Zuguang; Brydegaard, Mikkel; Lundin, Patrik; Wellenreuther, Maren; Runemark, Anna; Svensson, Erik I; Svanberg, Sune

    2010-09-20

    Results from field experiments using a fluorescence lidar system to monitor movements of insects are reported. Measurements over a river surface were made at distances between 100 and 300 m, detecting, in particular, damselflies entering the 355 nm pulsed laser beam. The lidar system recorded the depolarized elastic backscattering and two broad bands of laser-induced fluorescence, with the separation wavelength at 500 nm. Captured species, dusted with characteristic fluorescent dye powders, could be followed spatially and temporally after release. Implications for ecological research are discussed.

  13. 5-Aminolevulinic Acid-Induced Protoporphyrin IX Fluorescence in Meningioma: Qualitative and Quantitative Measurements In Vivo

    PubMed Central

    Valdes, Pablo A.; Bekelis, Kimon; Harris, Brent T.; Wilson, Brian C.; Leblond, Frederic; Kim, Anthony; Simmons, Nathan E.; Erkmen, Kadir; Paulsen, Keith D.; Roberts, David W.

    2014-01-01

    BACKGROUND The use of 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) fluorescence has shown promise as a surgical adjunct for maximizing the extent of surgical resection in gliomas. To date, the clinical utility of 5-ALA in meningiomas is not fully understood, with most descriptive studies using qualitative approaches to 5-ALA-PpIX. OBJECTIVE To assess the diagnostic performance of 5-ALA-PpIX fluorescence during surgical resection of meningioma. METHODS ALA was administered to 15 patients with meningioma undergoing PpIX fluorescence-guided surgery at our institution. At various points during the procedure, the surgeon performed qualitative, visual assessments of fluorescence by using the surgical microscope, followed by a quantitative fluorescence measurement by using an intra-operative probe. Specimens were collected at each point for subsequent neuropathological analysis. Clustered data analysis of variance was used to ascertain a difference between groups, and receiver operating characteristic analyses were performed to assess diagnostic capabilities. RESULTS Red-pink fluorescence was observed in 80% (12/15) of patients, with visible fluorescence generally demonstrating a strong, homogenous character. Quantitative fluorescence measured diagnostically significant PpIX concentrations (CPpIx) in both visibly and nonvisibly fluorescent tissues, with significantly higher CPpIx in both visibly fluorescent (P < .001) and tumor tissue (P = .002). Receiver operating characteristic analyses also showed diagnostic accuracies up to 90% for differentiating tumor from normal dura. CONCLUSION ALA-induced PpIX fluorescence guidance is a potential and promising adjunct in accurately detecting neoplastic tissue during meningioma resective surgery. These results suggest a broader reach for PpIX as a biomarker for meningiomas than was previously noted in the literature. PMID:23887194

  14. A unified planar measurement technique for compressible flows using laser-induced iodine fluorescence

    NASA Technical Reports Server (NTRS)

    Hartfield, Roy J., Jr.; Hollo, Steven D.; Mcdaniel, James C.

    1992-01-01

    A unified laser-induced fluorescence technique for conducting planar measurements of temperature, pressure and velocity in nonreacting, highly compressible flows has been developed, validated and demonstrated. Planar fluorescence from iodine, seeded into air, was induced by an argon-ion laser and collected using a liquid-nitrogen cooled CCD camera. In the measurement technique, temperature is determined from the fluorescence induced with the laser operated broad band. Pressure and velocity are determined from the shape and position of the fluorescence excitation spectrum which is measured with the laser operated narrow band. The measurement approach described herein provides a means of obtaining accurate, spatially-complete maps of the primary flow field parameters in a wide variety of cold supersonic and transonic flows.

  15. Remote Sensing Plant Stress Using Combined Fluorescence and Reflectance Measurements for Early Detection of Defoliants within the Battlefield Environment

    DTIC Science & Technology

    2012-10-02

    REPORT Remote sensing plant stress using combined fluorescence and reflectance measurements for early detection of defoliants within the battlefied...stress using combined fluorescence and reflectance measurements for early detection of defoliants within the battlefied environment Report Title...combined fluorescence and reflectance measurements for early detection of defoliants within the battlefield environment ARO Proposal # (49364-EV

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

    PubMed

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

    2011-11-04

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

  17. Intrinsic photosensitizer fluorescence measured using multi-diameter single-fiber spectroscopy in vivo

    NASA Astrophysics Data System (ADS)

    van Leeuwen-van Zaane, Floor; Gamm, Ute A.; van Driel, Pieter B. A. A.; Snoeks, Thomas J.; de Bruijn, Henriette S.; van der Ploeg-van den Heuvel, Angelique; Sterenborg, Henricus J. C. M.; Löwik, Clemens W.; Amelink, Arjen; Robinson, Dominic J.

    2014-01-01

    Quantification of fluorescence in vivo is complicated by the influence of tissue optical properties on the collected fluorescence signal. When tissue optical properties in the measurement volume are quantified, one can obtain the intrinsic fluorescence, which equals the product of fluorophore absorption coefficient and quantum yield. We applied this method to in vivo single-fiber fluorescence spectroscopy measurements on mouse tongue, skin, liver, and oral squamous cell carcinoma, where we detected intrinsic fluorescence spectra of the photosensitizers chlorin e6 and Bremachlorin at t=[3,4.5,6,24,48] h incubation time. We observed a tissue-dependent maximum of 35% variation in the total correction factor over the visible wavelength range. Significant differences in spectral shape over time between sensitizers were observed. Although the wavelength position of the fluorescence intensity maximum for ce6 shifted to the red, Bremachlorin showed a blue shift. Furthermore, the Bremachlorin peak appeared to be broader than the ce6 fluorescence peak. Intrinsic fluorescence intensity, which can be related to photosensitizer concentration, was decreasing for all time points but showed significantly more Bremachlorin present compared to ce6 at long incubation times. Results from this study can be used to define an optimal treatment protocol for Bremachlorin-based photodynamic therapy.

  18. Intrinsic photosensitizer fluorescence measured using multi-diameter single-fiber spectroscopy in vivo.

    PubMed

    van Leeuwen-van Zaane, Floor; Gamm, Ute A; van Driel, Pieter B A A; Snoeks, Thomas J; de Bruijn, Henriette S; van der Ploeg-van den Heuvel, Angelique; Sterenborg, Henricus J C M; Löwik, Clemens W; Amelink, Arjen; Robinson, Dominic J

    2014-01-01

    Quantification of fluorescence in vivo is complicated by the influence of tissue optical properties on the collected fluorescence signal. When tissue optical properties in the measurement volume are quantified, one can obtain the intrinsic fluorescence, which equals the product of fluorophore absorption coefficient and quantum yield. We applied this method to in vivo single-fiber fluorescence spectroscopy measurements on mouse tongue, skin, liver, and oral squamous cell carcinoma, where we detected intrinsic fluorescence spectra of the photosensitizers chlorin e6 and Bremachlorin at t=[3,4.5,6,24,48]  h incubation time. We observed a tissue-dependent maximum of 35% variation in the total correction factor over the visible wavelength range. Significant differences in spectral shape over time between sensitizers were observed. Although the wavelength position of the fluorescence intensity maximum for ce6 shifted to the red, Bremachlorin showed a blue shift. Furthermore, the Bremachlorin peak appeared to be broader than the ce6 fluorescence peak. Intrinsic fluorescence intensity, which can be related to photosensitizer concentration, was decreasing for all time points but showed significantly more Bremachlorin present compared to ce6 at long incubation times. Results from this study can be used to define an optimal treatment protocol for Bremachlorin-based photodynamic therapy.

  19. Planar Laser-Induced Fluorescence fuel concentration measurements in isothermal Diesel sprays.

    PubMed

    Pastor, José; López, José; Juliá, J; Benajes, Jesús

    2002-04-08

    This paper presents a complete methodology to perform fuel concentration measurements of Diesel sprays in isothermal conditions using the Planar Laser-Induced Fluorescence (PLIF) technique. The natural fluorescence of a commercial Diesel fuel is used with an excitation wavelength of 355 nm. The correction and calibration procedures to perform accurate measurements are studied. These procedures include the study of the fluorescence characteristics of the fuel as well as the correction of the laser sheet non-homogeneities and the losses due to Mie scattering, absorption and autoabsorption. The results obtained are compared with theoretical models and other experimental techniques.

  20. Note: Measurement of saturable absorption by intense vacuum ultraviolet free electron laser using fluorescent material.

    PubMed

    Inubushi, Y; Yoneda, H; Higashiya, A; Ishikawa, T; Kimura, H; Kumagai, T; Morimoto, S; Nagasono, M; Ohashi, H; Sato, F; Tanaka, T; Togashi, T; Tono, K; Yabashi, M; Yamaguchi, Y; Kodama, R

    2010-03-01

    Advances in free electron lasers (FELs) which generate high energy photons are expected to open novel nonlinear optics in the x-ray and vacuum ultraviolet (VUV) regions. In this paper, we report a new method for performing VUV-FEL focusing experiments. A VUV-FEL was focused with Kirkpatrick-Baez optics on a multilayer target, which contains fused silica as a fluorescent material. By measuring the fluorescence, a 5.6x4.9 microm(2) focal spot was observed in situ. Fluorescence was used to measure the saturable absorption of VUV pulses in the tin layer. The transmission increases nonlinearly higher with increasing laser intensity.

  1. Measuring reactive oxygen and nitrogen species with fluorescent probes: challenges and limitations

    PubMed Central

    Kalyanaraman, Balaraman; Darley-Usmar, Victor; Davies, Kelvin J.A.; Dennery, Phyllis A.; Forman, Henry Jay; Grisham, Matthew B.; Mann, Giovanni E.; Moore, Kevin; Roberts, L. Jackson; Ischiropoulos, Harry

    2013-01-01

    The purpose of this position paper is to present a critical analysis of the challenges and limitations of the most widely used fluorescent probes for detecting and measuring reactive oxygen and nitrogen species. Where feasible, we have made recommendations for the use of alternate probes and appropriate analytical techniques that measure the specific products formed from the reactions between fluorescent probes and reactive oxygen and nitrogen species. We have proposed guidelines that will help present and future researchers with regard to the optimal use of selected fluorescent probes and interpretation of results. PMID:22027063

  2. A Depolarizing Electrogenic Pump in Frog Muscle

    DTIC Science & Technology

    1975-08-01

    mw copy AFRRI SR75-20 AUGUST 1975 AFRRI SCIENTIFIC REPORT O ■ to A DEPOLARIZING ELECTROGENIC PUMP IN FROG MUSCLE D. Geduldig D. R...Academy of Sciences - National Research Council. AFRRI SR75-20 August 1975 A DEPOLARIZING ELECTROGENIC PUMP IN FROG MUSCLE D. GEDULDIG* D. R...INTRODUCTION When Na-enriched frog muscles are bathed in Na- and K-free saline, the small amount of potassium which could accumulate outside of the membrane

  3. Lidar ratio and depolarization ratio for cirrus clouds.

    PubMed

    Chen, Wei-Nai; Chiang, Chih-Wei; Nee, Jan-Bai

    2002-10-20

    We report on studies of the lidar and the depolarization ratios for cirrus clouds. The optical depth and effective lidar ratio are derived from the transmission of clouds, which is determined by comparing the backscattering signals at the cloud base and cloud top. The lidar signals were fitted to a background atmospheric density profile outside the cloud region to warrant the linear response of the return signals with the scattering media. An average lidar ratio, 29 +/- 12 sr, has been found for all clouds measured in 1999 and 2000. The height and temperature dependences ofthe lidar ratio, the optical depth, and the depolarization ratio were investigated and compared with results of LITE and PROBE. Cirrus clouds detected near the tropopause are usually optically thin and mostly subvisual. Clouds with the largest optical depths were found near 12 km with a temperature of approximately -55 degrees C. The multiple-scattering effect is considered for clouds with high optical depths, and this effect lowers the lidar ratios compared with a single-scattering condition. Lidar ratios are in the 20-40 range for clouds at heights of 12.5-15 km and are smaller than approximately 30 in height above 15 km. Clouds are usually optically thin for temperatures below approximately -65 degrees C, and in this region the optical depth tends to decrease with height. The depolarization ratio is found to increase with a height at 11-15 km and smaller than 0.3 above 16 km. The variation in the depolarization ratio with the lidar ratio was also reported. The lidar and depolarization ratios were discussed in terms of the types of hexagonal ice crystals.

  4. On the uncertainty in single molecule fluorescent lifetime and energy emission measurements

    NASA Technical Reports Server (NTRS)

    Brown, Emery N.; Zhang, Zhenhua; Mccollom, Alex D.

    1995-01-01

    Time-correlated single photon counting has recently been combined with mode-locked picosecond pulsed excitation to measure the fluorescent lifetimes and energy emissions of single molecules in a flow stream. Maximum likelihood (ML) and least square methods agree and are optimal when the number of detected photons is large however, in single molecule fluorescence experiments the number of detected photons can be less than 20, 67% of those can be noise and the detection time is restricted to 10 nanoseconds. Under the assumption that the photon signal and background noise are two independent inhomogeneous poisson processes, we derive the exact joint arrival time probably density of the photons collected in a single counting experiment performed in the presence of background noise. The model obviates the need to bin experimental data for analysis, and makes it possible to analyze formally the effect of background noise on the photon detection experiment using both ML or Bayesian methods. For both methods we derive the joint and marginal probability densities of the fluorescent lifetime and fluorescent emission. the ML and Bayesian methods are compared in an analysis of simulated single molecule fluorescence experiments of Rhodamine 110 using different combinations of expected background nose and expected fluorescence emission. While both the ML or Bayesian procedures perform well for analyzing fluorescence emissions, the Bayesian methods provide more realistic measures of uncertainty in the fluorescent lifetimes. The Bayesian methods would be especially useful for measuring uncertainty in fluorescent lifetime estimates in current single molecule flow stream experiments where the expected fluorescence emission is low. Both the ML and Bayesian algorithms can be automated for applications in molecular biology.

  5. On the Uncertainty in Single Molecule Fluorescent Lifetime and Energy Emission Measurements

    NASA Technical Reports Server (NTRS)

    Brown, Emery N.; Zhang, Zhenhua; McCollom, Alex D.

    1996-01-01

    Time-correlated single photon counting has recently been combined with mode-locked picosecond pulsed excitation to measure the fluorescent lifetimes and energy emissions of single molecules in a flow stream. Maximum likelihood (ML) and least squares methods agree and are optimal when the number of detected photons is large, however, in single molecule fluorescence experiments the number of detected photons can be less than 20, 67 percent of those can be noise, and the detection time is restricted to 10 nanoseconds. Under the assumption that the photon signal and background noise are two independent inhomogeneous Poisson processes, we derive the exact joint arrival time probability density of the photons collected in a single counting experiment performed in the presence of background noise. The model obviates the need to bin experimental data for analysis, and makes it possible to analyze formally the effect of background noise on the photon detection experiment using both ML or Bayesian methods. For both methods we derive the joint and marginal probability densities of the fluorescent lifetime and fluorescent emission. The ML and Bayesian methods are compared in an analysis of simulated single molecule fluorescence experiments of Rhodamine 110 using different combinations of expected background noise and expected fluorescence emission. While both the ML or Bayesian procedures perform well for analyzing fluorescence emissions, the Bayesian methods provide more realistic measures of uncertainty in the fluorescent lifetimes. The Bayesian methods would be especially useful for measuring uncertainty in fluorescent lifetime estimates in current single molecule flow stream experiments where the expected fluorescence emission is low. Both the ML and Bayesian algorithms can be automated for applications in molecular biology.

  6. Fluorescence measurement by a streak camera in a single-photon-counting mode.

    PubMed

    Komura, Masayuki; Itoh, Shigeru

    2009-01-01

    We describe here a recently developed fluorescence measurement system that uses a streak camera to detect fluorescence decay in a single photon-counting mode. This system allows for easy measurements of various samples and provides 2D images of fluorescence in the wavelength and time domains. The great advantage of the system is that the data can be handled with ease; furthermore, the data are amenable to detailed analysis. We describe the picosecond kinetics of fluorescence in spinach Photosystem (PS) II particles at 4-77 K as a typical experimental example. Through the global analysis of the data, we have identified a new fluorescence band (F689) in addition to the already established F680, F685, and F695 emission bands. The blue shift of the steady-state fluorescence spectrum upon cooling below 77 K can be interpreted as an increase of the shorter-wavelength fluorescence, especially F689, due to the slowdown of the excitation energy transfer process. The F685 and F695 bands seem to be thermally equilibrated at 77 K but not at 4 K. The simple and efficient photon accumulation feature of the system allows us to measure fluorescence from leaves, solutions, single colonies, and even single cells. The 2D fluorescence images obtained by this system are presented for isolated spinach PS II particles, intact leaves of Arabidopsis thaliana, the PS I super-complex of a marine centric diatom, Chaetoceros gracilis, isolated membranes of a purple photosynthetic bacterium, Acidiphilium rubrum, which contains Zn-BChl a, and a coral that contains a green fluorescent protein and an algal endosymbiont, Zooxanthella.

  7. Simulation of fluorescent measurements in the human skin

    NASA Astrophysics Data System (ADS)

    Meglinski, Igor V.; Sinichkin, Yurii P.; Utz, Sergei R.; Pilipenko, Helena A.

    1995-05-01

    Reflectance and fluorescence spectroscopy are successfully used for skin disease diagnostics. Human skin optical parameters are defined by its turbid, scattering properties with nonuniform absorption and fluorescence chromophores distribution, its multilayered structure, and variability under different physiological and pathological conditions. Theoretical modeling of light propagation in skin could improve the understanding of these condition and may be useful in the interpretation of in vivo reflectance and autofluorescence (AF) spectra. Laser application in medical optical tomography, tissue spectroscopy, and phototherapy stimulates the development of optical and mathematical light-tissue interaction models allowing to account the specific features of laser beam and tissue inhomogeneities. This paper presents the version of a Monte Carlo method for simulating of optical radiation propagation in biotissue and highly scattering media, allowing for 3D geometry of a medium. The simulation is based on use of Green's function of medium response to single external pulse. The process of radiation propagation is studied in the area with given boundary conditions, taking into account the processes of reflection and refraction at the boundaries of layers inside the medium under study. Results of Monte Carlo simulation were compared with experimental investigations and demonstrated good agreement.

  8. Tools and techniques to measure mitophagy using fluorescence microscopy.

    PubMed

    Dolman, Nick J; Chambers, Kevin M; Mandavilli, Bhaskar; Batchelor, Robert H; Janes, Michael S

    2013-11-01

    Mitophagy is a specialized form of autophagy that removes damaged mitochondria, thereby maintaining efficient cellular metabolism and reducing cellular stress caused by aberrant oxidative bursts. Deficits in mitophagy underlie several diseases, and a substantial body of research has elucidated key steps in the pathways that lead to and execute autophagic clearance of mitochondria. Many of these studies employ fluorescence microscopy to visualize mitochondrial morphology, mass, and functional state. Studies in this area also examine colocalization/recruitment of accessory factors, components of the autophagic machinery and signaling molecules to mitochondria. In this review, we provide a brief summary of the current understanding about the processes involved in mitophagy followed by a discussion of probes commonly employed and important considerations of the methodologies to study and analyze mitophagy using fluorescence microscopy. Representative data, where appropriate, are provided to highlight the use of key probes to monitor mitophagy. The review will conclude with a consideration of new possibilities for mitophagy research and a discussion of recently developed technologies for this emerging area of cell biology.

  9. Subcellular localization-dependent changes in EGFP fluorescence lifetime measured by time-resolved flow cytometry

    PubMed Central

    Gohar, Ali Vaziri; Cao, Ruofan; Jenkins, Patrick; Li, Wenyan; Houston, Jessica P.; Houston, Kevin D.

    2013-01-01

    Intracellular protein transport and localization to subcellular regions are processes necessary for normal protein function. Fluorescent proteins can be fused to proteins of interest to track movement and determine localization within a cell. Currently, fluorescence microscopy combined with image processing is most often used to study protein movement and subcellular localization. In this contribution we evaluate a high-throughput time-resolved flow cytometry approach to correlate intracellular localization of human LC3 protein with the fluorescence lifetime of enhanced green fluorescent protein (EGFP). Subcellular LC3 localization to autophagosomes is a marker of the cellular process called autophagy. In breast cancer cells expressing native EGFP and EGFP-LC3 fusion proteins, we measured the fluorescence intensity and lifetime of (i) diffuse EGFP (ii) punctate EGFP-LC3 and (iii) diffuse EGFP-ΔLC3 after amino acid starvation to induce autophagy-dependent LC3 localization. We verify EGFP-LC3 localization with low-throughput confocal microscopy and compare to fluorescence intensity measured by standard flow cytometry. Our results demonstrate that time-resolved flow cytometry can be correlated to subcellular localization of EGFP fusion proteins by measuring changes in fluorescence lifetime. PMID:24010001

  10. [The analysis of sinusoidal modulated method used for measuring fluorescence lifetime].

    PubMed

    Feng, Ying; Huang, Shi-hua

    2007-12-01

    This paper has built a system with a sinusoidal modulated LED as the excitation source. Such exciter was used upon the sample Eu2 L'3 x nH2O (L' = C4H4O4). Both the excitation light and the 5Do-7F2 emission of Eu3+ ion were measured. Fluorescence lifetime, which approximate to 0.680 ms, can then be obtained from the measured excitation and fluorescence waveforms by non-linear least square curve fitting based on the principle of phase-shift measurement of fluorescence lifetime. Data processing methods considering respectively the high order harmonics in the modulation and multi-exponential decay of the fluorescence were discussed. A method of utilizing Fourier series expandedness to amendatory the result was put forward. Accordingly, the applicability for phase-shift method was expanded as well as a more exact result was acquired.

  11. Detecting and Quantifying Biomolecular Interactions of a Dendritic Polyglycerol Sulfate Nanoparticle Using Fluorescence Lifetime Measurements.

    PubMed

    Boreham, Alexander; Pikkemaat, Jens; Volz, Pierre; Brodwolf, Robert; Kuehne, Christian; Licha, Kai; Haag, Rainer; Dernedde, Jens; Alexiev, Ulrike

    2015-12-24

    Interactions of nanoparticles with biomaterials determine the biological activity that is key for the physiological response. Dendritic polyglycerol sulfates (dPGS) were found recently to act as an inhibitor of inflammation by blocking selectins. Systemic application of dPGS would present this nanoparticle to various biological molecules that rapidly adsorb to the nanoparticle surface or lead to adsorption of the nanoparticle to cellular structures such as lipid membranes. In the past, fluorescence lifetime measurements of fluorescently tagged nanoparticles at a molecular and cellular/tissue level have been proven to reveal valuable information on the local nanoparticle environment via characteristic fluorescent lifetime signatures of the nanoparticle bound dye. Here, we established fluorescence lifetime measurements as a tool to determine the binding affinity to fluorescently tagged dPGS (dPGS-ICC; ICC: indocarbocyanine). The binding to a cell adhesion molecule (L-selectin) and a human complement protein (C1q) to dPGS-ICC was evaluated by the concentration dependent change in the unique fluorescence lifetime signature of dPGS-ICC. The apparent binding affinity was found to be in the nanomolar range for both proteins (L-selectin: 87 ± 4 nM and C1q: 42 ± 12 nM). Furthermore, the effect of human serum on the unique fluorescence lifetime signature of dPGS-ICC was measured and found to be different from the interactions with the two proteins and lipid membranes. A comparison between the unique lifetime signatures of dPGS-ICC in different biological environments shows that fluorescence lifetime measurements of unique dPGS-ICC fluorescence lifetime signatures are a versatile tool to probe the microenvironment of dPGS in cells and tissue.

  12. Stimulation of high affinity gamma-aminobutyric acidB receptors potentiates the depolarization-induced increase of intraneuronal ionized calcium content in cerebellar granule neurons.

    PubMed

    De Erausquin, G; Brooker, G; Costa, E; Wojcik, W J

    1992-09-01

    In the treatment of spasticity, the therapeutic cerebrospinal fluid levels of (+/-)-baclofen, a gamma-aminobutyric acid (GABA)B receptor agonist, are below 1 microM. However, the mechanism of the therapeutic action of (+/-)-baclofen remains unknown, because, for the most part, the action of (+/-)-baclofen on GABAB receptors requires micromolar concentrations. Using fura-2 fluorescence microscopy, intracellular ionized calcium was measured in cerebellar granule neurons. Stimulation of a high affinity GABAB receptor potentiated by 2-3-fold the rise in intracellular calcium observed after depolarization of the cell with a Krebs Ringer's buffered solution containing 40 mM K+. Both GABA (100 nM) and (+/-)-baclofen (10-100 nM) stimulated this high affinity receptor. The potentiation of the depolarization-induced rise in intracellular calcium by (+/-)-baclofen (100 nM) was completely blocked by the GABAB receptor antagonist CGP 35348 (200 microM). Also, the intracellular calcium response induced by the activation of high affinity GABAB receptors was prevented by dantrolene (10 microM). The cerebellar granule neurons contained calcium-induced calcium release (CICR) stores. Caffeine (3 mM) and ryanodine (100 microM) potentiated the depolarization-induced rise in intracellular calcium, and this response to both drugs was blocked by dantrolene (10 microM). Because dantrolene does not prevent the rise in intracellular calcium after cell depolarization (this calcium originated from the influx of extracellular calcium), (+/-)-baclofen acting via the high affinity GABAB receptor indirectly activates the CICR stores, allowing the influx of extracellular calcium to trigger the release of calcium from these dantrolene-sensitive CICR stores. Thus, this high affinity GABAB receptor might become activated during persistent depolarization caused by pathological states and could be a mechanism to be studied for the therapeutic action of (+/-)-baclofen in spasticity.

  13. Accurate modeling of fluorescence line narrowing difference spectra: Direct measurement of the single-site fluorescence spectrum

    NASA Astrophysics Data System (ADS)

    Reppert, Mike; Naibo, Virginia; Jankowiak, Ryszard

    2010-07-01

    Accurate lineshape functions for modeling fluorescence line narrowing (FLN) difference spectra (ΔFLN spectra) in the low-fluence limit are derived and examined in terms of the physical interpretation of various contributions, including photoproduct absorption and emission. While in agreement with the earlier results of Jaaniso [Proc. Est. Acad. Sci., Phys., Math. 34, 277 (1985)] and Fünfschilling et al. [J. Lumin. 36, 85 (1986)], the derived formulas differ substantially from functions used recently [e.g., M. Rätsep et al., Chem. Phys. Lett. 479, 140 (2009)] to model ΔFLN spectra. In contrast to traditional FLN spectra, it is demonstrated that for most physically reasonable parameters, the ΔFLN spectrum reduces simply to the single-site fluorescence lineshape function. These results imply that direct measurement of a bulk-averaged single-site fluorescence lineshape function can be accomplished with no complicated extraction process or knowledge of any additional parameters such as site distribution function shape and width. We argue that previous analysis of ΔFLN spectra obtained for many photosynthetic complexes led to strong artificial lowering of apparent electron-phonon coupling strength, especially on the high-energy side of the pigment site distribution function.

  14. DAPI-fluorescent fading: a problem in microscopy or a way to measure nuclear DNA content?

    NASA Astrophysics Data System (ADS)

    Gallardo-Escárate, Cristian; Álvarez-Borrego, Josué; Kober, V.; del Río-Portilla, Miguel Á.

    2006-01-01

    In observation by confocal or conventional fluorescence microscopy, the retardation of the lost in fluorescence, from highest signal of fluorescence to lowest intensity are important factors in order to obtain accurate images. This problem is very common in fluorochromes for nuclear DNA and especially for DAPI stain. The fluorescence of DAPI is rapidly lost when it is exposure to excitation by ultra violet (UV) light, and especially under optimal condition of observation. Although the fading process could be retardate by using of mounting medium with antifading solutions, the photochemical process underlying the fluorescence decay has not yet been fully explained. In addiction, neither relationship has been tested between the fluorescence fading and nuclear DNA content. However, the capacity of the DNA to absorb UV light is knows. In order to test this relationship we measured by means of image analysis the fluorescence intensity in several nuclei types during a fading period. The analysis was performed by an algorithm specifically built in MATLAB software. The relationship between nuclear DNA content and DAPI-fluorescence fading was found equal to 99%. This study demonstrates the feasibility for estimates genome size by quantification of fluorescence fading. In this context, the present method allows to measure nuclear DNA content in several medical applications (cancer, HIV, organ transplants, etc). Nowadays, for measuring DNA content, flow cytometry is widely used; however, with the flow cytometry method it is not possible to select a specific group of cells, such as from a specific region of a tumor. Moreover, the using of image analysis allows automatizing diagnostics procedures.

  15. Fluorescence techniques used to measure interactions between hydroxyapatite nanoparticles and epidermal growth factor receptors.

    PubMed

    Kathawala, Mustafa H; Khoo, Stella P K; Sudhaharan, Thankiah; Zhao, Xinxin; Say Chye Loo, Joachim; Ahmed, Sohail; Woei Ng, Kee

    2015-01-01

    The potential applications of nanomaterials in therapeutics are immense and to fully explore this potential, it is important to understand the interaction of nanoparticles with cellular components. To examine the interaction between nanoparticles and cell membrane receptors, this report describes the use of advanced fluorescence techniques to measure interactions between hydroxyapatite (HA) nanoparticles and epidermal growth factor receptors (EGFRs), as a model system. FITC-labelled HA nanoparticles and monomeric red fluorescent protein (mRFP)-conjugated EGFRs expressed in Chinese hamster ovary cells (CHO-K1) were generated and their interaction measured using acceptor photobleaching-fluorescence resonance energy transfer (AP-FRET) and fluorescence lifetime imaging microscopy-fluorescence resonance energy transfer (FLIM-FRET). Results confirmed that hydroxyapatite nanoparticles not only interacted with EGFR but also attenuated downstream EGFR signalling, possibly by hindering normal dimerization of EGFR. Furthermore, the extent of signal attenuation suggested correlation with specific surface area of the nanoparticles, whereby greater specific surface area resulted in greater downstream signal attenuation. This novel demonstration establishes fluorescence techniques as a viable method to study nanoparticle interactions with proteins such as cell surface receptors. The approach described herein can be extended to study interactions between any fluorescently labelled nanoparticle-biomolecule pair.

  16. Depolarization of Cellular Resting Membrane Potential Promotes Neonatal Cardiomyocyte Proliferation In Vitro

    PubMed Central

    Lan, Jen-Yu; Williams, Corin; Levin, Michael; Black, Lauren Deems

    2014-01-01

    Cardiomyocytes (CMs) undergo a rapid transition from hyperplastic to hypertrophic growth soon after birth, which is a major challenge to the development of engineered cardiac tissue for pediatric patients. Resting membrane potential (Vmem) has been shown to play an important role in cell differentiation and proliferation during development. We hypothesized that depolarization of neonatal CMs would stimulate or maintain CM proliferation in vitro. To test our hypothesis, we isolated postnatal day 3 neonatal rat CMs and subjected them to sustained depolarization via the addition of potassium gluconate or Ouabain to the culture medium. Cell density and CM percentage measurements demonstrated an increase in mitotic CMs along with a ~2 fold increase in CM numbers with depolarization. In addition, depolarization led to an increase in cells in G2 and S phase, indicating increased proliferation, as measured by flow cytometry. Surprisingly depolarization of Vmem with either treatment led to inhibition of proliferation in cardiac fibroblasts. This effect is abrogated when the study was carried out on postnatal day 7 neonatal CMs, which are less proliferative, indicating that the likely mechanism of depolarization is the maintenance of the proliferating CM population. In summary, our findings suggest that depolarization maintains postnatal CM proliferation and may be a novel approach to encourage growth of engineered tissue and cardiac regeneration in pediatric patients. PMID:25295125

  17. Direct measurement of S2 -- S0 fluorescence lifetimes and anisotropy of tetraphenylporphyrins

    NASA Astrophysics Data System (ADS)

    Gurzadyan, Gagik G.; Tran-Thi, Thu-Hoa; Gustavsson, Thomas

    1999-12-01

    Various tetraphenylporphyrins (zinc, magnesium, free base) were excited to the upper electronic levels of the Soret band with the second harmonic of a mode-locked Ti-sapphire laser (394 nm). An up-conversion fluorescence set-up with the time resolution of 120 fs was used to measure the decay times of the S2 fluorescence in conjunction with the risetime of the S1 fluorescence. The depopulation of the excited electronic state S2 was studied as a function of the metal ion and the solvent. The lifetimes of the electronic S2 level, measured for ZnTPP and MgTPP in different solvents were (tau) equals 1.4 - 3.4 ps. The depopulation channel from S2 to S1 was studied by measuring simultaneously the decay of S2 and the rise of S1 fluorescence. The rate constant of the process can be correlated to the energy gap between the S2 and S1 levels, which depends on the nature of the metal ions and solvents. The rotational dynamics in the Soret band was also studied by measuring the anisotropy of S2 ---> S0 fluorescence. The anisotropy decay of S2 fluorescence was found to be biexponential, with a fast component around 100 fs and a slow one (t >> 10 ps), attributed to the partial dephasing of the degenerate energy levels of the S2 state and to rotational diffusion, respectively.

  18. Quantification of in vivo fluorescence decoupled from the effects of tissue optical properties using fiber-optic spectroscopy measurements

    NASA Astrophysics Data System (ADS)

    Kim, Anthony; Khurana, Mamta; Moriyama, Yumi; Wilson, Brian C.

    2010-11-01

    We present a method for tissue fluorescence quantification in situ using a handheld fiber optic probe that measures both the fluorescence and diffuse reflectance spectra. A simplified method to decouple the fluorescence spectrum from distorting effects of the tissue optical absorption and scattering is developed, with the objective of accurately quantifying the fluorescence in absolute units. The primary motivation is measurement of 5-aminolevulinic acid-induced protoporphyrin IX (ALA-PpIX) concentration in tissue during fluorescence-guided resection of malignant brain tumors. This technique is validated in phantoms and ex vivo mouse tissues, and tested in vivo in a rabbit brain tumor model using ALA-PpIX fluorescence contrast.

  19. Non-intrusive temperature measurements using three-color laser-induced fluorescence

    NASA Astrophysics Data System (ADS)

    Lavieille, P.; Delconte, A.; Blondel, D.; Lebouché, M.; Lemoine, F.

    This paper presents a new temperature measurement technique in a liquid, based on laser-induced fluorescence of rhodamine B. The fluorescence intensity is detected on three spectral bands, where the ratios between the emission of each band determine the temperature while correcting for the effects of fluorescent re-absorption. In addition, the influence of parameters such as probe volume size, dye concentration, and Beer's absorption is removed. The principles of the technique are described in this paper, and the technique is demonstrated on a heated liquid jet studied under a constant and a spatially variable dye concentration.

  20. A fluorescence-based assay for measuring the redox potential of 5-lipoxygenase inhibitors.

    PubMed

    Lee, Sangchul; Park, Youngsam; Kim, Junghwan; Han, Sung-Jun

    2014-01-01

    The activities and side effects of 5-lipoxygenase (5-LO) inhibitors can be predicted by identifying their redox mechanisms. In this study, we developed a fluorescence-based method to measure the redox potential of 5-LO inhibitors and compared it to the conventional, absorbance-based method. After the pseudo-peroxidase reaction, the amount of remaining lipid peroxide was quantified using the H2DCFDA (2',7'-dichlorodihydrofluorescein diacetate) fluorescence dye. Our method showed large signal windows and provided comparable redox potential values. Importantly, the redox mechanisms of known inhibitors were accurately measured with the fluorescence assay, whereas the conventional, absorbance-based method showed contradictory results. Our findings suggest that our developed method is a better alternative for classifying the redox potential of 5-LO inhibitors, and the fluorescence assay can be effectively used to study the mechanisms of action that are related to redox cycling.

  1. On-line measurement of lignin in wood pulp by color shift of fluorescence

    DOEpatents

    Jeffers, Larry A.; Malito, Michael L.

    1996-01-01

    Lignin concentrations from wood pulp samples are measured by applying an excitation light at a selected wavelength to the samples in order to cause the lignin to emit fluorescence. A spectral distribution of the fluorescence emission is then determined. The lignin concentration is then calculated based on the spectral distribution signal. The spectral distribution is quantified by either a wavelength centroid method or a band ratio method.

  2. On-line measurement of lignin in wood pulp by color shift of fluorescence

    DOEpatents

    Jeffers, L.A.; Malito, M.L.

    1996-01-23

    Lignin concentrations from wood pulp samples are measured by applying an excitation light at a selected wavelength to the samples in order to cause the lignin to emit fluorescence. A spectral distribution of the fluorescence emission is then determined. The lignin concentration is then calculated based on the spectral distribution signal. The spectral distribution is quantified by either a wavelength centroid method or a band ratio method. 6 figs.

  3. Absolute fluorescence measurements > 1000 nm: setup design, calibration and standards (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Resch-Genger, Ute; Würth, Christian; Pauli, Jutta; Hatami, Soheil; Kaiser, Martin

    2016-03-01

    There is an increasing interest in optical reporters like semiconductor quantum dots and upconversion nanophosphors with emission < 1000 nm for bioanalysis, medical diagnostics, and safety barcodes and hence, in reliable fluorescence measurements in this wavelength region, e.g., for the comparison of material performance and the rational design of new nanomaterials with improved properties [1-4]. The performance of fluorescence measurements < 800 nm and especially < 1000 nm is currently hampered by the lack of suitable methods and standards for the simple determination of the wavelength-dependent spectral responsivity of fluorescence measuring systems and the control of measured emission spectra and intensities [3-5]. This is of special relevance for nanocrystalline emitters like quantum dots and rods as well as for upconversion nanocrystals, where surface states and the accessibility of emissive states by quenchers largely control accomplishable quantum yields and hence, signal sizes and detection sensitivities from the reporter side. Here, we present the design of an integrating sphere setup for the absolute measurement of emission spectra and quantum yields in the wavelength region of 650 to 1600 nm and its calibration as well as examples for potential fluorescence standards from different reporter classes for the control of the reliability of such measurements [5]. This includes new spectral fluorescence standards for the wavelength region of 650 nm to 1000 nm as well as a set of quantum yield standards covering the wavelength region from 400 nm to 1000 nm.

  4. Longitudinal polarization periodicity of unpolarized light passing through a double wedge depolarizer.

    PubMed

    de Sande, Juan Carlos G; Santarsiero, Massimo; Piquero, Gemma; Gori, Franco

    2012-12-03

    The polarization characteristics of unpolarized light passing through a double wedge depolarizer are studied. It is found that the degree of polarization of the radiation propagating after the depolarizer is uniform across transverse planes after the depolarizer, but it changes from one plane to another in a periodic way giving, at different distances, unpolarized, partially polarized, or even perfectly polarized light. An experiment is performed to confirm this result. Measured values of the Stokes parameters and of the degree of polarization are in complete agreement with the theoretical predictions.

  5. Characterization of homogenous depolarizing media based on Mueller matrix differential decomposition.

    PubMed

    Arteaga, Oriol; Kahr, Bart

    2013-04-01

    In a depolarizing medium in which the optical properties are uniformly distributed, the logarithm of the Mueller matrix can be used to calculate the differential Mueller matrix. From the differential Mueller matrix, the 10 optical properties of a homogeneous depolarizing medium are recovered. A modified calculation is introduced for media showing small time-irreversal depolarization events. The benefits of this method are illustrated in the determination of circular dichroism and circular birefringence of a nickel sulfate hexahydrate crystal from spectroscopic Mueller matrix measurements.

  6. Quenching-independent measurement of species concentrations in flames by laser-induced fluorescence

    SciTech Connect

    Salmon, J.T.; Carter, C.D.; Laurendeau, N.M.

    1990-09-01

    This report describes work accomplished in the last two years on measurement of species concentrations in flames via laser-induced fluorescence. During this period, we have published absolute number densities of atomic hydrogen in subatmospheric, premixed C{sub 2}H{sub 4}/O{sub 2}/Ar flames at equivalence ratios of 1.0 and 1.7 via two-photon excited fluorescence. This work has led to the development of a new single-laser, two-step fluorescence method for the detection of atomic hydrogen in flames. Using photoionization controlled-loss spectroscopy (PICLS), we have verified the T{sup {minus}1/2} dependence of quenching on temperature for atomic hydrogen, in agreement with kinetic theory. Previous work on pyrometry using laser-saturated fluorescence (LSF) and the anomalous fluorescence from pyrene has evolved into publication of a major review paper on temperature measurements by light-scattering methods. Finally, we have demonstrated the feasibility of quantitative LSF measurements of NO concentration by obtaining relative saturation curves and NO fluorescence profiles. 25 refs.

  7. Dynamic measurement of fluorescent proteins spectral distribution on virus infected cells

    NASA Astrophysics Data System (ADS)

    Lee, Ja-Yun; Wu, Ming-Xiu; Kao, Chia-Yun; Wu, Tzong-Yuan; Hsu, I.-Jen

    2006-09-01

    We constructed a dynamic spectroscopy system that can simultaneously measure the intensity and spectral distributions of samples with multi-fluorophores in a single scan. The system was used to monitor the fluorescence distribution of cells infected by the virus, which is constructed by a recombinant baculoviruses, vAcD-Rhir-E, containing the red and green fluorescent protein gene that can simultaneously produce dual fluorescence in recombinant virus-infected Spodoptera frugiperda 21 cells (Sf21) under the control of a polyhedrin promoter. The system was composed of an excitation light source, a scanning system and a spectrometer. We also developed an algorithm and fitting process to analyze the pattern of fluorescence distribution of the dual fluorescence produced in the recombinant virus-infected cells. All the algorithm and calculation are automatically processed in a visualized scanning program and can monitor the specific region of sample by calculating its intensity distribution. The spectral measurement of each pixel was performed at millisecond range and the two dimensional distribution of full spectrum was recorded within several seconds. We have constructed a dynamic spectroscopy system to monitor the process of virus-infection of cells. The distributions of the dual fluorescence were simultaneously measured at micrometer resolution.

  8. Spectrally resolved fluorescence lifetime imaging of Nile red for measurements of intracellular polarity

    NASA Astrophysics Data System (ADS)

    Levitt, James A.; Chung, Pei-Hua; Suhling, Klaus

    2015-09-01

    Spectrally resolved confocal microscopy and fluorescence lifetime imaging have been used to measure the polarity of lipid-rich regions in living HeLa cells stained with Nile red. The emission peak from the solvatochromic dye in lipid droplets is at a shorter wavelength than other, more polar, stained internal membranes, and this is indicative of a low polarity environment. We estimate that the dielectric constant, ɛ, is around 5 in lipid droplets and 25<ɛ<40 in other lipid-rich regions. Our spectrally resolved fluorescence lifetime imaging microscopy (FLIM) data show that intracellular Nile red exhibits complex, multiexponential fluorescence decays due to emission from a short lifetime locally excited state and a longer lifetime intramolecular charge transfer state. We measure an increase in the average fluorescence lifetime of the dye with increasing emission wavelength, as shown using phasor plots of the FLIM data. We also show using these phasor plots that the shortest lifetime decay components arise from lipid droplets. Thus, fluorescence lifetime is a viable contrast parameter for distinguishing lipid droplets from other stained lipid-rich regions. Finally, we discuss the FLIM of Nile red as a method for simultaneously mapping both polarity and relative viscosity based on fluorescence lifetime measurements.

  9. Proximity to Intrinsic Depolarizing Resonances with a Partial Siberian Snake

    NASA Astrophysics Data System (ADS)

    Crandell, D. A.; Alexeeva, L. V.; Anferov, V. A.; Blinov, B. B.; Chu, C. M.; Caussyn, D. D.; Courant, E. D.; Gladycheva, S. E.; Hu, S.; Krisch, A. D.; Nurushev, T. S.; Phelps, R. A.; Ratner, L. G.; Varzar, S. M.; Wong, V. K.; Derbenev, Ya. S.; Lee, S. Y.; Rinckel, T.; Schwandt, P.; Sperisen, F.; Stephenson, E. J.; von Przewoski, B.; Baiod, R.; Russell, A. D.; Ohmori, C.; Sato, H.

    1996-05-01

    Partial Siberian snakes are effective in overcoming imperfection depolarizing resonances, but they may also change the crossing energy for intrinsic depolarizing resonances. We experimentally investigated the effect of a partial Siberian snake near intrinsic depolarizing resonances with stored 140 MeV and 160 MeV polarized proton beams. Using various partial Siberian snake strengths up to 30%, depolarization was observed; this may be due to a change in the spin precession frequency which moves the energy of nearby intrinsic depolarizing resonances.

  10. Validation of fluorescent-labeled microspheres for measurement of relative blood flow in severely injured lungs

    NASA Technical Reports Server (NTRS)

    Hubler, M.; Souders, J. E.; Shade, E. D.; Hlastala, M. P.; Polissar, N. L.; Glenny, R. W.

    1999-01-01

    The aim of the study was to validate a nonradioactive method for relative blood flow measurements in severely injured lungs that avoids labor-intensive tissue processing. The use of fluorescent-labeled microspheres was compared with the standard radiolabeled-microsphere method. In seven sheep, lung injury was established by using oleic acid. Five pairs of radio- and fluorescent-labeled microspheres were injected before and after established lung injury. Across all animals, 175 pieces were selected randomly. The radioactivity of each piece was determined by using a scintillation counter. The fluorescent dye was extracted from each piece with a solvent without digestion or filtering. The fluorescence was determined with an automated fluorescent spectrophotometer. Perfusion was calculated for each piece from both the radioactivity and fluorescence and volume normalized. Correlations between flow determined by the two methods were in the range from 0.987 +/- 0.007 (SD) to 0.991 +/- 0.002 (SD) after 9 days of soaking. Thus the fluorescent microsphere technique is a valuable tool for investigating regional perfusion in severely injured lungs and can replace radioactivity.

  11. DEVELOPMENT OF EVALUATION OF A QUANTITATIVE VIDEO-FLUORESCENCE IMAGING SYSTEM AND FLUORESCENT TRACER FOR MEASURING TRANSFER OF PESTICIDE RESIDUES FROM SURFACES TO HANDS WITH REPEATED CONTACTS

    EPA Science Inventory

    A video imaging system and the associated quantification methods have been developed for measurement of the transfers of a fluorescent tracer from surfaces to hands. The highly fluorescent compound riboflavin (Vitamin B2), which is also water soluble and non-toxic, was chosen as...

  12. An analysis of long term temperature measurement using laser induced fluorescence

    NASA Astrophysics Data System (ADS)

    Jaszczur, M.; Styszko, K.; Tomaszek, J.; Żurawska, K.

    2016-09-01

    The temperature measurement is extremely important because it occurs in many technical and engineering processes, including combustion chambers, mixers or chemical reactors as well as environmental flows. In contrast to the point measurement method, Laser Induced Fluorescence (LIF) allows temperature determination in the whole plain 2D, or even 3D, domain. A major advantage of LIF is also its relatively high accuracy. This technique involves dissolving a temperature- sensitive fluorescence dye to a fluid. It is known that in LIF the fluorescent reemission is a function of temperature but, in many cases, it can also be a function of time, due to dye properties degradation. In the present research, a long-term temperature measurement using LIF was performed in order to analyse the method uncertainty related to time. The results of the stability of Rhodamine-B in nonisothermal experimental measurements in water solution, together with the chemical analysis using spectrophotometry, are presented.

  13. Measurement of Pressure Dependent Fluorescence Yield of Air: Calibration Factor for UHECR Detectors

    SciTech Connect

    Belz, J.W.; Burt, G.W.; Cao, Z.; Chang, F.Y.; Chen, C.C.; Chen, C.W.; Chen, P.; Field, C.; Findlay, J.; Huntemeyer, Petra; Huang, M.A.; Hwang, W.-Y.P.; Iverson, R.; Jones, B.F.; Jui, C.C.H.; Kirn, M.; Lin, G.-L.; Loh, E.C.; Maestas, M.M.; Manago, N.; Martens, K.; /Montana U. /Utah U. /Taiwan, Natl. Taiwan U. /SLAC /Rutgers U., Piscataway

    2005-07-06

    In a test experiment at the Final Focus Test Beam of the Stanford Linear Accelerator Center, the fluorescence yield of 28.5 GeV electrons in air and nitrogen was measured. The measured photon yields between 300 and 400 nm at 1 atm and 29 C are Y(760 Torr){sup air} = 4.42 {+-} 0.73 and Y(760 Torr){sup N{sub 2}} = 29.2 {+-} 4.8 photons per electron per meter. Assuming that the fluorescence yield is proportional to the energy deposition of a charged particle traveling through air, good agreement with measurements at lower particle energies is observed.

  14. UV light phototransduction depolarizes human melanocytes.

    PubMed

    Bellono, Nicholas W; Oancea, Elena

    2013-01-01

    Exposure of human skin to low doses of solar UV radiation (UVR) causes increased pigmentation, while chronic exposure is a powerful risk factor for skin cancers. The mechanisms mediating UVR detection in skin, however, remain poorly understood. Our recent studies revealed that UVR activates a retinal-dependent G protein-coupled signaling pathway in melanocytes. This phototransduction pathway leads to the activation of transient receptor potential A1 (TRPA1) ion channels, elevation of intracellular calcium (Ca( 2+)) and rapid increase in cellular melanin content. Here we report that physiological doses of solar-like UVR elicit a retinal-dependent membrane depolarization in human epidermal melanocytes. This transient depolarization correlates with delayed inactivation time of the UVR-evoked photocurrent and with sustained Ca( 2+) responses required for early melanin synthesis. Thus, the cellular depolarization induced by UVR phototransduction in melanocytes is likely to be a critical signaling mechanism necessary for the protective response represented by increased melanin content.

  15. Zinc depolarized electrochemical CO2 concentration

    NASA Technical Reports Server (NTRS)

    Woods, R. R.; Marshall, R. D.; Schubert, F. H.

    1975-01-01

    Two zinc depolarized electrochemical carbon dioxide concentrator concepts were analytically and experimentally evaluated for portable life support system carbon dioxide (CO2) removal application. The first concept, referred to as the zinc hydrogen generator electrochemical depolarized CO2 concentrator, uses a ZHG to generate hydrogen for direct use in an EDC. The second concept, referred to as the zinc/electrochemical depolarized concentrator, uses a standard EDC cell construction modified for use with the Zn anode. The Zn anode is consumed and subsequently regenerated, thereby eliminating the need to supply H2 to the EDC for the CO2 removal process. The evaluation was based primarily on an analytical evaluation of the two ZnDCs at projected end item performance and hardware design levels. Both ZnDC concepts for PLSS CO2 removal application were found to be noncompetitive in both total equivalent launch weight and individual extravehicular activity mission volume when compared to other candidate regenerable PLSS CO2 scrubbers.

  16. Simultaneous one-dimensional fluorescence lifetime measurements of OH and CO in premixed flames

    NASA Astrophysics Data System (ADS)

    Jonsson, Malin; Ehn, Andreas; Christensen, Moah; Aldén, Marcus; Bood, Joakim

    2014-04-01

    A method for simultaneous measurements of fluorescence lifetimes of two species along a line is described. The experimental setup is based on picosecond laser pulses from two tunable optical parametric generator/optical parametric amplifier systems together with a streak camera. With an appropriate optical time delay between the two laser pulses, whose wavelengths are tuned to excite two different species, laser-induced fluorescence can be both detected temporally and spatially resolved by the streak camera. Hence, our method enables one-dimensional imaging of fluorescence lifetimes of two species in the same streak camera recording. The concept is demonstrated for fluorescence lifetime measurements of CO and OH in a laminar methane/air flame on a Bunsen-type burner. Measurements were taken in flames with four different equivalence ratios, namely ϕ = 0.9, 1.0, 1.15, and 1.25. The measured one-dimensional lifetime profiles generally agree well with lifetimes calculated from quenching cross sections found in the literature and quencher concentrations predicted by the GRI 3.0 mechanism. For OH, there is a systematic deviation of approximately 30 % between calculated and measured lifetimes. It is found that this is mainly due to the adiabatic assumption regarding the flame and uncertainty in H2O quenching cross section. This emphasizes the strength of measuring the quenching rates rather than relying on models. The measurement concept might be useful for single-shot measurements of fluorescence lifetimes of several species pairs of vital importance in combustion processes, hence allowing fluorescence signals to be corrected for quenching and ultimately yield quantitative concentration profiles.

  17. Ruby crystal for demonstrating time- and frequency-domain methods of fluorescence lifetime measurements.

    PubMed

    Chandler, Danielle E; Majumdar, Zigurts K; Heiss, Gregor J; Clegg, Robert M

    2006-11-01

    We present experiments that are convenient and educational for measuring fluorescence lifetimes with both time- and frequency-domain methods. The sample is ruby crystal, which has a lifetime of about 3.5 milliseconds, and is easy to use as a class-room demonstration. The experiments and methods of data analysis are used in the lab section of a class on optical spectroscopy, where we go through the theory and applications of fluorescence. Because the fluorescence decay time of ruby is in the millisecond region, the instrumentation for this experiment can be constructed easily and inexpensively compared to the nanosecond-resolved instrumentation required for most fluorescent compounds, which have nanosecond fluorescence lifetimes. The methods are applicable to other luminescent compounds with decay constants from microseconds and longer, such as transition metal and lanthanide complexes and phosphorescent samples. The experiments, which clearly demonstrate the theory and methods of measuring temporally resolved fluorescence, are instructive and demonstrate what the students have learned in the lectures without the distraction of highly sophisticated instrumentation.

  18. Fluorescence based imaging for M-band drive symmetry measurement in hohlraum

    NASA Astrophysics Data System (ADS)

    Li, Qi; Yao, Li; Jing, Longfei; Hu, Zhimin; Huang, Chengwu; Yuan, Zheng; Zhao, Yang; Xiong, Gang; Qing, Bo; Lv, Min; Zhu, Tuo; Deng, Bo; Li, Jin; Wei, Minxi; Zhan, Xiayu; Li, Jun; Yang, Yimeng; Su, Chunxiao; Yang, Guohong; Zhang, Jiyan; Li, Sanwei; Yang, Jiamin; Ding, Yongkun

    2016-11-01

    We describe an experimental technique to measure the drive symmetry of M-band radiation on the capsule in hohlraum. M-band radiation from the corona of the laser-produced gold plasma, especially the laser spot regions in the cavity, was used to pump x-ray fluorescence of a thin layer of Si-tracer coated on a solid CH-ball. The fluorescence images were time resolvedly recorded by an x-ray framing camera and the drive asymmetry due to M-band radiation was deduced from these fluorescence images. Moreover, a Si-doped gold cavity was used with the initial purpose of maximizing the fluorescence signal through resonance transitions. Since the Si-plasma expands more rapidly than the gold-plasma, the evolution of drive asymmetry was accelerated in Si-doped hohlraum.

  19. Aerosol-fluorescence spectrum analyzer: real-time measurement of emission spectra of airborne biological particles

    NASA Astrophysics Data System (ADS)

    Hill, Steven C.; Pinnick, Ronald G.; Nachman, Paul; Chen, Gang; Chang, Richard K.; Mayo, Michael W.; Fernandez, Gilbert L.

    1995-10-01

    We have assembled an aerosol-fluorescence spectrum analyzer (AFS), which can measure the fluorescence spectra and elastic scattering of airborne particles as they flow through a laser beam. The aerosols traverse a scattering cell where they are illuminated with intense (50 kW/cm 2) light inside the cavity of an argon-ion laser operating at 488 nm. This AFS can obtain fluorescence spectra of individual dye-doped polystyrene microspheres as small as 0.5 mu m in diameter. The spectra obtained from microspheres doped with pink and green-yellow dyes are clearly different. We have also detected the fluorescence spectra of airborne particles (although not single particles) made from various

  20. Fluorescence Spectrum and Decay Measurement for Hsil VS Normal Cytology Differentiation in Liquid Pap Smear Supernatant

    NASA Astrophysics Data System (ADS)

    Vaitkuviene, A.; Gegzna, V.; Juodkazis, S.; Jursenas, S.; Miasojedovas, S.; Kurtinaitiene, R.; Rimiene, J.; Vaitkus, J.

    2009-06-01

    Cervical smear material contains endo and exocervical cells, mucus and inflammative, immune cells in cases of pathology. Just not destroyed keratinocytes lay on the glass for microscopy. Liquid cytology supernatant apart other diagnostics could be used for photodiagnostic. The spectroscopic parameters suitable for Normal and HSIL cytology groups supernatant differentiation are demonstrated. The dried liquid PAP supernatant fractions—sediment and liquid were investigated. Excitation and emission matrices (EEM), supernatant fluorescence decay measured under 280 nm diode short pulse excitation and fluorescence spectroscopy by excitation with 355 nm laser light were analyzed. The differences between Normal and HSIL groups were statistically proven in the certain spectral regions. Fluorescence decay peculiarities show spectral regions consisting of few fluorophores. Obtained results on fluorescence differences in Normal and HSIL groups' supernatant shows the potency of photodiagnosis application in cervical screening.

  1. Laser measurement of the spectral extinction coefficients of fluorescent, highly absorbing liquids. [crude petroleum oils

    NASA Technical Reports Server (NTRS)

    Hoge, F. E.

    1982-01-01

    A conceptual method is developed to deduce rapidly the spectral extinction coefficient of fluorescent, highly absorbing liquids, such as crude or refined petroleum oils. The technique offers the advantage of only requiring one laser wavelength and a single experimental assembly and execution for any specific fluorescent liquid. The liquid is inserted into an extremely thin wedge-shaped cavity for stimulation by a laser from one side and flurescence measurement on the other side by a monochromator system. For each arbitrarily selected extinction wavelength, the wedge is driven slowly to increasing thicknesses until the fluorescence extinguishes. The fluorescence as a function of wedge thickness permits a determination of the extinction coefficient using an included theoretical model. When the monochromator is set to the laser emission wavelength, the extinction coefficient is determined using the usual on-wavelength signal extinction procedure.

  2. Laser-induced fluorescence measurement of the dynamics of a pulsed planar sheath

    SciTech Connect

    Goeckner, M.J.; Malik, S.M. ); Conrad, J.R. ); Breun, R.A. )

    1994-04-01

    Using laser-induced fluorescence (LIF) the ion density near the edge of an expanding plasma sheath has been measured. These measurements utilized a transition of N[sup +][sub 2] [the P12 component of the [ital X] [sup 2][Sigma][sup +][sub [ital g

  3. Capturing Depolarization Information in GPS Reflections

    NASA Technical Reports Server (NTRS)

    Kelly, Kenneth C.

    2000-01-01

    The state of the surface of the ocean has a prominent effect on the depolarization of the circularly polarized emissions of the GPS satellites. The system designers election to capture the important information carries with it the need to implement the data extraction in a cost efficient manner. Antenna components, and associated networks for deriving depolarization information are described. For typical sea states the polarization characteristics of the reflected GPS signal vary rapidly with time so various methods for recording the changes are discussed.

  4. Measurement of diffusion of fluorescent compounds and autofluorescence in skin in vivo using a confocal instrument

    NASA Astrophysics Data System (ADS)

    Buttenschoen, K. K.; Sutton, E. E.; Daly, D.; Girkin, J. M.

    2016-02-01

    Using compact and affordable instrumentation based upon fluorescent confocal imaging we have tracked the movement of autofluorescent compounds through skin in near real time with high temporal and spatial resolution and sensitivity. The ability to measure the diffusion of compounds through skin with such resolution plays an important role for applications such as monitoring the penetration of pharmaceuticals applied to skin and assessing the integrity of the skin barrier. Several measurement methods exist, but they suffer from a number of problems such as being slow, expensive, non-portable and lacking sensitivity. To address these issues, we adapted a technique that we previously developed for tracking fluorescent compounds in the eye to measure the autofluorescence and the diffusion of externally applied fluorescent compounds in skin in vivo. Results are presented that show the change in autofluorescence of the volar forearm over the course of a week. We furthermore demonstrate the ability of the instrument to measure the diffusion speed and depth of externally applied fluorescent compounds both in healthy skin and after the skin barrier function has been perturbed. The instrument is currently being developed further for increased sensitivity and multi-wavelength excitation. We believe that the presented instrument is suitable for a large number of applications in fields such as assessment of damage to the skin barrier, development of topical and systemic medication and tracking the diffusion of fluorescent compounds through skin constructs as well as monitoring effects of skin products and general consumer products which may come into contact with the skin.

  5. Fluorescence Quantum Yield Measurements of Fluorescent Proteins: A Laboratory Experiment for a Biochemistry or Molecular Biophysics Laboratory Course

    ERIC Educational Resources Information Center

    Wall, Kathryn P.; Dillon, Rebecca; Knowles, Michelle K.

    2015-01-01

    Fluorescent proteins are commonly used in cell biology to assess where proteins are within a cell as a function of time and provide insight into intracellular protein function. However, the usefulness of a fluorescent protein depends directly on the quantum yield. The quantum yield relates the efficiency at which a fluorescent molecule converts…

  6. Peptide-based fluorescence biosensors for detection/measurement of nanoparticles.

    PubMed

    Akinloye, Oluyemi; Krishnamurthy, Ramanarayan; Wishart, David; Goss, Greg G

    2017-02-01

    The ability to detect and quantify nanoparticles is essential but there is currently no simple, sensitive, and rapid method for the detection of nanomaterials. We have developed a novel peptide-based fluorescence-based biosensor for detection and measurement of negatively charged engineered nanoparticles (ENPs). A peptide biosensor (seven lysine residues linked to a cysteine through a three glycine residue linker) with attached fluorescent probes-fluorescein-5-maleimide (F5M) and tetramethylrhodamine-5-maleimide (TMR5M)-was constructed. The fluorescent probes allow close monitoring of the molecular interaction of the labeled peptide with ENPs. The ENP-peptide interaction induces the formation of agglomerates that can be detected and measured by changes in the fluorescence intensities of the labeled peptides or/and by differential light scattering. The relative fluorescence intensities of F5M and TMR5M decreased in a concentration-dependent manner on interaction with various types of negatively charged ENPs (ZnO, Fe3O4, CeO, and single-walled carbon nanotubes). Differential light scattering measurements also showed increases in the hydrodynamic size of the complex. The interactions were not affected by the pH of aqueous media, where humic acid (1 μg/mL) quenched the fluorescence intensity of F5M by approximately 25 %, whereas that of TMR5M was completely quenched. Interference by humic acid at lower concentrations was less prevalent. This novel method is a simple, rapid, and inexpensive in situ assay that shows promise as a primary-level testing technique for detection of ENPs in environmental samples. Graphical Abstract Detection of nanomaterials in aqueous solutions using fluorescently-labeled designer peptides.

  7. Fast repetition rate (FRR) fluorometer and method for measuring fluorescence and photosynthetic parameters

    DOEpatents

    Kolber, Zbigniew; Falkowski, Paul

    1995-06-20

    A fast repetition rate fluorometer device and method for measuring in vivo fluorescence of phytoplankton or higher plants chlorophyll and photosynthetic parameters of phytoplankton or higher plants by illuminating the phytoplankton or higher plants with a series of fast repetition rate excitation flashes effective to bring about and measure resultant changes in fluorescence yield of their Photosystem II. The series of fast repetition rate excitation flashes has a predetermined energy per flash and a rate greater than 10,000 Hz. Also, disclosed is a flasher circuit for producing the series of fast repetition rate flashes.

  8. Fast repetition rate (FRR) fluorometer and method for measuring fluorescence and photosynthetic parameters

    DOEpatents

    Kolber, Z.; Falkowski, P.

    1995-06-20

    A fast repetition rate fluorometer device and method for measuring in vivo fluorescence of phytoplankton or higher plants chlorophyll and photosynthetic parameters of phytoplankton or higher plants is revealed. The phytoplankton or higher plants are illuminated with a series of fast repetition rate excitation flashes effective to bring about and measure resultant changes in fluorescence yield of their Photosystem II. The series of fast repetition rate excitation flashes has a predetermined energy per flash and a rate greater than 10,000 Hz. Also, disclosed is a flasher circuit for producing the series of fast repetition rate flashes. 14 figs.

  9. Planar temperature measurement in compressible flows using laser-induced iodine fluorescence

    NASA Technical Reports Server (NTRS)

    Hartfield, Roy J., Jr.; Hollo, Steven D.; Mcdaniel, James C.

    1991-01-01

    A laser-induced iodine fluorescence technique that is suitable for the planar measurement of temperature in cold nonreacting compressible air flows is investigated analytically and demonstrated in a known flow field. The technique is based on the temperature dependence of the broadband fluorescence from iodine excited by the 514-nm line of an argon-ion laser. Temperatures ranging from 165 to 245 K were measured in the calibration flow field. This technique makes complete, spatially resolved surveys of temperature practical in highly three-dimensional, low-temperature compressible flows.

  10. Noninvasive measurement of pharmacokinetics by near-infrared fluorescence imaging in the eye of mice

    NASA Astrophysics Data System (ADS)

    Dobosz, Michael; Strobel, Steffen; Stubenrauch, Kay-Gunnar; Osl, Franz; Scheuer, Werner

    2014-01-01

    Purpose: For generating preclinical pharmacokinetics (PKs) of compounds, blood is drawn at different time points and levels are quantified by different analytical methods. In order to receive statistically meaningful data, 3 to 5 animals are used for each time point to get serum peak-level and half-life of the compound. Both characteristics are determined by data interpolation, which may influence the accuracy of these values. We provide a method that allows continuous monitoring of blood levels noninvasively by measuring the fluorescence intensity of labeled compounds in the eye and other body regions of anesthetized mice. Procedures: The method evaluation was performed with four different fluorescent compounds: (i) indocyanine green, a nontargeting dye; (ii) OsteoSense750, a bone targeting agent; (iii) tumor targeting Trastuzumab-Alexa750; and (iv) its F(-alxea750 fragment. The latter was used for a direct comparison between fluorescence imaging and classical blood analysis using enzyme-linked immunosorbent assay (ELISA). Results: We found an excellent correlation between blood levels measured by noninvasive eye imaging with the results generated by classical methods. A strong correlation between eye imaging and ELISA was demonstrated for the F( fragment. Whole body imaging revealed a compound accumulation in the expected regions (e.g., liver, bone). Conclusions: The combination of eye and whole body fluorescence imaging enables the simultaneous measurement of blood PKs and biodistribution of fluorescent-labeled compounds.

  11. Fluorescent reports for detection and measurement of DNA damage

    SciTech Connect

    Uziel, M.; Houck, K. )

    1993-01-01

    Epidemiological studies of real populations are complicated by the inevitable coexistence of exposure to multiple agents within the target population. An alternative method for characterizing these types of exposures is to use the reactive chemical functional group as the toxic agent identify the corresponding classes (families) of damage as markers of effects. We have begun studies to develop spectrometric reporters of DNA damage that can be measured on intact DNA. The direct measurement of adducts on microgram levels of DNA from tissue biopsy may succeed because of the high sensitivity and selectivity of different reporter compounds. While one cannot readily distinguish between recent or persistent exposures, baseline values for individuals may be constructed. For example, normal oxidative metabolism and environmental radiation create oxidation processes that continually damage DNA. These reactions create lesions that can be measured with the reporter compound FABA [N- (5- fluoresceinyl), N[prime]-(3-boronatophenyl)thioureal]. We report preliminary observations with binding FABA (selective for cis, vicdiol structures) to damage sites present on intact nonirradiated and irradiated DNA from C3H10T[sub 1/2] cells. We have observed binding of 42,000 FABA per mouse tetraploid genome (9 billion base pairs) to the putative thymidylic glycol resulting from normal oxidative processes in nonirradiated DNA. Additional binding of FABA to DNA from cells exposed to 100, 300, and 500 rad shows an exponential increase in binding sites of up to 140,000 with 500 rad exposure. This damage reporter may prove useful in characterizing levels of nonovert and overt oxidative damage to DNA.

  12. Laser induced fluorescence measurements and modeling of nitric oxide in high-pressure premixed flames

    NASA Technical Reports Server (NTRS)

    Reisel, John R.; Laurendeau, Normand M.

    1994-01-01

    Laser-induced fluorescence (LIF) has been applied to the quantitative measurement of nitric oxide (NO) in premixed, laminar, high-pressure flames. Their chemistry was also studied using three current kinetics schemes to determine the predictive capabilities of each mechanism with respect to NO concentrations. The flames studied were low-temperature (1600 less than T less than 1850K) C2H6/O2/N2 and C2H6/O2/N2 flames, and high temperature (2100 less than T less than 2300K) C2H6/O2/N2 flames. Laser-saturated fluorescence (LSF) was initially used to measure the NO concentrations. However, while the excitation transition was well saturated at atmospheric pressure, the fluorescence behavior was basically linear with respect to laser power at pressures above 6 atm. Measurements and calculations demonstrated that the fluorescence quenching rate variation is negligible for LIF measurements of NO at a given pressure. Therefore, linear LIF was used to perform quantitative measurements of NO concentration in these high-pressure flames. The transportability of a calibration factor from one set of flame conditions to another also was investigated by considering changes in the absorption and quenching environment for different flame conditions. The feasibility of performing LIF measurements of (NO) in turbulent flames was studied; the single-shot detection limit was determined to be 2 ppm.

  13. Composition measurement of bicomponent droplets using laser-induced fluorescence of acetone

    NASA Astrophysics Data System (ADS)

    Maqua, C.; Depredurand, V.; Castanet, G.; Wolff, M.; Lemoine, F.

    2007-12-01

    Commercial fuels are complex mixtures, the evaporation of which remains particularly difficult to model. Experimental characterization of the differential vaporization of the components is a problem that is seldom addressed. In this paper, the evaporation of binary droplets made of ethyl-alcohol and acetone is investigated using a technique of measurement of the droplet composition developed in purpose. This technique exploits the laser induced fluorescence of acetone which acts as a fluorescent tracer as well as the more volatile component of the fuel associated with an accurate measurement of the droplet diameter by forward scattering interferometry. A model of the fluorescence intensity of the binary mixture, taking into account the absorption of the acetone molecules, is proposed and validated. The sensitivity of the technique is discussed. Finally, the reliability of the technique is demonstrated on binary combusting droplets in linear stream.

  14. Dualex: A New Instrument for Field Measurements of Epidermal Ultraviolet Absorbance by Chlorophyll Fluorescence

    NASA Astrophysics Data System (ADS)

    Goulas, Yves; Cerovic, Zoran G.; Cartelat, Aurélie; Moya, Ismaël

    2004-08-01

    Dualex (dual excitation) is a field-portable instrument, hereby described, for the assessment of polyphenolic compounds in leaves from the measurement of UV absorbance of the leaf epidermis by double excitation of chlorophyll fluorescence. The instrument takes advantage of a feedback loop that equalizes the fluorescence level induced by a reference red light to the UV-light-induced fluorescence level. This allows quick measurement from attached leaves even under field conditions. The use of light-emitting diodes and of a leaf-clip configuration makes Dualex a user-friendly instrument with potential applications in ecophysiological research, light climate analysis, agriculture, forestry, horticulture, pest management, selection of medicinal plants, and wherever accumulation of leaf polyphenolics is involved in plant responses to the environment.

  15. Depolarized light scattering studies of the collision induced polarizability anisotropy of atoms and spherical top molecules

    NASA Astrophysics Data System (ADS)

    Dunmur, D. A.; Manterfield, M. R.; Robinson, D. J.

    New measurements are reported of the density dependent depolarization ratio for argon, krypton, xenon, methane and sulphur hexafluoride, and the results are analysed to provide values for the second and third depolarization virial coefficients. The relationships between the second depolarization virial coefficient, the zeroth moment of the two-body Rayleigh spectrum and the second Kerr virial coefficient are considered, and it is shown that they now provide consistent results for the collision-induced pair polarizability anisotropy. Former inconsistencies are attributed to insufficient allowance for the effects of three-body interactions. Calculations of the second and third depolarization virial coefficients based on the DID model and using the Maitland-Smith potential are in excellent agreement with the experimental results for argon, krypton and xenon.

  16. A low cost fluorescence lifetime measurement system based on SPAD detectors and FPGA processing

    NASA Astrophysics Data System (ADS)

    Franch, N.; Alonso, O.; Canals, J.; Vilà, A.; Dieguez, A.

    2017-02-01

    This work presents a low cost fluorescence life time measurement system, aimed at carrying out fast diagnostic tests through label detection in a portable system so it can be used in a medical consultation, within a short time span. The system uses Time Correlated Single Photon Counting (TCSPC), measuring the arrival time of individual photons and building a histogram of those times, showing the fluorescence decay of the label which is characteristic of each fluorescent substance. The system is implemented using a Xilinx FPGA which controls the experiment and includes a Time to Digital Converter (TDC) to perform measurements with a resolution in the order of tenths of picoseconds. Also included are a laser diode and the driving electronics to generate short pulses as well as a HV-CMOS implemented Single Photon Avalanche Diode (SPAD) as a high gain sensor. The system is entirely configurable so it can easily be adapted to the target label molecule and measurement needs. The histogram is constructed within the FPGA and can then be read as convenient. Various performance parameters are also shown, as well as experimental measurements of a quantum dot fluorescence decay as a proof of concept.

  17. Monitoring dynamical vegetation processes with solar-induced chlorophyll fluorescence measurements from space (Invited)

    NASA Astrophysics Data System (ADS)

    Moreno, J. F.; Guanter, L.; Alonso, L.; Gomez-Chova, L.; Drusch, M.; Kraft, S.; Carnicero, B.; Bezy, J.

    2009-12-01

    Fluorescence is a powerful non-invasive tool to track the status, resilience, and recovery of photochemical processes and moreover provides important information on overall vegetation photosynthetic performance with implications for related carbon sequestration, allowing to measure planetary photosynthesis by means of a global monitoring of steady-state chlorophyll fluorescence in terrestrial vegetation. The FLuorescence EXperiment (FLEX) is designed to observe the photosynthetic activity of the vegetation layer, by using a completely novel technique measuring the chlorophyll fluorescence signal that originates from the core of the photosynthetic machinery, i.e. the ‘breathing’ of the vegetation layer of the living planet. Conceived as a technology demonstration mission, it proposes a set of instruments for the measurement of the interrelated features of fluorescence, spectral reflectance, and canopy temperature, by using a dedicated small satellite flying in tandem with GMES Sentinel-3. This will provide a completely new possibility to quantify the photosynthetic efficiency of terrestrial ecosystems at the global scale, to improve the predictability of dynamical vegetation models on scales comprising canopies and biomes, and to provide an improved estimate of GPP for a better understanding of the global carbon cycle. It will also improve understanding of the role of vegetation in the coupled global carbon / water cycles, the global assessment of the vegetation health conditions and vegetation stress and the support the development of future crop production strategies in a changing climate. The measurement represent a challenge: the weak fluorescence signal is masked by the reflected background radiance, and accurate compensation of all perturbing effects becomes essential. Recent developments have demonstrated the feasibility of the measurements of canopy fluorescence from space. Recent model developments and data processing tools have made possible to

  18. High microvascular endothelial water permeability in mouse lung measured by a pleural surface fluorescence method.

    PubMed Central

    Carter, E P; Olveczky, B P; Matthay, M A; Verkman, A S

    1998-01-01

    Transport of water between the capillary and airspace compartments in lung encounters serial barriers: the alveolar epithelium, interstitium, and capillary endothelium. We previously reported a pleural surface fluorescence method to measure net capillary-to-airspace water transport. To measure the osmotic water permeability across the microvascular endothelial barrier in intact lung, the airspace was filled with a water-immiscible fluorocarbon. The capillaries were perfused via the pulmonary artery with solutions of specified osmolalites containing a high-molecular-weight fluorescent dextran. An increase in perfusate osmolality produced a prompt decrease in surface fluorescence due to dye dilution in the capillaries, followed by a slower return to initial fluorescence as capillary and lung interstitial osmolality equilibrate. A mathematical model was developed to determine the osmotic water permeability coefficient (Pf) of lung microvessels from the time course of pleural surface fluorescence. As predicted, the magnitude of the prompt change in surface fluorescence increased with decreased pulmonary artery perfusion rate and increased osmotic gradient size. With raffinose used to induce the osmotic gradient, Pf was 0.03 cm/s at 23 degrees C and was reduced 54% by 0.5 mM HgCl2. Temperature dependence measurements gave an Arrhenius activation energy (Ea) of 5.4 kcal/mol (12-37 degrees C). The apparent Pf induced by the smaller osmolytes mannitol and glycine was 0.021 and 0.011 cm/s (23 degrees C). Immunoblot analysis showed approximately 1.4 x 10(12) aquaporin-1 water channels/cm2 of capillary surface, which accounted quantitatively for the high Pf. These results establish a novel method for measuring osmotically driven water permeability across microvessels in intact lung. The high Pf, low Ea, and mercurial inhibition indicate the involvement of molecular water channels in water transport across the lung endothelium. PMID:9545071

  19. Anisole fluorescence spectroscopy for temperature measurements with a Hg (Xe) arc lamp excitation

    NASA Astrophysics Data System (ADS)

    Guibert, P.; Kanumuri, S. S.; Bonnety, J.; Tran, K.-H.; Serio, B.; Bonnet, D.; Luc, J.; Lavayssiere, M.

    2017-04-01

    The main contribution of this study is to propose time-resolved measurements to determine temperature with a novel source of continuous excitation for an induced fluorescence technique with laser diagnosis based on tracer-induced fluorescence, which has become a major tool for experimental studies of fluid dynamics in reaction flows. We use a Hg (Xe) arc lamp as a continuous light source that has a wide range of emissions in wavelength. With this setup, one can reach high spatial and temporal resolution (temperature, pressure, species concentration, and velocity) to acquire quantitative data for the control of fluid thermal systems, such as engines, combustion chambers, furnaces, and reactors. A fluorescence study was performed on various tracers and their configurations. We focus on an anisole tracer using a broad wavelength of excitations. We propose a calibration to achieve temperature measurements in the range of 493-773 K and from 0.2 to 3.5 MPa of pressure. The temperature-dependent fluorescence is based on a two-line technique. The results give a better understanding of the influence of temperature and pressure in a nitrogen bath gas on the fluorescence photophysics in the UV domain. High temporal resolution was acquired using a high-speed intensified camera setup. The application of the photomultipliers manages the time-scale evolution of the flow in continuous emission and this eliminates the signal-to-noise ratio impact.

  20. The retrieval of the Asian dust depolarization ratio in Korea with the correction of the polarization-dependent transmission

    NASA Astrophysics Data System (ADS)

    Shin, Sungkyun; Müller, Detlef; Kim, Y. J.; Tatarov, Boyan; Shin, Dongho; Seifert, Patric; Noh, Young Min

    2013-01-01

    The linear particle depolarization ratios were retrieved from the observation with a multiwavelength Raman lidar at the Gwangju Institute of Science and Technology (GIST), Korea (35.11°N, 126.54°E). The measurements were carried out in spring (March to May) 2011. The transmission ratio measurements were performed to solve problems of the depolarization-dependent transmission at a receiver of the lidar and applied to correct the retrieved depolarization ratio of Asian dust at first time in Korea. The analyzed data from the GIST multiwavelength Raman lidar were classified into three categories according to the linear particle depolarization ratios, which are pure Asian dust on 21 March, the intermediate case which means Asian dust mixed with urban pollution on 13 May, and haze case on 10 April. The measured transmission ratios were applied to these cases respectively. We found that the transmission ratio is needed to be used to retrieve the accurate depolarization ratio of Asian dust and also would be useful to distinguish the mixed dust particles between intermediate case and haze. The particle depolarization ratios of pure Asian dust were approximately 0.25 at 532 nm and 0.14 at 532 nm for the intermediate case. The linear particle depolarization ratios of pure Asian dust observed with the GIST multiwavelength Raman lidar were compared to the linear particle depolarization ratios of Saharan dust observed in Morocco and Asian dust observed both in Japan and China.

  1. Fluorescence lifetime measurements of NADH and tryptophan in intact ischemic, intact rabbit myocardium

    NASA Astrophysics Data System (ADS)

    Hamburger, Adrian; Gryczynski, Zygmunt; Lakowicz, Joseph R.; Sommers, Keith

    1999-07-01

    Ischemia-reperfusion injury is the leading cause of early dysfunction following transplantation. Currently, there are no techniques available to accurately measure ischemic changes during organ storage. Therefore, the interest exists in developing non-invasive monitoring techniques. We used NADH and tryptophan as fluorescent markers, since both are intrinsic fluorophores and excellent indicators for levels of hypoxia and protein denaturation, respectively.

  2. Filter-fluorescer measurement of low-voltage simulator x-ray energy spectra

    SciTech Connect

    Baldwin, G.T.; Craven, R.E.

    1986-01-01

    X-ray energy spectra of the Maxwell Laboratories MBS and Physics International Pulserad 737 were measured using an eight-channel filter-fluorescer array. The PHOSCAT computer code was used to calculate channel response functions, and the UFO code to unfold spectrum.

  3. Winter wheat GPC estimation with fluorescence-based sensor measurements of canopy

    NASA Astrophysics Data System (ADS)

    Song, Xiaoyu; Wang, Jihua; Gu, Xiaohe; Xu, Xingang

    2015-10-01

    This study focused on the wheat grain protein content (GPC) estimation based on wheat canopy chlorophyll parameters which acquired by hand-held instrument, Multiplex 3. Nine fluorescence spectral indices from Multiplex sensor were used in this study. The wheat GPC estimation experiment was conducted in 2012 at the National Experiment Station for Precision Agriculture in Changping district, Beijing. A square with area of 1.1 ha was selected and divided to 110 small plots by 10×10m in this study. In each plot, four 1-m2 area distributed in the square were selected for canopy fluorescence spectral measurements, physiological and biochemical analyses. Measurements were performed five times at wheat raising, jointing, heading stage, milking and ripening stage, respectively. The wheat plant samples for each plot were then collected after the measurement and sent to Lab for leaf N concentration (LNC) and canopy nitrogen density (CND) analyzed. GPC sampling for each plot was collected manually during the harvested season. Then, statistical analysis were performed to detect the correlation between fluorescence spectral indices and wheat CND for each growth stage, as well as GPC. The results indicate that two Nitrogen Balance Indices, NBI_G and NBI_R were more sensitive to wheat GPC than other fluorescence spectral indices at milking stage and ripening stage. Five linear regression models with GPC and fluorescence indices at different winter wheat growth stages were then established. The R2 of GPC estimated model increased form 0.312 at raising stage to 0.686 at ripening stage. The study reveals that canopy-level fluorescence spectral parameters were better indicators for the wheat group activity and could be demonstrated to be good indicators for winter wheat GPC estimation.

  4. A new method for measuring concentration of a fluorescent tracer in bubbly gas-liquid flows

    NASA Astrophysics Data System (ADS)

    Moghaddas, J. S.; Trägårdh, C.; Kovacs, T.; Östergren, K.

    2002-06-01

    A new experimental model, the two-tracer method (TTM), based on the planar laser-induced fluorescence technique (PLIF), is presented for the measurement of the local concentration of a fluorescent tracer in the liquid phase of a bubbly two-phase system. Light scattering and shading effects due to the bubbles were compensated for using the new model. The TTM results were found to give more accurate predictions of the local concentration than the normal PLIF method in a bubbly two-phase system.

  5. Tumor detection in mice by measurement of fluorescence decay time matrices

    NASA Astrophysics Data System (ADS)

    Cubeddu, R.; Pifferi, A.; Taroni, P.; Valentini, G.; Canti, G.

    1995-12-01

    An intensified CCD video camera has been used to measure the spatial distribution of the fluorescence decay time in tumor-bearing mice sensitized with hematoporphyrin derivative. Mice were injected with five doses of sensitizer, ranging from 0.1 to 10 mg / kg body weight. For any drug dose the decay time of the exogenous fluorescence in the tumor is always significantly longer than in normal tissues. The image created by associating a gray-shade scale to the decay time matrix of each mouse permits a reliable and precise detection of the neoplasia.

  6. Molecular diffusivity measurement through an alumina membrane using time-resolved fluorescence imaging

    NASA Astrophysics Data System (ADS)

    Kennard, Raymond; DeSisto, William J.; Mason, Michael D.

    2010-11-01

    We present a simple fluorescence imaging method for measuring the time-resolved concentration of a fluorescent molecule diffusing through an anodic alumina membrane with a pore diameter of 20 nm. From the concentration breakthrough curve, the molecular diffusivity of the fluorophore was extracted. The experimentally determined diffusivity was three orders of magnitude lower than reported bulk values. Due to the relative simplicity and ease of use, this method can be applied to provide fundamental information for biomolecular separations applications. One feature of this method is the high sensitivity at intercellular volumes broadening its application to drug delivery and controlled cell growth.

  7. Diffusivity of asphaltene molecules by fluorescence correlation spectroscopy.

    PubMed

    Andrews, A Ballard; Guerra, Rodrigo E; Mullins, Oliver C; Sen, Pabitra N

    2006-07-06

    Using fluorescence correlation spectroscopy (FCS) we measure the translational diffusion coefficient of asphaltene molecules in toluene at extremely low concentrations (0.03-3.0 mg/L): where aggregation does not occur. We find that the translational diffusion coefficient of asphaltene molecules in toluene is about 0.35 x 10(-5) cm(2)/s at room temperature. This diffusion coefficient corresponds to a hydrodynamic radius of approximately 1 nm. These data confirm previously estimated size from rotational diffusion studied using fluorescence depolarization. The implication of this concurrence is that asphaltene molecular structures are monomeric, not polymeric.

  8. Mueller matrix signature in advanced fluorescence microscopy imaging

    NASA Astrophysics Data System (ADS)

    Mazumder, Nirmal; Qiu, Jianjun; Kao, Fu-Jen; Diaspro, Alberto

    2017-02-01

    We have demonstrated the measurement and characterization of the polarization properties of a fluorescence signal using four-channel photon counting based Stokes-Mueller polarization microscopy. Thus, Lu-Chipman decomposition was applied to extract the critical polarization properties such as depolarization, linear retardance and the optical rotation of collagen type I fiber. We observed the spatial distribution of anisotropic and helical molecules of collagen from the reconstructed 2D Mueller images based on the fluorescence signal in a pixel-by-pixel manner.

  9. Experimental feasibility of the airborne measurement of absolute oil fluorescence spectral conversion efficiency

    NASA Technical Reports Server (NTRS)

    Hoge, F. E.; Swift, R. N.

    1983-01-01

    Airborne lidar oil spill experiments carried out to determine the practicability of the AOFSCE (absolute oil fluorescence spectral conversion efficiency) computational model are described. The results reveal that the model is suitable over a considerable range of oil film thicknesses provided the fluorescence efficiency of the oil does not approach the minimum detection sensitivity limitations of the lidar system. Separate airborne lidar experiments to demonstrate measurement of the water column Raman conversion efficiency are also conducted to ascertain the ultimate feasibility of converting such relative oil fluorescence to absolute values. Whereas the AOFSCE model is seen as highly promising, further airborne water column Raman conversion efficiency experiments with improved temporal or depth-resolved waveform calibration and software deconvolution techniques are thought necessary for a final determination of suitability.

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

    SciTech Connect

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

    2015-01-01

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

  11. Quantitative measurement of intracellular protein dynamics using photobleaching or photoactivation of fluorescent proteins.

    PubMed

    Matsuda, Tomoki; Nagai, Takeharu

    2014-12-01

    Unlike in vitro protein dynamics, intracellular protein dynamics are intricately regulated by protein-protein interactions or interactions between proteins and other cellular components, including nucleic acids, the plasma membrane and the cytoskeleton. Alteration of these dynamics plays a crucial role in physiological phenomena such as gene expression and cell division. Live-cell imaging via microscopy with the inherent properties of fluorescent proteins, i.e. photobleaching and photoconversion, or fluorescence correlation spectroscopy, provides insight into the movement of proteins and their interactions with cellular components. This article reviews techniques based on photo-induced changes in the physicochemical properties of fluorescent proteins to measure protein dynamics inside living cells, and it also discusses the strengths and weaknesses of these techniques.

  12. Fluorescence measurements for evaluating the application of multivariate analysis techniques to optically thick environments.

    SciTech Connect

    Reichardt, Thomas A.; Timlin, Jerilyn Ann; Jones, Howland D. T.; Sickafoose, Shane M.; Schmitt, Randal L.

    2010-09-01

    Laser-induced fluorescence measurements of cuvette-contained laser dye mixtures are made for evaluation of multivariate analysis techniques to optically thick environments. Nine mixtures of Coumarin 500 and Rhodamine 610 are analyzed, as well as the pure dyes. For each sample, the cuvette is positioned on a two-axis translation stage to allow the interrogation at different spatial locations, allowing the examination of both primary (absorption of the laser light) and secondary (absorption of the fluorescence) inner filter effects. In addition to these expected inner filter effects, we find evidence that a portion of the absorbed fluorescence is re-emitted. A total of 688 spectra are acquired for the evaluation of multivariate analysis approaches to account for nonlinear effects.

  13. Mechanisms of fluorescence decays of colloidal CdSe-CdS/ZnS quantum dots unraveled by time-resolved fluorescence measurement.

    PubMed

    Xu, Hao; Chmyrov, Volodymyr; Widengren, Jerker; Brismar, Hjalmar; Fu, Ying

    2015-11-07

    By narrowing the detection bandpass and increasing the signal-to-noise ratio in measuring the time-resolved fluorescence decay spectrum of colloidal CdSe-CdS/ZnS quantum dots (QDs), we show that directly after the photoexcitation, the fluorescence decay spectrum is characterized by a single exponential decay, which represents the energy relaxation of the photogenerated exciton from its initial high-energy state to the ground exciton state. The fluorescence decay spectrum of long decay time is in the form of β/t(2), where β is the radiative recombination time of the ground-state exciton and t is the decay time. Our findings provide us with a direct and quantitative link between fluorescence decay measurement data and fundamental photophysics of QD exciton, thereby leading to a novel way of applying colloidal QDs to study microscopic, physical and chemical processes in many fields including biomedicine.

  14. In vitro fluorescence measurements and Monte Carlo simulation of laser irradiation propagation in porcine skin tissue.

    PubMed

    Drakaki, E; Makropoulou, M; Serafetinides, A A

    2008-07-01

    In dermatology, the in vivo spectral fluorescence measurements of human skin can serve as a valuable supplement to standard non-invasive techniques for diagnosing various skin diseases. However, quantitative analysis of the fluorescence spectra is complicated by the fact that skin is a complex multi-layered and inhomogeneous organ, with varied optical properties and biophysical characteristics. In this work, we recorded, in vitro, the laser-induced fluorescence emission signals of healthy porcine skin, one of the animals, which is considered as one of the most common models for investigations related to medical diagnostics of human cutaneous tissues. Differences were observed in the form and intensity of the fluorescence signal of the porcine skin, which can be attributed to the different concentrations of the native fluorophores and the variable physical and biological conditions of the skin tissue. As the light transport in the tissue target is directly influencing the absorption and the fluorescence emission signals, we performed Monte Carlo simulation of the light distribution in a five-layer model of human skin tissue, with a pulsed ultraviolet laser beam.

  15. Measurement of fluorescent probes concentration ratio in the cerebrospinal fluid for early detection of Alzheimer's disease

    NASA Astrophysics Data System (ADS)

    Harbater, Osnat; Gannot, Israel

    2014-03-01

    The pathogenic process of Alzheimer's Disease (AD), characterized by amyloid plaques and neurofibrillary tangles in the brain, begins years before the clinical diagnosis. Here, we suggest a novel method which may detect AD up to nine years earlier than current exams, minimally invasive, with minimal risk, pain and side effects. The method is based on previous reports which relate the concentrations of biomarkers in the Cerebrospinal Fluid (CSF) (Aβ and Tau proteins) to the future development of AD in mild cognitive impairment patients. Our method, which uses fluorescence measurements of the relative concentrations of the CSF biomarkers, replaces the lumbar puncture process required for CSF drawing. The process uses a miniature needle coupled trough an optical fiber to a laser source and a detector. The laser radiation excites fluorescent probes which were prior injected and bond to the CSF biomarkers. Using the ratio between the fluorescence intensities emitted from the two biomarkers, which is correlated to their concentration ratio, the patient's risk of developing AD is estimated. A theoretical model was developed and validated using Monte Carlo simulations, demonstrating the relation between fluorescence emission and biomarker concentration. The method was tested using multi-layered tissue phantoms simulating the epidural fat, the CSF in the sub-arachnoid space and the bone. These phantoms were prepared with different scattering and absorption coefficients, thicknesses and fluorescence concentrations in order to simulate variations in human anatomy and in the needle location. The theoretical and in-vitro results are compared and the method's accuracy is discussed.

  16. Reflectance and fluorescence characterization of maize species using field laboratory measurements and lidar remote sensing.

    PubMed

    Zhao, Guangyu; Duan, Zheng; Ming, Lian; Li, Yiyun; Chen, Ruipeng; Hu, Jiandong; Svanberg, Sune; Han, Yanlai

    2016-07-01

    Laser-induced fluorescence is an important technique to study photosynthesis and plants. Information on chlorophyll and other pigments can be obtained. We have been using a mobile laboratory in a Chinese experimental farm setting to study maize (Zea mays L.) leaves by reflectance and fluorescence measurements and correlated the spectroscopic signals to the amount of fertilizer supplied. Further, we studied five different species of maize using the remote monitoring of the fluorescence signatures obtained with the same mobile laboratory, but now in a laser radar remote-sensing configuration. The system separation from the target area was 50 m, and 355 nm pulsed excitation using the frequency-tripled output from an Nd:YAG laser was employed. Principal component analysis and linear discriminant analysis were combined to identify the different maize species using their fluorescence spectra. Likewise, the spectral signatures in reflectance and fluorescence frequently allowed us to separate different fertilizer levels applied to plants of the same species.

  17. Method for qualitative determination of measurement errors caused by sample fluorescence

    NASA Astrophysics Data System (ADS)

    Spooner, David L.

    1995-04-01

    Many of the papers and inks used to produce color hard copy products contain fluorescent materials. Most density and/or color instruments use the ratio of value of the light returned by the sample at each wavelength relative to that returned by a white calibration standard to derive the measurement value. Generally, no effort is made to differentiate between light reflected by the sample and light emitted by the sample due to fluorescence. The blue emitted light resulting from the inclusion of fluorescent whitening agents (FWA) in graphic arts materials is excited by violet and ultraviolet (UV) light in the instrument illumination source. Differences in instrument source UV intensity can cause significant differences in the blue reflectance values of FWA containing samples reported by the instrument measurement system. Standard reference materials (SRM) which contain known amounts of FWA are commercially available. These SRMs allow a semiquantitative assessment of the UV content of an instrument's illuminating source. A further refinement, using thin UV cutoff filters, allows the qualitative determination of the presence or absence of FWA in paper samples. We anticipate that with the use of other thin filters, measurement errors caused by visible light excited fluorescence of inks, particular yellows, will be possible.

  18. Quasi-continuous combined scattered light and fluorescence measurements: a novel measurement technique for shaken microtiter plates.

    PubMed

    Samorski, M; Müller-Newen, G; Büchs, J

    2005-10-05

    A novel quasi-continuous on-line measuring technique for shaken microtiter plates is presented. Light scattering as well as intracellular and/or protein fluorescence (e.g. NADH, YFP) is measured during the shaking procedure, thus allowing a process monitoring of 96 different simultaneous cultures in a microtiter plate. In contrast to existing measurement techniques, the shaking process does not have to be stopped to take the measurements, thus avoiding the corresponding interruption of the cultures' oxygen supply and any unpredictable effects on the cultures. Experiments were conducted with E. coli in LB, TB, and MOPS minimal medium and V. natriegens in modified LB and TB media. Intensity curves of scattered light and NADH fluorescence were used to distinguish different lag phases, growth velocities, or inoculation densities. Data from this new method corresponded well to the off-line measured optical densities and to the oxygen transfer rates of cultures run in simultaneously conducted shake flask experiments at equivalent oxygen transfer capacities. With the aid of yellow fluorescence protein fused to interleukin-6 the optimal induction time of an expressing E. coli strain could be determined by on-line monitoring of product formation. Thus, this measuring technique enables the researcher to evaluate and to discriminate different cultures on a screening level and to improve screening conditions, process development and scale-up.

  19. Air fluorescence efficiency measurements for AIRWATCH based mission: Experimental set-up

    SciTech Connect

    Biondo, B.; Catalano, O.; Celi, F.; Fazio, G.; Giarrusso, S.; La Rosa, G.; Mangano, A.; Bonanno, G.; Cosentino, R.; Di Benedetto, R.; Scuderi, S.; Richiusa, G.; Gregorio, A.

    1998-06-15

    In the framework of the AIRWATCH project we present an experimental set-up to measure the efficiency of the UV fluorescence production of the air using hard X-ray stimulus. The measures will be carried out at different pressure and temperature to emulate the same condition of the upper layers of the atmosphere where X-ray and gamma ray photons of Gamma Ray Bursts are absorbed.

  20. Time-Resolved Fluorescence Anisotropy of Bicyclo[1.1.1]pentane/Tolane-Based Molecular Rods Included in Tris(o-phenylenedioxy)cyclotriphosphazene (TPP)

    PubMed Central

    2015-01-01

    We examine the fluorescence anisotropy of rod-shaped guests held inside the channels of tris(o-phenylenedioxy)cyclotriphosphazene (TPP) host nanocrystals, characterized by powder X-ray diffraction and solid state NMR spectroscopy. We address two issues: (i) are light polarization measurements on an aqueous colloidal solution of TPP nanocrystals meaningful, or is depolarization by scattering excessive? (ii) Can measurements of the rotational mobility of the included guests be performed at low enough loading levels to suppress depolarization by intercrystallite energy transfer? We find that meaningful measurements are possible and demonstrate that the long axis of molecular rods included in TPP channels performs negligible vibrational motion. PMID:25937858

  1. Time-Resolved Fluorescence Anisotropy of Bicyclo[1.1.1]pentane/Tolane-Based Molecular Rods Included in Tris(o-phenylenedioxy)cyclotriphosphazene (TPP).

    PubMed

    Cipolloni, Marco; Kaleta, Jiří; Mašát, Milan; Dron, Paul I; Shen, Yongqiang; Zhao, Ke; Rogers, Charles T; Shoemaker, Richard K; Michl, Josef

    2015-04-23

    We examine the fluorescence anisotropy of rod-shaped guests held inside the channels of tris(o-phenylenedioxy)cyclotriphosphazene (TPP) host nanocrystals, characterized by powder X-ray diffraction and solid state NMR spectroscopy. We address two issues: (i) are light polarization measurements on an aqueous colloidal solution of TPP nanocrystals meaningful, or is depolarization by scattering excessive? (ii) Can measurements of the rotational mobility of the included guests be performed at low enough loading levels to suppress depolarization by intercrystallite energy transfer? We find that meaningful measurements are possible and demonstrate that the long axis of molecular rods included in TPP channels performs negligible vibrational motion.

  2. Electron beam depolarization in a damping ring

    SciTech Connect

    Minty, M.

    1993-04-01

    Depolarization of a polarized electron beam injected into a damping ring is analyzed by extending calculations conventionally applied to proton synchrotrons. Synchrotron radiation in an electron ring gives rise to both polarizing and depolarizing effects. In a damping ring, the beam is stored for a time much less than the time for self polarization. Spin flip radiation may therefore be neglected. Synchrotron radiation without spin flips, however, must be considered as the resonance strength depends on the vertical betatron oscillation amplitude which changes as the electron beam is radiation damped. An expression for the beam polarization at extraction is derived which takes into account radiation damping. The results are applied to the electron ring at the Stanford Linear Collider and are compared with numerical matrix formalisms.

  3. A scattering model for rain depolarization

    NASA Technical Reports Server (NTRS)

    Wiley, P. H.; Stutzman, W. L.; Bostian, C. W.

    1973-01-01

    A method is presented for calculating the amount of depolarization caused by precipitation for a propagation path. In the model the effects of each scatterer and their interactions are accounted for by using a series of simplifying steps. It is necessary only to know the forward scattering properties of a single scatterer. For the case of rain the results of this model for attenuation, differential phase shift, and cross polarization agree very well with the results of the only other model available, that of differential attenuation and differential phase shift. Calculations presented here show that horizontal polarization is more sensitive to depolarization than is vertical polarization for small rain drop canting angle changes. This effect increases with increasing path length.

  4. Aquatic and terrestrial optical measurements - laser induced fluorescence technique (ATOM-LIFT): Summer 1997 field measurement campaign

    NASA Astrophysics Data System (ADS)

    McMurtrey, James E., III; Cecchi, Giovanna; Chappelle, Emmett W.; Kim, Moon S.; Bazzani, Marco; Corp, Lawrence A.

    1998-07-01

    A joint IROE-CNR, NASA/GSFC, and USDA/ARS measurement campaign was conducted in Italy for a three week period in July, 1997. The campaign was split into two parts: the first part for aquatic vegetation studies and the second part for terrestrial vegetation studies. The main objective of the campaign was to study optical properties of intact plant material as it relates to photosynthetic activity of living vegetation. The aquatic studies were carried out at an aquarium-laboratory in the seashore city of Livorno on the West coast of Italy. The investigations involved an important sea grass species that is native to the Mediterranean Sea. The terrestrial studies were carried out Northeast of the Town of St. Stefano di Cadore (Belluno), Italy. Measurements were taken in a wooded site at an Italian Department of Forestry Station on species of natural alpine vegetation. Instrumentation available for the studies were the Italian Fluorescence Light Detection And Ranging (FLIDAR) System, the NASA/USDA Fluorescence Imaging System (FIS), the Perkin Elmer Spectrofluorometer and LI-COR 6400 infrared gas exchange analyzer for photosynthesis measurements. Preliminary evaluations, analysis, and summaries were made by personnel from both Italian and United Sates groups on data collected during the measurement campaign. The joint Italian/American data collection effort with Aquatic and Terrestrial Optical Measurements produced a range of data for characterizing the relationships between fluorescence and the photosynthetic potentials of vegetative scenes.

  5. Quantitative correlation between light depolarization and transport albedo of various porcine tissues.

    PubMed

    Alali, Sanaz; Ahmad, Manzoor; Kim, Anthony; Vurgun, Nasit; Wood, Michael F G; Vitkin, I Alex

    2012-04-01

    We present a quantitative study of depolarization in biological tissues and correlate it with measured optical properties (reduced scattering and absorption coefficients). Polarized light imaging was used to examine optically thick samples of both isotropic (liver, kidney cortex, and brain) and anisotropic (cardiac muscle, loin muscle, and tendon) pig tissues in transmission and reflection geometries. Depolarization (total, linear, and circular), as derived from polar decomposition of the measured tissue Mueller matrix, was shown to be related to the measured optical properties. We observed that depolarization increases with the transport albedo for isotropic and anisotropic tissues, independent of measurement geometry. For anisotropic tissues, depolarization was higher compared to isotropic tissues of similar transport albedo, indicating birefringence-caused depolarization effects. For tissues with large transport albedos (greater than ~0.97), backscattering geometry was preferred over transmission due to its greater retention of light polarization; this was not the case for tissues with lower transport albedo. Preferential preservation of linearly polarized light over circularly polarized light was seen in all tissue types and all measurement geometries, implying the dominance of Rayleigh-like scattering. The tabulated polarization properties of different tissue types and their links to bulk optical properties should prove useful in future polarimetric tissue characterization and imaging studies.

  6. Time-autocorrelated two-photon counting technique for time-resolved fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Borst, Walter L.; Liu, Lin-I.

    1999-01-01

    We describe a new instrumental technique for the excitation, acquisition, and analysis of fluorescence decays from a variety of substances, in the present case plastic scintillators. The fluorescence is excited by β particles from a radioactive source (100 μCi Sr-90). A random photon from the resulting fluorescence decay provides a trigger pulse to start a time-to-amplitude converter (TAC), while another random photon from the same β-excitation event provides the stop pulse. The optical components and geometry for detecting these two photons, i.e., the two photomultipliers (PMT), the filters, and the pulse counting system, are identical. As a consequence, the measured fluorescence signal is the autocorrelation function of the fluorescence decay from the sample. A delay line of 50 ns is inserted between the "stop" signal PMT and the TAC so that those "stop" pulses which arrive before "start pulses" also are recorded. Thus the acquired fluorescence signal versus time is symmetric about the delay time and contains twice as many counts as without delay. We call the new technique the "time-autocorrelated two-photon counting technique" (TATPC) in distinction to the conventional "time-correlated single-photon counting technique" (TCSPC). We compared both techniques with the same equipment and scintillators, where in the TCSPC case, a β particle is used for the start of the TAC instead of a random photon in the TATPC technique. We find that under similar experimental circumstances, the signal count rate with TATPC is about 50 times larger than with TCSPC. The new method is well suited for obtaining fluorescence decay times from plastic scintillators, which we use in this article to exemplify the technique. More generally, β-particle excitation in combination with TATPC should prove useful for materials with high energy levels or band gaps, which cannot be excited with pulsed lasers in the visible region. The length of our excitation pulse is less than 20 ps and is

  7. Fluid shear stress induces calcium transients in osteoblasts through depolarization of osteoblastic membrane.

    PubMed

    Sun, Junqing; Liu, Xifang; Tong, Jie; Sun, Lijun; Xu, Hao; Shi, Liang; Zhang, Jianbao

    2014-12-18

    Intracellular calcium transient ([Ca(2+)]i transient) induced by fluid shear stress (FSS) plays an important role in osteoblastic mechanotransduction. Changes of membrane potential usually affect [Ca(2+)]i level. Here, we sought to determine whether there was a relationship between membrane potential and FSS-induced [Ca(2+)]i transient in osteoblasts. Fluorescent dyes DiBAC4(3) and fura-2AM were respectively used to detect membrane potential and [Ca(2+)]i. Our results showed that FSS firstly induced depolarization of membrane potential and then a transient rising of [Ca(2+)]i in osteoblasts. There was a same threshold for FSS to induce depolarization of membrane potential and [Ca(2+)]i transients. Replacing extracellular Na(+) with tetraethylammonium or blocking stretch-activated channels (SACs) with gadolinium both effectively inhibited FSS-induced membrane depolarization and [Ca(2+)]i transients. However, voltage-activated K(+) channel inhibitor, 4-Aminopyridine, did not affect these responses. Removing extracellular Ca(2+) or blocking of L-type voltage-sensitive Ca(2+) channels (L-VSCCs) with nifedipine inhibited FSS-induced [Ca(2+)]i transients in osteoblasts too. Quantifying membrane potential with patch clamp showed that the resting potential of osteoblasts was -43.3mV and the depolarization induced by FSS was about 44mV. Voltage clamp indicated that this depolarization was enough to activated L-VSCCs in osteoblasts. These results suggested a time line of Ca(2+) mobilization wherein FSS activated SACs to promote Na(+) entry to depolarize membrane that, in turn, activated L-VSCCs and Ca(2+) influx though L-VSCCs switched on [Ca(2+)]i response in osteoblasts.

  8. Role of membrane depolarization and extracellular calcium in increased complement receptor expression during neutrophil (PMN) activation

    SciTech Connect

    Berger, M.; Wetzler, E.; Birx, D.L.

    1986-03-05

    During PMN activation the surface expression of receptors (R) for C3b and C3bi increases rapidly. This is necessary for optimal cell adhesion, migration, and phagocytosis. Following stimulation with fMLP or LTB-4, the increased expression of C3bR depends only on the Ca/sup + +/ released from intracellular stores and is not inhibited by 5mM EDTA, while the increase in C3biR also requires extracellular Ca/sup + +/. CR expression also increases when the PMN are depolarized with 140 mM K/sup +/, but with this stimulus, EDTA inhibits C3bR by 67% and C3biR 100%, suggesting that intracellular Ca/sup + +/ stores may not be released. Pertussis toxin caused dose-dependent inhibition of both CR responses to fMLP and also inhibited the increases in both CR induced by K/sup +/. Membrane depolarization (monitored by di-O-C5 fluorescence) due to fMLP was similarly inhibited by toxin but the depolarization due to K/sup +/ was not. The dose of phorbol myristate acetate that maximally increased CR expression, 0.1 ng/ml, did not depolarize the membrane. These results suggest that membrane depolarization is neither necessary nor sufficient for increased CR expression. A Ca/sup + +/ and GTP binding protein-dependent enzyme such as phospholipase C is necessary to the amplify initial signals generated either by release of Ca/sup + +/ stores or by opening voltage dependent Ca/sup + +/ channels following membrane depolarization.

  9. Development and use of chlorotetracycline fluorescence as a measurement assay of chloroplast envelope-bound mg.

    PubMed

    Gupta, A S; Berkowitz, G A

    1989-03-01

    Experiments were conducted to develop chlorotetracycline (CTC) fluorescence as an assay of Mg(2+) bound to the envelope of the intact chloroplast. This assay technique has been widely used to measure envelope associated divalent cations in animal cell and subcellular systems, but has not been used with chloroplasts. Chloroplast envelope-associated Mg(2+) was altered by pretreatment with Mg(2+) and divalent cation chelating agents and by additions of Mg(2+) to the CTC assay medium. Results indicated that for a given chloroplast preparation, relative changes in envelope-associated Mg(2+) can be effectively monitored with CTC fluorescence. It was concluded that the limitations of this assay system are: (a) chlorophyll strongly quenches CTC fluorescence signal, so a constant chlorophyll concentration must be maintained, (b) measurements must be made quickly, and (c) use of the technique to compare different chloroplast preparations may not be valid. Studies with (28)Mg(2+) confirmed our interpretation of the fluorescence results, and also suggested that the chloroplast envelope is fairly impermeable to Mg(2+). It was concluded that changes in Mg(2+) associated with the chloroplast due to incubation of plastids in solutions containing up to 5 millimolar Mg(2+) may be exclusively due to increased envelope-associated Mg(2+). The CTC assay was used in experiments to demonstrate that increases in chloroplast envelope-associated Mg(2+) inhibit photosynthetic capacity. This inhibition can be partially overcome by the presence of K(+) in the photosynthetic reaction media.

  10. Fluorescence lifetime imaging to quantify sub-cellular oxygen measurements in live macrophage during bacterial invasion

    NASA Astrophysics Data System (ADS)

    Dragavon, Joe; Amiri, Megdouda; Marteyn, Benoit; Sansonetti, Philipe; Shorte, Spencer

    2011-03-01

    Fluorophore concentration, the surrounding microenvironment, and photobleaching greatly influence the fluorescence intensity of a fluorophore, increasing the difficulty to directly observe micro-environmental factors such as pH and oxygen. However, the fluorescence lifetime of a fluorophore is essentially independent of both the fluorophore concentration and photobleaching, providing a viable alternative to intensity measurements. The development of fluorescence lifetime imaging (FLI) allows for the direct measurement of the microenvironment surrounding a fluorophore. Pt-porphyrin is a fluorophore whose optical properties include a very stable triplet excited state. This energy level overlaps strongly with the ground triplet state of oxygen, making the phosphorescent lifetime directly proportional to the surrounding oxygen concentration. Initial experiments using this fluorophore involved the use of individual microwells coated with the porphyrin. Cells were allowed to enter the micro-wells before being sealed to create a diffusionally isolated volume. The decrease in the extracellular oxygen concentration was observed using FLI. However, this isolation technique provides only the consumption rate but cannot indicate the subcellular oxygen distribution. To improve upon this, live macrophages are loaded with the porphyrin and the fluorescence lifetime determined using a Lambert Instruments Lifa-X FLI system. Initial results indicate that an increase in subcellular oxygen is observed upon initial exposure to invasive bacteria. A substantial decrease in oxygen is observed after about 1 hour of exposure. The cells remain in this deoxygenated state until the bacteria are removed or cell death occurs.

  11. Dual-emissive fluorescence measurements of hydroxyl radicals using a coumarin-activated silica nanohybrid probe.

    PubMed

    Liu, Saisai; Zhao, Jun; Zhang, Kui; Yang, Lei; Sun, Mingtai; Yu, Huan; Yan, Yehan; Zhang, Yajun; Wu, Lijun; Wang, Suhua

    2016-04-07

    This work reports a novel dual-emissive fluorescent probe based on dye hybrid silica nanoparticles for ratiometric measurement of the hydroxyl radical (˙OH). In the probe sensing system, the blue emission of coumarin dye (coumarin-3-carboxylic acid, CCA) immobilized on the nanoparticle surface is selectively enhanced by ˙OH due to the formation of a coumarin hydroxylation product with strong fluorescence, whereas the emission of red fluorescent dye encapsulated in the silica nanoparticle is insensitive to ˙OH as a self-referencing signal, and so the probe provides a good quantitative analysis based on ratiometric fluorescence measurement with a detection limit of 1.65 μM. Moreover, the probe also shows high selectivity for ˙OH determination against metal ions, other reactive oxygen species and biological species. More importantly, it exhibits low cytotoxicity and high biocompatibility in living cells, and has been successfully used for cellular imaging of ˙OH, showing its promising application for monitoring of intracellular ˙OH signaling events.

  12. Microlensed dual-fiber probe for depth-resolved fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Choi, Hae Young; Ryu, Seon Young; Kim, Jae Young; Kim, Geon Hee; Park, Seong Jun; Lee, Byeong Ha; Chang, Ki Soo

    2011-07-01

    We propose and demonstrate a compact microlensed dual-fiber probe that has a good collection efficiency and a high depth-resolution ability for fluorescence measurements. The probe is formed with a conventional fusion splicer creating a common focusing lens on two fibers placed side by side. The collection efficiency of the fabricated probe was evaluated by measuring the fluorescence signal of a fresh ginkgo leaf. It was shown experimentally that the proposed probe could effectively collect the fluorescence signal with a six-fold increase compared to that of a general flat-tipped probe. The beam propagation method was used to design a probe with an optimized working distance and an improved resolving depth. It was found that the working distance depends mainly on the radius of curvature of the lens, whereas the resolving depth is determined by the core diameters of the illumination and collection fibers. The depth-resolved ability of probes with working distances of ~100 μm and 300 μm was validated by using a two-layer tissue phantom. The experimental results demonstrate that the microlensed dual-fiber probe has the potential to facilitate depth-resolved fluorescence detection of epithelial tissue.

  13. Spatially resolved density and ionization measurements of shocked foams using x-ray fluorescence

    DOE PAGES

    MacDonald, M. J.; Keiter, P. A.; Montgomery, D. S.; ...

    2016-09-22

    We present experiments at the Trident laser facility demonstrating the use of x-ray fluorescence (XRF) to simultaneously measure density, ionization state populations, and electron temperature in shocked foams. An imaging x-ray spectrometer was used to obtain spatially-resolved measurements of Ti K-more » $$\\alpha$$ emission. Density profiles were measured from K-$$\\alpha$$ intensity. Ti ionization state distributions and electron temperatures were inferred by fitting K-$$\\alpha$$ spectra to spectra from CRETIN simulations. This study shows that XRF provides a powerful tool to complement other diagnostics to make equation of state measurements of shocked materials containing a suitable tracer element.« less

  14. Spatially resolved density and ionization measurements of shocked foams using x-ray fluorescence

    SciTech Connect

    MacDonald, M. J.; Keiter, P. A.; Montgomery, D. S.; Scott, H. A.; Biener, M. M.; Fein, J. R.; Fournier, K. B.; Gamboa, E. J.; Kemp, G. E.; Klein, S. R.; Kuranz, C. C.; LeFevre, H. J.; Manuel, M. J. -E.; Wan, W. C.; Drake, R. P.

    2016-09-22

    We present experiments at the Trident laser facility demonstrating the use of x-ray fluorescence (XRF) to simultaneously measure density, ionization state populations, and electron temperature in shocked foams. An imaging x-ray spectrometer was used to obtain spatially-resolved measurements of Ti K-$\\alpha$ emission. Density profiles were measured from K-$\\alpha$ intensity. Ti ionization state distributions and electron temperatures were inferred by fitting K-$\\alpha$ spectra to spectra from CRETIN simulations. This study shows that XRF provides a powerful tool to complement other diagnostics to make equation of state measurements of shocked materials containing a suitable tracer element.

  15. Molecular Diffusion in Plasma Membranes of Primary Lymphocytes Measured by Fluorescence Correlation Spectroscopy.

    PubMed

    Staaf, Elina; Bagawath-Singh, Sunitha; Johansson, Sofia

    2017-02-01

    Fluorescence correlation spectroscopy (FCS) is a powerful technique for studying the diffusion of molecules within biological membranes with high spatial and temporal resolution. FCS can quantify the molecular concentration and diffusion coefficient of fluorescently labeled molecules in the cell membrane. This technique has the ability to explore the molecular diffusion characteristics of molecules in the plasma membrane of immune cells in steady state (i.e., without processes affecting the result during the actual measurement time). FCS is suitable for studying the diffusion of proteins that are expressed at levels typical for most endogenous proteins. Here, a straightforward and robust method to determine the diffusion rate of cell membrane proteins on primary lymphocytes is demonstrated. An effective way to perform measurements on antibody-stained live cells and commonly occurring observations after acquisition are described. The recent advancements in the development of photo-stable fluorescent dyes can be utilized by conjugating the antibodies of interest to appropriate dyes that do not bleach extensively during the measurements. Additionally, this allows for the detection of slowly diffusing entities, which is a common feature of proteins expressed in cell membranes. The analysis procedure to extract molecular concentration and diffusion parameters from the generated autocorrelation curves is highlighted. In summary, a basic protocol for FCS measurements is provided; it can be followed by immunologists with an understanding of confocal microscopy but with no other previous experience of techniques for measuring dynamic parameters, such as molecular diffusion rates.

  16. Measuring protein dynamics in live cells: protocols and practical considerations for fluorescence fluctuation microscopy

    PubMed Central

    Youker, Robert T.; Teng, Haibing

    2014-01-01

    Abstract. Quantitative analysis of protein complex stoichiometries and mobilities are critical for elucidating the mechanisms that regulate cellular pathways. Fluorescence fluctuation spectroscopy (FFS) techniques can measure protein dynamics, such as diffusion coefficients and formation of complexes, with extraordinary precision and sensitivity. Complete calibration and characterization of the microscope instrument is necessary in order to avoid artifacts during data acquisition and to capitalize on the full capabilities of FFS techniques. We provide an overview of the theory behind FFS techniques, discuss calibration procedures, provide protocols, and give practical considerations for performing FFS experiments. One important parameter recovered from FFS measurements is the relative molecular brightness that can correlate with oligomerization. Three methods for measuring molecular brightness (fluorescence correlation spectroscopy, photon-counting histogram, and number and brightness analysis) recover similar values when measuring samples under ideal conditions in vitro. However, examples are given illustrating that these different methods used for calculating molecular brightness of fluorescent molecules in cells are not always equivalent. Methods relying on spot measurements are more prone to bleaching and movement artifacts that can lead to underestimation of brightness values. We advocate for the use of multiple FFS techniques to study molecular brightnesses to overcome and compliment limitations of individual techniques. PMID:25260867

  17. Concentration Measurements in a Cold Flow Model Annular Combustor Using Laser Induced Fluorescence

    NASA Technical Reports Server (NTRS)

    Morgan, Douglas C.

    1996-01-01

    A nonintrusive concentration measurement method is developed for determining the concentration distribution in a complex flow field. The measurement method consists of marking a liquid flow with a water soluble fluorescent dye. The dye is excited by a two dimensional sheet of laser light. The fluorescent intensity is shown to be proportional to the relative concentration level. The fluorescent field is recorded on a video cassette recorder through a video camera. The recorded images are analyzed with image processing hardware and software to obtain intensity levels. Mean and root mean square (rms) values are calculated from these intensity levels. The method is tested on a single round turbulent jet because previous concentration measurements have been made on this configuration by other investigators. The previous results were used to comparison to qualify the current method. These comparisons showed that this method provides satisfactory results. 'Me concentration measurement system was used to measure the concentrations in the complex flow field of a model gas turbine annular combustor. The model annular combustor consists of opposing primary jets and an annular jet which discharges perpendicular to the primary jets. The mixing between the different jet flows can be visualized from the calculated mean and rms profiles. Concentration field visualization images obtained from the processing provide further qualitative information about the flow field.

  18. Excitation-emission matrices measurements of human cutaneous lesions: tool for fluorescence origin

    NASA Astrophysics Data System (ADS)

    Zhelyazkova, A.; Borisova, E.; Angelova, L.; Pavlova, E.; Keremedchiev, M.

    2013-11-01

    The light induced fluorescence (LIF) technique has the potential of providing real-time diagnosis of malignant and premalignant skin tissue; however, human skin is a multilayered and inhomogeneous organ with different optical properties that complicate the analysis of cutaneous fluorescence spectra. In spite of the difficulties related to the detection and analysis of fluorescent data from skin lesions, this technique is among the most widely applied techniques in laboratorial and pre-clinical investigations for early skin neoplasia diagnosis. The important point is to evaluate all sources of intrinsic fluorescence and find any significant alterations distinguishing the normal skin from a cancerous state of the tissue; this would make the autofluorescence signal obtained useful for the development of a non-invasive diagnostic tool for the dermatological practice. Our investigations presented here were based on ex vivo point-by-point measurements of excitation-emission matrices (EEM) from excised tumor lesions and the surrounding skin taken during the daily clinical practice of Queen Jiovanna- ISUL University Hospital, Sofia, the local Ethical Committee's approval having already been obtained. The fluorescence emission was measured between 300 nm and 800 nm using excitation in the 280-440 nm spectral range. In the process of excitation-emission matrices (EEM) measurements we could establish the origin of the autofluorescence and the compounds related by assigning the excitation and emission maxima obtained during the experiments. The EEM were compared for normal human skin, basal cell carcinoma, squamous cell carcinoma, benign nevi and malignant melanoma lesions to obtain information for the most common skin malignancies and their precursors. The main spectral features and the applicability of the technique of autofluorescent spectroscopy of human skin in general as an initial diagnostic tool are discussed as well.

  19. Understanding Solar Induced Fluorescence: Building up from Leaf Scale Measurements (Invited)

    NASA Astrophysics Data System (ADS)

    Berry, J. A.; Van der Tol, C.; Frankenberg, C.; Joiner, J.; Guanter, L.

    2013-12-01

    Measurements of chlorophyll fluorescence have long been a key method for probing the mechanisms of photosynthesis in laboratory studies. Recent advances in satellite spectroscopy have enabled retrieval of chlorophyll fluorescence from terrestrial ecosystems at a global scale. Analyses of these retrievals show promising potential as an indicator of photosynthetic rate and of its response to environmental stress. This talk will explore the mechanistic basis for interpreting and modeling of solar induced chlorophyll fluorescence ( SIF). SIF is essentially a leak of photons from photosynthetic membranes, and it is, therefore, related to the flux of photons absorbed by chlorophyll and to biochemical processes that regulate the processing of these photons in macromolecuar complexes associated with photosystem II. Thus: SIF = aPAR * φF, where aPAR is the flux of absorbed photosynthetically active radiation and φF, is the yield (light-use efficiency) of fluorescence. (For simplicity we will ignore the transport of fluorescence from its sources to the sensor for the moment). This expression for SIF is similar to a common expression for photosynthesis or gross primary productivity, GPP = aPAR * LUE, where LUE, is the light-use-efficiency for CO2 uptake. These equations can be combined and simplified to illustrate the relationship between SIF and GPP; GPP =SIF *LUE / φF. The extent to which GPP is proportional to SIF hinges on the stability of the ratio, LUE / φF, and it leads to the key question to be considered here. What is the relationship between the light-use-efficiency for photosynthesis and that for fluorescence? Satellite retrievals of SIF occur at mid-day, conditions where the capacity for CO2 fixation usually limits the rate of photosynthesis. Under this condition the rate of the photo-acts must be down-regulated to protect from photo-damage. This balancing the source with the sink is accomplished by opening non-photochemical trapping centers that compete with

  20. An Intelligent Optical Dissolved Oxygen Measurement Method Based on a Fluorescent Quenching Mechanism.

    PubMed

    Li, Fengmei; Wei, Yaoguang; Chen, Yingyi; Li, Daoliang; Zhang, Xu

    2015-12-09

    Dissolved oxygen (DO) is a key factor that influences the healthy growth of fishes in aquaculture. The DO content changes with the aquatic environment and should therefore be monitored online. However, traditional measurement methods, such as iodometry and other chemical analysis methods, are not suitable for online monitoring. The Clark method is not stable enough for extended periods of monitoring. To solve these problems, this paper proposes an intelligent DO measurement method based on the fluorescence quenching mechanism. The measurement system is composed of fluorescent quenching detection, signal conditioning, intelligent processing, and power supply modules. The optical probe adopts the fluorescent quenching mechanism to detect the DO content and solves the problem, whereas traditional chemical methods are easily influenced by the environment. The optical probe contains a thermistor and dual excitation sources to isolate visible parasitic light and execute a compensation strategy. The intelligent processing module adopts the IEEE 1451.2 standard and realizes intelligent compensation. Experimental results show that the optical measurement method is stable, accurate, and suitable for online DO monitoring in aquaculture applications.

  1. An Intelligent Optical Dissolved Oxygen Measurement Method Based on a Fluorescent Quenching Mechanism

    PubMed Central

    Li, Fengmei; Wei, Yaoguang; Chen, Yingyi; Li, Daoliang; Zhang, Xu

    2015-01-01

    Dissolved oxygen (DO) is a key factor that influences the healthy growth of fishes in aquaculture. The DO content changes with the aquatic environment and should therefore be monitored online. However, traditional measurement methods, such as iodometry and other chemical analysis methods, are not suitable for online monitoring. The Clark method is not stable enough for extended periods of monitoring. To solve these problems, this paper proposes an intelligent DO measurement method based on the fluorescence quenching mechanism. The measurement system is composed of fluorescent quenching detection, signal conditioning, intelligent processing, and power supply modules. The optical probe adopts the fluorescent quenching mechanism to detect the DO content and solves the problem, whereas traditional chemical methods are easily influenced by the environment. The optical probe contains a thermistor and dual excitation sources to isolate visible parasitic light and execute a compensation strategy. The intelligent processing module adopts the IEEE 1451.2 standard and realizes intelligent compensation. Experimental results show that the optical measurement method is stable, accurate, and suitable for online DO monitoring in aquaculture applications. PMID:26690176

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

    PubMed

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

    2015-11-01

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

  3. Measurement of Small Molecule Diffusion in Carbon Dioxide Swollen Polymers using Fluorescence Nonradiative Energy Transfer

    NASA Astrophysics Data System (ADS)

    Watkins, James; Gupta, Ravi; Ramachandrarao, Vijay

    2001-03-01

    Diffusion coefficients of molecular probes in CO2-swollen polystyrene films were measured in situ using high-pressure fluorescence nonradiative energy transfer (NRET). Specifically, the diffusivities of decacyclene and BPEA (9,10- bis phenyl ethnyl Anthracene), relatively large fluorescence acceptor probes, were determined in real time using pyrene-labeled polystyrene as the corresponding energy donor in carbon dioxide-plasticized films. Decacyclene diffusivities were measured at 65 and 80 C and carbon dioxide pressures ranging from 62 to 144 bar, conditions near and well above the previously reported, solvent-depressed glass transition of polystyrene. Decacyclene diffusivity shows an increase of over 5 orders of magnitude upon CO2 sorption relative to the PS glass at ambient pressure and equivalent temperatures. BPEA exhibits similar behavior but diffuses about an order of magnitude faster than decacyclene in CO2 plasticized polystyrene under similar conditions.

  4. [Measurement and analysis of lead in soil using X-ray fluorescence spectroscopy].

    PubMed

    Zhang, Rong; Zhang, Yu-Jun; Zhang, Wei; Chen, Dong; Yu, Xiao-Ya; Gao, Yan-Wei

    2013-02-01

    The present paper analyzed the characteristics of X-ray fluorescence spectroscopy (XRF) of metal element lead in soil using the NITON XLt793 portable X-ray fluorescence spectra of heavy metal analyzer under laboratory conditions. The characteristic spectral lines of L(alpha) (energy: 10. 55 keV) and L(beta) (energy: 12. 61 keV) with different matrix elements were selected respectively for lead in the experiment. By measuring the intensities of the characteristic spectral line with different Pb concentration, the results demonstrate that the relation between concentration [mass fraction 10 x 10(-6) - 1 800 x 10(-6)] of Pb element and the intensity of the characteristic spectrum is well linear. The calibration curve of Pb was plotted based on the different concentration measurement results, and the limit of detection of 7.89 x 10(-6) was obtained for Pb in soil.

  5. In-Situ Silver Acetylide Silver Nitrate Explosive Deposition Measurements Using X-Ray Fluorescence.

    SciTech Connect

    Covert, Timothy Todd

    2014-09-01

    The Light Initiated High Explosive facility utilized a spray deposited coating of silver acetylide - silver nitrate explosive to impart a mechanical shock into targets of interest. A diagnostic was required to measure the explosive deposition in - situ. An X - ray fluorescence spectrometer was deployed at the facility. A measurement methodology was developed to measure the explosive quantity with sufficient accuracy. Through the use of a tin reference material under the silver based explosive, a field calibration relationship has been developed with a standard deviation of 3.2 % . The effect of the inserted tin material into the experiment configuration has been explored.

  6. Temperature measurements in hypersonic air flows using laser-induced O2 fluorescence

    NASA Technical Reports Server (NTRS)

    Laufer, Gabriel; Mckenzie, Robert L.

    1988-01-01

    An investigation is reported of the use of laser-induced fluorescence on oxygen for the measurement of air temperature and its fluctuations owing to turbulence in hypersonic wind tunnel flows. The results show that for temperatures higher than 60 K and densities higher than 0.01 amagat, the uncertainty in the temperature measurement can be less than 2 percent if it is limited by photon-statistical noise. The measurement is unaffected by collisional quenching and, if the laser fluence is kept below 1.5 J/sq cm, it is also unaffected by nonlinear effects which are associated with depletion of the absorbing states.

  7. Laser-Induced Fluorescence Measurements for Optical Single Atom Detection for Nuclear Astrophysics

    NASA Astrophysics Data System (ADS)

    Parzuchowski, Kristen; Singh, Jaideep; Wenzl, Jennifer; Frisbie, Dustin; Johnson, Maegan

    2016-09-01

    We propose a new highly selective detector to measure rare nuclear reactions relevant for nuclear astrophysics. Our primary interest is the 22Ne(α , n) 25Mg reaction, which is a primary source of neutrons for the s-process. Our proposed detector, in conjunction with a recoil separator, captures the recoil products resulting from the reaction in a cryogenically frozen thin film of solid neon. The fluorescence spectra of the captured atoms is shifted from the absorption spectra by hundreds of nanometers. This allows for the optical detection of individual fluorescence photons against a background of intense excitation light. We will describe our initial studies of laser-induced fluorescence of Yb and Mg in solid Ne. Neon is an attractive medium because it is optically transparent and provides efficient, pure, stable, & chemically inert confinement for a wide variety of atomic and molecular species. Yb is used as a test atom because of its similar atomic structure to Mg and much brighter fluorescence signal. This work is supported by funds from Michigan State University.

  8. Molecular rotor measures viscosity of live cells via fluorescence lifetime imaging.

    PubMed

    Kuimova, Marina K; Yahioglu, Gokhan; Levitt, James A; Suhling, Klaus

    2008-05-28

    The fluorescence intensity and lifetime of the 4,4'-difluoro-4-bora-5-(p-oxoalkyl)phenyl-3a,4a-diaza-s-indacene (1) show a strong correlation with the viscosity of the medium due to the viscosity-dependent twisting of the 5-phenyl group, which gives access to the dark nonemissive excited state. We propose a sensitive and versatile method for measuring the local microviscosity in biological systems, based on the determination of the fluorescence lifetime of 1. Fluorescence lifetime imaging (FLIM) performed on live cells incubated with 1 demonstrates the distinct intracellular lifetime of the molecular rotor of 1.6 +/- 0.2 ns corresponding to the intracellular viscosity of ca. 140 cP. Time-resolved fluorescence anisotropy of 1 in cells confirms insignificant binding of the fluorophore. The viscosity value obtained in the present study is considerably higher than that of water and of cellular cytoplasm. The high viscosity of intracellular compartments is likely to play an important role in vital intracellular processes, including the rate of diffusion of reactive oxygen species, causing programmed cell destruction.

  9. Delayed fluorescence spectra of intact leaves photoexcited by sunlight measured with a multichannel Fourier-transform chemiluminescence spectrometer

    NASA Astrophysics Data System (ADS)

    Akita, Saeka; Yano, Ayako; Ishii, Hiroshi; Satoh, Chikahiro; Akai, Nobuyuki; Nakata, Munetaka

    2013-06-01

    Delayed fluorescence spectra of intact leaves of Green pak choi (Brassica rapa var. chinensis) were measured with a multichannel Fourier-transform chemiluminescence spectrometer, which we developed recently. The intact samples, photoexcited by sunlight without artificial light sources, showed delayed fluorescence around 740 nm with a lifetime of ˜6 s. The observed spectra were deconvoluted into two Gaussian bands: the delayed fluorescence from photosystem II and photosystem I complexes. Their relative intensities depended on the chlorophyll concentration, but their wavelengths were unchanged.

  10. Evaluation of Portable X-Ray Fluorescence (XRF) Analyzer for Zirconium-Thickness Measurements

    SciTech Connect

    Glenn Moore

    2013-09-01

    This Technical Evaluation Report provides details of preliminary testing/experiments performed using a handheld X-ray fluorescence analyzer. The analyzer will be utilized in upcoming fuel-foil-rolling optimization studies at the INL. The studies are being performed in support of DOE’s Office of Global Threat Reduction -- Reactor Conversion Subprogram. Details of the equipment used, operating parameters, and measurement results are provided in this report.

  11. Planar laser-induced fluorescence measurements of high-enthalpy free jet flow with nitric oxide

    NASA Technical Reports Server (NTRS)

    Palmer, Jennifer L.; Mcmillin, Brian K.; Hanson, Ronald K.

    1992-01-01

    Planar laser-induced fluorescence (PLIF) measurements of property fields in a high-enthalpy, supersonic, underexpanded free jet generated in a reflection-type shock tunnel are reported. PLIF images showing velocity and temperature sensitivity are presented. The inferred radial velocity and relative rotational temperature fields are found to be in agreement with those predicted by a numerical simulation of the flowfield using the method of characteristics.

  12. Study of excitation transfer in laser dye mixtures by direct measurement of fluorescence lifetime

    NASA Technical Reports Server (NTRS)

    Lin, C.; Dienes, A.

    1973-01-01

    By directly measuring the donor fluorescence lifetime as a function of acceptor concentration in the laser dye mixture Rhodamine 6G-Cresyl violet, we found that the Stern-Volmer relation is obeyed, from which the rate of excitation transfer is determined. The experimental results indicate that the dominant mechanism responsible for the efficient excitation transfer is that of resonance transfer due to long range dipole-dipole interaction.

  13. Retrievals of Aerosol and Cloud Particle Microphysics Using Polarization and Depolarization Techniques

    NASA Technical Reports Server (NTRS)

    Mishchenko, Michael; Hansen, James E. (Technical Monitor)

    2001-01-01

    The recent availability of theoretical techniques for computing single and multiple scattering of light by realistic polydispersions of spherical and nonspherical particles and the strong dependence of the Stokes scattering matrix on particle size, shape, and refractive index make polarization and depolarization measurements a powerful particle characterization tool. In this presentation I will describe recent applications of photopolarimetric and lidar depolarization measurements to remote sensing characterization of tropospheric aerosols, polar stratospheric clouds (PSCs), and contrails. The talk will include (1) a short theoretical overview of the effects of particle microphysics on particle single-scattering characteristics; (2) the use of multi-angle multi-spectral photopolarimetry to retrieve the optical thickness, size distribution, refractive index, and number concentration of tropospheric aerosols over the ocean surface; and (3) the application of the T-matrix method to constraining the PSC and contrail particle microphysics using multi-spectral measurements of lidar backscatter and depolarization.

  14. Front-face fluorescence measurement of photosensitizers and lipid oxidation products during the photooxidation of butter.

    PubMed

    Veberg, A; Olsen, E; Nilsen, A N; Wold, J P

    2007-05-01

    This paper shows that fluorescence spectroscopy can measure both degradation of photosensitizers and formation of lipid oxidation products in light-exposed butter. The photosensitizers were already notably degraded after 4 h of light exposure, whereas fluorescent lipid oxidation products were detected after 5 d. The fluorescence measurements were highly correlated with sensory assessments of acidic and rancid flavor. Photosensitizer degradation is therefore a promising indirect indicator of the onset of lipid oxidation in butter. Sensory analysis and measurement of peroxide value showed that the level of lipid oxidation was significantly higher for butter stored in air compared with butter stored in nitrogen (N2). This might be explained by the formation of singlet oxygen from direct photooxidation and type II photosensitized oxidation. Addition of the singlet oxygen quencher beta-carotene reduced the rancid flavor intensity in the air and N2 packages from 9.0 to 4.9 and from 6.5 to 4.7, respectively. Results indicate that lipid oxidation in the butter stored in N2 was mainly caused by type I photosensitized reactions, because addition of beta-carotene had little effect on the rancid flavor intensity.

  15. Measuring precise diffusion coefficients with two-focus fluorescence correlation spectroscopy

    NASA Astrophysics Data System (ADS)

    Dertinger, Thomas; Gregor, Ingo; von der Hocht, Iris; Erdmann, Rainer; Krämer, Benedikt; Koberling, Felix; Hartmann, Rudolf; Enderlein, Jörg

    2006-02-01

    We present a new method for precisely measuring diffusion coefficients of fluorescent molecules at nanomolar concentrations. The method is based on a modified Fluorescence Correlation Spectroscopy (FCS)-setup which is robust against many artifacts that are inherent to standard FCS 1, 2. The core idea of the new method is the introduction of an external ruler by generating two laterally shifted and overlapping laser foci at a fixed and known distance. Data fitting is facilitated by ab initio calculations of resulting correlation curves and subsequent affine transformation of these curves to match the measured auto- and cross-correlation functions. The affine transformation coefficient along the time axis then directly yields the correct diffusion coefficient. This method is not relying on the rather inexact assumption of a 3D Gaussian shaped detection volume. We measured the diffusion coefficient of the red fluorescent dye Atto-655 (Atto-Tec GmbH) in water and compared the obtained value with results from Gradient Pulsed Field NMR (GPF-NMR).

  16. Fluorescence measurements of activity associated with a molecularly imprinted polymer imprinted to dipicolinic acid

    NASA Astrophysics Data System (ADS)

    Anderson, John; Pestov, Dmitry; Fischer, Robert L.; Webb, Stanley; Tepper, Gary C.

    2004-03-01

    Steady state and lifetime fluorescence measurements were acquired to measure the binding activity associated with molecularly imprinted polymer (MIP) microparticles imprinted to dipicolinic acid. Dipicolinic acid is a unique compound associated with the sporulation phase of spore-forming bacteria (e.g., genus Bacillus and Clostridium). Vinylic monomers were polymerized in a dimethylformamide solution containing the dipicolinic acid as a template. The resulting MIP was then pulverized and size selected into small microscale particles. Samplers were adapted incorporating the MIP particles within a dialyzer (500 MW). Tests were run on replicate samples of biologically active cultures representing both stationary phase and sporulation post fermentation products in standard media. The permeability of the membrane permitted diffusion of lighter molecular weight constituents from media effluents to enter the dialyzer chamber and contact the MIP. Extractions of the media were measured using steady state and lifetime fluorescence. Results showed dramatic steady state fluorescence changes as a function of excitation, emission and intensity and an estimated lifetime of 5.8 ns.

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

    PubMed

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

    2008-09-15

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

  18. Linear depolarization of lidar returns by aged smoke particles.

    PubMed

    Mishchenko, Michael I; Dlugach, Janna M; Liu, Li

    2016-12-10

    We use the numerically exact (superposition) T-matrix method to analyze recent measurements of the backscattering linear depolarization ratio (LDR) for a plume of aged smoke at lidar wavelengths ranging from 355 to 1064 nm. We show that the unique spectral dependence of the measured LDRs can be modeled, but only by assuming expressly nonspherical morphologies of smoke particles containing substantial amounts of nonabsorbing (or weakly absorbing) refractory materials such as sulfates. Our results demonstrate that spectral backscattering LDR measurements can be indicative of the presence of morphologically complex smoke particles, but additional (e.g., passive polarimetric or bistatic lidar) measurements may be required for a definitive characterization of the particle morphology and composition.

  19. Mathematical analysis of depolarization block mediated by slow inactivation of fast sodium channels in midbrain dopamine neurons

    PubMed Central

    Qian, Kun; Yu, Na; Tucker, Kristal R.; Levitan, Edwin S.

    2014-01-01

    Dopamine neurons in freely moving rats often fire behaviorally relevant high-frequency bursts, but depolarization block limits the maximum steady firing rate of dopamine neurons in vitro to ∼10 Hz. Using a reduced model that faithfully reproduces the sodium current measured in these neurons, we show that adding an additional slow component of sodium channel inactivation, recently observed in these neurons, qualitatively changes in two different ways how the model enters into depolarization block. First, the slow time course of inactivation allows multiple spikes to be elicited during a strong depolarization prior to entry into depolarization block. Second, depolarization block occurs near or below the spike threshold, which ranges from −45 to −30 mV in vitro, because the additional slow component of inactivation negates the sodium window current. In the absence of the additional slow component of inactivation, this window current produces an N-shaped steady-state current-voltage (I-V) curve that prevents depolarization block in the experimentally observed voltage range near −40 mV. The time constant of recovery from slow inactivation during the interspike interval limits the maximum steady firing rate observed prior to entry into depolarization block. These qualitative features of the entry into depolarization block can be reversed experimentally by replacing the native sodium conductance with a virtual conductance lacking the slow component of inactivation. We show that the activation of NMDA and AMPA receptors can affect bursting and depolarization block in different ways, depending upon their relative contributions to depolarization versus to the total linear/nonlinear conductance. PMID:25185810

  20. Measurement and quantification of fluorescent changes in ocular tissue using a novel confocal instrument

    NASA Astrophysics Data System (ADS)

    Buttenschoen, Kim K.; Girkin, John M.; Daly, Daniel J.

    2014-05-01

    Our sight is a major contributor to our quality of life. The treatment of diseases like macular degeneration and glaucoma, however, presents a challenge as the delivery of medication to ocular tissue is not well understood. The instrument described here will help quantify targeted delivery by non-invasively and simultaneously measuring light reflected from and fluorescence excited in the eye, used as position marker and to track compounds respectively. The measurement concept has been proven by monitoring the diffusion of fluorescein and a pharmaceutical compound for treating open angle glaucoma in vitro in a cuvette and in ex vivo porcine eyes. To obtain a baseline of natural fluorescence we measured the change in corneal and crystalline lens autofluorescence in volunteers over a week. We furthermore present data on 3D ocular autofluorescence. Our results demonstrate the capability to measure the location and concentration of the compound of interest with high axial and temporal resolution of 178 μm and 0.6 s respectively. The current detection limit is 2 nM for fluorescein, and compounds with a quantum yield as low as 0.01 were measured to concentrations below 1 μM. The instrument has many applications in assessing the diffusion of fluorescent compounds through the eye and skin in vitro and in vivo, measuring autofluorescence of ocular tissues and reducing the number of animals needed for research. The instrument has the capability of being used both in the clinical and home care environment opening up the possibility of measuring controlled drug release in a patient friendly manner.

  1. Fluorescence dilution technique for measurement of albumin reflection coefficient in isolated glomeruli

    PubMed Central

    Fan, Fan; Chen, Chun Cheng Andy; Zhang, Jin; Schreck, Carlos M. N.; Williams, Jan M.; Hirata, Takashi; Sharma, Mukut; Beard, Daniel A.; Savin, Virginia J.; Roman, Richard J.

    2015-01-01

    This study describes a high-throughput fluorescence dilution technique to measure the albumin reflection coefficient (σAlb) of isolated glomeruli. Rats were injected with FITC-dextran 250 (75 mg/kg), and the glomeruli were isolated in a 6% BSA solution. Changes in the fluorescence of the glomerulus due to water influx in response to an imposed oncotic gradient was used to determine σAlb. Adjustment of the albumin concentration of the bath from 6 to 5, 4, 3, and 2% produced a 10, 25, 35, and 50% decrease in the fluorescence of the glomeruli. Pretreatment of glomeruli with protamine sulfate (2 mg/ml) or TGF-β1 (10 ng/ml) decreased σAlb from 1 to 0.54 and 0.48, respectively. Water and solute movement were modeled using Kedem-Katchalsky equations, and the measured responses closely fit the predicted behavior, indicating that loss of albumin by solvent drag or diffusion is negligible compared with the movement of water. We also found that σAlb was reduced by 17% in fawn hooded hypertensive rats, 33% in hypertensive Dahl salt-sensitive (SS) rats, 26% in streptozotocin-treated diabetic Dahl SS rats, and 21% in 6-mo old type II diabetic nephropathy rats relative to control Sprague-Dawley rats. The changes in glomerular permeability to albumin were correlated with the degree of proteinuria in these strains. These findings indicate that the fluorescence dilution technique can be used to measure σAlb in populations of isolated glomeruli and provides a means to assess the development of glomerular injury in hypertensive and diabetic models. PMID:26447220

  2. Spontaneous mechanical activity in depolarized frog ventricle

    PubMed Central

    1976-01-01

    Spontaneous mechanical activity can be produced in depolarized frog ventricle by bathing the tissue in a solution with low Na, Iow Ca, and high K+. The contractions can be inhibited by depleting the tissue of Ca first, but they are relatively insensitive to changes in either extracellular [Ca++] or [Ca++]/[Na+]2. They are terminated very rapidly by raising [Na+] to 40 mM. Local anesthetics enhance the spontaneous activity in proportion to the concentration of their free base form. These contractions occur relatively rhythmically for several hours. Since the preparation is multicellular, this suggests a mechanism for intercellular communication without change in membrane potential. PMID:822122

  3. Application of dual-focus fluorescence correlation spectroscopy to microfluidic flow-velocity measurement.

    PubMed

    Arbour, Tyler J; Enderlein, Jörg

    2010-05-21

    Several methods exist to measure and map fluid velocities in microfluidic devices, which are vital to understanding properties on the micro- and nano-scale. Fluorescence correlation spectroscopy (FCS) is a method traditionally exploited for its ability to measure molecular diffusion coefficients. However, several reports during the past decade have shown that FCS can also be successfully used to measure precise flow rates in microfluidics with very high spatial resolution, making it a competitive alternative to other common flow-measurement methods. In 2007 we introduced a modified version of conventional FCS that overcomes many of the artifacts troubling the standard technique. Here we show how the advantages of this method, called dual-focus FCS, extend to flow measurements. To do so, we have measured the velocity flow profile along the cross-section of a square-bore microfluidic channel and compared the result to the theoretical prediction.

  4. Mechanism of blue-light-induced plasma-membrane depolarization in etiolated cucumber hypocotyls

    NASA Technical Reports Server (NTRS)

    Spalding, E. P.; Cosgrove, D. J.

    1992-01-01

    A large, transient depolarization of the plasma membrane precedes the rapid blue-light (BL)-induced growth suppression in etiolated seedlings of Cucumis sativus L. The mechanism of this voltage transient was investigated by applying inhibitors of ion channels and the plasma-membrane H(+)-ATPase, by manipulating extracellular ion concentrations, and by measuring cell input resistance and ATP levels. The depolarizing phase was not affected by Ca(2+)-channel blockers (verapamil, La3+) or by reducing extracellular free Ca2+ by treatment with ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA). However, these treatments did reduce the rate of repolarization, indicating an inward movement of Ca2+ is involved. No effects of the K(+)-channel blocker tetraethylammonium (TEA+) were detected. Vanadate and KCN, used to inhibit the H(+)-ATPase, reduced or completely inhibited the BL-induced depolarization. Levels of ATP increased by 11-26% after 1-2 min of BL. Input resistance of trichrome cells, measured with double-barreled microelectrodes, remained constant during the onset of the depolarization but decreased as the membrane voltage became more positive than -90 mV. The results indicate that the depolarization mechanism initially involves inactivation of the H(+)-ATPase with subsequent transient activation of one or more types of ion channels.

  5. Resolution of heterogeneous fluorescence emission signals and decay lifetime measurement on fluorochrome-labeled cells by phase-sensitive FCM

    SciTech Connect

    Steinkamp, J.A.; Crissman, H.A.

    1993-02-01

    A phase-sensitive flow cytometer has been developed to resolve signals from heterogeneous fluorescence emission spectra and quantify fluorescence decay times on cells labeled with fluorescent dyes. This instrument combines flow cytometry (FCM) and fluorescence spectroscopy measurement principles to provide unique capabilities for making phase-resolved measurements on single cells in flow, while preserving conventional FCM measurement capabilities. Stained cells are analyzed as they pass through an intensity-modulated (sinusoid) laser excitation beam. Fluorescence is measured orthogonally using a s barrier filter to block scattered laser excitation light, and a photomultiplier tube detector output signals, which are shifted in phase from a reference signal and amplitude demodulated, are processed by phase-sensitive detection electronics to resolve signals from heterogeneous emissions and quantify decay lifetimes directly. The output signals are displayed as frequency distribution histograms and bivariate diagrams using a computer-based data acquisition system. Results have demonstrated signal phase shift, amplitude demodulation, and average measurement of fluorescence lifetimes on stained cells; a detection limit threshold of 300 to 500 fluorescein isothiocyanate (FITC); fluorescence measurement precision of 1.3% on alignment fluorospheres and 3.4% on propidium iodide (PI)-stained cells; the resolution of PI and FITC signals from cells stainedin combination with PI and FITC, based on differences in their decay lifetimes; and the ability to measure single decay nines by the two-phase, phase comparator, method.

  6. Resolution of heterogeneous fluorescence emission signals and decay lifetime measurement on fluorochrome-labeled cells by phase-sensitive FCM

    SciTech Connect

    Steinkamp, J.A.; Crissman, H.A.

    1993-01-01

    A phase-sensitive flow cytometer has been developed to resolve signals from heterogeneous fluorescence emission spectra and quantify fluorescence decay times on cells labeled with fluorescent dyes. This instrument combines flow cytometry (FCM) and fluorescence spectroscopy measurement principles to provide unique capabilities for making phase-resolved measurements on single cells in flow, while preserving conventional FCM measurement capabilities. Stained cells are analyzed as they pass through an intensity-modulated (sinusoid) laser excitation beam. Fluorescence is measured orthogonally using a s barrier filter to block scattered laser excitation light, and a photomultiplier tube detector output signals, which are shifted in phase from a reference signal and amplitude demodulated, are processed by phase-sensitive detection electronics to resolve signals from heterogeneous emissions and quantify decay lifetimes directly. The output signals are displayed as frequency distribution histograms and bivariate diagrams using a computer-based data acquisition system. Results have demonstrated signal phase shift, amplitude demodulation, and average measurement of fluorescence lifetimes on stained cells; a detection limit threshold of 300 to 500 fluorescein isothiocyanate (FITC); fluorescence measurement precision of 1.3% on alignment fluorospheres and 3.4% on propidium iodide (PI)-stained cells; the resolution of PI and FITC signals from cells stainedin combination with PI and FITC, based on differences in their decay lifetimes; and the ability to measure single decay nines by the two-phase, phase comparator, method.

  7. Fluorescence Recovery after Merging a Droplet to Measure the Two-dimensional Diffusion of a Phospholipid Monolayer

    PubMed Central

    Jeong, Dae-Woong; Kim, KyuHan; Choi, Myung Chul; Choi, Siyoung Q.

    2015-01-01

    We introduce a new method to measure the lateral diffusivity of a surfactant monolayer at the fluid-fluid interface, called fluorescence recovery after merging (FRAM). FRAM adopts the same principles as the fluorescence recovery after photobleaching (FRAP) technique, especially for measuring fluorescence recovery after bleaching a specific area, but FRAM uses a drop coalescence instead of photobleaching dye molecules to induce a chemical potential gradient of dye molecules. Our technique has several advantages over FRAP: it only requires a fluorescence microscope rather than a confocal microscope equipped with high power lasers; it is essentially free from the selection of fluorescence dyes; and it has far more freedom to define the measured diffusion area. Furthermore, FRAM potentially provides a route for studying the mixing or inter-diffusion of two different surfactants, when the monolayers at a surface of droplet and at a flat air/water interface are prepared with different species, independently. PMID:26556128

  8. Quantification of fluorophore concentration in tissue-simulating media by fluorescence measurements with a single optical fiber.

    PubMed

    Diamond, Kevin R; Patterson, Michael S; Farrell, Thomas J

    2003-05-01

    Quantifying fluorescent compounds in turbid media such as tissue is made difficult by the effects of multiple scattering and absorption of the excitation and emission light. The approach that we used was to measure fluorescence using a single 200-microm optical fiber as both the illumination source and the detector. Fluorescence of aluminum phthalocyanine tetrasulfonate (AlPcS4) was measured over a wide range of fluorophore concentrations and optical properties in tissue-simulating phantoms. A root-mean-square accuracy of 10.6% in AlPcS4 concentration was attainable when fluorescence was measured either interstitially or at the phantom surface. The individual effects of scattering, absorption, and the scattering phase function on the fluorescence signal were also studied by experiments and Monte Carlo simulations.

  9. Fluorescence Recovery after Merging a Droplet to Measure the Two-dimensional Diffusion of a Phospholipid Monolayer.

    PubMed

    Jeong, Dae-Woong; Kim, KyuHan; Choi, Myung Chul; Choi, Siyoung Q

    2015-10-15

    We introduce a new method to measure the lateral diffusivity of a surfactant monolayer at the fluid-fluid interface, called fluorescence recovery after merging (FRAM). FRAM adopts the same principles as the fluorescence recovery after photobleaching (FRAP) technique, especially for measuring fluorescence recovery after bleaching a specific area, but FRAM uses a drop coalescence instead of photobleaching dye molecules to induce a chemical potential gradient of dye molecules. Our technique has several advantages over FRAP: it only requires a fluorescence microscope rather than a confocal microscope equipped with high power lasers; it is essentially free from the selection of fluorescence dyes; and it has far more freedom to define the measured diffusion area. Furthermore, FRAM potentially provides a route for studying the mixing or inter-diffusion of two different surfactants, when the monolayers at a surface of droplet and at a flat air/water interface are prepared with different species, independently.

  10. Fluorescence of tryptophan in designed hairpin and Trp-cage miniproteins: measurements of fluorescence yields and calculations by quantum mechanical molecular dynamics simulations.

    PubMed

    McMillan, Andrew W; Kier, Brandon L; Shu, Irene; Byrne, Aimee; Andersen, Niels H; Parson, William W

    2013-02-14

    The quantum yield of tryptophan (Trp) fluorescence was measured in 30 designed miniproteins (17 β-hairpins and 13 Trp-cage peptides), each containing a single Trp residue. Measurements were made in D(2)O and H(2)O to distinguish between fluorescence quenching mechanisms involving electron and proton transfer in the hairpin peptides, and at two temperatures to check for effects of partial unfolding of the Trp-cage peptides. The extent of folding of all the peptides also was measured by NMR. The fluorescence yields ranged from 0.01 in some of the Trp-cage peptides to 0.27 in some hairpins. Fluorescence quenching was found to occur by electron transfer from the excited indole ring of the Trp to a backbone amide group or the protonated side chain of a nearby histidine, glutamate, aspartate, tyrosine, or cysteine residue. Ionized tyrosine side chains quenched strongly by resonance energy transfer or electron transfer to the excited indole ring. Hybrid classical/quantum mechanical molecular dynamics simulations were performed by a method that optimized induced electric dipoles separately for the ground and excited states in multiple π-π* and charge-transfer (CT) excitations. Twenty 0.5 ns trajectories in the tryptophan's lowest excited singlet π-π* state were run for each peptide, beginning by projections from trajectories in the ground state. Fluorescence quenching was correlated with the availability of a CT or exciton state that was strongly coupled to the π-π* state and that matched or fell below the π-π* state in energy. The fluorescence yields predicted by summing the calculated rates of charge and energy transfer are in good accord with the measured yields.

  11. "Open-Box" Approach to Measuring Fluorescence Quenching Using an iPad Screen and Digital SLR Camera

    ERIC Educational Resources Information Center

    Koenig, Michael H.; Yi, Eun P.; Sandridge, Matthew J.; Mathew, Alexander S.; Demas, James N.

    2015-01-01

    Fluorescence quenching is an analytical technique and a common undergraduate laboratory exercise. Unfortunately, a typical quenching experiment requires the use of an expensive fluorometer that measures the relative fluorescence intensity of a single sample in a closed compartment unseen by the experimenter. To overcome these shortcomings, we…

  12. Nonlinear reconstruction of absorption and fluorescence contrast from measured diffuse transmittance and reflectance of a compressed-breast-simulating phantom.

    PubMed

    Ziegler, Ronny; Nielsen, Tim; Koehler, Thomas; Grosenick, Dirk; Steinkellner, Oliver; Hagen, Axel; Macdonald, Rainer; Rinneberg, Herbert

    2009-08-20

    We report on the nonlinear reconstruction of local absorption and fluorescence contrast in tissuelike scattering media from measured time-domain diffuse reflectance and transmittance of laser as well as laser-excited fluorescence radiation. Measurements were taken at selected source-detector offsets using slablike diffusely scattering and fluorescent phantoms containing fluorescent heterogeneities. Such measurements simulate in vivo data that would be obtained employing a scanning, time-domain fluorescence mammograph, where the breast is gently compressed between two parallel glass plates, and source and detector optical fibers scan synchronously at various source-detector offsets, allowing the recording of laser and fluorescence mammograms. The diffusion equations modeling the propagation of the laser and fluorescence radiation were solved in frequency domain by the finite element method simultaneously for several modulation frequencies using Fourier transformation and preprocessed experimental data. To reconstruct the concentration of the fluorescent contrast agent, the Born approximation including higher-order reconstructed photon densities at the excitation wavelength was used. Axial resolution was determined that can be achieved by various detection schemes. We show that remission measurements increase the depth resolution significantly.

  13. Measurement of plutonium in spent nuclear fuel by self-induced x-ray fluorescence

    SciTech Connect

    Hoover, Andrew S; Rudy, Cliff R; Tobin, Steve J; Charlton, William S; Stafford, A; Strohmeyer, D; Saavadra, S

    2009-01-01

    Direct measurement of the plutonium content in spent nuclear fuel is a challenging problem in non-destructive assay. The very high gamma-ray flux from fission product isotopes overwhelms the weaker gamma-ray emissions from plutonium and uranium, making passive gamma-ray measurements impossible. However, the intense fission product radiation is effective at exciting plutonium and uranium atoms, resulting in subsequent fluorescence X-ray emission. K-shell X-rays in the 100 keV energy range can escape the fuel and cladding, providing a direct signal from uranium and plutonium that can be measured with a standard germanium detector. The measured plutonium to uranium elemental ratio can be used to compute the plutonium content of the fuel. The technique can potentially provide a passive, non-destructive assay tool for determining plutonium content in spent fuel. In this paper, we discuss recent non-destructive measurements of plutonium X-ray fluorescence (XRF) signatures from pressurized water reactor spent fuel rods. We also discuss how emerging new technologies, like very high energy resolution microcalorimeter detectors, might be applied to XRF measurements.

  14. Numerical analysis of quantitative measurement of hydroxyl radical concentration using laser-induced fluorescence in flame

    NASA Astrophysics Data System (ADS)

    Shuang, Chen; Tie, Su; Yao-Bang, Zheng; Li, Chen; Ting-Xu, Liu; Ren-Bing, Li; Fu-Rong, Yang

    2016-06-01

    The aim of the present work is to quantitatively measure the hydroxyl radical concentration by using LIF (laser-induced fluorescence) in flame. The detailed physical models of spectral absorption lineshape broadening, collisional transition and quenching at elevated pressure are built. The fine energy level structure of the OH molecule is illustrated to understand the process with laser-induced fluorescence emission and others in the case without radiation, which include collisional quenching, rotational energy transfer (RET), and vibrational energy transfer (VET). Based on these, some numerical results are achieved by simulations in order to evaluate the fluorescence yield at elevated pressure. These results are useful for understanding the real physical processes in OH-LIF technique and finding a way to calibrate the signal for quantitative measurement of OH concentration in a practical combustor. Project supported by the National Natural Science Foundation of China (Grant No. 11272338) and the Fund from the Science and Technology on Scramjet Key Laboratory, China (Grant No. STSKFKT2013004).

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

    NASA Astrophysics Data System (ADS)

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

    2010-08-01

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

  16. A method of measuring gold nanoparticle concentrations by x-ray fluorescence for biomedical applications

    SciTech Connect

    Wu Di; Li Yuhua; Wong, Molly D.; Liu Hong

    2013-05-15

    Purpose: This paper reports a technique that enables the quantitative determination of the concentration of gold nanoparticles (GNPs) through the accurate detection of their fluorescence radiation in the diagnostic x-ray spectrum. Methods: Experimentally, x-ray fluorescence spectra of 1.9 and 15 nm GNP solutions are measured using an x-ray spectrometer, individually and within chicken breast tissue samples. An optimal combination of excitation and emission filters is determined to segregate the fluorescence spectra at 66.99 and 68.80 keV from the background scattering. A roadmap method is developed that subtracts the scattered radiation (acquired before the insertion of GNP solutions) from the signal radiation acquired after the GNP solutions are inserted. Results: The methods effectively minimize the background scattering in the spectrum measurements, showing linear relationships between GNP solutions from 0.1% to 10% weight concentration and from 0.1% to 1.0% weight concentration inside a chicken breast tissue sample. Conclusions: The investigation demonstrated the potential of imaging gold nanoparticles quantitatively in vivo for in-tissue studies, but future studies will be needed to investigate the ability to apply this method to clinical applications.

  17. Measuring incidence angle for through-the-objective total internal reflection fluorescence microscopy

    PubMed Central

    2012-01-01

    Abstract. Total internal reflection fluorescence (TIRF) microscopy has the exciting laser beam incident beyond critical angle from the glass side of a glass/aqueous interface formed by the coverslip and aqueous sample. The aqueous side evanescent field decays exponentially with distance from the interface with penetration depth depending on incidence angle. Through-the-objective TIRF has the exciting laser focused at the back focal plane (BFP) creating a refracted parallel beam approaching the interface in the small gap between objective and coverslip, making incidence angle challenging to measure. Objective axial scanning does not affect incidence angle but translates beam and interface intersection detected by the fluorescence center of mass from fluorescent spheres attached to the aqueous side of the interface. Center of mass translation divided by the axial translation is the tangent of the incidence angle that is sampled repeatedly over objective trajectory to obtain a best estimate. Incidence angle is measured for progressively larger radial positions of the focused beam on the BFP. A through-the-objective TIRF microscope, utilizing a micrometer and relay lenses to position the focused beam at the BFP, is calibrated for incidence angle. Calibration depends on microscope characteristics and TIRF objective and is applicable to any interface or sample. PMID:23208218

  18. SmartFluo: A Method and Affordable Adapter to Measure Chlorophyll a Fluorescence with Smartphones.

    PubMed

    Friedrichs, Anna; Busch, Julia Anke; van der Woerd, Hendrik Jan; Zielinski, Oliver

    2017-03-25

    In order to increase the monitoring capabilities of inland and coastal waters, there is a need for new, affordable, sensitive and mobile instruments that could be operated semi-automatically in the field. This paper presents a prototype device to measure chlorophyll a fluorescence: the SmartFluo. The device is a combination of a smartphone offering an intuitive operation interface and an adapter implying a cuvette holder, as well as a suitable illumination source. SmartFluo is based on stimulated fluorescence of water constituents such as chlorophyll a. The red band of the digital smartphone camera is sensitive enough to detect quantitatively the characteristic red fluorescence emission. The adapter contains a light source, a strong light emitting diode and additional filters to enhance the signal-to-noise ratio and to suppress the impact of scattering. A novel algorithm utilizing the red band of the camera is provided. Laboratory experiments of the SmartFluo show a linear correlation (R 2 = 0.98) to the chlorophyll a concentrations measured by reference instruments, such as a high-performance benchtop laboratory fluorometer (LS 55, PerkinElmer).

  19. Relation between circular and linear depolarization ratios under multiple-scattering conditions.

    PubMed

    Roy, Gilles; Roy, Nathalie

    2008-12-10

    A simple relationship is established between the linear and the circular depolarization ratios averaged over the azimuth angle of clouds made of spherical particles. The relationship is validated theoretically using double-scattering calculations; in the framework, the measurements are performed with a multiple-field-of-view lidar (MFOV) lidar. The relationship is also validated using data obtained with MFOV lidar equipped with linear and circular polarization measurement capabilities. The experimental data support theoretical results for small optical depths. At higher optical depths and large fields of view, the contribution of multiple scatterings is important; experimental data suggest that the relationship established between the linear and circular depolarization stays valid as long as the main depolarization mechanism comes from one scattering (most likely a backscattering a few degrees away from 180 degrees ).

  20. Determination of Enantiomeric Compositions of Analytes Using Novel Fluorescent Chiral Molecular Micelles and Steady State Fluorescence Measurements

    PubMed Central

    Williams, Alicia A.; Fakayode, Sayo O.; Alptürk, Onur; Jones, Christina M.; Lowry, Mark; Strongin, Robert M.

    2009-01-01

    Novel fluorescent chiral molecular micelles (FCMMs) were synthesized, characterized, and employed as chiral selectors for enantiomeric recognition of non-fluorescent chiral molecules using steady state fluorescence spectroscopy. The sensitivity of the fluorescence technique allowed for investigation of low concentrations of chiral selector (3.0×10−5 M) and analyte (5.0×10−6 M) to be used in these studies. The chiral interactions of glucose, tartaric acid, and serine in the presence of FCMMs poly(sodium N-undecanoyl-L-tryptophanate) [poly-L-SUW], poly(sodium N-undecanoyl-L-tyrosinate) [poly-L-SUY], and poly(sodium N-undecanoyl-L-phenylalininate) [poly-SUF] were based on diastereomeric complex formation. Poly-L-SUW had a significant fluorescence emission spectral difference as compared to poly-L-SUY and poly-L-SUF for the enantiomeric recognition of glucose, tartaric acid, and serine. Studies with the hydrophobic molecule α-pinene suggested that poly-L-SUY and poly-L-SUF had better chiral discrimination ability for hydrophobic analytes as compared to hydrophilic analytes. Partial-least-squares regression modeling (PLS-1) was used to correlate changes in the fluorescence emission spectra of poly-L-SUW due to varying enantiomeric compositions of glucose, tartaric acid, and serine for a set of calibration samples. Validation of the calibration regression models was determined by use of a set of independently prepared samples of the same concentration of chiral selector and analyte with varying enantiomeric composition. Prediction ability was evaluated by use of the root-mean-square percent relative error (RMS%RE) and was found to range from 2.04 to 4.06%. PMID:17985217

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

    PubMed

    Broeken, Jordi; Rieger, Bernd; Stallinga, Sjoerd

    2014-06-01

    We propose a method for simultaneously measuring the position and emission color of single fluorescent emitters based on the use of a large pitch diffraction grating in the emission light path. The grating produces satellite spots adjacent to the main spot; the relative distance between the spots is a measure for the emission wavelength. We present proof-of-principle experiments on beads and mixtures of quantum dots using a spatial light modulator for making a programmable diffraction grating. A wavelength precision of around 10 nm can be achieved for 1000 signal photons and practical background levels, while maintaining a localization precision of around 10 nm.

  2. Application of a pulsed laser for measurements of bathymetry and algal fluorescence.

    NASA Technical Reports Server (NTRS)

    Hickman, G. D.; Hogg, J. E.; Friedman, E. J.; Ghovanlou, A. H.

    1973-01-01

    The technique of measuring water depths with an airborne pulsed dye laser is studied, with emphasis on the degrading effect of some environmental and operational parameters on the transmitted and reflected laser signals. Extrapolation of measurements of laser stimulated fluorescence, performed as a function of both the algal cell concentration and the distance between the algae and the laser/receiver, indicate that a laser system operating from a height of 500 m should be capable of detecting chlorophyll concentrations as low as 1.0 mg/cu m.-

  3. On the measurement of particle number and mobility in nonideal solutions by fluorescence correlation spectroscopy.

    PubMed Central

    Abney, J R; Scalettar, B A; Hackenbrock, C R

    1990-01-01

    Interparticle interactions are incorporated into the theoretical description of the initial amplitude, G(0), of the normalized fluorescence correlation spectroscopy autocorrelation function. Measurements of particle number, aggregate size, and interaction-dependent diffusion are then analyzed in the context of this generalized theory. It is shown that the neglect of interactions can introduce order-of-magnitude errors into estimates of particle number and aggregate size. It is also shown that measurement of G(0) provides an essentially unique method for testing the validity of theories of interaction-dependent membrane protein diffusion. PMID:2383634

  4. Investigation of laser-induced iodine fluorescence for the measurement of density in compressible flows

    NASA Technical Reports Server (NTRS)

    Mcdaniel, J. C., Jr.

    1982-01-01

    Laser induced fluorescence is an attractive nonintrusive approach for measuring molecular number density in compressible flows although this technique does not produce a signal that is directly related to the number density. Saturation and frequency detuned excitation are explored as means for minimizing the quenching effect using iodine as the molecular system because of its convenient absorption spectrum. Saturation experiments indicate that with available continuous wave laser sources of Gaussian transverse intensity distribution only partial saturation could be achieved in iodine at the pressures of interest in gas dynamics. Using a fluorescence lineshape theory, it is shown that for sufficiently large detuning of a narrow bandwidth laser from a molecular transition, the quenching can be cancelled by collisional broadening over a large range of pressures and temperatures. Experimental data obtained in a Mach 4.3 underexpanded jet of nitrogen seeded with iodine for various single mode argon laser detunings from a strong iodine transition at 5145 A are discussed.

  5. Laser-induced fluorescence diagnostic for temperature and velocity measurements in a hydrogen arcjet plume.

    PubMed

    Liebeskind, J G; Hanson, R K; Cappelli, M A

    1993-10-20

    A diagnostic has been developed to measure velocity and translational temperature in the plume of a 1-kW-class arcjet thruster operating on hydrogen. Laser-induced fluorescence with a narrow-band cw laser is used to probe the Balmer α transition of excited atomic hydrogen. The velocity is determined from the Doppler shift of the fluorescence excitation spectrum, whereas the temperature is inferred from the lineshape. Analysis shows that although Doppler broadening is the only significant broadening mechanism, the fine structure of the transition must be taken into account. Near the exit plane, axial velocities vary from 4 to 14 km/s, radial velocities vary from 0 to 4 km/s, and swirl velocities are shown to be relatively small. Temperatures from 1000 to 5000 K indicate high dissociation fractions.

  6. Immobilized fluorescent dyes for sensitive pH measurements on enamel surfaces with fiber optics

    NASA Astrophysics Data System (ADS)

    Rumphorst, A.; Seeger, Stefan; Duschner, H.

    1996-01-01

    Information on the pH directly on surfaces of dental enamel is an important aspect in research on tooth decay. As an alternative to pH-electrodes our approach to the problem is the optical determination of pH by pH sensitive fluorescent dyes immobilized to tooth surfaces. In this study a model for measuring pH either on aminated cellulose substrates or on enamel (in vitro) with a fluorescein type dye is presented. The experimental realization is a fiber optic sensor with a nitrogen-pumped dye laser system and photodiode for the detection of the emitted fluorescence light. The surface pH values in the range between 4 and 7 were derived from the ratios of the excitation bands at 490 nm and 460 nm.

  7. Determining a fluorophore's transition dipole moment from fluorescence lifetime measurements in solvents of varying refractive index.

    PubMed

    Chung, Pei-Hua; Tregidgo, Carolyn; Suhling, Klaus

    2016-11-11

    The transition dipole moment of organic dyes PM546 and rhodamine 123 is calculated from fluorescence lifetime measurements in solutions of different refractive index. A model proposed by Toptygin et al (2002 J. Phys. Chem. B 106 3724-34) provides a relationship between the radiative rate constant and refractive index of the solvent, and allows the electronic transition dipole moments to be found: it is (7.1  ±  1.1) D for PM546 which matches that found in the literature, and (8.1  ±  0.1) D for rhodamine 123. Toptygin's model goes further in predicting the shape of the fluorescent dye and here we predict the shape of PM546 and rhodamine 123 to be ellipsoidal.

  8. Depolarization Lidar Determination Of Cloud-Base Microphysical Properties

    NASA Astrophysics Data System (ADS)

    Donovan, D. P.; Klein Baltink, H.; Henzing, J. S.; de Roode, S.; Siebesma, A. P.

    2016-06-01

    The links between multiple-scattering induced depolarization and cloud microphysical properties (e.g. cloud particle number density, effective radius, water content) have long been recognised. Previous efforts to use depolarization information in a quantitative manner to retrieve cloud microphysical cloud properties have also been undertaken but with limited scope and, arguably, success. In this work we present a retrieval procedure applicable to liquid stratus clouds with (quasi-)linear LWC profiles and (quasi-)constant number density profiles in the cloud-base region. This set of assumptions allows us to employ a fast and robust inversion procedure based on a lookup-table approach applied to extensive lidar Monte-Carlo multiple-scattering calculations. An example validation case is presented where the results of the inversion procedure are compared with simultaneous cloud radar observations. In non-drizzling conditions it was found, in general, that the lidar- only inversion results can be used to predict the radar reflectivity within the radar calibration uncertainty (2-3 dBZ). Results of a comparison between ground-based aerosol number concentration and lidar-derived cloud base number considerations are also presented. The observed relationship between the two quantities is seen to be consistent with the results of previous studies based on aircraft-based in situ measurements.

  9. Biodegradability of anthropogenic organic matter in polluted rivers using fluorescence, UV, and BDOC measurements.

    PubMed

    Knapik, Heloise G; Fernandes, Cristovão V S; de Azevedo, Julio Cesar R; dos Santos, Mauricius M; Dall'Agnol, Patrícia; Fontane, Darrell G

    2015-03-01

    The presence of highly urbanized and polluted areas affects both the quantity and the composition of organic matter in rivers through effluent loads and urban runoff discharges in watersheds. In such context, this paper aims to evaluate the biodegradability of anthropogenic organic matter in polluted rivers. Stream water samples were collected in three different sites considering a non-impacted area, a highly urbanized site located after a sewage treatment plant, and a site downstream of the watershed. For the biodegradation experiment, two adaptations of biodegradable dissolved organic carbon (BDOC) essay were evaluated to assess the decomposition rates between 10 days, with added nutrients, in the dark at 20 °C. The organic matter biodegradation was monitored by distinct parameters such as dissolved organic carbon (DOC), total organic carbon (TOC), particulate organic carbon (POC), fluorescence excitation-emission matrix (EEM), and UV absorbance measurements. The measured BDOC ranged from 0.8 mg/L at site IG01 (low anthropogenic occupation) to 4.2 mg/L at site IG02 (high impacted area), with averaged percentage of initial DOC ranging from 20 to 56 %, while an average of 28 % up to 95 % of POC can be considered as biodegradable. This pattern of biodegradation of fluorescent components was also observed through a decrease of tryptophan-like and tyrosine-like fluorescence peak intensity during the incubation time. The results also showed a higher decrease of humic-like fluorescence peak intensity at polluted sites (IG02 and IG05). Our experimental approach and monitoring strategy suggests that the evaluation of the organic matter biodegradability is essential to understand the fate and transformation mechanism of organic matter in urbanized and polluted rivers. And, considering a water quality planning and management perspective, this approach is important to identify the presence and location of organic compounds potentially important for dissolved oxygen

  10. In situ measurement of airway surface liquid [K+] using a ratioable K+-sensitive fluorescent dye.

    PubMed

    Namkung, Wan; Song, Yuanlin; Mills, Aaron D; Padmawar, Prashant; Finkbeiner, Walter E; Verkman, A S

    2009-06-05

    The airway surface liquid (ASL) is the thin fluid layer lining airway surface epithelial cells, whose volume and composition are tightly regulated and may be abnormal in cystic fibrosis (CF). We synthesized a two-color fluorescent dextran to measure ASL [K(+)], TAC-Lime-dextran-TMR, consisting of a green-fluorescing triazacryptand K(+) ionophore-Bodipy conjugate, coupled to dextran, together with a red fluorescing tetramethylrhodamine reference chromophore. TAC-Lime-dextran-TMR fluorescence was K(+)-selective, increasing >4-fold with increasing [K(+)] from 0 to 40 mm. In well differentiated human airway epithelial cells, ASL [K(+)] was 20.8 +/- 0.3 mm and decreased by inhibition of the Na(+)/K(+) pump (ouabain), ENaC (amiloride), CF transmembrane conductance regulator (CFTR(inh)-172), or K(+) channels (TEA or XE991). ASL [K(+)] was increased by forskolin but not affected by Na(+)/K(+)/2Cl(-) cotransporter inhibition (bumetanide). Functional and expression studies indicated the involvement of [K(+)] channels KCNQ1, KCNQ3, and KCNQ5 as determinants of ASL [K(+)]. [K(+)] in CF cultures was similar to that in non-CF cultures, suggesting that abnormal ASL [K(+)] is not a factor in CF lung disease. In intact airways, ASL [K(+)] was also well above extracellular [K(+)]: 22 +/- 1 mm in pig trachea ex vivo and 16 +/- 1 mm in mouse trachea in vivo. Our results provide the first noninvasive measurements of [K(+)] in the ASL and indicate the involvement of apical and basolateral membrane ion transporters in maintaining a high ASL [K(+)].

  11. Multifaceted roles for astrocytes in spreading depolarization

    PubMed Central

    Seidel, Jessica L.; Escartin, Carole; Ayata, Cenk; Bonvento, Gilles; Shuttleworth, C. William

    2015-01-01

    Spreading depolarizations (SD) are coordinated waves of synchronous depolarization, involving large numbers of neurons and astrocytes as they spread slowly through brain tissue. The recent identification of SDs as likely contributors to pathophysiology in human subjects has led to a significant increase in interest in SD mechanisms, and possible approaches to limit the numbers of SDs or their deleterious consequences in injured brain. Astrocytes regulate many events associated with SD. SD initiation and propagation is dependent on extracellular accumulation of K+ and glutamate, both of which involve astrocytic clearance. SDs are extremely metabolically demanding events, and signaling through astrocyte networks is likely central to the dramatic increase in regional blood flow that accompanies SD in otherwise healthy tissues. Astrocytes may provide metabolic support to neurons following SD, and may provide a source of adenosine that inhibits neuronal activity following SD. It is also possible that astrocytes contribute to the pathophysiology of SD, as a consequence of excessive glutamate release, facilitation of NMDA receptor activation, brain edema due to astrocyte swelling, or disrupted coupling to appropriate vascular responses after SD. Direct or indirect evidence has accumulated implicating astrocytes in many of these responses, but much remains unknown about their specific contributions, especially in the context of injury. Conversion of astrocytes to a reactive phenotype is a prominent feature of injured brain, and recent work suggests that the different functional properties of reactive astrocytes could be targeted to limit SDs in pathophysiological conditions. PMID:26301517

  12. Diode laser atomic fluorescence temperature measurements in low-pressure flames

    NASA Astrophysics Data System (ADS)

    Burns, I. S.; Lamoureux, N.; Kaminski, C. F.; Hult, J.; Desgroux, P.

    2008-12-01

    Temperature measurements have been performed in a low-pressure flame by the technique of diode laser induced atomic fluorescence. The experiments were done in a near-stoichiometric flat-flame of premixed methane, oxygen and nitrogen, at a pressure of 5.3 kPa. Indium atoms were seeded to the flame and probed using blue diode lasers; the lineshapes of the resulting fluorescence spectra were used to determine the flame temperature at a range of heights above the burner plate. The particular issues associated with the implementation of this measurement approach at low pressure are discussed, and it is shown to work especially well under these conditions. The atomic fluorescence lineshape thermometry technique is quicker to perform and requires less elaborate equipment than other methods that have previously been implemented in low-pressure flames, including OH-LIF and NO-LIF. There was sufficient indium present to perform measurements at all locations in the flame, including in the pre-heat zone close to the burner plate. Two sets of temperature measurements have been independently performed by using two different diode lasers to probe two separate transitions in atomic indium. The good agreement between the two sets of data provides a validation of the technique. By comparing thermocouple profiles recorded with and without seeding of the flame, we demonstrate that any influence of seeding on the flame temperature is negligible. The overall uncertainty of the measurements reported here is estimated to be ±2.5% in the burnt gas region.

  13. Upgrade of goniospectrophtometer GEFE for near-field scattering and fluorescence radiance measurements

    NASA Astrophysics Data System (ADS)

    Bernad, Berta; Ferrero, Alejandro; Pons, Alicia; Hernanz, M. L.; Campos, Joaquín.

    2015-03-01

    The goniospectrophotometer GEFE, designed and developed at IO-CSIC (Instituto de Optica, Agencia Estatal Consejo Superior de Investigaciones Cientificas), was conceived to measure the spectral Bidirectional Reflectance Distribution Function (BRDF) at any pair of irradiation and detection directions. Although the potential of this instrument has largely been proved, it still required to be upgraded to deal with some important scattering features for the assessment of the appearance. Since it was not provided with a detector with spatial resolution, it simply could not measure spectrophotometric quantities to characterize texture through the Bidirectional Texture Function (BTF) or translucency through the more complex Bidirectional Scattering-Surface Reflectance Distribution Function (BSSRDF). Another requirement in the GEFE upgrading was to provide it with the capability of measuring fluorescence at different geometries, since some of the new pigments used in industry are fluorescent, which can have a non-negligible impact in the color of the product. Then, spectral resolution at irradiation and detection had to be available in GEFE. This paper describes the upgrading of the goniospectrophotometer GEFE, and its new capabilities through the presentation of sparkle and goniofluorescence measurements. In addition, the potential of the instrument to evaluate translucency by the measurement of the BSSRDF is briefly discussed.

  14. High speed velocimetry and concentration measurements in a microfluidic mixer using fluorescence confocal microscopy

    NASA Astrophysics Data System (ADS)

    Inguva, Venkatesh; Perot, Blair; Kathuria, Sagar; Rothstein, Jonathan; Bilsel, Osman

    2016-11-01

    This work experimentally examines the performance of a quasi-turbulent micro-mixer that was designed to produce rapid mixing for protein-folding experiments. The original design of the mixer was performed using Direct Numerical Simulation (DNS) of the flow field and LES of the high Sc number scalar field representing the protein. The experimental work is designed to validate the DNS results. Both the velocity field and the protein concentration require validation. Different experiments were carried out to measure these two quantities. Concentration measurements are performed using a 488nm continuous wave laser coupled with a confocal microscope to measure fluorescence intensity during mixing. This is calibrated using the case where no mixing occurs. The velocity measurements use a novel high speed velocimetry technique capable of measuring speeds on the order of 10 m/s in a micro channel. The technique involves creating a pulsed confocal volume from a Ti-Sapphire laser with a pulse width of 260ns and observing the decay of fluorescence due to the fluid motion. Results from both experiments will be presented along with a comparison to the DNS results. The work is supported by NSF IDBR Award No. 1353942.

  15. Shifts in the fluorescence lifetime of EGFP during bacterial phagocytosis measured by phase-sensitive flow cytometry

    NASA Astrophysics Data System (ADS)

    Li, Wenyan; Houston, Kevin D.; Houston, Jessica P.

    2017-01-01

    Phase-sensitive flow cytometry (PSFC) is a technique in which fluorescence excited state decay times are measured as fluorescently labeled cells rapidly transit a finely focused, frequency-modulated laser beam. With PSFC the fluorescence lifetime is taken as a cytometric parameter to differentiate intracellular events that are challenging to distinguish with standard flow cytometry. For example PSFC can report changes in protein conformation, expression, interactions, and movement, as well as differences in intracellular microenvironments. This contribution focuses on the latter case by taking PSFC measurements of macrophage cells when inoculated with enhanced green fluorescent protein (EGFP)-expressing E. coli. During progressive internalization of EGFP-E. coli, fluorescence lifetimes were acquired and compared to control groups. It was hypothesized that fluorescence lifetimes would correlate well with phagocytosis because phagosomes become acidified and the average fluorescence lifetime of EGFP is known to be affected by pH. We confirmed that average EGFP lifetimes consistently decreased (3 to 2 ns) with inoculation time. The broad significance of this work is the demonstration of how high-throughput fluorescence lifetime measurements correlate well to changes that are not easily tracked by intensity-only cytometry, which is affected by heterogeneous protein expression, cell-to-cell differences in phagosome formation, and number of bacterium engulfed.

  16. Shifts in the fluorescence lifetime of EGFP during bacterial phagocytosis measured by phase-sensitive flow cytometry

    PubMed Central

    Li, Wenyan; Houston, Kevin D.; Houston, Jessica P.

    2017-01-01

    Phase-sensitive flow cytometry (PSFC) is a technique in which fluorescence excited state decay times are measured as fluorescently labeled cells rapidly transit a finely focused, frequency-modulated laser beam. With PSFC the fluorescence lifetime is taken as a cytometric parameter to differentiate intracellular events that are challenging to distinguish with standard flow cytometry. For example PSFC can report changes in protein conformation, expression, interactions, and movement, as well as differences in intracellular microenvironments. This contribution focuses on the latter case by taking PSFC measurements of macrophage cells when inoculated with enhanced green fluorescent protein (EGFP)-expressing E. coli. During progressive internalization of EGFP-E. coli, fluorescence lifetimes were acquired and compared to control groups. It was hypothesized that fluorescence lifetimes would correlate well with phagocytosis because phagosomes become acidified and the average fluorescence lifetime of EGFP is known to be affected by pH. We confirmed that average EGFP lifetimes consistently decreased (3 to 2 ns) with inoculation time. The broad significance of this work is the demonstration of how high-throughput fluorescence lifetime measurements correlate well to changes that are not easily tracked by intensity-only cytometry, which is affected by heterogeneous protein expression, cell-to-cell differences in phagosome formation, and number of bacterium engulfed. PMID:28091553

  17. Tunable depolarized light scattering from gold and gold/silver nanorods.

    PubMed

    Khlebtsov, Boris; Khanadeev, Vitaly; Khlebtsov, Nikolai

    2010-04-07

    We combine the fabrication of Au and Au/Ag core/shell nanorods (NRs), as well as experimental measurements of vis-NIR extinction, unpolarized, and cross-polarized light scattering spectra, with T-matrix and separation-of-variables simulations, to gain insight into the relation between the morphology/composition of nanoparticles and their isotropic and anisotropic optical responses. Using several Au and Au/Ag core/shell NR samples, we present unambiguous experimental evidence of a tunable correlation between the longitudinal plasmon resonances of NRs and their spectral depolarization maxima. For gold NRs, the depolarization maxima follow the extinction plasmon resonances blue-shifted from 80 to 270 nm. In contrast, the depolarization maximum of Au/Ag NRs is located just near the longitudinal resonance, and the spectral shift of the depolarization maximum is about 10 to 20 nm. The experimental extinction and depolarization spectra of gold NRs are in good agreement with T-matrix simulations based on TEM-fitted models that account for the aspect ratio polydispersity and byproduct contributions. For composite Au/Ag NRs, the separation-of-variables simulations provide a calibration curve that correlates the relative spectral shift of the extinction resonance with the silver shell thicknesses and generates experimental data that are in good agreement with estimations based on the Ag/Au mass ratio.

  18. Use of a laser-induced fluorescence thermal imaging system for film cooling heat transfer measurement

    SciTech Connect

    Chyu, M.K.

    1996-04-01

    This paper describes a novel approach based on fluorescence imaging of thermographic phosphor that enables the simultaneous determination of both local film effectiveness and local heat transfer on a film-cooled surface. The film cooling model demonstrated consists of a single row of three discrete holes on a flat plate. The transient temperature measurement relies on the temperature-sensitive fluorescent properties of europium-doped lanthanum oxysulfide (La{sub 2}O{sub 2}S:Eu{sup +3}) thermographic phosphor. A series of full-field surface temperatures, mainstream temperatures, and coolant film temperatures were acquired during the heating of a test surface. These temperatures are used to calculate the heat transfer coefficients and the film effectiveness simultaneously. Because of the superior spatial resolution capability for the heat transfer data reduced from these temperature frames, the laser-induced fluorescence (LIF) imaging system, the present study observes the detailed heat transfer characteristics over a film-protected surface. The trend of the results agrees with those obtained using other conventional thermal methods, as well as the liquid crystal imaging technique. One major advantage of this technique is the capability to record a large number of temperature frames over a given testing period. This offers multiple-sample consistency.

  19. Use of a laser-induced fluorescence thermal imaging system for film cooling heat transfer measurement

    SciTech Connect

    Chyu, M.K.

    1995-10-01

    This paper describes a novel approach based on fluorescence imaging of thermographic phosphor that enables the simultaneous determination of both local film effectiveness and local heat transfer on a film-cooled surface. The film cooling model demonstrated consists of a single row of three discrete holes on a flat plate. The transient temperature measurement relies on the temperature-sensitive fluorescent properties of europium-doped lanthanum oxysulfide (La{sub 2}O{sub 2}S:EU{sup 3+}) thermographic phosphor. A series of full-field surface temperatures, mainstream temperatures, and coolant film temperatures were acquired during the heating of a test surface. These temperatures are used to calculate the heat transfer coefficients and the film effectiveness simultaneously. Because of the superior spatial resolution capability for the heat transfer data reduced from these temperature frames, the laser-induced fluorescence (LIF) imaging system, the present study observes the detailed heat transfer characteristics over a film-protected surface. The trend of the results agrees with those obtained using other conventional thermal methods, as well as the liquid crystal imaging technique. One major advantage of this technique is the capability to record a large number of temperature frames over a given testing period. This offers multiple-sample consistency.

  20. Effects of isoflurane on measurement of fluorescence spectra and CLSM imaging in Acetabularia acetabulum

    NASA Astrophysics Data System (ADS)

    Chen, WenLi; Quan, Zhou; Xing, Da

    2007-05-01

    The volatile halogenated methyl ethyl ether is used as anesthetic to inhibit actin-based dynamics directly or indirectly in animal cells. In plant cells, most intracellular movements are related with actin pathways too. We utilized isoflurane to study the dynamic choloroplast organization in unicellular baby and adult alga Acetabularia acetabulum. Fluorescence spectra were measured and choloroplast movements were recorded by confocal laser scanning microscope (CLSM) imaging in in Acetabularia acetabulum. Isoflurane was effective in the unicellular baby and adult organisms and showed time- dependent actin-inhibition patterns. Acetabularia acetabulum cells were treated for different times with isoflurane saturated solutions in artificial seawater (it was defined to be 100% isoflurane). The intensity of fluorescence at 680nm and 730nm were progressively decreased at 100% isoflurane. It was remarkable difference between fluorescence spectra of baby and adult Acetabularia were inhibited by isoflurance, adult Acetabularia cells showed more sensitive. Whereas the choloroplast in Acetabularia acetabulum was commendably imaged by CLSM at 20 and 40 zoom.

  1. Effects of signal corrections on measurements of temperature and OH concentrations using laser-induced fluorescence

    NASA Astrophysics Data System (ADS)

    Yin, Zhiyao; Carter, Campbell D.; Lempert, Walter R.

    2014-07-01

    Temperature and OH concentrations derived from OH laser-induced fluorescence (LIF) are known to be susceptible to effects such as collisional quenching, laser absorption, and fluorescence trapping. In this paper, a set of analytical and easy-to-implement methods is presented for treating these effects. The significance of these signal corrections on inferred temperature and absolute OH concentration is demonstrated in an atmospheric-pressure, near-stoichiometric CH4-air flame stabilized on a Hencken burner, for laser excitation of both the A2Σ+←X2Π (0,0) and (1,0) bands. It is found that the combined effect of laser attenuation and fluorescence trapping can cause considerable error in the OH number density and temperature if not accounted for, even with A-X(1,0) excitation. The validity of the assumptions used in signal correction (that the excited-state distribution is either thermalized or frozen) is examined using time-dependent modeling of the ro-vibronic states during and after laser excitation. These assumptions are shown to provide good bounding approximations for treating transition-dependent issues in OH LIF, especially for an unknown collisional environment, and it is noted that the proposed methods are generally applicable to LIF-based measurements.

  2. A novel fluorescence imaging approach for comparative measurements of pancreatic islet function in vitro.

    PubMed

    Corbin, Kathryn L; Hall, Thomas E; Haile, Ruth; Nunemaker, Craig S

    2011-01-01

    Pancreatic islet dysfunction is a key element in the development of type 2 diabetes. Determining possible early warning signs of dysfunction is thus important to determining the underlying causes of diabetes. We describe an improved fluorescent imaging approach to detect potential islet dysfunction. Using Cell Tracker Red (CTR, a mildly thiol-reactive fluorescent probe) to positively label particular islets, we measured intracellular free calcium with fura-2 AM in both CTR-labeled and unlabeled sets of pancreatic islets simultaneously in vitro. This approach enhances sensitivity by controlling for differences in background fluorescence, temperature, and perifusion dynamics. We confirmed that 200 nM CTR produced no spectral overlap with fura-2 and no significant physiological effects in selective tests of islet function. To demonstrate the utility of dual-labeling, we compared untreated islets with islets pretreated with low-dose pro-inflammatory cytokines (IL-6 + IL-1B) to induce mild dysfunction. We alternated CTR-labeling between control and test islets and identified consistent reductions in the amplitude and trajectory of glucose-stimulated calcium responses (GSCa) among cytokine-treated islets that were independent of labeling. Observations were verified using a MATLAB program specifically designed to identify key features in the GSCa. Our findings thus demonstrate the utility of CTR-labeling in identifying islet dysfunction and propose that this technique can be adapted for other cells and tissues.

  3. Quantitative Laser-Saturated Fluorescence Measurements of Nitric Oxide in a Heptane Spray Flame

    NASA Technical Reports Server (NTRS)

    Cooper, Clayton S.; Laurendeau, Normand M.; Lee, Chi (Technical Monitor)

    1997-01-01

    We report spatially resolved laser-saturated fluorescence measurements of NO concentration in a pre-heated, lean-direct injection (LDI) spray flame at atmospheric pressure. The spray is produced by a hollow-cone, pressure-atomized nozzle supplied with liquid heptane. NO is excited via the Q2(26.5) transition of the gamma(0,0) band. Detection is performed in a 2-nm region centered on the gamma(0,1) band. Because of the relatively close spectral spacing between the excitation (226 nm) and detection wavelengths (236 nm), the gamma(0,1) band of NO cannot be isolated from the spectral wings of the Mie scattering signal produced by the spray. To account for the resulting superposition of the fluorescence and scattering signals, a background subtraction method has been developed that utilizes a nearby non-resonant wavelength. Excitation scans have been performed to locate the optimum off-line wavelength. Detection scans have been performed at problematic locations in the flame to determine possible fluorescence interferences from UHCs and PAHs at both the on-line and off-line excitation wavelengths. Quantitative radial NO profiles are presented and analyzed so as to better understand the operation of lean-direct injectors for gas turbine combustors.

  4. A fluorescent assay to quantitatively measure in vitro acyl CoA:diacylglycerol acyltransferase activity.

    PubMed

    McFie, Pamela J; Stone, Scot J

    2011-09-01

    Triacylglycerols (TG) are the major storage form of energy in eukaryotic organisms and are synthesized primarily by acyl CoA:1,2-diacylglycerol acyltransferase (DGAT) enzymes. In vitro DGAT activity has previously been quantified by measuring the incorporation of either radiolabeled fatty acyl CoA or diacylglycerol (DG) into TG. We developed a modified acyltransferase assay using a fluorescent fatty acyl CoA substrate to accurately quantify in vitro DGAT activity. In the modified assay, radioactive fatty acyl CoA is replaced with fluorescent NBD-palmitoyl CoA, which is used as a substrate by DGAT with DG to produce NBD-TG. After extraction with organic solvents and separation by thin layer chromatography, NBD-TG formation can be detected and accurately quantified using a fluorescent imaging system. We demonstrate that this method can be adapted to detect other acyltransferase activities. Because NBD-palmitoyl CoA is commercially available at a much lower cost compared with radioactive acyl CoA substrates, it is a more economical alternative to radioactive tracers. In addition, the exposure of laboratory personnel to radioactivity is greatly reduced.

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

    NASA Technical Reports Server (NTRS)

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

    2010-01-01

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

  6. Automatic measurement of compression wood cell attributes in fluorescence microscopy images.

    PubMed

    Selig, B; Luengo Hendriks, C L; Bardage, S; Daniel, G; Borgefors, G

    2012-06-01

    This paper presents a new automated method for analyzing compression wood fibers in fluorescence microscopy. Abnormal wood known as compression wood is present in almost every softwood tree harvested. Compression wood fibers show a different cell wall morphology and chemistry compared to normal wood fibers, and their mechanical and physical characteristics are considered detrimental for both construction wood and pulp and paper purposes. Currently there is the need for improved methodologies for characterization of lignin distribution in wood cell walls, such as from compression wood fibers, that will allow for a better understanding of fiber mechanical properties. Traditionally, analysis of fluorescence microscopy images of fiber cross-sections has been done manually, which is time consuming and subjective. Here, we present an automatic method, using digital image analysis, that detects and delineates softwood fibers in fluorescence microscopy images, dividing them into cell lumen, normal and highly lignified areas. It also quantifies the different areas, as well as measures cell wall thickness. The method is evaluated by comparing the automatic with a manual delineation. While the boundaries between the various fiber wall regions are detected using the automatic method with precision similar to inter and intra expert variability, the position of the boundary between lumen and the cell wall has a systematic shift that can be corrected. Our method allows for transverse structural characterization of compression wood fibers, which may allow for improved understanding of the micro-mechanical modeling of wood and pulp fibers.

  7. Temperature and density measurement by electron beam fluorescence technique in rocket experiment

    NASA Astrophysics Data System (ADS)

    Kurihara, J.; Oyama, K.-I.

    The Electron Beam Fluorescence (EBF) technique has been widely used in the field of rarefied gas dynamics for over 40 years and applied to measurements for a variety of gases and flow conditions in the laboratory experiment. The EBF technique uses a high-energy electron beam to excite a gas molecule by an inelastic collision with an electron. Spectrum of subsequent fluorescence by the excited molecule consists of many vibrational bands, and each band has a fine rotational structure. If the excitation-emission process is known precisely, the analysis of the vibrational-rotational band provides properties of the initial state of molecules. We applied the EBF technique to an in-situ measurement in the lower thermosphere and the vibrational temperature, the rotational temperature, and the number density of atmospheric molecular nitrogen between 100 - 150 km altitudes were observed by the sounding rocket experiment. Aerodynamic effects on the measurement caused by the rocket flight are corrected quantitatively using Direct Simulation Monte Carlo (DSMC) method. The great advantage of this type of instrument is that temperature and density are observed simultaneously and the consistency between the two measurements can be checked assuming hydrostatic equilibrium.

  8. Plant-Stress Measurements Using Laser-Induced Fluorescence Excitation: Poland Experiment

    SciTech Connect

    Gene Capelle; Steve Jones

    1999-05-01

    Bechtel Nevada's Special Technologies Laboratory (STL) has been involved in remote sensing for many years, and in April 1995 STL began to study the use of active remote sensing for detecting plant stress. This work was motivated by the need to detect subsurface contamination, with the supposition that this could be accomplished by remote measurement of optical signatures from the overgrowing vegetation. The project has been a cooperative DOE/Disney effort, in which basic optical signature measurements (primarily fluorescence) were done at the Disney greenhouse facilities at Epcot Center in Florida, using instrumentation developed by STL on DOE funding. The primary instrument is a LIFI system, which had originally been developed for detection of surface uranium contamination at DOE sites. To deal specifically with the plant stress measurements, a LIFS system was built that utilizes the same laser, but captures the complete fluorescence spectrum from blue to red wavelengths. This system had continued to evolve, and the version in existence in September 1997 was sent to Poland, accompanied by two people from STL, for the purpose of making the measurements described in this report.

  9. Measurement of buried undercut structures in microfluidic devices by laser fluorescent confocal microscopy

    SciTech Connect

    Li Shiguang; Liu Jing; Nguyen, Nam-Trung; Fang Zhongping; Yoon, Soon Fatt

    2009-11-20

    Measuring buried, undercut microstructures is a challenging task in metrology. These structures are usually characterized by measuring their cross sections after physically cutting the samples. This method is destructive and the obtained information is incomplete. The distortion due to cutting also affects the measurement accuracy. In this paper, we first apply the laser fluorescent confocal microscopy and intensity differentiation algorithm to obtain the complete three-dimensional profile of the buried, undercut structures in microfluidic devices, which are made by the soft lithography technique and bonded by the oxygen plasma method. The impact of material wettability and the refractive index (n) mismatch among the liquid, samples, cover layer, and objective on the measurement accuracy are experimentally investigated.

  10. Absolute Density Calibration Cell for Laser Induced Fluorescence Erosion Rate Measurements

    NASA Technical Reports Server (NTRS)

    Domonkos, Matthew T.; Stevens, Richard E.

    2001-01-01

    Flight qualification of ion thrusters typically requires testing on the order of 10,000 hours. Extensive knowledge of wear mechanisms and rates is necessary to establish design confidence prior to long duration tests. Consequently, real-time erosion rate measurements offer the potential both to reduce development costs and to enhance knowledge of the dependency of component wear on operating conditions. Several previous studies have used laser-induced fluorescence (LIF) to measure real-time, in situ erosion rates of ion thruster accelerator grids. Those studies provided only relative measurements of the erosion rate. In the present investigation, a molybdenum tube was resistively heated such that the evaporation rate yielded densities within the tube on the order of those expected from accelerator grid erosion. This work examines the suitability of the density cell as an absolute calibration source for LIF measurements, and the intrinsic error was evaluated.

  11. Transmission Nuclear Resonance Fluorescence Measurements of 238U in Thick Targets

    SciTech Connect

    Quiter, Brian J.; Ludewigt, Bernhard A.; Mozin, Vladimir V.; Wilson, Cody; Korbly, Steve

    2010-08-31

    Transmission nuclear resonance fluorescence measurements were made on targets consisting of Pb and depleted U with total areal densities near 86 g/cm2. The 238U content n the targets varied from 0 to 8.5percent (atom fraction). The experiment demonstrates the capability of using transmission measurements as a non-destructive technique to identify and quantify the presence of an isotope in samples with thicknesses comparable to he average thickness of a nuclear fuel assembly. The experimental data also appear to demonstrate the process of notch refilling with a predictable intensity. Comparison of measured spectra to previous backscatter 238U measurements indicates general agreement in observed excited states. Two new 238U excited states and possibly a third state have also been observed.

  12. Measuring the fluorescent quantum efficiency of indocyanine green encapsulated in nanocomposite particulates.

    PubMed

    Russin, T J; Altınoğlu, E İ; Adair, J H; Eklund, P C

    2010-08-25

    We present results of a fluorescent quantum efficiency (Φ(F)) study on the encapsulation of the near-infrared dye indocyanine green (ICG) in bioresorbable calcium phosphate nanoparticles (CPNPs). The Φ(F) (described as the ratio of photons emitted to photons absorbed) provides a quantitative means of describing the fluorescence of an arbitrary molecule. However, standard quantum efficiency measurement techniques provide only the Φ(F) of the smallest fluorescing unit-in the case of a nanoparticle suspension, the nanoparticle itself. This presents a problem in accurately describing the Φ(F) of fluorophores embedded in an inorganic nanoparticle. Combining the incidence of scattering with an evaluation of the differences in local electric field and photochemical environment, we have developed a method to determine the Φ(F) of the constituent fluorescent molecules embedded in such a nanoparticle, which provides a more meaningful comparison with the unencapsulated fluorophore. While applicable to generic systems, we present results obtained by our method for the ICG-CPNP in a phosphate buffered 0.15 M saline solution (PBS, pH 7.4)--specifically, Φ(F, free dye) = 0.027 ± 0.001, Φ(F, particle) = 0.053 ± 0.003, and for the individual encapsulated molecules, Φ(F, molecule) = 0.066 ± 0.004. The method developed also provides insight into the influences of encapsulation and key parameters to engineer resonant enhancement effects from the emission of the encapsulated fluorophores corresponding to an eigenmode of the embedding particle for tailored optical properties.

  13. Temperature measurements of micro-droplets using pulsed 2-color laser-induced fluorescence with MDR-enhanced energy transfer

    NASA Astrophysics Data System (ADS)

    Palmer, Johannes; Reddemann, Manuel A.; Kirsch, Valeri; Kneer, Reinhold

    2016-12-01

    In this work, a new measurement system is presented for studying temperature of micro-droplets by pulsed 2-color laser-induced fluorescence. Pulsed fluorescence excitation allows motion blur suppression and thus simultaneous measurements of droplet size, velocity and temperature. However, high excitation intensities of pulsed lasers lead to morphology-dependent resonances inside micro-droplets, which are accompanied by disruptive stimulated emission. Investigations showed that stimulated emission can be avoided by enhanced energy transfer via an additional dye. The suitability and accuracy of the new pulsed method are verified on the basis of a spectroscopic analysis and comparison to continuously excited 2-color laser-induced fluorescence.

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

    PubMed Central

    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. PMID:25798136

  15. Particle velocity measurements with macroscopic fluorescence imaging in lymph tissue mimicking microfluidic phantoms

    NASA Astrophysics Data System (ADS)

    Hennessy, Ricky; Koo, Chiwan; Ton, Phuc; Han, Arum; Righetti, Raffaella; Maitland, Kristen C.

    2011-03-01

    Ultrasound poroelastography can quantify structural and mechanical properties of tissues such as stiffness, compressibility, and fluid flow rate. This novel ultrasound technique is being explored to detect tissue changes associated with lymphatic disease. We have constructed a macroscopic fluorescence imaging system to validate ultrasonic fluid flow measurements and to provide high resolution imaging of microfluidic phantoms. The optical imaging system is composed of a white light source, excitation and emission filters, and a camera with a zoom lens. The field of view can be adjusted from 100 mm x 75 mm to 10 mm x 7.5 mm. The microfluidic device is made of polydimethylsiloxane (PDMS) and has 9 channels, each 40 μm deep with widths ranging from 30 μm to 200 μm. A syringe pump was used to propel water containing 15 μm diameter fluorescent microspheres through the microchannels, with flow rates ranging from 0.5 μl/min to 10 μl/min. Video was captured at a rate of 25 frames/sec. The velocity of the microspheres in the microchannels was calculated using an algorithm that tracked the movement of the fluorescent microspheres. The imaging system was able to measure particle velocities ranging from 0.2 mm/sec to 10 mm/sec. The range of flow velocities of interest in lymph vessels is between 1 mm/sec to 10 mm/sec; therefore our imaging system is sufficient to measure particle velocity in phantoms modeling lymphatic flow.

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

    PubMed

    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.

  17. Fluorescence measurement of chloride transport in monolayer cultured cells. Mechanisms of chloride transport in fibroblasts.

    PubMed

    Chao, A C; Dix, J A; Sellers, M C; Verkman, A S

    1989-12-01

    The methodology has been developed to measure Cl activity and transport in cultured cells grown on a monolayer using the entrapped Cl-sensitive fluorophore 6-methoxy-N-[3-sulfopropyl] quinolinium (SPQ). The method was applied to a renal epithelial cell line, LLC-PKI, and a nonepithelial cell line, Swiss 3T3 fibroblasts. SPQ was nontoxic to cells when present for greater than h in the culture media. To load with SPQ (5 mM), cells were made transiently permeable by exposure to hypotonic buffer (150 mOsm, 4 min). Intracellular fluorescence was monitored continuously by epifluorescence microscopy using low illumination intensity at 360 +/- 5 nm excitation wavelength and photomultiplier detection at greater than 410 nm. Over 60 min at 37 degrees C, there was no photobleaching and less than 10% leakage of SPQ out of cells; intracellular SPQ fluorescence was uniform. SPQ fluorescence was calibrated against intracellular [Cl] using high K solutions containing the ionophores nigericin and tributyltin. The Stern-Volmer constant (Kq) for quenching of intracellular SPQ by Cl was 13 M-1 for fibroblasts and LLC-PKl cells. In the absence of Cl, SPQ lifetime was 26 ns in aqueous solution and 3.7 +/- 0.6 ns in cells, showing that the lower Kq in cells than in free solution (Kq = 118 M-1) was due to SPQ quenching by intracellular anions. To examine Cl transport mechanisms, the time course of intracellular [Cl] was measured in response to rapid Cl addition and removal in the presence of ion or pH gradients. In fibroblasts, three distinct Cl transporting systems were identified: a stilbeneinhibitable Cl/HCO3 exchanger, a furosemide-sensitive Na/K/2Cl cotransporter, and a Ca-regulated Cl conductance. These results establish a direct optical method to measure intracellular [Cl] continuously in cultured cells.

  18. SU-C-303-05: Photosensitizer Determination for PDT Using Interstitial and Surface Measurements of Fluorescence

    SciTech Connect

    Kim, M; Finlay, J; Zhu, T

    2015-06-15

    Purpose: Photosensitizer concentration during photodynamic therapy (PDT) is an important parameter for accurate dosimetry. Fluorescence signal can be used as a measure of photosensitizer concentration. Two methods of data acquisition were compared to an ex vivo study both for in vivo and phantom models. Methods: Fluorescence signal of commonly used photosensitizer benzoporphyrin derivative monoacid ring A (BPD) was obtained in phantoms and mouse tumors using an excitation light of 405 nm. Interstitial fluorescence signal was obtained using a side-cut fiber inserted into the tumor tissue of interest. Using a previously developed multi-fiber probe, tumor surface fluorescence measurements were also collected. Signals were calibrated according to optical phantoms with known sensitizer fluorescence. Optical properties for each sample were determined and the influence of different absorption and scattering properties on the fluorescence signals was investigated. Using single value decomposition of the spectra, the sensitizer concentration was determined using the two different measurement geometries. An ex vivo analysis was also performed for tumor samples to determine the sensitizer concentration. Results: The two fluorescence signals obtained from the surface multi-fiber probe and the interstitial measurements were compared and were corresponding for both phantoms and mouse models. The values obtained were comparable to the ex vivo measurements as well. Despite the difference in geometry, the surface probe measurements can still be used as a metric for determining the presence of sensitizer in small volume tumors. Conclusion: The multi-fiber contact probe can be used as a tool to measure fluorescence at the surface of the treatment area for PDT and predict sensitizer concentration throughout the tumor. This is advantageous in that the measurement does not damage any tissue. Future work will include investigating the dependence of these results on intratumor sensitizer

  19. Measuring Fluorescent Dye in the Bubbly and Sediment-Laden Surfzone

    DTIC Science & Technology

    2009-04-16

    Water Air Soil Pollut DOI 10.1007/s11270-009-0030-z Measuring Fluorescent Dye in the Bubbly and Sediment-Laden Surfzone David B. Clark · Falk...Feddersen · Melissa M. Omand · R. T. Guza Received: 25 September 2008 / Accepted: 10 February 2009 © The Author(s) 2009. This article is published with open...concentrations (e.g., cascading rivers and streams). D. B. Clark (B) · F. Feddersen · M. M. Omand · R. T. Guza Scripps Institution of Oceanography, UCSD, 9500

  20. Experimental determination of the spring constant of an individual multiwalled carbon nanotube cantilever using fluorescence measurement

    NASA Astrophysics Data System (ADS)

    Kwon, Soongeun; Park, Hyojun; Shim, Hyung Cheoul; Lee, Hyung Woo; Kwak, Yoon Keun; Kim, Soohyun

    2009-07-01

    We report an experimental method to determine the spring constant of a multiwalled carbon nanotube (MWNT) cantilever as a mechanical piconewton force transducer. Electrostatic actuation was employed to investigate the mechanical properties of a MWNT cantilever. In order to measure nanotube's deflection during actuation, fluorescent dyes were noncovalently attached to the end of the nanotubes. Also, the length dependence of the spring constant is studied by adjusting the length of MWNT via electrochemical etching. The results show that the spring constant of a MWNT cantilever is as small as 0.001 N/m and tunable in the range of 0.001-0.05 N/m.

  1. Valley depolarization in monolayer WSe2

    PubMed Central

    Yan, Tengfei; Qiao, Xiaofen; Tan, Pingheng; Zhang, Xinhui

    2015-01-01

    We have systematically examined the circular polarization of monolayer WSe2 at different temperature, excitation energy and exciton density. The valley depolarization in WSe2 is experimentally confirmed to be governed by the intervalley electron-hole exchange interaction. More importantly, a non-monotonic dependence of valley circular polarization on the excitation power density has been observed, providing the experimental evidence for the non-monotonic dependence of exciton intervalley scattering rate on the excited exciton density. The physical origination of our experimental observations has been proposed to be in analogy to the D′yakonov-Perel′ mechanism that is operative in conventional GaAs quantum well systems. Our experimental results are fundamentally important for well understanding the valley pseudospin relaxation in atomically thin transition metal dichalcogenides. PMID:26490157

  2. Image reconstruction for diagnosis and prognosis of breast cancer using fluorescence measurements: phantom studies

    NASA Astrophysics Data System (ADS)

    Roy, R.; Godavarty, A.; Thompson, A. B., Jr.; Sevick-Muraca, E. M.

    2005-04-01

    Fluorescence-enhance optical tomography is performed using (i) point illumination and point collection and (ii) area illumination and area collection geometrics in 3D. In both measurement techniques, an image-intensified charge-coupled (ICCD) imaging system is used in the frequency-domain to image near-infrared contrast agents. The experimental measurements are compared to diffusion model predictions in least squares form in the inverse problem. For image recovery for both area and point illumination geometries, an efficient gradient-based optimization technique based on the Penalty/modified barrier function (PMBF) method and the constrained truncated Newton with trust region (CONTN) method is developed. Targets in 3D were reconstructed from experimental data under two conditions of (i) perfect uptake (1:0, target to background ratio) and (ii) imperfect uptake (100:1, target to background ratio). Parameters of absorption cross section due to fluorophore and lifetimes are reconstructed. The present work demonstrates that 3D fluorescence enhanced optical tomography reconstructions can be successfully performed from both point/area illumination and collection experimental measurements of the time-dependent light propagation on clinically relevant tissue phantoms using a gain-modulated ICCD camera.

  3. Stark broadening corrections to laser-induced fluorescence temperature measurements in a hydrogen arcjet plume.

    PubMed

    Storm, P V; Cappelli, M A

    1996-08-20

    Laser-induced fluorescence of the H(α) transition of atomic hydrogen has previously been performed in the plume of a hydrogen arcjet thruster. Measurements of plasma velocity and temperature, based on the Doppler shift and broadening of the H(α) line shape, were previously published [Appl. Opt. 32, 6117 (1993)]. In that paper the Stark broadening of the H(α) transition was estimated from static-ion calculations performed in the early 1970's and found to be negligible in comparison with the Doppler broadening. However, more recent dynamic-ion calculations have shown the Stark broadening to be considerably larger than was previously assumed, resulting in inaccurate temperature measurements. We present a reanalysis of the fluorescence data, taking into account the improved Stark broadening calculations. The correct atomic hydrogen translation temperature and electron number density are obtained from the Doppler and Stark broadening components of the measured line shape. The results indicate a substantial drop in temperature from those previously reported.

  4. Fluorescence-based blood coagulation assay device for measuring activated partial thromboplastin time.

    PubMed

    Dudek, Magdalena M; Kent, Nigel; Gustafsson, Kerstin M; Lindahl, Tomas L; Killard, Anthony J

    2011-01-01

    The measurement of blood clotting time is important in a range of clinical applications such as assessing coagulation disorders and controlling the effect of various anticoagulant drug therapies. Clotting time tests essentially measure the onset of clot formation which results from the formation of fibrin fibers in the blood sample. However, such assays are inherently imprecise due to the highly variable nature of the clot formation process and the sample matrix. This work describes a clotting time measurement assay which uses a fluorescent probe to very precisely detect the onset of fibrin clot formation. It uses a microstructured surface which enhances the formation of multiple localized clot loci and which results in the abrupt redistribution of the fluorescent label at the onset of clot formation in both whole blood and plasma. This methodology was applied to the development of an activated partial thromboplastin time (aPTT) test in a lateral flow microfluidic platform and used to monitor the effect of heparin dosage where it showed linearity from 0 to 2 U/mL in spiked plasma samples (R(2)=0.996, n = 3), correlation against gold standard coagulometry of 0.9986, and correlation against standard hospital aPTT in 32 patient samples of 0.78.

  5. Development of a fluorescent method for simultaneous measurement of glucose concentrations in interstitial fluid and blood

    NASA Astrophysics Data System (ADS)

    Shi, Ting; Li, Dachao; Li, Guoqing; Chen, Limin; Lin, Yuan; Xu, Kexin; Lu, Luo

    2013-12-01

    Continuous blood glucose monitoring is of great clinical significance to patients with diabetes. One of the effective methods to monitor blood glucose is to measure glucose concentrations of interstitial fluid (ISF). However, a time-delay problem exists between ISF and blood glucose concentrations, which results in difficulty in indicating real-time blood glucose concentrations. Therefore, we developed a fluorescent method to verify the accuracy and reliability of simultaneous ISF and blood glucose measurement, especially incorporating it into research on the delay relationship between blood and ISF glucose changes. This method is based on a competitive reaction among borate polymer, alizarin and glucose. When glucose molecules combine with borate polymers in alizarin-borate polymer competitively, changes in fluorescence intensity demonstrate changes in glucose concentrations. By applying the measured results to the blood and ISF glucose delay relationship, we were able to calculate the time delay as an average of 2.16 ± 2.05 min for ISF glucose changes with reference to blood glucose concentrations.

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

    NASA Technical Reports Server (NTRS)

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

    2010-01-01

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

  7. Demonstration of a transmission nuclear resonance fluorescence measurement for a realistic radioactive waste canister scenario

    NASA Astrophysics Data System (ADS)

    Angell, C. T.; Hajima, R.; Hayakawa, T.; Shizuma, T.; Karwowski, H. J.; Silano, J.

    2015-03-01

    Transmission nuclear resonance fluorescence (NRF) is a promising method for precision non-destructive assay (NDA) of fissile isotopes-including 239Pu-in spent fuel while inside a storage canister. The assay, however, could be confounded by the presence of overlapping resonances from competing isotopes in the canister. A measurement is needed to demonstrate that transmission NRF is unaffected by the shielding material. To this end, we carried out a transmission NRF measurement using a mono-energetic γ-ray beam on a proxy target (Al) and absorbing material simulating a realistic spent fuel storage canister. Similar amounts of material as would be found in a possible spent fuel storage canister were placed upstream: concrete, stainless steel (SS 304), lead (as a proxy for U), and water. An Al absorption target was also used as a reference. These measurements demonstrated that the canister material should not significantly influence the non-destructive assay.

  8. A study of density measurements in hypersonic helium tunnels using an electron beam fluorescence technique

    NASA Technical Reports Server (NTRS)

    Honaker, W. C.; Hunter, W. W., Jr.; Woods, W. C.

    1979-01-01

    A series of experiments have been conducted at Langley Research Center to determine the feasibility of using electron-beam fluorescence to measure the free-stream static density of gaseous helium flow over a wide range of conditions. These experiments were conducted in the Langley hypersonic helium tunnel facility and its 3-inch prototype. Measurements were made for a range of stagnation pressures and temperatures and produced free-stream number densities of 1.53 x 10 to the 23rd to 1.25 x 10 to the 24th molecules/cu m and static temperatures from 2 K to 80 K. The results showed the collision quenching cross section to be 4.4 x 10 to the -15th sq cm at 1 K and to have a weak temperature dependence of T to the 1/6. With knowledge of these two values, the free-stream number density can be measured quite accurately.

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

    NASA Technical Reports Server (NTRS)

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

    1983-01-01

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

  10. Characterization of electric thruster plumes using multiplexed laser induced fluorescence measurements

    NASA Technical Reports Server (NTRS)

    Ruyten, W. M.; Keefer, D.

    1992-01-01

    The use of laser-induced fluorescence to obtain spatially resolved measurements of propellant velocities and temperatures in electric thruster plumes is discussed, with emphasis on two innovations of the technique, namely simultaneous recording of the optogalvanic signal in a hollow cathode lamp for the purpose of calibrating Doppler shifts, and two-beam multiplexing to allow the measurement of two velocity components at once. It is also shown how information on plume fluctuations can be obtained from the multiplxed LIF data. The techniques are demonstrated on the plume from a low power arcjet, operated on argon, and its extension to the measurement of ion velocities in electrostatic ion thrusters and stationary plasma thrusters is discussed.

  11. Depolarization of radiation from high-power neodymium lasers and second harmonic generation of partly depolarized radiation

    SciTech Connect

    Arifzhanov, S.B.; Gulamov, A.A.; Redkorechev, V.I.; Usmanov, T.

    1985-07-01

    A theoretical analysis is made of two mechanisms (linear and nonlinear) of depolarization of radiation emitted by high-power solid-state glass lasers: these mechanisms are anisotropy-induced in the active elements and a nonlinear rotation of the polarization ellipse. In the cases of linear and circular polarization of practical interest, a qualitative and numerical analysis is made of the influence of depolarization of the radiation on second harmonic generation by the second type of interaction in KDP crystals. It is shown that in a field of partly depolarized radiation the second harmonic generation efficiency is limited mainly by depolarization of the radiation.

  12. Light stress effect and by nitrogen deficiency in plants of Petiveria alliacea measured with two-chlorophyll-fluorescence technique

    NASA Astrophysics Data System (ADS)

    Zuluaga, H.; Oviedo, A.; Solarte, Efrain; Pena, E. J.

    2004-10-01

    The chlorophyll fluorescence was studied in Petiveria alliacea plants exposed to different nitrogen concentrations and light radiation, the response was measured by two different forms; (1) measuring the photosynthetic efficiency with a pulse amplitude modulated fluorometro (PAM) emitted by a 650 nm diode and (2) measuring the fluorescence spectra caused by high power 452 nm diode with a SD2000 spectrometer. It was found out that the photosynthetic efficiency decreased in the plants exposed to high radiance and low nitrogen. Two chlorophyll fluorescence peaks were observed on 684 nm and 739 nm, the intensities in this wavelengths are inversely related with the light radiance. The correlation between the FIR and photosynthetic efficiency was very strong (r2 = -0.809, p <<0.01) this let us conclude that the fluorescence spectral analysis induced by the diode (excitation at 452 nm) is an efficient technique to detect stress by high light intensity and nitrogen in P. Alliacea plants.

  13. [Application of PARAFAC method and 3-D fluorescence spectra in petroleum pollutant measurement and analysis].

    PubMed

    Pan, Zhao; Wang, Yu-tian; Shao, Xiao-qing; Wu, Xi-jun; Yang, Li-li

    2012-03-01

    A method for identification and concentration measurement of petroleum pollutant by combining three-dimensional (3-D) fluorescence spectra with parallel factor analysis (PARAFAC) was proposed. The main emphasis of research was the measurement of coexisting different kinds of petroleum. The CCl4 solutions of a 0# diesel sample, a 97# gasoline sample, and a kerosene sample were used as measurement objects. The condition of multiple petroleum coexistence was simulated by petroleum solutions with different mixed ratios. The character of PARAFAC in complex mixture coexisting system analysis was studied. The spectra of three kinds of solutions and the spectra of gasoline-diesel mixed samples, diesel-kerosene mixed samples, and gas oline-diesel mixed with small counts of kerosene interference samples were analyzed respectively. The core consistency diagnostic method and residual sum of squares method were applied to calculate the number of factors in PARAFAC. In gasoline-diesel experiment, gasoline or diesel can be identified and measured as a whole respectively by 2-factors parallel factors analysis. In diesel-kerosene experiment, 2-factors parallel factors analysis can only obtain the characters of diesel, and the 3rd factor is needed to separate the kerosene spectral character from the mixture spectrum. When small counts of kerosene exist in gasoline-diesel solution, gasoline and diesel still can be identified and measured as principal components by a 2-factors parallel factor analysis, and the effect of interference on qualitative analysis is not significant. The experiment verified that the PARAFAC method can obtain characteristic spectrum of each kind of petroleum, and the concentration of petroleum in solutions can be predicted simultaneously, with recoveries shown in the paper. The results showed the possibility of petroleum pollutant identification and concentration measurement based on the 3-D fluorescence spectra and PARAFAC.

  14. Automated sorting of polymer flakes: fluorescence labeling and development of a measurement system prototype.

    PubMed

    Brunner, S; Fomin, P; Kargel, Ch

    2015-04-01

    The extensive demand and use of plastics in modern life is associated with a significant economical impact and a serious ecological footprint. The production of plastics involves a high energy consumption and CO2 emission as well as the large need for (limited) fossil resources. Due to the high durability of plastics, large amounts of plastic garbage is mounting in overflowing landfills (plus 9.6 million tons in Europe in the year 2012) and plastic debris is floating in the world oceans or waste-to-energy combustion releases even more CO2 plus toxic substances (dioxins, heavy metals) to the atmosphere. The recycling of plastic products after their life cycle can obviously contribute a great deal to the reduction of the environmental and economical impacts. In order to produce high-quality recycling products, mono-fractional compositions of waste polymers are required. However, existing measurement technologies such as near infrared spectroscopy show limitations in the sorting of complex mixtures and different grades of polymers, especially when black plastics are involved. More recently invented technologies based on mid-infrared, Raman spectroscopy or laser-aided spectroscopy are still under development and expected to be rather expensive. A promising approach to put high sorting purities into practice is to label plastic resins with unique combinations of fluorescence markers (tracers). These are incorporated into virgin resins during the manufacturing process at the ppm (or sub ppm) concentration level, just large enough that the fluorescence emissions can be detected with sensitive instrumentation but neither affect the visual appearance nor the mechanical properties of the polymers. In this paper we present the prototype of a measurement and classification system that identifies polymer flakes (mill material of a few millimeters size) located on a conveyor belt in real time based on the emitted fluorescence of incorporated markers. Classification performance

  15. A two-photon laser induced fluorescence diagnostic with improved sensitivity, localization, and measurement rate

    NASA Astrophysics Data System (ADS)

    Elliott, Drew; Scime, Earl; Short, Zachary

    2016-10-01

    A two-photon absorption laser induced fluorescence diagnostic has been developed for measuring neutrals in fusion plasmas. Implementation of this diagnostic on the HIT-SI3 spheromak has demonstrated the sensitivity of the diagnostic and shown that measurements taken over several plasma pulses are possible. These measurements yielded an unexpected loss of signal when complex collection optics were utilized. Simulations show that this loss of signal can be explained by chromatic aberrations caused by the disparate Kr and D emission. This loss of signal has been addressed with the development of a new calibration scheme involving xenon gas. The Xe calibration scheme emission occurs at 656.00 nm while the deuterium emission is 656.09 nm. This nearly identical emission allows for advanced optical techniques such as confocal collection/injection and spatial filtering to be employed without loss of signal. Spatial filtering has been demonstrated to decrease noise while improving measurement localization, while confocal collection/injection allows for probing and measuring to occur through one viewport. The Xe scheme also allows for a Doppler-free hydrogen measurement. Doppler-free measurements eliminate the need to scan the laser spectrally thus greatly increasing the rate of measurement.

  16. Measurement of Retinal Blood Flow Using Fluorescently Labeled Red Blood Cells1,2,3

    PubMed Central

    Kornfield, Tess E.

    2015-01-01

    Abstract Blood flow is a useful indicator of the metabolic state of the retina. However, accurate measurement of retinal blood flow is difficult to achieve in practice. Most existing optical techniques used for measuring blood flow require complex assumptions and calculations. We describe here a simple and direct method for calculating absolute blood flow in vessels of all sizes in the rat retina. The method relies on ultrafast confocal line scans to track the passage of fluorescently labeled red blood cells (fRBCs). The accuracy of the blood flow measurements was verified by (1) comparing blood flow calculated independently using either flux or velocity combined with diameter measurements, (2) measuring total retinal blood flow in arterioles and venules, (3) measuring blood flow at vessel branch points, and (4) measuring changes in blood flow in response to hyperoxic and hypercapnic challenge. Confocal line scans oriented parallel and diagonal to vessels were used to compute fRBC velocity and to examine velocity profiles across the width of vessels. We demonstrate that these methods provide accurate measures of absolute blood flow and velocity in retinal vessels of all sizes. PMID:26082942

  17. Depolarization ratio, SNR estimation, and polarization sensitivity analysis for a commercial Raman depolarization lidar system

    NASA Astrophysics Data System (ADS)

    Avdikos, George; Georgoussis, George

    2016-05-01

    In this paper we focus on the estimation of the Signal-to-Noise (SNR) ratio of a 3-channel commercial (Raymetics) volcanic ash detection system, (LR111-D300), already operating in UK, and also, we perform a basic lidar polarization sensitivity analysis. The results show that SNR values are higher than 10 for ranges up to 13 km for daytime conditions. This is a quite good result compared with other values presented in bibliography and prove that such system is able to detect volcanic ash detection over a range of 20 km. We also assess the lidar polarization sensitivity and then, we estimate the linear depolarization ratio. By careful choice of the optical components (emitting and receiving optics), it has been shown that uncertainties of polarization states at receiver (and thus too depolarization ratio estimation) can be much reduced.

  18. Dynamic fluorescence anisotropy imaging microscopy in the frequency domain (rFLIM).

    PubMed Central

    Clayton, Andrew H A; Hanley, Quentin S; Arndt-Jovin, Donna J; Subramaniam, Vinod; Jovin, Thomas M

    2002-01-01

    We describe a novel variant of fluorescence lifetime imaging microscopy (FLIM), denoted anisotropy-FLIM or rFLIM, which enables the wide-field measurement of the anisotropy decay of fluorophores on a pixel-by-pixel basis. We adapted existing frequency-domain FLIM technology for rFLIM by introducing linear polarizers in the excitation and emission paths. The phase delay and intensity ratios (AC and DC) between the polarized components of the fluorescence signal are recorded, leading to estimations of rotational correlation times and limiting anisotropies. Theory is developed that allows all the parameters of the hindered rotator model to be extracted from measurements carried out at a single modulation frequency. Two-dimensional image detection with a sensitive CCD camera provides wide-field imaging of dynamic depolarization with parallel interrogation of different compartments of a complex biological structure such as a cell. The concepts and technique of rFLIM are illustrated with a fluorophore-solvent (fluorescein-glycerol) system as a model for isotropic rotational dynamics and with bacteria expressing enhanced green fluorescent protein (EGFP) exhibiting depolarization due to homotransfer of electronic excitation energy (emFRET). The frequency-domain formalism was extended to cover the phenomenon of emFRET and yielded data consistent with a concentration depolarization mechanism resulting from the high intracellular concentration of EGFP. These investigations establish rFLIM as a powerful tool for cellular imaging based on rotational dynamics and molecular proximity. PMID:12202387

  19. Depolarization of light backscattered by randomly oriented nonspherical particles.

    PubMed

    Mishchenko, M I; Hovenier, J W

    1995-06-15

    We derive theoretically and validate numerically general relationships for the elements of the backscattering matrix and for the linear, delta(L), and circular, delta(C), backscattering depolarization ratios for nonspherical particles in random orientation. For the practically important case of randomly oriented particles with a plane of symmetry or particles and their mirror particles occurring in equal numbers and in random orientation, delta(C) = 2delta(L)/(1 - delta(L)). Extensive T-matrix computations for randomly oriented spheroids demonstrate that, although both delta(L) and delta(C) are indicators of particle nonsphericity, they cannot be considered a universal measure of the departure of particle shape from that of a sphere and have no simple dependence on particle size and refractive index.

  20. Single action potentials and subthreshold electrical events imaged in neurons with a fluorescent protein voltage probe.

    PubMed

    Jin, Lei; Han, Zhou; Platisa, Jelena; Wooltorton, Julian R A; Cohen, Lawrence B; Pieribone, Vincent A

    2012-09-06

    Monitoring neuronal electrical activity using fluorescent protein-based voltage sensors has been limited by small response magnitudes and slow kinetics of existing probes. Here we report the development of a fluorescent protein voltage sensor, named ArcLight, and derivative probes that exhibit large changes in fluorescence intensity in response to voltage changes. ArcLight consists of the voltage-sensing domain of Ciona intestinalis voltage-sensitive phosphatase and super ecliptic pHluorin that carries the point mutation A227D. The fluorescence intensity of ArcLight A242 decreases by 35% in response to a 100 mV depolarization when measured in HEK293 cells, which is more than five times larger than the signals from previously reported fluorescent protein voltage sensors. We show that the combination of signal size and response speed of these new probes allows the reliable detection of single action potentials and excitatory potentials in individual neurons and dendrites.

  1. AUV Measured Variability in Phytoplankton Fluorescence within the ETM of the Columbia River during Summer 2013

    NASA Astrophysics Data System (ADS)

    McNeil, C. L.; Shcherbina, A.; Litchendorf, T. M.; Sanford, T. B.; Martin, D.; Baptista, A. M.; Lopez, J.; Crump, B. C.; Peterson, T. D.; Prahl, F. G.; Cravo, A.

    2014-12-01

    We present highly resolved observations of fluorescence and optical backscatter taken in the estuarine turbidity maxima (ETM) of the North Channel of the Columbia River estuary (USA) during summer 2013. Measurements were made using two REMUS-100 autonomous underwater vehicles (AUVs) equipped with ECO Puck triplets. Concentrations of three phytoplankton pigments were measured by fluorescence emission at wavelengths of 695 nm for chlorophyll, 570 nm for phycoerythrin, and 680 nm for phycocyanin. We use phycocyanin to indicate the presence of freshwater phytoplankton. Optical backscatter at wavelengths of 700 nm and 880 nm are used to characterize turbidity. During flood tide, high phycocyanin concentrations were associated with a strong ETM event which had relatively low salinity waters of approximately 6 psu. These data indicate that this low salinity ETM event contained large concentrations of freshwater phytoplankton. Since freshwater phytoplankton are known to lyse in saltwater, the brackish ETM event may have formed by the accumulation of lysed freshwater phytoplankton that settled out from the river as it mixed in the lower estuary. As the flood tide proceeded, it brought high concentrations of marine phytoplankton into the north channel at mid-depth as indicated by high chlorophyll levels with significantly lower phycoerythrin concentrations in high salinity waters of approximately 30 psu. The data set highlights the potential for large variability in phytoplankton species composition and concentrations within the ETM depending on mixing rates and phytoplankton bloom dynamics. Visualization of the 4-D data is aided by generating interpolated data movies.

  2. Distance measurements near the myosin head-rod junction using fluorescence spectroscopy.

    PubMed

    Kekic, M; Huang, W; Moens, P D; Hambly, B D; dos Remedios, C G

    1996-07-01

    We reacted a fluorescent probe, N-methyl-2-anilino-6-naphthalenesulfonyl chloride (MNS-Ci), with a specific lysine residue of porcine cardiac myosin located in the S-2 region of myosin. We performed fluorescence resonance energy transfer (FRET) spectroscopy measurements between this site and three loci (Cys109, Cys125, and Cys154) located within different myosin light-chain 2s (LC2) bound to the myosin "head". We used LC2s from rabbit skeletal muscle myosin (Cys125), chicken gizzard smooth muscle myosin (Cys109), or a genetically engineered mutant of chicken skeletal muscle myosin (Cys154). The atomic coordinates of these LC2 loci can be closely approximated, and the FRET measurements were used to determine the position of the MNS-labeled lysine with respect to the myosin head. The C-terminus of myosin subfragment-1 determined by Rayment et al. ends abruptly after a sharp turn of its predominantly alpha-helical structure. We have constructed a model based on our FRET distance data combined with the known structure of chicken skeletal muscle myosin subfragment-1. This model suggests that the loci that bracket the head-rod junction will be useful for evaluating dynamic changes in this region.

  3. Distance measurements near the myosin head-rod junction using fluorescence spectroscopy.

    PubMed Central

    Kekic, M; Huang, W; Moens, P D; Hambly, B D; dos Remedios, C G

    1996-01-01

    We reacted a fluorescent probe, N-methyl-2-anilino-6-naphthalenesulfonyl chloride (MNS-Ci), with a specific lysine residue of porcine cardiac myosin located in the S-2 region of myosin. We performed fluorescence resonance energy transfer (FRET) spectroscopy measurements between this site and three loci (Cys109, Cys125, and Cys154) located within different myosin light-chain 2s (LC2) bound to the myosin "head". We used LC2s from rabbit skeletal muscle myosin (Cys125), chicken gizzard smooth muscle myosin (Cys109), or a genetically engineered mutant of chicken skeletal muscle myosin (Cys154). The atomic coordinates of these LC2 loci can be closely approximated, and the FRET measurements were used to determine the position of the MNS-labeled lysine with respect to the myosin head. The C-terminus of myosin subfragment-1 determined by Rayment et al. ends abruptly after a sharp turn of its predominantly alpha-helical structure. We have constructed a model based on our FRET distance data combined with the known structure of chicken skeletal muscle myosin subfragment-1. This model suggests that the loci that bracket the head-rod junction will be useful for evaluating dynamic changes in this region. Images FIGURE 4 FIGURE 5 PMID:8804587

  4. Modeling of dual emission laser induced fluorescence for slurry thickness measurements in chemical mechanical polishing

    NASA Astrophysics Data System (ADS)

    Gray, Caprice; Rogers, Chris B.; Manno, Vincent P.; White, Robert D.

    2011-07-01

    Dual emission laser induced fluorescence (DELIF) is a technique for measuring the instantaneous thin fluid film thickness in dynamic systems. Two fluorophores within the system produce laser induced emissions that are filtered and captured by two cameras. The ratio of the images from these cameras is used to cancel the effect of the laser beam profile on the image intensity. The resultant intensity ratio can be calibrated to a fluid film thickness. The utilization of a 2-dye system when applied to Chemical Mechanical Polishing (CMP) is complicated by the fluorescence of the polymeric polishing pad and the light scattering particles in the polishing slurry. We have developed a model of DELIF for CMP with 1-dye employing the polishing pad as the second fluorophore. While scattering particles in the slurry decrease the overall intensity of the individual images, the contrast in the image ratio increases. Using the 1-dye DELIF system to measure thin slurry films, our model results indicate that a cubic calibration may be needed. However, experimental results suggest a linear calibration is achieved for slurry films between 0 and 133 μm thick with scattering coefficients as high as 8.66 mm-1 at a wavelength equal to 410 nm.

  5. Three-dimensional printed miniaturized spectral system for collagen fluorescence lifetime measurements

    NASA Astrophysics Data System (ADS)

    Zou, Luwei; Koslakiewicz, Ronald; Mahmoud, Mohamad; Fahs, Mehdi; Liu, Rui; Lo, Joe Fujiou

    2016-07-01

    Various types of collagens, e.g., type I and III, represent the main load-bearing components in biological tissues. Their composition changes during processes such as wound healing and fibrosis. When excited by ultraviolet light, collagens exhibit autofluorescence distinguishable by their unique fluorescent lifetimes across a range of emission wavelengths. Here, we designed a miniaturized spectral-lifetime detection system as a noninvasive probe for monitoring tissue collagen compositions. A sine-modulated LED illumination was applied to enable frequency domain fluorescence lifetime measurements under three wavelength bands, separated via a series of longpass dichroics at 387, 409, and 435 nm. We employed a lithography-based three-dimensional (3-D) printer with <50 μm resolution to create a custom designed optomechanics in a handheld form factor. We examined the characteristics of the optomechanics with finite element modeling to simulate the effect of thermal (from LED) and mechanical (from handling) strain on the optical system. The geometry was further optimized with ray tracing to form the final 3-D printed structure. Using this device, the phase shift and demodulation of collagen types were measured, where the separate spectral bands enhanced the differentiation of their lifetimes. This system represents a low cost, handheld probe for clinical tissue monitoring applications.

  6. 3D printed miniaturized spectral system for tissue fluorescence lifetime measurements

    NASA Astrophysics Data System (ADS)

    Zou, Luwei; Mahmoud, Mohamad; Fahs, Mehdi; Liu, Rui; Lo, Joe F.

    2016-04-01

    Various types of collagens, e.g. type I and III, represent the main load-bearing components in biological tissues. Their composition changes during processes like wound healing and fibrosis. Collagens exhibit autofluorescence when excited by ultra-violet light, distinguishable by their unique fluorescent lifetimes across a range of emission wavelengths. Therefore, we designed a miniaturized spectral-lifetime detection system for collagens as a non-invasive probe for monitoring tissue in wound healing and scarring applications. A sine modulated LED illumination was applied to enable frequency domain (FD) fluorescence lifetime measurements under different wavelengths bands, separated via a series of longpass dichroics at 387nm, 409nm and 435nm. To achieve the minute scale of optomechanics, we employed a stereolithography based 3D printer with <50 μm resolution to create a custom designed optical mount in a hand-held form factor. We examined the characteristics of the 3D printed optical system with finite element modeling to simulate the effect of thermal (LED) and mechanical (handling) strain on the optical system. Using this device, the phase shift and demodulation of collagen types were measured, where the separate spectral bands enhanced the differentiation of their lifetimes.

  7. In vitro quantitative light-induced fluorescence to measure changes in enamel mineralization.

    PubMed

    Gmür, Rudolf; Giertsen, Elin; van der Veen, Monique H; de Josselin de Jong, Elbert; ten Cate, Jacob M; Guggenheim, Bernhard

    2006-09-01

    A sensitive, quantitative method for investigating changes in enamel mineralization of specimens subjected to in vitro or in situ experimentation is presented. The fluorescence-detecting instrument integrates a Xenon arc light source and an object positioning stage, which makes it particularly suitable for the nondestructive assessment of demineralized or remineralized enamel. We demonstrate the ability of in vitro quantitative light-induced fluorescence (QLF) to quantify changes in mineralization of bovine enamel discs that had been exposed in vitro to a demineralizing gel (n=36) or biofilm-mediated demineralization challenges (n=10), or were carried in situ by three volunteers during a 10-day experiment (n=12). Further experiments show the technique's value for monitoring the extent of remineralization in 36 specimens exposed in vitro to oral multispecies biofilms and document the repeatability of in vitro QLF measurements (n=10) under standardized assay conditions. The validity of the method is illustrated by comparison with transversal microradiography (TMR), the invasive current gold standard for assessing experimental changes in enamel mineralization. Ten discs with 22 measurement areas for comparison demonstrated a positive correlation between TMR and QLF (r=0.82). Filling a technological gap, this QLF system is a promising tool to assay in vitro nondestructively localized changes in mineralization of enamel specimens.

  8. Quantitative Affinity Determination by Fluorescence Anisotropy Measurements of Individual Nanoliter Droplets

    PubMed Central

    2017-01-01

    Fluorescence anisotropy measurements of reagents compartmentalized into individual nanoliter droplets are shown to yield high-resolution binding curves from which precise dissociation constants (Kd) for protein–peptide interactions can be inferred. With the current platform, four titrations can be obtained per minute (based on ∼100 data points each), with stoichiometries spanning more than 2 orders of magnitude and requiring only tens of microliters of reagents. In addition to affinity measurements with purified components, Kd values for unpurified proteins in crude cell lysates can be obtained without prior knowledge of the concentration of the expressed protein, so that protein purification can be avoided. Finally, we show how a competition assay can be set up to perform focused library screens, so that compound labeling is not required anymore. These data demonstrate the utility of droplet compartments for the quantitative characterization of biomolecular interactions and establish fluorescence anisotropy imaging as a quantitative technique in a miniaturized droplet format, which is shown to be as reliable as its macroscopic test tube equivalent. PMID:28192993

  9. Guided fluorescence diagnosis of childhood caries: preliminary measures correlate with depth of carious decay

    NASA Astrophysics Data System (ADS)

    Timoshchuk, Mari-Alina; Zhang, Liang; Dickinson, Brian A.; Ridge, Jeremy S.; Kim, Amy S.; Baltuck, Camille T.; Nelson, Leonard Y.; Berg, Joel H.; Seibel, Eric J.

    2014-02-01

    The current rise in childhood caries worldwide has increased the demand for portable technologies that can quickly and accurately detect and diagnose early stage carious lesions. These lesions, if identified at an early stage, can be reversed with remineralization treatments, education, and improvements in home care. A multi-modal optical prototype for detecting and diagnosing occlusal caries demineralization in vivo has been developed and pilot tested. The device uses a 405-nm laser as a scanned illumination source to obtain high resolution and high surface contrast reflectance images, which allows the user to quickly image and screen for any signs of demineralized enamel. When a suspicious region is located, the device can be switched to perform dual laser fluorescence spectroscopy using 405-nm and 532-nm laser excitations. These spectra are used to compute an auto-fluorescence (AF) ratio of the suspicious region and the percent difference of AF ratios from a healthy region of the same tooth. The device was tested on 7 children's teeth in vivo with clinically diagnosed carious lesions. Lesion depth was then visually estimated from the video image using the 405-nm scanned light source, and within a month the maximum drill depth was assessed by a clinician. The researcher and clinicians were masked from previous measurements in a blinded study protocol. Preliminary results show that the ratiometric percent difference measurement of the AF spectrum of the tooth correlates with the severity of the demineralization as assessed by the clinician after drilling.

  10. Assessment of lidar depolarization uncertainty by means of a polarimetric lidar simulator

    NASA Astrophysics Data System (ADS)

    Bravo-Aranda, Juan Antonio; Belegante, Livio; Freudenthaler, Volker; Alados-Arboledas, Lucas; Nicolae, Doina; José Granados-Muñoz, María; Guerrero-Rascado, Juan Luis; Amodeo, Aldo; D'Amico, Giusseppe; Engelmann, Ronny; Pappalardo, Gelsomina; Kokkalis, Panos; Mamouri, Rodanthy; Papayannis, Alex; Navas-Guzmán, Francisco; José Olmo, Francisco; Wandinger, Ulla; Amato, Francesco; Haeffelin, Martial

    2016-10-01

    Lidar depolarization measurements distinguish between spherical and non-spherical aerosol particles based on the change of the polarization state between the emitted and received signal. The particle shape information in combination with other aerosol optical properties allows the characterization of different aerosol types and the retrieval of aerosol particle microphysical properties. Regarding the microphysical inversions, the lidar depolarization technique is becoming a key method since particle shape information can be used by algorithms based on spheres and spheroids, optimizing the retrieval procedure. Thus, the identification of the depolarization error sources and the quantification of their effects are crucial. This work presents a new tool to assess the systematic error of the volume linear depolarization ratio (δ), combining the Stokes-Müller formalism and the complete sampling of the error space using the lidar model presented in Freudenthaler (2016a). This tool is applied to a synthetic lidar system and to several EARLINET lidars with depolarization capabilities at 355 or 532 nm. The lidar systems show relative errors of δ larger than 100 % for δ values around molecular linear depolarization ratios (˜ 0.004 and up to ˜ 10 % for δ = 0.45). However, one system shows only relative errors of 25 and 0.22 % for δ = 0.004 and δ = 0.45, respectively, and gives an example of how a proper identification and reduction of the main error sources can drastically reduce the systematic errors of δ. In this regard, we provide some indications of how to reduce the systematic errors.

  11. Laser-Induced Fluorescence Measurements and Modeling of Nitric Oxide in Counterflow Diffusion Flames

    NASA Technical Reports Server (NTRS)

    Ravikrishna, Rayavarapu V.

    2000-01-01

    The feasibility of making quantitative nonintrusive NO concentration ([NO]) measurements in nonpremixed flames has been assessed by obtaining laser-induced fluorescence (LIF) measurements of [NO] in counterflow diffusion flames at atmospheric and higher pressures. Comparisons at atmospheric pressure between laser-saturated fluorescence (LSF) and linear LIF measurements in four diluted ethane-air counterflow diffusion flames with strain rates from 5 to 48/s yielded excellent agreement from fuel-lean to moderately fuel-rich conditions, thus indicating the utility of a model-based quenching correction technique, which was then extended to higher pressures. Quantitative LIF measurements of [NO] in three diluted methane-air counterflow diffusion flames with strain rates from 5 to 35/s were compared with OPPDIF model predictions using the GRI (version 2.11) chemical kinetic mechanism. The comparisons revealed that the GRI mechanism underpredicts prompt-NO by 30-50% at atmospheric pressure. Based on these measurements, a modified reaction rate coefficient for the prompt-NO initiation reaction was proposed which causes the predictions to match experimental data. Temperature measurements using thin filament pyrometry (TFP) in conjunction with a new calibration method utilizing a near-adiabatic H2-air Hencken burner gave very good comparisons with model predictions in these counterflow diffusion flames. Quantitative LIF measurements of [NO] were also obtained in four methane-air counterflow partially-premixed flames with fuel-side equivalence ratios (phi(sub B)) of 1.45, 1.6, 1.8 and 2.0. The measurements were in excellent agreement with model predictions when accounting for radiative heat loss. Spatial separation between regions dominated by the prompt and thermal NO mechanisms was observed in the phi(sub B) = 1.45 flame. The modified rate coefficient proposed earlier for the prompt-NO initiation reaction improved agreement between code predictions and measurements in the

  12. Intercomparison of Hantzsch and fiber-laser-induced-fluorescence formaldehyde measurements

    NASA Astrophysics Data System (ADS)

    Kaiser, J.; Li, X.; Tillmann, R.; Acir, I.; Holland, F.; Rohrer, F.; Wegener, R.; Keutsch, F. N.

    2014-06-01

    Two gas-phase formaldehyde (HCHO) measurement techniques, a modified commercial wet-chemical instrument based on Hantzsch fluorimetry and a custom-built instrument based on fiber laser-induced fluorescence (FILIF), were deployed at the atmospheric simulation chamber SAPHIR (Simulation of Atmospheric PHotochemistry In a large Reaction Chamber) to compare the instruments' performances under a range of conditions. Thermolysis of para-HCHO and ozonolysis of 1-butene were used as HCHO sources, allowing for calculations of theoretical HCHO mixing ratios. Calculated HCHO mixing ratios are compared to measurements, and the two measurements are also compared. Experiments were repeated under dry and humid conditions (RH < 2% and RH > 60%) to investigate the possibility of a water artifact in the FILIF measurements. The ozonolysis of 1-butene also allowed for the investigation of an ozone artifact seen in some Hantzsch measurements in previous intercomparisons. Results show that under all conditions the two techniques are well correlated (R2 ≥ 0.997), and linear regression statistics show measurements agree with within stated uncertainty (15% FILIF + 5% Hantzsch). No water or ozone artifacts are identified. While a slight curvature is observed in some Hantzsch vs. FILIF regressions, the potential for variable instrument sensitivity cannot be attributed to a single instrument at this time. Measurements at low concentrations highlight the need for a secondary method for testing the purity of air used in instrument zeroing and the need for further FILIF White cell outgassing experiments.

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

    NASA Astrophysics Data System (ADS)

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

    2012-12-01

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

  14. Q(M) and the depolarization index scalar metrics.

    PubMed

    Espinosa-Luna, Rafael; Bernabeu, Eusebio; Atondo-Rubio, Gelacio

    2008-04-01

    A depolarization scalar metric for Mueller matrices, named Q(M), is derived from the degree of polarization. Q(M) has been recently reported, and it has been deduced from the nine bilinear constraints between the sixteen elements of the Mueller-Jones matrix. We discuss the relations between Q(M) and the depolarization index.

  15. Simple analytic formula for the strength of spin depolarizing resonance

    SciTech Connect

    Lee, S.Y.

    1985-01-01

    A simple analytic formula is derived to explain the periodicity of spin depolarizing resonance. The spin depolarizing resonance strengths of CPS and SPS at CERN and the lattices of meson factory at TRIUMF are used to compare with the analytic formula.

  16. [Cortical spreading depolarization: a new pathophysiological mechanism in neurological diseases].

    PubMed

    Sánchez-Porras, Renán; Robles-Cabrera, Adriana; Santos, Edgar

    2014-05-20

    Cortical spreading depolarization is a wave of almost complete depolarization of the neuronal and glial cells that occurs in different neurological diseases such as migraine with aura, subarachnoid hemorrhage, intracerebral hemorrhage, head trauma and stroke. These depolarization waves are characterized by a change in the negative potential with an amplitude between -10 and -30mV, duration of ∼1min and changes in the ion homeostasis between the intra- and extracellular space. This results in neuronal edema and dendritic distortion. Under pathologic states of hypoperfusion, cortical spreading depolarization can produce oxidative stress, worsen hypoxia and induce neuronal death. This is due to intense arterial vasoconstriction produced by an inverse response called spreading ischemia. Only in the last years there has been an electrophysiological confirmation of cortical spreading depolarization in human brains. Occurrence of cortical spreading depolarization has been associated with worse outcome in patients. Currently, increased knowledge regarding the pathophysiologic mechanisms supports the hypothetical correlation of cortical spreading depolarization with brain damage in humans. There are diverse therapeutic alternatives that promise inhibition of cortical spreading depolarization and subsequent better outcomes.

  17. Saturated fluorescence measurements of the hydroxyl radical in laminar high-pressure flames

    NASA Technical Reports Server (NTRS)

    Carter, Campbell D.; King, Galen B.; Laurendeau, Normand M.

    1990-01-01

    The efficacy of laser saturated fluorescence (LSF) for OH concentration measurements in high pressure flames was studied theoretically and experimentally. Using a numerical model describing the interaction of hydroxyl with nonuniform laser excitation, the effect of pressure on the validity of the balanced cross-rate model was studied along with the sensitivity of the depopulation of the laser-coupled levels to the ratio of rate coefficients describing: (1) electronic quenching to (sup 2) Sigma (+) (v double prime greater than 0), and (2) vibrational relaxation from v double prime greater than 0 to v double prime = 0. At sufficiently high pressures and near-saturated conditions, the total population of the laser-coupled levels reaches an asymptotic value, which is insensitive to the degree of saturation. When the ratio of electronic quenching to vibrational relaxation is small and the rate of coefficients for rotational transfer in the ground and excited electronic states are nearly the same, the balanced cross-rate model remains a good approximation for all pressures. When the above ratio is large, depopulation of the laser-coupled levels becomes significant at high pressures, and thus the balanced cross-rate model no longer holds. Under these conditions, however, knowledge of the depletion of the laser-coupled levels can be used to correct the model. A combustion facility for operation up to 20 atm was developed to allow LSF measurements of OH in high pressure flames. Using this facility, partial saturation in laminar high pressure (less than or equal to 12.3 atm) C2H6/O2/N2 flames was achieved. To evaluate the limits of the balanced cross-rate model, absorption and calibrated LSF measurements at 3.1 and 6.1 atm were compared. The fluorescence voltages were calibrated with absorption measurements in an atmospheric flame and corrected for their finite sensitivity to quenching with: (1) estimated quenching rate coefficients, and (2) an in situ measurement from a

  18. Lymphocyte fluorescence polarization measurements with the cellscan system: application to the SCM cancer test.

    PubMed

    Deutsch, M; Ron, I; Weinreb, A; Tirosh, R; Chaitchik, S

    1996-02-01

    The SCM (Structuredness of Cytoplasmic Matrix) cancer test, a procedure based on detection of differences in lymphocyte activation between individuals with and without cancer, has remained controversial with inconsistent results reported by different authors. As originally described, the test includes two technically demanding steps, the first a lymphocyte separation procedure and the second a series of fluorescence polarization measurements. The Cellscan, a high-precision static cytometer system has been configured to perform the SCM test. The apparatus facilitates the polarization measurements and can analyze cells separated using simpler procedures than were originally described. Using methods and diagnostic criteria adapted for the Cellscan system, the SCM test correctly classified > 90% of patients with cancer and > 90% of individuals without cancer.

  19. Measuring Membrane Protein Dimerization Equilibrium in Lipid Bilayers by Single-Molecule Fluorescence Microscopy.

    PubMed

    Chadda, R; Robertson, J L

    2016-01-01

    Dimerization of membrane protein interfaces occurs during membrane protein folding and cell receptor signaling. Here, we summarize a method that allows for measurement of equilibrium dimerization reactions of membrane proteins in lipid bilayers, by measuring the Poisson distribution of subunit capture into liposomes by single-molecule photobleaching analysis. This strategy is grounded in the fact that given a comparable labeling efficiency, monomeric or dimeric forms of a membrane protein will give rise to distinctly different photobleaching probability distributions. These methods have been used to verify the dimer stoichiometry of the Fluc F(-) ion channel and the dimerization equilibrium constant of the ClC-ec1 Cl(-)/H(+) antiporter in lipid bilayers. This approach can be applied to any membrane protein system provided it can be purified, fluorescently labeled in a quantitative manner, and verified to be correctly folded by functional assays, even if the structure is not yet known.

  20. Line scan fluorescence correlation spectroscopy for three-dimensional microfluidic flow velocity measurements

    NASA Astrophysics Data System (ADS)

    Pan, Xiaotao; Shi, Xianke; Korzh, Vladimir; Yu, Hanry; Wohland, Thorsten

    2009-03-01

    The flow direction of microfluidics in biological applications is not limited to two dimensions, but often extends to three dimensions. Currently there are optical methods available for the measurement of 3-D microfluidic flow vectors, but with low spatial resolution. Line scan fluorescence correlation spectroscopy (FCS) was proposed to determine flow directions in 2-D within microchannels and small blood vessels in our previous work. Importantly, its spatial resolution was demonstrated to be as good as 0.5 μm. In this work, we extend line scan FCS to the third dimension for the characterization of 3-D flow velocity vectors. The spatial resolution is close to the diffraction limit using a scan length of 0.5 μm in all three dimensions. The feasibility of line scan FCS for 3-D microfluidic flow is verified by measurements in microchannels and small blood vessels of zebrafish embryos.

  1. Chromosome translocations measured by fluorescence in-situ hybridization: A promising biomarker

    SciTech Connect

    Lucas, J.N.; Straume, T.

    1995-10-01

    A biomarker for exposure and risk assessment would be most useful if it employs an endpoint that is highly quantitative, is stable with time, and is relevant to human risk. Recent advances in chromosome staining using fluorescence in situ hybridization (FISH) facilitate fast and reliable measurement of reciprocal translocations, a kind of DNA damage linked to both prior exposure and risk. In contrast to other biomarkers available, the frequency of reciprocal translocations in individuals exposed to whole-body radiation is stable with time post exposure, has a rather small inter-individual variability, and can be measured accurately at the low levels. Here, the authors discuss results from their studies demonstrating that chromosome painting can be used to reconstruct radiation dose for workers exposed within the dose limits, for individuals exposed a long time ago, and even for those who have been diagnosed with leukemia but not yet undergone therapy.

  2. Spectral fluorescence signature techniques and absorption measurements for continuous monitoring of biofuel-producing microalgae cultures

    NASA Astrophysics Data System (ADS)

    Martín de la Cruz, M. C.; Gonzalez Vilas, L.; Yarovenko, N.; Spyrakos, E.; Torres Palenzuela, J. M.

    2013-08-01

    Biofuel production from microalgae can be both sustainable and economically viable. Particularly in the case of algal growth in wastewater an extra benefit is the removal or biotransformation of pollutants from these types of waters. A continuous monitoring system of the microalgae status and the concentration of different wastewater contaminants could be of great help in the biomass production and the water characterisation. In this study we present a system where spectral fluorescence signature (SFS) techniques are used along with absorption measurements to monitor microalgae cultures in wastewater and other mediums. This system aims to optimise the microalgae production for biofuel applications or other uses and was developed and tested in prototype indoor photo-bioreactors at the University of Vigo. SFS techniques were applied using the fluorescence analyser INSTAND-SCREENER developed by Laser Diagnostic Instruments AS. INSTAND-SCREENER permits wavelength scanning in two modes, one in UV and another in VIS. In parallel, it permits the on-line monitoring and rapid analysis of both water quality and phytoplankton status without prior treatment of the sample. Considering that different contaminants and microalgae features (density, status etc.) have different spectral signatures of fluorescence and absorption properties, it is possible to characterise them developing classification libraries. Several algorithms were used for the classification. The implementation of this system in an outdoor raceway reactor in a Spanish wastewater treatment plant is also discussed. This study was part of the Project EnerBioAlgae (http://www.enerbioalgae.com/), which was funded by the Interreg SUDOE and led by the University of Vigo.

  3. Real-time quantitative fluorescence measurement of microscale cell culture analog systems

    NASA Astrophysics Data System (ADS)

    Oh, Taek-il; Kim, Donghyun; Tatosian, Daniel; Sung, Jong Hwan; Shuler, Michael

    2007-02-01

    A microscale cell culture analog (μCCA) is a cell-based lab-on-a-chip assay that, as an animal surrogate, is applied to pharmacological studies for toxicology tests. A μCCA typically comprises multiple chambers and microfluidics that connect the chambers, which represent animal organs and blood flow to mimic animal metabolism more realistically. A μCCA is expected to provide a tool for high-throughput drug discovery. Previously, a portable fluorescence detection system was investigated for a single μCCA device in real-time. In this study, we present a fluorescence-based imaging system that provides quantitative real-time data of the metabolic interactions in μCCAs with an emphasis on measuring multiple μCCA samples simultaneously for high-throughput screening. The detection system is based on discrete optics components, with a high-power LED and a charge-coupled device (CCD) camera as a light source and a detector, for monitoring cellular status on the chambers of each μCCA sample. Multiple samples are characterized mechanically on a motorized linear stage, which is fully-automated. Each μCCA sample has four chambers, where cell lines MES-SA/DX- 5, and MES-SA (tumor cells of human uterus) have been cultured. All cell-lines have been transfected to express the fusion protein H2B-GFP, which is a human histone protein fused at the amino terminus to EGFP. As a model cytotoxic drug, 10 μM doxorubicin (DOX) was used. Real-time quantitative data of the intensity loss of enhanced green fluorescent protein (EGFP) during cell death of target cells have been collected over several minutes to 40 hours. Design issues and improvements are also discussed.

  4. Measurement of OH reactivity by laser flash photolysis coupled with laser-induced fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Stone, Daniel; Whalley, Lisa K.; Ingham, Trevor; Edwards, Peter M.; Cryer, Danny R.; Brumby, Charlotte A.; Seakins, Paul W.; Heard, Dwayne E.

    2016-07-01

    OH reactivity (k'OH) is the total pseudo-first-order loss rate coefficient describing the removal of OH radicals to all sinks in the atmosphere, and is the inverse of the chemical lifetime of OH. Measurements of ambient OH reactivity can be used to discover the extent to which measured OH sinks contribute to the total OH loss rate. Thus, OH reactivity measurements enable determination of the comprehensiveness of measurements used in models to predict air quality and ozone production, and, in conjunction with measurements of OH radical concentrations, to assess our understanding of OH production rates. In this work, we describe the design and characterisation of an instrument to measure OH reactivity using laser flash photolysis coupled to laser-induced fluorescence (LFP-LIF) spectroscopy. The LFP-LIF technique produces OH radicals in isolation, and thus minimises potential interferences in OH reactivity measurements owing to the reaction of HO2 with NO which can occur if HO2 is co-produced with OH in the instrument. Capabilities of the instrument for ambient OH reactivity measurements are illustrated by data collected during field campaigns in London, UK, and York, UK. The instrumental limit of detection for k'OH was determined to be 1.0 s-1 for the campaign in London and 0.4 s-1 for the campaign in York. The precision, determined by laboratory experiment, is typically < 1 s-1 for most ambient measurements of OH reactivity. Total uncertainty in ambient measurements of OH reactivity is ˜ 6 %. We also present the coupling and characterisation of the LFP-LIF instrument to an atmospheric chamber for measurements of OH reactivity during simulated experiments, and provide suggestions for future improvements to OH reactivity LFP-LIF instruments.

  5. Electron Beam Fluorescence Temperature Measurements of N2 in a SemiConductor Plasma Reactor

    NASA Astrophysics Data System (ADS)

    Shimada, Masashi; Cattolica, Robert; Tynan George, R.

    2003-10-01

    The rotational temperature of nitrogen molecules in inductively coupled plasma (ICP) discharge has been measured using the electron beam fluorescence (EBF) technique. The neutral gas temperature is an important parameter in understanding the energy balance and neutral radical uniformity in plasma processing. The EBF technique has been used to study rarefied flows, but has not been applied previously to characterize a semiconductor plasma reactor. In this work an electron beam was integrated into an inductively coupled semiconductor plasma reactor, and was used to excite neutral nitrogen molecules from the N2X1 state into the excited molecular ion N2+B2 state. The electron-beam excitation process maps the rotational population distribution of the original ground state into the excited molecular ion state following a dipole model with deltaK=+/-1. The rotational temperature, which is equivalent to the neutral gas translational temperature under plasma reactor conditions, can be determined from the rotational intensity distribution of the fluorescence spectrum from the N2+B2 state. For the range of neutral gas temperatures found in the plasma reactor a dipole model of the excitation-emission process provides good agreement with the measured spectrum. A least-square fitting procedure comparing the model and measured emission spectra is used to determine the rotation temperature. In this poster the first EBF measurements of gas heating in an ICP reactor at different gas pressures (20, 50 mTorr) and plasma source input powers (250, 500, 1000W) are reported and compared with the plasma emission data by using a insertable optical fiber.

  6. Electron beam fluorescence temperature measurements of N2 in a semiconductor plasma reactor

    NASA Astrophysics Data System (ADS)

    Shimada, M.; Cattolica, R.; Tynan, G. R.

    2004-03-01

    The rotational temperature of nitrogen molecules in inductively coupled plasma (ICP) discharge has been measured using the electron beam fluorescence (EBF) technique. The neutral gas temperature is an important parameter in understanding the energy balance and neutral radical uniformity in plasma processing. The EBF technique has been used to study rarefied flows, but has not been applied previously to characterize a semiconductor plasma reactor. In this work an electron beam was integrated into an inductively coupled semiconductor plasma reactor, and was used to excite neutral nitrogen molecules from the N2X 1Σg+ state into the excited molecular ion N2+B 2Σu+ state. The electron beam excitation process maps the rotational population distribution of the original ground state into the excited molecular ion state following a dipole model with ΔK=+/-1. The rotational temperature, which is thought to be equivalent to the neutral gas translational temperature under plasma reactor conditions, can be determined from the rotational intensity distribution of the fluorescence spectrum from the N2+B 2Σu+ state. For the range of neutral gas temperatures found in the plasma reactor a dipole model of the excitation-emission process provides good agreement with the measured spectrum. A least-square fitting procedure comparing the model and measured emission spectra is used to determine the rotation temperature. In this article the first EBF measurements of gas heating in an ICP reactor at different gas pressures (20, 50 mTorr) and plasma source input powers (250, 500, and 1000 W) are reported. .

  7. Intercomparison of Hantzsch and fiber-laser-induced-fluorescence formaldehyde measurements

    NASA Astrophysics Data System (ADS)

    Kaiser, J.; Li, X.; Tillmann, R.; Acir, I.; Rohrer, F.; Wegener, R.; Keutsch, F. N.

    2014-01-01

    Two gas-phase formaldehyde (HCHO) measurement techniques, a modified commercial wet-chemical instrument based on Hantzsch Fluorimetry and a custom-built instrument based on Fiber-Laser Induced Fluorescence (FILIF), were deployed at the atmospheric simulation chamber SAPHIR to compare the instruments' performances under a range of conditions. Thermolysis of para-HCHO and ozonolysis of 1-butene were used as HCHO sources, allowing for calculations of theoretical HCHO mixing ratios. Calculated HCHO mixing ratios are compared to measurements, and the two measurements are also compared. Experiments were repeated under dry and humid conditions (RH < 2% and RH > 60%) to investigate the possibility of a water artifact in the FILIF measurements. The ozonolysis of 1-butene also allowed for the investigation of an ozone artifact seen in some Hantzsch measurements in previous intercomparisons. Results show that under all conditions the two techniques are well correlated (R2 ≥ 0.997), and linear regression statistics show measurements agree with within stated uncertainty (15% FILIF + 5% Hantzsch). No water or ozone artifacts are identified.

  8. Comparison of NO and OH planar fluorescence temperature measurements in scramjet model flowfields

    SciTech Connect

    Mcmillin, B.K.; Seitzman, J.M.; Hanson, R.K.

    1994-10-01

    The use of nitric oxide (NO) and the hydroxyl radical (OH) as temperature tracers, in a two-line planar laser-induced fluorescence technique, is examined in the context of a supersonic mixing and combustion flowfield. The temperature measurements were based on the sequential excitation of two transitions, either in the A implied by X (0,0) band of NO near 226 nm or the A implied by X (1,0) band of OH near 283 nm. The measurements were obtained for each species through the use of two lasers and two cameras, with each camera integrating signal induced from only one of the lasers. Both temporally resolved and frame-averaged temperature measurements of each species are presented. Additional results include simultaneous NO and OH visualizations, in which seeded NO marks the fuel jet fluid and nascent OH marks the reaction zones and convected combustion gases. A detailed temperature comparison shows good agreement in the common measurement regions and indicates that shot noise is the largest source of uncertainty. The comparison also illustrates the importance of a careful interpretation of the measurements, since, depending on the origin of the tracer and the degree of mixing, the measurements may be biased toward the fuel, freestream, or reaction zone temperatures. 33 refs.

  9. Plant chlorophyll fluorescence: active and passive measurements at canopy and leaf scales with different nitrogen treatments

    PubMed Central

    Cendrero-Mateo, M. Pilar; Moran, M. Susan; Papuga, Shirley A.; Thorp, K.R.; Alonso, L.; Moreno, J.; Ponce-Campos, G.; Rascher, U.; Wang, G.

    2016-01-01

    Most studies assessing chlorophyll fluorescence (ChlF) have examined leaf responses to environmental stress conditions using active techniques. Alternatively, passive techniques are able to measure ChlF at both leaf and canopy scales. However, the measurement principles of both techniques are different, and only a few datasets concerning the relationships between them are reported in the literature. In this study, we investigated the potential for interchanging ChlF measurements using active techniques with passive measurements at different temporal and spatial scales. The ultimate objective was to determine the limits within which active and passive techniques are comparable. The results presented in this study showed that active and passive measurements were highly correlated over the growing season across nitrogen treatments at both canopy and leaf-average scale. At the single-leaf scale, the seasonal relation between techniques was weaker, but still significant. The variability within single-leaf measurements was largely related to leaf heterogeneity associated with variations in CO2 assimilation and stomatal conductance, and less so to variations in leaf chlorophyll content, leaf size or measurement inputs (e.g. light reflected and emitted by the leaf and illumination conditions and leaf spectrum). This uncertainty was exacerbated when single-leaf analysis was limited to a particular day rather than the entire season. We concluded that daily measurements of active and passive ChlF at the single-leaf scale are not comparable. However, canopy and leaf-average active measurements can be used to better understand the daily and seasonal behaviour of passive ChlF measurements. In turn, this can be used to better estimate plant photosynthetic capacity and therefore to provide improved information for crop management. PMID:26482242

  10. A human CXCL13-induced actin polymerization assay measured by fluorescence plate reader.

    PubMed

    Alley, Jennifer; Bloom, Laird; Kasaian, Marion; Gao, Huilan; Berstein, Gabriel; Clark, James D; Miao, Wenyan

    2010-02-01

    The chemokine receptor CXCR5 is predominantly expressed on mature B cells and follicular T-helper cells. CXCR5 and its ligand CXCL13 participate in ectopic germinal center formation at the inflammatory sites of multiple immune diseases such as rheumatoid arthritis, multiple sclerosis, and Sjogren's syndrome. Therefore, disrupting CXCL13-induced chemotaxis may be a fruitful approach for developing therapeutics in treating these diseases. Cells undergo cytoskeletal rearrangement prior to chemotaxis, and therefore actin polymerization can be used as a surrogate readout more proximal to chemokine receptor activation than chemotaxis. Conventionally, actin polymerization is measured by fluorescence microscopy or flow cytometry, which are either of low throughput or in need of special instruments. We developed a 96-well actin polymerization assay that can process 1,000 to 1,500 samples a day. This assay uses a standard laboratory fluorescence microplate reader as the detection instrument and was optimized for various experimental conditions such as cell density, actin filament staining reagent, staining buffer, and cell culture conditions. We demonstrate that this actin polymerization assay in 96-well format exhibits the expected pharmacology for human CXCR5 and is suitable as a primary functional assay to screen neutralizing scFv in crude bacterial peri-preps and a secondary assay for small compound collections.

  11. Robust measurement of membrane bending moduli using light sheet fluorescence imaging of vesicle fluctuations.

    PubMed

    Loftus, Andrew F; Noreng, Sigrid; Hsieh, Vivian L; Parthasarathy, Raghuveer

    2013-11-26

    The mechanical rigidity of lipid membranes is a key determinant of the energetics of cellular membrane deformation. Measurements of membrane bending moduli remain rare, however, and show a large variance, a situation that can be addressed by the development of improved techniques and by comparisons between disparate techniques applied to the same systems. We introduce here the use of selective plane illumination microscopy (SPIM, also known as light sheet fluorescence microscopy) to image thermal fluctuations of giant vesicles. The optical sectioning of SPIM enables high-speed fluorescence imaging of freely suspended vesicles and quantification of edge localization precision, yielding robust fluctuation spectra and rigidity estimates. For both lipid-only membranes and membranes bound by the intracellular trafficking protein Sar1p, which lowers membrane rigidity in a concentration-dependent manner, we show that the resulting bending modulus values are in close agreement with those derived from an independent assay based on membrane tether pulling. We also show that the fluctuation spectra of vesicles bound by the mammalian Sar1A protein, which stiffens membranes at high concentrations, are not well fit by a model of homogeneous quasi-spherical vesicles, suggesting that SPIM-based analysis can offer insights into spatially inhomogeneous properties induced by protein assemblies.

  12. Two-Beam multiplexed laser-induced fluorescence measurements of an argon arcjet plume

    NASA Astrophysics Data System (ADS)

    Ruyten, Wilhelmus M.; Keefer, Dennis

    1993-11-01

    We describe a multiplexed, laser-induced fluorescence (LIF) technique with which radial and axial profiles of vector velocities of excited propellant species were obtained in the exhaust plume from a 300-W argon arcjet. Although the arcjet is a prototype, and although argon is not an interesting propellant from a propulsion perspective, the technique clearly demonstrates how a narrowband, frequency-stabilized ring-dye laser can be used to obtain simultaneous measurements of two velocity components in an arcjet plume and how a third signal from an optogalvanic cell can be used as a frequency reference. We also show that much information on the flow can be obtained by analyzing the Doppler widths and fluorescence intensities of the LIF data. Specifically, the data identify a boundary layer in the radial direction of the plume and a shock in the downstream region of the flow. Also, some flow anisotropy is observed, consistent with the assumption that the magnitude of the mean flow velocity fluctuates. The peak velocity on centerline remains roughly constant at 3 km/s throughout the expansion.

  13. Development of a canopy Solar-induced chlorophyll fluorescence measurement instrument

    NASA Astrophysics Data System (ADS)

    Sun, G.; Wang, X.; Niu, Zh; Chen, F.

    2014-02-01

    A portable solar-induced chlorophyll fluorescence detecting instrument based on Fraunhofer line principle was designed and tested. The instrument has a valid survey area of 1.3 × 1.3 meter when the height was fixed to 1.3 meter. The instrument uses sunlight as its light source. The instrument is quipped with two sets of special photoelectrical detectors with the centre wavelength at 760 nm and 771 nm respectively and bandwidth less than 1nm. Both sets of detectors are composed of an upper detector which are used for detecting incidence sunlight and a bottom detector which are used for detecting reflex light from the canopy of crop. This instrument includes photoelectric detector module, signal process module, A/D convert module, the data storage and upload module and human-machine interface module. The microprocessor calculates solar-induced fluorescence value based on the A/D values get from detectors. And the value can be displayed on the instrument's LCD, stored in the flash memory of instrument and can also be uploaded to PC through the PC's serial interface. The prototype was tested in the crop field and the results demonstrate that the instrument can measure the solar-induced chlorophyll value exactly with the correlation coefficients was 0.9 compared to the values got from Analytical Spectral Devices FieldSpec Pro spectrometer. This instrument can diagnose the plant growth status by the acquired spectral response.

  14. Effects of fosmidomycin on plant photosynthesis as measured by gas exchange and chlorophyll fluorescence.

    PubMed

    Possell, Malcolm; Ryan, Annette; Vickers, Claudia E; Mullineaux, Philip M; Hewitt, C Nicholas

    2010-04-01

    In higher plants, many isoprenoids are synthesised via the chloroplastic 1-deoxy-D-xylulose 5-phosphate/2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. Attempts to elucidate the function of individual isoprenoids have used the antibiotic/herbicidal compound fosmidomycin (3-[N-formyl-N-hydroxy amino] propyl phosphonic acid) to inhibit this pathway. Examination of the effect of fosmidomycin on the major components of photosynthesis in leaves of white poplar (Populus alba) and tobacco (Nicotiana tabacum) was made. Fosmidomycin reduced net photosynthesis in both species within 1 h of application, but only when photosynthesis was light-saturated. In P. alba, these reductions were confounded by high light and fosmidomycin inducing stomatal patchiness. In tobacco, this was caused by significant reductions in PSII chlorophyll fluorescence and reductions in V(cmax) and J(max). Our data indicate that the diminution of photosynthesis is likely a complex effect resulting from the inhibition of multiple MEP pathway products, resulting in photoinhibition and photo-damage. These effects should be accounted for in experimental design and analysis when using fosmidomycin to avoid misinterpretation of results as measured by gas exchange and chlorophyll fluorescence.

  15. Development of an X-ray fluorescence holographic measurement system for protein crystals

    NASA Astrophysics Data System (ADS)

    Sato-Tomita, Ayana; Shibayama, Naoya; Happo, Naohisa; Kimura, Koji; Okabe, Takahiro; Matsushita, Tomohiro; Park, Sam-Yong; Sasaki, Yuji C.; Hayashi, Kouichi

    2016-06-01

    Experimental procedure and setup for obtaining X-ray fluorescence hologram of crystalline metalloprotein samples are described. Human hemoglobin, an α2β2 tetrameric metalloprotein containing the Fe(II) heme active-site in each chain, was chosen for this study because of its wealth of crystallographic data. A cold gas flow system was introduced to reduce X-ray radiation damage of protein crystals that are usually fragile and susceptible to damage. A χ-stage was installed to rotate the sample while avoiding intersection between the X-ray beam and the sample loop or holder, which is needed for supporting fragile protein crystals. Huge hemoglobin crystals (with a maximum size of 8 × 6 × 3 mm3) were prepared and used to keep the footprint of the incident X-ray beam smaller than the sample size during the entire course of the measurement with the incident angle of 0°-70°. Under these experimental and data acquisition conditions, we achieved the first observation of the X-ray fluorescence hologram pattern from the protein crystals with minimal radiation damage, opening up a new and potential method for investigating the stereochemistry of the metal active-sites in biomacromolecules.

  16. Time-resolved measurements of short-wavelength fluorescence from x-ray-excited ions.

    PubMed

    Kapteyn, H C; Murnane, M M; Falcone, R W

    1987-09-01

    We demonstrate a novel technique for time-resolved spectroscopic studies of highly excited ions. The technique uses a laser-produced plasma as a short-pulse, soft-x-ray light source with a high repetition rate. A Nd:YAG laser with a pulse duration of 90 psec, a pulse energy of 70 microJ, and repetition rate of 10(4) pulses per second is focused onto a rotating metal target. Soft x rays from the resulting plasma photoionize a gas surrounding the target, and fluorescence from the gas is detected by using a spectrometer and a high-speed photodetector. Using the technique of time-correlated photon counting, we determined the radiative lifetime and collisional quenching rate of the Xe III 5s(0)5p(6)(1)S(0) state by observing its fluorescence at 108.9 nm. A time resolution of better than 400 psec was obtained. We also measured relative Auger decay yields of a core hole state in xenon using a higher-energy laser-produced plasma light source at a lower repetition rate.

  17. Fluorescence emission spectral measurements for the detection of oil on shore

    SciTech Connect

    Balick, L.K.; Di Benedetto, J.A.; Lutz, S.S.

    1996-12-31

    The U.S. DOE Special Technologies Laboratory is developing an airborne Laser-Induced Fluorescence Imaging (LIFI) system to support environmental management of government facilities. This system, or a system to be derived from it, is being evaluated for its potential to detect spilled oils oN shore, in wetlands, and on ice. To more fully understand the detectivity of oil spills, emphasis has been placed on the spectral contrast between the oil signatures and signatures associated with the natural backgrounds (sand, vegetation, etc.). To support this evaluation, two series of controlled measurements have been performed to provide rigorous characterization of the excitation-emission properties of some oils and background materials, and to look at the effects of weathering of oil on terrestrial background materials. Oil targets included a heavy crude oil, diesel, kerosene, and aviation fuel and backgrounds included beach sand, straw, mud, tar and kelp. Fluorescence of oil on background materials decreases rapidly over the first few days of exposure to the environment and is more rapid than for neat oil samples.

  18. Fluorescence emission spectral measurements for the detection of oil on shore

    SciTech Connect

    Balick, L.K.; Di Benedetto, J.A.; Lutz, S.S.

    1997-06-01

    The US DOE Special Technologies Laboratory is developing an airborne Laser-Induced Fluorescence Imaging (LIFI) system to support environmental management of government Utilities. This system, or a system to be derived from it, is being evaluated for its potential to detect spilled oils on shore, in wetlands, and on ice. To more fully understand the detectivity of oil spills, emphasis has been placed on the spectral contrast between the oil signatures and signatures associated with the natural backgrounds (sand, vegetation, etc.). To support this evaluation, two series of controlled measurements have been performed to provide rigorous characterization of the excitation-emission properties of some oils and background materials, and to look at the effects of weathering of oil on terrestrial background materials. Oil targets included a heavy crude oil, diesel, kerosene, and aviation fuel and backgrounds included beach sand, straw, mud, tar and kelp. Fluorescence of oil on background materials decreases rapidly over the first few days of exposure to the environment and is more rapid than for neat oil samples.

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

    NASA Astrophysics Data System (ADS)

    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.

  20. Heavy Metals Effect on Cyanobacteria Synechocystis aquatilis Study Using Absorption, Fluorescence, Flow Cytometry, and Photothermal Measurements

    NASA Astrophysics Data System (ADS)

    Dudkowiak, A.; Olejarz, B.; Łukasiewicz, J.; Banaszek, J.; Sikora, J.; Wiktorowicz, K.

    2011-04-01

    The toxic effect of six heavy metals on cyanobacteria Synechocystis aquatilis was studied by absorption, fluorescence, flow cytometry, and photothermal measurements. This study indicates that at the concentration used, the cyanobacteria are more sensitive to silver, copper, and mercury than to cadmium, lead, and zinc metals. Disregarding the decrease in the yields of the related radiative processes caused by photochemical processes and/or damage to phycobilisomes, no changes were detected in the efficiency of thermal deactivation processes within a few microseconds, which can indicate the lack of disturbances in the photosynthetic light reaction and the lack of damage to the photosystem caused by the heavy metal ions in the concentrations used. The results demonstrate that the relative values of fluorescence yield as well as promptly generated heat calculated for the metal-affected and unaffected (reference) bacteria are sensitive indicators of environmental pollution with heavy metal ions, whereas the complementary methods proposed could be used as a noninvasive and fast procedure for in vivo assessment of their toxicity.

  1. Performance evaluation of principal component analysis for dynamic fluorescence tomographic imaging in measurement space

    NASA Astrophysics Data System (ADS)

    Liu, Xin; He, Xiaowei; Yan, Zhuangzhi

    2015-05-01

    Challenges remain in resolving drug (fluorescent biomarkers) distributions within small animals by fluorescence diffuse optical tomography (FDOT). Principal component analysis (PCA) provides the capability of detecting organs (functional structures) from dynamic FDOT images. However, the resolving performance of PCA may be affected by various experimental factors, e.g., the noise levels in measurement data, the variance in optical properties, the number of acquired frames, and so on. To address the problem, based on a simulation model, we analyze and compare the performance of PCA when applied to three typical sets of experimental conditions (frames number, noise level, and optical properties). The results show that the noise is a critical factor affecting the performance of PCA. When input data containing a low noise (<5%), by a short (e.g., 6 frame) projection sequence, we can resolve the poly(DL-lactic-coglycolic acid)/indocynaine green (PLGA/ICG) distributions in heart and lungs, even though there are great variances in optical properties. In contrast, when 20% Gaussian noise is added to the input data, it hardly resolves the distributions of PLGA/ICG in heart and lungs even though accurate optical properties are used. However, with an increased number of frames, the resolving performance of PCA may gradually recover.

  2. Two-Beam multiplexed laser-induced fluorescence measurements of an argon arcjet plume

    NASA Technical Reports Server (NTRS)

    Ruyten, Wilhelmus M.; Keefer, Dennis

    1993-01-01

    We describe a multiplexed, laser-induced fluorescence (LIF) technique with which radial and axial profiles of vector velocities of excited propellant species were obtained in the exhaust plume from a 300-W argon arcjet. Although the arcjet is a prototype, and although argon is not an interesting propellant from a propulsion perspective, the technique clearly demonstrates how a narrowband, frequency-stabilized ring-dye laser can be used to obtain simultaneous measurements of two velocity components in an arcjet plume and how a third signal from an optogalvanic cell can be used as a frequency reference. We also show that much information on the flow can be obtained by analyzing the Doppler widths and fluorescence intensities of the LIF data. Specifically, the data identify a boundary layer in the radial direction of the plume and a shock in the downstream region of the flow. Also, some flow anisotropy is observed, consistent with the assumption that the magnitude of the mean flow velocity fluctuates. The peak velocity on centerline remains roughly constant at 3 km/s throughout the expansion.

  3. Macromolecular diffusion of biological polymers measured by confocal fluorescence recovery after photobleaching.

    PubMed Central

    Gribbon, P; Hardingham, T E

    1998-01-01

    Fluorescence recovery after photobleaching with an unmodified confocal laser scanning microscope (confocal FRAP) was used to determine the diffusion properties of network forming biological macromolecules such as aggrecan. The technique was validated using fluorescein isothiocyanate (FITC)-labeled dextrans and proteins (molecular mass 4-2000 kDa) at 25 degrees C and with fluorescent microspheres (207 nm diameter) over a temperature range of 5-50 degrees C. Lateral diffusion coefficients (D) were independent of the focus position, and the degree and extent of bleach. The free diffusion coefficient (Do) of FITC-aggrecan determined by confocal FRAP was 4.25 +/- 0.6 x 10(-8) cm2 s-1, which is compatible with dynamic laser light scattering measurements. It appeared to be independent of concentration below 2.0 mg/ml, but at higher concentrations (2-20 mg/ml) the self-diffusion coefficient followed the function D = Do(e)(-Bc). The concentration at which the self-diffusion coefficient began to fall corresponded to the concentration predicted for domain overlap. Multimolecular aggregates of aggrecan ( approximately 30 monomers) had a much lower free diffusion coefficient (Do = 6.6 +/- 1.0 x 10(-9) cm2 s-1) but showed a decrease in mobility with concentration of a form similar to that of the monomer. The method provides a technique for investigating the macromolecular organization in glycan-rich networks at concentrations close to those found physiologically. PMID:9675204

  4. Fluorescence excitation and propagation through brain phantom gelatins: measurements and potential applications

    SciTech Connect

    Allison, Stephen W; Gillies, George

    2010-01-01

    We have investigated the utility of 0.6% agarose gels as surrogate materials for brain tissues in optical propagation studies for possible diagnostic and therapeutic applications. Centimeter-scale layers of the gel exhibited a Beer's law attenuation factor, , of 0.2 mm 1 for incident illumination via a pulsed LED (100 Hz) at 405 nm. This result was different by only about a factor of 3 from the effective penetration depth at similar wavelengths through in vitro samples of the gray (cortical) matter of human brain, as measured by others. Then, films of the thermographic phosphors La2O2S:Eu, Mg4FGeO6:Mn, YAG:Cr and variants of the latter were formed on aluminum substrates and the fluorescence of these samples was stimulated and observed through layers of the gel up to 4 cm thick. In all cases, the fluorescence was easily excited and distinguishable above the background. The results demonstrate that this gel might serve as an inexpensive and robust test bed for exploratory studies of neurological modalities involving propagation of optical signals within brain tissues.

  5. Application of fluorescent tracer agent technology to point-of-care gastrointestinal permeability measurement

    NASA Astrophysics Data System (ADS)

    Dorshow, Richard B.; Shieh, Jeng-Jong; Rogers, Thomas E.; Hall-Moore, Carla; Shaikh, Nurmohammad; Talcott, Michael; Tarr, Phillip I.

    2016-03-01

    Gut dysfunction, often accompanied by increased mucosal permeability to gut contents, frequently accompanies a variety of human intestinal inflammatory conditions. These disorders include inflammatory bowel diseases (e.g., Crohn's Disease) and environmental enteropathy and enteric dysfunction, a condition strongly associated with childhood malnutrition and stunting in resource poor areas of the world. The most widely used diagnostic assay for gastrointestinal permeability is the lactulose to mannitol ratio (L:M) measurement. These sugars are administered orally, differentially absorbed by the gut, and then cleared from the body by glomerular filtration in the kidney. The amount of each sugar excreted in the urine is measured. The larger sugar, lactulose, is minimally absorbed through a healthy gut. The smaller sugar, mannitol, in contrast, is readily absorbed through both a healthy and injured gut. Thus a higher ratio of lactulose to mannitol reflects increased intestinal permeability. However, several issues prevent widespread use of the L:M ratio in clinical practice. Urine needs to be collected over time intervals of several hours, the specimen then needs to be transported to an analytical laboratory, and sophisticated equipment is required to measure the concentration of each sugar in the urine. In this presentation we show that fluorescent tracer agents with molecular weights similar to those of the sugars, selected from our portfolio of biocompatible renally cleared fluorophores, mimic the L:M ratio test for gut permeability. This fluorescent tracer agent detection technology can be used to overcome the limitations of the L:M assay, and is amenable to point-of-care clinical use.

  6. Membrane order parameters for interdigitated lipid bilayers measured via polarized total-internal-reflection fluorescence microscopy.

    PubMed

    Ngo, An T; Jakubek, Zygmunt J; Lu, Zhengfang; Joós, Béla; Morris, Catherine E; Johnston, Linda J

    2014-11-01

    Incorporating ethanol in lipid membranes leads to changes in bilayer structure, including the formation of an interdigitated phase. We have used polarized total-internal-reflection fluorescence microscopy (pTIRFM) to measure the order parameter for Texas Red DHPE incorporated in the ethanol-induced interdigitated phase (LβI) formed from ternary lipid mixtures comprising dioleoylphosphatidylcholine, cholesterol and egg sphingomyelin or dipalmitoylphosphatidylcholine. These lipid mixtures have 3 co-existing phases in the presence of ethanol: liquid-ordered, liquid-disordered and LβI. pTIRFM using Texas Red DHPE shows a reversal in fluorescence contrast between the LβI phase and the surrounding disordered phase with changes in the polarization angle. The contrast reversal is due to changes in the orientation of the dye, and provides a rapid method to identify the LβI phase. The measured order parameters for the LβI phase are consistent with a highly ordered membrane environment, similar to a gel phase. An acyl-chain labeled BODIPY-FL-PC was also tested for pTIRFM studies of ethanol-treated bilayers; however, this probe is less useful since the order parameters of the interdigitated phase are consistent with orientations that are close to random, either due to local membrane disorder or to a mixture of extended and looping conformations in which the fluorophore is localized in the polar headgroup region of the bilayer. In summary, we demonstrate that order parameter measurements via pTIRFM using Texas Red-DHPE can rapidly identify the interdigitated phase in supported bilayers. We anticipate that this technique will aid further research in the effects of alcohols and other additives on membranes.

  7. Combined fluorescence, reflectance, and ground measurements of a stressed Norway spruce forest for forest damage assessment

    NASA Technical Reports Server (NTRS)

    Banninger, C.

    1991-01-01

    The detection and monitoring of stress and damage in forested areas is of utmost importance to forest managers for planning purposes. Remote sensing are the most suitable means to obtain this information. This requires that remote sensing data employed in a forest survey be properly chosen and utilized for their ability to measure canopy spectral features directly related to key tree and canopy properties that are indicators of forest health and vitality. Plant reflectance in the visible to short wave IR regions (400 to 2500 nm) provides information on its biochemical, biophysical, and morphological make up, whereas plant fluorescence in the 400 to 750 nm region is more indicative of the capacity and functioning of its photosynthetic apparatus. A measure of both these spectral properties can be used to provide an accurate assessment of stress and damage within the forest canopy. Foliar chlorophyll and nitrogen are essential biochemical constituents required for the proper functioning and maintenance of a plant's biological processes. Chlorophyll-a is the prime reactive center for photosynthesis, by which a plant converts CO2 and H2O into necessary plant products. Nitrogen forms an important component of the amino-acids, enzymes, proteins, alkaloids, and cyanogenic compounds that make up a plant, including its pigments. Both chlorophyll and nitrogen have characteristic absorption features in the visible to short wave IR region. By measuring the wavelength position and depth of these features and the fluorescence response of the foliage, the health and vitality of a canopy can be ascertained. Examples for a stressed Norway spruce forest in south-eastern Austria are presented.

  8. The Growth and Mechanical Properties of Living Neurons Measured via Atomic Force and Fluorescence Microscopy

    NASA Astrophysics Data System (ADS)

    Spedden, Elise

    In this thesis we explore specific properties of the cytoskeleton and growth of living neurons via atomic force and fluorescence microscopies. We make the first comparative elastic modulus measurements on three types of neuronal cells plated on three types of substrate adhesion factors. We discover that during phases of active neurite extension the soma of cortical neurons stiffens reversibly due to changes in microtubule aggregation. Additionally, we demonstrate that mechanical properties of cortical neurons measured near physiological temperatures are primarily dependent on the microtubule component of the cytoskeleton. We further explore the response of the neuronal cytoskeleton to changes in ambient temperature. The elastic modulus of cortical neuron somas is discovered to increase dramatically upon a drop in ambient temperature. We determine through fluorescent staining and chemical modification of the cytoskeleton that this stiffening is due primarily to a change in the mechanically dominant component of the cytoskeleton from microtubules at 37ºC to actin at 25ºC precipitated by changes in myosin II dynamics within the cell. We make the first direct mechanical measurements of the pericellular brush layer on living neurons, demonstrating that the traditionally observed viscoelastic behavior of the neuronal soma is due to the properties of this brush layer. When the brush layer is excluded, the underlying soma is discovered to be both stiffer than previously observed, and elastic, with no loading-speed dependence to the elastic modulus under the test conditions. We additionally demonstrate that the soma elastic modulus, brush length, and brush density are all dependent on the ambient temperature. Finally, through fluorescent and bright field microscopies we track the outgrowth of living neurons on patterned directional surfaces, demonstrating that asymmetrical ratchet topographies unidirectionally bias axonal outgrowth. We model the outgrowth of the neurons

  9. Airborne vacuum ultraviolet resonance fluorescence instrument for in situ measurement of CO

    NASA Astrophysics Data System (ADS)

    Takegawa, N.; Kita, K.; Kondo, Y.; Matsumi, Y.; Parrish, D. D.; Holloway, J. S.; Koike, M.; Miyazaki, Y.; Toriyama, N.; Kawakami, S.; Ogawa, T.

    2001-10-01

    An airborne instrument for fast-response, high-precision measurement of tropospheric carbon monoxide (CO) was developed using a vacuum ultraviolet (VUV) resonance fluorescence technique. The excitation radiation is obtained by a DC discharge CO resonance lamp combined with an optical filter for the CO fourth positive band emission around 150 nm. The optical filter consists of a VUV monochromator and a crystalline quartz window (<147-nm cutoff). The crystalline quartz window ensures a sharp discrimination against wavelengths below 135.7 nm that yield a positive interference from water vapor. Laboratory tests showed that the optical system achieved a precision of 1.1 parts per billion by volume (ppbv) at a CO concentration of 100 ppbv for a 1-s integration period, and the flow system provided a response time (1/e time constant) of ˜2 s. The aircraft measurement campaign Biomass Burning and Lightning Experiment-phase B (BIBLE-B) was conducted between August and September 1999 over the western Pacific and Australia. The flight data obtained during this campaign were used to demonstrate the high precision and fast response of the instrument. An intercomparison of the VUV CO measurement and a gas chromatographic CO measurement was conducted during BIBLE-B. Overall, these two independent measurements showed good agreement, within the experimental uncertainties.

  10. Structural relaxation mechanisms in liquid Eugenol. A depolarized light scattering study

    NASA Astrophysics Data System (ADS)

    Bezot, P.; Hesse-Bezot, C.; Roynard, D.; Jeanneaux, F.

    1988-07-01

    A depolarized light scattering study of liquid Eugenol, over a large temperature range including the supercooled region, is proposed. Comparisons with shear mechanical impedance measurements, obtained at lower frequencies, lead to more precise information on the viscoelastic parameters in the supercooled region. The structural relaxation process measurements by means of the photon correlation technique are compared to the dielectric and mechanical measurements. Molecular mechanisms are proposed.

  11. Temperature modulation of cerebral depolarization during focal cerebral ischemia in rats: correlation with ischemic injury.

    PubMed

    Chen, Q; Chopp, M; Bodzin, G; Chen, H

    1993-05-01

    The role of cerebral depolarizations in focal cerebral ischemia is unknown. We therefore measured the direct current (DC) electrical activity in the cortex of Wistar rats subjected to transient occlusion of the middle cerebral artery (MCA). Focal ischemia was induced for 90 min by insertion of an intraluminal filament to occlude the MCA. To modulate cell damage, we subjected the rats to hypothermic (30 degrees C, n = 4), normothermic (37 degrees C, n = 4), and hyperthermic (40 degrees C, n = 6) ischemia. Controlled temperatures were also maintained during 1 h of reperfusion. Continuous cortical DC potential changes were measured using two active Ag-AgCl electrodes placed in the cortical lesion. Animals were killed 1 week after ischemia. The brains were sectioned and stained with hematoxylin and eosin, for evaluation of neuronal damage, and calculation of infarct volume. All animals exhibited an initial depolarization within 30 min of ischemia, followed by a single depolarization event in hypothermic animals, and multiple periodic depolarization events in both normothermic and hyperthermic animals. Hyperthermic animals exhibited significantly more (p < 0.05) DC potential deflections (n = 6.17 +/- 0.67) than normothermic animals (n = 2.75 +/- 0.96). The ischemic infarct volume (% of hemisphere) was significantly different for the various groups; hypothermic animals exhibited no measurable infarct volume, while the ischemic infarct volume was 10.2 +/- 12.3% in normothermic animals and 36.5 +/- 3.4% in hyperthermic animals (p < 0.05). A significant correlation was detected between the volume of infarct and number of depolarization events (r = 0.90, p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

  12. Two photon absorption laser induced fluorescence measurements of neutral density in a helicon plasma

    SciTech Connect

    Galante, M. E.; Magee, R. M.; Scime, E. E.

    2014-05-15

    We have developed a new diagnostic based on two-photon absorption laser induced fluorescence (TALIF). We use a high intensity (5 MW/cm{sup 2}), narrow bandwidth (0.1 cm{sup −1}) laser to probe the ground state of neutral hydrogen, deuterium and krypton with spatial resolution better than 0.2 cm, a time resolution of 10 ns, and a measurement cadence of 20 Hz. Here, we describe proof-of-principle measurements in a helicon plasma source that demonstrate the TALIF diagnostic is capable of measuring neutral densities spanning four orders of magnitude; comparable to the edge neutral gradients predicted in the DIII-D tokamak pedestal. The measurements are performed in hydrogen and deuterium plasmas and absolute calibration is accomplished through TALIF measurements in neutral krypton. The optical configuration employed is confocal, i.e., both light injection and collection are accomplished with a single lens through a single optical port in the vacuum vessel. The wavelength resolution of the diagnostic is sufficient to separate hydrogen and deuterium spectra and we present measurements from mixed hydrogen and deuterium plasmas that demonstrate isotopic abundance measurements are feasible. Time resolved measurements also allow us to explore the evolution of the neutral hydrogen density and temperature and effects of wall recycling. We find that the atomic neutral density grows rapidly at the initiation of the discharge, reaching the steady-state value within 1 ms. Additionally, we find that neutral hydrogen atoms are born with 0.08 eV temperatures, not 2 eV as is typically assumed.

  13. The influence of DNA shape fluctuations on fluorescence resonance energy transfer efficiency measurements in nucleosomes

    NASA Astrophysics Data System (ADS)

    Lenz, Lucia; Hoenderdos, Maurice; Prinsen, Peter; Schiessel, Helmut

    2015-02-01

    Fluorescence resonance energy transfer (FRET) measurements allow one to observe site exposure in nucleosomes, i.e. the transient unwrapping of a part of the wrapped DNA from the histone octamer. In such experiments one can typically distinguish between a closed state and an open state but in principle one might hope to detect several states, each corresponding to a certain number of open binding sites. Here we show that even in an ideal FRET setup it would be hard to detect unwrapping states with intermediate levels of FRET efficiencies. As the unwrapped DNA molecule, modelled here as a wormlike chain, has a finite stiffness, shape fluctuations smear out FRET signals completely from such intermediate states.

  14. Thyroid iodine content measured by x-ray fluorescence in amiodarone-induced thyrotoxicosis: concise communication

    SciTech Connect

    Leger, A.F.; Fragu, P.; Rougier, P.; Laurent, M.F.; Tubiana, M.; Savole, J.C.

    1983-07-01

    Iodine-induced thyrotoxicosis (IiT) is characterized by (a) a low radioiodine uptake, increased by exogenous TSH, and (b) a spontaneous evolution towards cure within a few months. An hypothetical pathogenesis of IiT is an initial inflation in the stores of thyroid hormones during iodine excess, followed by their sudden discharge into the circulation. Thyroid iodine content was measured by fluorescent scanning in 10 patients with amiodarone-induced thyrotoxicosis and in various control groups. Results were found to be high at the onset of the disease and to decrease during its course. The data agree with the hypothetical pathogenesis. Furthermore they may permit exclusion of a painless subacute thyroiditis, which is the main differential diagnosis of IiT.

  15. Downsizing of Georgia Tech's Airborne Fluorescence Spectrometer (AFS) for the Measurement of Nitrogen Oxides

    NASA Technical Reports Server (NTRS)

    Sandholm, Scott

    1998-01-01

    This report addresses the Tropospheric Trace Gas and Airborne Measurements (TTGAMG) endeavors to further downsize and stabilize the Georgia Institute of Technology's Airborne Laser Induced Fluorescence Experiment (GITALIFE). It will mainly address the TTGAMG successes and failures as participants in the summer 1998 Wallops Island test flights on board the P3-B. Due to the restructuring and reorganization of the TTGAMG since the original funding of this grant, some of the objectives and time lines of the deliverables have been changed. Most of these changes have been covered in the preceding annual report. We are anticipating getting back on track with the original proposal's downsizing effort this summer, culminating in the GITALIFE no longer occupying a high bay rack and the loss of several hundred pounds.

  16. Organization of FtsZ filaments in the bacterial division ring measured from polarized fluorescence microscopy.

    PubMed

    Si, Fangwei; Busiek, Kimberly; Margolin, William; Sun, Sean X

    2013-11-05

    Cytokinesis in bacteria is accomplished by a ring-shaped cell-division complex (the Z-ring). The primary component of the Z-ring is FtsZ, a filamentous tubulin homolog that serves as a scaffold for the recruitment of other cell-division-related proteins. FtsZ forms filaments and bundles. In the cell, it has been suggested that FtsZ filaments form the arcs of the ring and are aligned in the cell-circumferential direction. Using polarized fluorescence microscopy in live Escherichia coli cells, we measure the structural organization of FtsZ filaments in the Z-ring. The data suggest a disordered organization: a substantial portion of FtsZ filaments are aligned in the cell-axis direction. FtsZ organization in the Z-ring also appears to depend on the bacterial species. Taken together, the unique arrangement of FtsZ suggests novel unexplored mechanisms in bacterial cell division.

  17. Feasibility of hydroxyl concentration measurements by laser-saturated fluorescence in high-pressure flames

    NASA Technical Reports Server (NTRS)

    Carter, Campbell D.; King, Galen B.; Laurendeau, Normand M.; Salmon, J. Thaddeus

    1987-01-01

    The effect of pressure on the laser-saturated fluorescence method for measuring OH concentration in high-pressure flames is studied using calculations for the burned-gas region of a stoichiometric H2-O2 flame at 2000 K. A numerical model of the excitation dynamics of OH is developed to explore the validity of the balanced cross-rate model at higher pressures. It is shown that depopulation of the laser-coupled levels is sensitive to collisions which depopulate v-double-prime (VDP) = 0 and to rate coefficients for rotational transfer in the ground state which are smaller than those in the excited state. In particular, it is shown that the depopulation of VDP = 0, and hence the laser-coupled levels, depends on the probability of electronic quenching to vibrational levels for which VDP is greater than 0 and vibrational relaxation to VDP = 0.

  18. An in situ antimicrobial susceptibility testing method based on in vivo measurements of chlorophyll α fluorescence.

    PubMed

    Heliopoulos, Nikolaos S; Galeou, Angeliki; Papageorgiou, Sergios K; Favvas, Evangelos P; Katsaros, Fotios K; Stamatakis, Kostas

    2015-05-01

    Up to now antimicrobial susceptibility testing (AST) methods are indirect and generally involve the manual counting of bacterial colonies following the extraction of microorganisms from the surface under study and their inoculation in a separate procedure. In this work, an in situ, direct and instrumental method for the evaluation and assessment of antibacterial properties of materials and surfaces is proposed. Instead of indirectly determining antibacterial activity using the typical gram(-) test organisms with the subsequent manual colony count or inhibition zone measurement, the proposed procedure, employs photosynthetic gram(-) cyanobacteria deposited directly onto the surface under study and assesses cell proliferation and viability by a quick, accurate and reproducible instrumental chlorophyll fluorescence spectrophotometric technique. In contrast with existing methods of determination of antibacterial properties, it produces high resolution and quantitative results and is so versatile that it could be used to evaluate the antibacterial properties of any compound (organic, inorganic, natural or man-made) under any experimental conditions, depending on the targeted application.

  19. Nanoscale diffusion in the synaptic cleft and beyond measured with time-resolved fluorescence anisotropy imaging

    PubMed Central

    Zheng, Kaiyu; Jensen, Thomas P.; Savtchenko, Leonid P.; Levitt, James A.; Suhling, Klaus; Rusakov, Dmitri A.

    2017-01-01

    Neural activity relies on molecular diffusion within nanoscopic spaces outside and inside nerve cells, such as synaptic clefts or dendritic spines. Measuring diffusion on this small scale in situ has not hitherto been possible, yet this knowledge is critical for understanding the dynamics of molecular events and electric currents that shape physiological signals throughout the brain. Here we advance time-resolved fluorescence anisotropy imaging combined with two-photon excitation microscopy to map nanoscale diffusivity in ex vivo brain slices. We find that in the brain interstitial gaps small molecules move on average ~30% slower than in a free medium whereas inside neuronal dendrites this retardation is ~70%. In the synaptic cleft free nanodiffusion is decelerated by ~46%. These quantities provide previously unattainable basic constrains for the receptor actions of released neurotransmitters, the electrical conductance of the brain interstitial space and the limiting rate of molecular interactions or conformational changes in the synaptic microenvironment. PMID:28181535

  20. Improving the modeling of the seasonal carbon cycle of the boreal forest with chlorophyll fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Thum, Tea; Aalto, Tuula; Aurela, Mika; Laurila, Tuomas; Zaehle, Sönke

    2014-05-01

    The boreal ecosystems are characterized a very strong seasonal cycle and they are very sensitive to the climatic variables. The vegetation's deep wintertime dormancy requires a long recovery time during spring before the plants reach their full photosynthetic capacity. During this recovery time the plants are highly susceptible the night frosts. The transition period is different during spring and autumn for the evergreen plants. During spring there is plenty of light, but cold air temperatures inhibit the photosynthesis. The plants therefore experience to high stress levels, as they need to protect their photosynthetic apparatus from intense light. In autumn the air temperature and light level decrease more concurrently. To have a realistic presentation of the carbon cycle in boreal forests it is important to have these characteristics properly modeled, so that also the implications of changing seasonality under climate change can be more reliably predicted. In this study, we focus on the CO2 exchange of a Scots pine forest Sodankylä located in Finnish Lapland, 100 km north from the Arctic Circle. Micrometeorological flux measurements provide information about the exchanges of carbon, energy and water between atmosphere and vegetation. To complement these fluxes, we use dark-adapted chlorophyll fluorescence (CF) measurements, which is an optical measurement and tracks the development of the photosynthetic capacity. These two approaches combined together are very useful when we want to improve the modeling of the forest's CO2 exchange. We used two models that describe the photosynthesis with the biochemical model of Farquhar et al. The FMI-CANOPY is a canopy level model that is feasible to use in parameter estimation. We used the CF measurements of Fv/Fm, that is a measure of the maximum photosynthetic capacity, to include a seasonal development in the base rate of the maximum carboxylation rate (Vc(max)) in FMI-CANOPY. The simulation results matched the

  1. Planar Laser-Induced Iodine Fluorescence Measurements in Rarefied Hypersonic Flow

    NASA Astrophysics Data System (ADS)

    Cecil, Eric; McDaniel, James C.

    2005-05-01

    A planar laser-induced fluorescence (PLIF) technique is discussed and applied to measurement of time-averaged values of velocity and temperature in an I2-seeded N2 hypersonic free jet facility. Using this technique, a low temperature, non-reacting, hypersonic flow over a simplified model of a reaction control system (RCS) was investigated. Data are presented of rarefied Mach 12 flow over a sharp leading edge flat plate at zero incidence, both with and without an interacting jet issuing from a nozzle built into the plate. The velocity profile in the boundary layer on the plate was resolved. The slip velocity along the plate, extrapolated from the velocity profile data, varied from nearly 100% down to 10% of the freestream value. These measurements are compared with results of a DSMC solution. The velocity variation along the centerline of a jet issuing from the plate was measured and found to match closely with the correlation of Ashkenas and Sherman. The velocity variation in the oblique shock terminating the jet was resolved sufficiently to measure the shock wave thickness.

  2. Planar Laser-Induced Iodine Fluorescence Measurements in Rarefied Hypersonic Flow

    NASA Technical Reports Server (NTRS)

    Cecil, Eric; McDaniel, James C.

    2005-01-01

    A planar laser-induced fluorescence (PLIF) technique is discussed and applied to measurement of time-averaged values of velocity and temperature in an I(sub 2)-seeded N(sub 2) hypersonic free jet facility. Using this technique, a low temperature, non-reacting, hypersonic flow over a simplified model of a reaction control system (RCS) was investigated. Data are presented of rarefied Mach 12 flow over a sharp leading edge flat plate at zero incidence, both with and without an interacting jet issuing from a nozzle built into the plate. The velocity profile in the boundary layer on the plate was resolved. The slip velocity along the plate, extrapolated from the velocity profile data, varied from nearly 100% down to 10% of the freestream value. These measurements are compared with results of a DSMC solution. The velocity variation along the centerline of a jet issuing from the plate was measured and found to match closely with the correlation of Ashkenas and Sherman. The velocity variation in the oblique shock terminating the jet was resolved sufficiently to measure the shock wave thickness.

  3. Spectrum measurement with the Telescope Array Low Energy Extension (TALE) fluorescence detector

    NASA Astrophysics Data System (ADS)

    Zundel, Zachary James

    The Telescope Array (TA) experiment is the largest Ultra High Energy cosmic ray observatory in the northern hemisphere and is designed to be sensitive to cosmic ray air showers above 1018eV. Despite the substantial measurements made by TA and AUGER (the largest cosmic ray observatory in the southern hemisphere), there remains uncertainty about whether the highest energy cosmic rays are galactic or extragalactic in origin. Locating features in the cosmic ray energy spectrum below 1018eV that indicate a transition from galactic to extragalactic sources would clarify the interpretation of measurements made at the highest energies. The Telescope Array Low Energy Extension (TALE) is designed to extend the energy threshold of the TA observatory down to 1016.5eV in order to make such measurements. This dissertation details the construction, calibration, and operation of the TALE flu- orescence detector. A measurement of the flux of cosmic rays in the energy range of 1016.5 -- 1018.5eV is made using the monocular data set taken between September 2013 and January 2014. The TALE fluorescence detector observes evidence for a softening of the cosmic spectrum at 1017.25+/-0.5eV. The evidence of a change in the spectrum motivates continued study of 1016.5 -- 1018.5eV cosmic rays.

  4. Laser Induced Fluorescence Measurements in a Hall Thruster Plume as a Function of Background Pressure

    NASA Technical Reports Server (NTRS)

    Spektor, R.; Tighe, W. G.; Kamhawi, H.

    2016-01-01

    A set of Laser Induced Fluorescence (LIF) measurements in the near-field region of the NASA- 173M Hall thruster plume is presented at four background pressure conditions varying from 9.4 x 10(exp -6) torr to 3.3 x 10(exp -5) torr. The xenon ion velocity distribution function was measured simultaneously along the axial and radial directions. An ultimate exhaust velocity of 19.6+/-0.25 km/s achieved at a distance of 20 mm was measured, and that value was not sensitive to pressure. On the other hand, the ion axial velocity at the thruster exit was strongly influenced by pressure, indicating that the accelerating electric field moved inward with increased pressure. The shift in electric field corresponded to an increase in measured thrust. Pressure had a minor effect on the radial component of ion velocity, mainly affecting ions exiting close to the channel inner wall. At that radial location the radial component of ion velocity was approximately 1000 m/s greater at the lowest pressure than at the highest pressure. A reduction of the inner magnet coil current by 0.6 A resulted in a lower axial ion velocity at the channel exit while the radial component of ion velocity at the channel inner wall location increased by 1300 m/s, and at the channel outer wall location the radial ion velocity remained unaffected. The ultimate exhaust velocity was not significantly affected by the inner magnet current.

  5. Chlorophyll-a determination via continuous measurement of plankton fluorescence: methodology development.

    PubMed

    Pinto, A M; Von Sperling, E; Moreira, R M

    2001-11-01

    A methodology is presented for the continuous measurement of chlorophyll-a concentration due to plankton, in surface water environments. A Turner 10-AU fluorometer equipped with the F4T5.B2/BP lamp (blue lamp), a Cs 5-60 equivalent excitation path filter, and a 680 nm emission filter, has been used. This configuration allows the in vivo, in situ determination of chlorophyll-a by measuring the fluorescence due to the pigments. In field work the fluorometer, data logging and positioning equipment were placed aboard a manageable boat which navigated following a scheme of regularly spaced crossings. Some water samples were collected during the measurement for laboratory chlorophyll-a measurements by the spectrophotometric method, thus providing for calibration and comparison. Spatial chlorophyll-a concentration distributions can be easily defined in large volumes, such as reservoirs, etc. Two distinct environments have been monitored: in the Vargem das Flores reservoir chlorophyll-a concentrations varied between 0.7 and 2.6 mg/m3, whereas in the Lagoa Santa lake these values lied in the 12 to 18 mg/m3 range. The simplicity, versatility and economy of the method, added to the large amount of data that can be gathered in a single run, clearly justify its use in field environmental studies.

  6. Optimization of advanced Wiener estimation methods for Raman reconstruction from narrow-band measurements in the presence of fluorescence background.

    PubMed

    Chen, Shuo; Ong, Yi Hong; Lin, Xiaoqian; Liu, Quan

    2015-07-01

    Raman spectroscopy has shown great potential in biomedical applications. However, intrinsically weak Raman signals cause slow data acquisition especially in Raman imaging. This problem can be overcome by narrow-band Raman imaging followed by spectral reconstruction. Our previous study has shown that Raman spectra free of fluorescence background can be reconstructed from narrow-band Raman measurements using traditional Wiener estimation. However, fluorescence-free Raman spectra are only available from those sophisticated Raman setups capable of fluorescence suppression. The reconstruction of Raman spectra with fluorescence background from narrow-band measurements is much more challenging due to the significant variation in fluorescence background. In this study, two advanced Wiener estimation methods, i.e. modified Wiener estimation and sequential weighted Wiener estimation, were optimized to achieve this goal. Both spontaneous Raman spectra and surface enhanced Raman spectra were evaluated. Compared with traditional Wiener estimation, two advanced methods showed significant improvement in the reconstruction of spontaneous Raman spectra. However, traditional Wiener estimation can work as effectively as the advanced methods for SERS spectra but much faster. The wise selection of these methods would enable accurate Raman reconstruction in a simple Raman setup without the function of fluorescence suppression for fast Raman imaging.

  7. Measurement of resistance to solute transport across surfactant-laden interfaces using a Fluorescence Recovery After Photobleaching (FRAP) technique

    NASA Technical Reports Server (NTRS)

    Browne, Edward P.; Nivaggioli, Thierry; Hatton, T. Alan

    1994-01-01

    A noninvasive fluorescence recovery after photobleaching (FRAP) technique is under development to measure interfacial transport in two phase systems without disturbing the interface. The concentration profiles of a probe solute are measured in both sides of the interface by argon-ion laser, and the system relaxation is then monitored by a microscope-mounted CCD camera.

  8. Surfactant modulates calcium response of neutrophils to physiologic stimulation via cell membrane depolarization.

    PubMed

    Chacon-Cruz, E; Buescher, E S; Oelberg, D G

    2000-03-01

    Pulmonary surfactant (PS) reduces inflammation in the lung by poorly understood mechanisms. We have observed that surfactant-associated proteins (SAP) insert monovalent cation channels in artificial membranes. Neutrophils are primary mediators of acute pulmonary inflammation, and their functions are activated by increases in cytosolic ionized calcium concentration ([Ca2+]) and by changes in membrane potential. We hypothesize that PS inserts SAP-dependent cation channels in neutrophils, causing membrane depolarization, altered [Ca2+] response, and depressed activation. Human neutrophils were isolated, exposed to PS+SAP (1% Survanta), PS-SAP (1% Exosurf), or buffer, and washed before activating with selected stimulants. PS+SAP reduced phorbol ester- and formyl peptide-stimulated adherence and aggregation by 38% (p < 0.05) and 54% (p < 0.02), respectively. PS+SAP also inhibited the formyl peptide-induced [Ca2+] response of neutrophils (p < 0.01), but only in the presence of external Ca2+. Further characterization of this inhibition demonstrated that PS+SAP blocked formyl peptide-induced influx of both Ca2+ and Mn2+, and that this inhibition was present during activation by other neutrophil stimulants (IL-8, immune complexes). Prior depolarization of neutrophils with gramicidin-D similarly inhibited the [Ca2+] response of neutrophils to formyl peptide, and analysis of neutrophil membrane potential by 3,3'-dipentyloxaearbocyanine iodide (diOC5(3)) fluorescence revealed that PS+SAP induced rapid neutrophil depolarization. In contrast, PS-SAP exhibited little effect on neutrophil function, [Ca2+], or membrane potential. We conclude that PS+SAP decreases neutrophil adherence and aggregation responses, blocks Ca2+ influx after physiologic stimulation, and decreases membrane potential. We speculate that these effects are caused by membrane depolarization via SAP-dependent cation channel insertion, and that all of these effects contribute to the antiinflammatory properties of

  9. Fluorescent sensors for the basic metabolic panel enable measurement with a smart phone device over the physiological range.

    PubMed

    Awqatty, Becker; Samaddar, Shayak; Cash, Kevin J; Clark, Heather A; Dubach, J Matthew

    2014-10-21

    The advanced functionality of portable devices such as smart phones provides the necessary hardware to potentially perform complex diagnostic measurements in any setting. Recent research and development have utilized cameras and data acquisition properties of smart phones to create diagnostic approaches for a variety of diseases or pollutants. However, in concentration measurements, such as blood glucose, the performance of handheld diagnostic devices depends largely on the sensing mechanism. To expand measurements to multiple components, often necessary in medical tests, with a single diagnostic device, robust platform based sensors are needed. Here, we developed a suite of dual wavelength fluorescent sensors with response characteristics necessary to measure each component of a basic metabolic panel, a common clinical measurement. Furthermore, the response of these sensors could be measured with a simple optical setup to convert a smart phone into a fluorescence measurement instrument. This approach could be used as a mobile basic metabolic panel measurement system for point of care diagnostics.

  10. Fluorescence ratio imaging microscopy: temporal and spatial measurements of cytoplasmic pH

    PubMed Central

    1987-01-01

    Fluorescence ratio imaging microscopy (Tanasugarn, L., P. McNeil, G. Reynolds, and D. L. Taylor, 1984, J. Cell Biol., 98:717-724) has been used to measure the spatial variations in cytoplasmic pH of individual quiescent and nonquiescent Swiss 3T3 cells. Fundamental issues of ratio imaging that permit precise and accurate temporal and spatial measurements have been addressed including: excitation light levels, lamp operation, intracellular probe concentrations, methods of threshold selection, photobleaching, and spatial signal-to-noise ratio measurements. Subcellular measurements can be measured accurately (less than 3% coefficient of variation) in an area of 3.65 microns 2 with the present imaging system. Quiescent Swiss 3T3 cells have a measured cytoplasmic pH of 7.09 (0.01 SEM), whereas nonquiescent cells have a pH of 7.35 (0.01 SEM) in the presence of bicarbonate buffer. A unimodal distribution of mean cytoplasmic pH in both quiescent and nonquiescent cells was identified from populations of cells measured on a cell by cell basis. Therefore, unlike earlier studies based on cell population averages, it can be stated that cells in each population exhibit a narrow range of cytoplasmic pH. However, the mean cytoplasmic pH can change based on the physiological state of the cells. In addition, there appears to be little, if any, spatial variation in cytoplasmic pH in either quiescent or nonquiescent Swiss 3T3 cells. The pH within the nucleus was always the same as the surrounding cytoplasm. These values will serve as a reference point for investigating the role of temporal and spatial variations in cytoplasmic pH in a variety of cellular processes including growth control and cell movement. PMID:3558476

  11. Measurements of IO in the Tropical Marine Boundary Layer using Laser-Induced Fluorescence Spectroscopy

    NASA Astrophysics Data System (ADS)

    Walker, H.; Ingham, T.; Heard, D. E.

    2012-12-01

    Halogenated short-lived substances (VSLS) are emitted from the oceans by marine species such as macroalgae and phytoplankton and contribute to halogen loading in the troposphere and lower stratosphere. Transport of halogenated VSLS into the stratosphere occurs mainly in the tropics, where ascending warm air carries them aloft, and leads to catalytic depletion of stratospheric ozone on a global scale and formation of the Antarctic ozone hole. The tropical marine environment is therefore an important region in which to study the effects of these short-lived halogen species on ozone depletion. The SHIVA (Stratospheric Ozone: Halogen Impacts in a Varying Atmosphere) project combines ship-borne, aircraft-based and ground-based measurements in and over the South China Sea and the Sulu Sea, and around the coast of Malaysian Borneo, to reduce uncertainties in the amount of halogenated VSLS reaching the stratosphere, the associated ozone depletion, and the effects of a changing climate on these processes. In this work we present measurements of IO radicals made onboard the German research vessel Sonne during SHIVA, between Singapore and Manila. IO is formed via photolysis of iodine-containing source gases (e.g. I2, CH3I) to produce I atoms, which react with ozone. It is therefore an important species to consider when assessing the impacts of halogen chemistry on ozone depletion. Measurements of IO were made over a two-week period by the University of Leeds Laser-Induced Fluorescence (LIF) instrument, which excites IO radicals at λ ~ 445 nm and detects the resultant fluorescence at λ ~ 512 nm. A suite of supporting gas- and aqueous-phase measurements were also made, including concentrations of halocarbons (e.g. CHBr3, CH3I), trace pollutant gases (e.g. CO, O3, NOx), and biological parameters (e.g. abundance and speciation of phytoplankton). Preliminary data analysis indicates that IO was detected above the instrumental limit of detection (0.3 pptv for a 30 minute averaging

  12. Two Photon Absorption Laser Induced Fluorescence for Neutral Hydrogen Profile Measurements

    SciTech Connect

    Scime, Earl E.

    2016-09-23

    The magnitude and spatial dependence of neutral density in magnetic confinement fusion experiments is a key physical parameter, particularly in the plasma edge. Modeling codes require precise measurements of the neutral density to calculate charge-exchange power losses and drag forces on rotating plasmas. However, direct measurements of the neutral density are problematic. In this work, we proposed to construct a laser-based diagnostic capable of providing spatially resolved measurements of the neutral density in the edge of plasma in the DIII-D tokamak. The diagnostic concept is based on two-photon absorption laser induced fluorescence (TALIF). By injecting two beams of 205 nm light (co or counter propagating), ground state hydrogen (or deuterium or tritium) can be excited from the n = 1 level to the n = 3 level at the location where the two beams intersect. Individually, the beams experience no absorption, and therefore have no difficulty penetrating even dense plasmas. After excitation, a fraction of the hydrogen atoms decay from the n = 3 level to the n = 2 level and emit photons at 656 nm (the Hα line). Calculations based on the results of previous TALIF experiments in magnetic fusion devices indicated that a laser pulse energy of approximately 3 mJ delivered in 5 ns would provide sufficient signal-to-noise for detection of the fluorescence. In collaboration with the DIII-D engineering staff and experts in plasma edge diagnostics for DIII-D from Oak Ridge National Laboratory (ORNL), WVU researchers designed a TALIF system capable of providing spatially resolved measurements of neutral deuterium densities in the DIII-D edge plasma. The laser systems were specified, purchased, and assembled at WVU. The TALIF system was tested on a low-power hydrogen discharge at WVU and the plan was to move the instrument to DIII-D for installation in collaboration with ORNL researchers. After budget cuts at DIII-D, the DIII-D facility declined to support

  13. Depolarization of MNOS structures in the regime of dispersion transport

    NASA Astrophysics Data System (ADS)

    Raikerus, P. A.; Gurtov, V. A.

    1991-01-01

    A theory of isothermal depolarization of insulators based on dispersive transport through traps (exponentially distributed in energy) is developed. Equations for the dependence of flat-band voltage on time have been obtained. It is shown that the depolarization process takes place during a sufficiently long time (over about six decades of time). Equations for the "energy activation" of the depolarization process increasing with time and decreasing with temperature and electric field have been obtained. Experimental data for MNOS structures are explained on the basis of the dispersive transport theory.

  14. Realizing controllable depolarization in photonic quantum-information channels

    SciTech Connect

    Shaham, A.; Eisenberg, H. S.

    2011-02-15

    Controlling the depolarization of light is a long-standing open problem. In recent years, many demonstrations have used the polarization of single photons to encode quantum information. The depolarization of these photons is equivalent to the decoherence of the quantum information they encode. We present schemes for building various depolarizing channels with controlled properties using birefringent crystals. Three such schemes are demonstrated, and their effects on single photons are shown by quantum process tomography to be in good agreement with a theoretical model.

  15. Nuclear Resonance Fluorescence to Measure Plutonium Mass in Spent Nuclear Fuel

    SciTech Connect

    Ludewigt, Bernhard A; Quiter, Brian J.; Ambers, Scott D.

    2011-01-14

    The Next Generation Safeguard Initiative (NGSI) of the U.S Department of Energy is supporting a multi-lab/university collaboration to quantify the plutonium (Pu) mass in spent nuclear fuel (SNF) assemblies and to detect the diversion of pins with non-destructive assay (NDA) methods. The following 14 NDA techniques are being studied: Delayed Neutrons, Differential Die-Away, Differential Die-Away Self-Interrogation, Lead Slowing Down Spectrometer, Neutron Multiplicity, Passive Neutron Albedo Reactivity, Total Neutron (Gross Neutron), X-Ray Fluorescence, {sup 252}Cf Interrogation with Prompt Neutron Detection, Delayed Gamma, Nuclear Resonance Fluorescence, Passive Prompt Gamma, Self-integration Neutron Resonance Densitometry, and Neutron Resonance Transmission Analysis. Understanding and maturity of the techniques vary greatly, ranging from decades old, well-understood methods to new approaches. Nuclear Resonance Fluorescence (NRF) is a technique that had not previously been studied for SNF assay or similar applications. Since NRF generates isotope-specific signals, the promise and appeal of the technique lies in its potential to directly measure the amount of a specific isotope in an SNF assay target. The objectives of this study were to design and model suitable NRF measurement methods, to quantify capabilities and corresponding instrumentation requirements, and to evaluate prospects and the potential of NRF for SNF assay. The main challenge of the technique is to achieve the sensitivity and precision, i.e., to accumulate sufficient counting statistics, required for quantifying the mass of Pu isotopes in SNF assemblies. Systematic errors, considered a lesser problem for a direct measurement and only briefly discussed in this report, need to be evaluated for specific instrument designs in the future. Also, since the technical capability of using NRF to measure Pu in SNF has not been established, this report does not directly address issues such as cost, size

  16. Spatially Multiplexed Imaging: Fluorescence Correlation Spectroscopy for Efficient Measurement of Molecular Diffusion at Solid-Liquid Interfaces.

    PubMed

    Cooper, Justin T; Harris, Joel M

    2016-04-01

    Fluorescence correlation spectroscopy (FCS) has become an important technique for the characterization of molecular dynamics, especially at interfaces. Fluorescence correlation spectroscopy provides both temporal and spatial resolution for measuring fast processes at equilibrium through analysis of noise in fluorescence intensities from the statistical fluctuations in a small number of molecules. The small molecular populations produce very low-level fluorescence signals, where time-averaging the fluorescence autocorrelation function is needed to generate reasonable signal-to-noise (S/N) ratios. Recently imaging cameras have been adapted to FCS measurements of molecular dynamics at interfaces (membranes and surfaces) through the use of electron-multiplying charge-coupled device (EM-CCD) detectors for acquisition of fluorescence from addressable areas on the detector. This approach provides a major advantage over traditional focused-spot FCS by allowing electronic control over the location and area of the acquired region on the sample surface. Imaging-FCS can also provide a spatial multiplexing advantage through its ability to measure intensity data from larger areas in parallel with no loss of time resolution. In this work, this multiplexing advantage is exploited to determine molecular diffusion rates from the simultaneous measurement of multiple areas on a surface, the autocorrelation traces from which are averaged to improve the S/N ratio. As proof of concept, the diffusion of 1,1'-dioctadecyl-3,3,3'3'-tetramethylindocarbocyanine perchlorate (DiI) on a C18-modified interface was measured using this multiplexed method and compared to autocorrelation data acquired from a single spot. Due to the slow thermal recovery of the EM-CCD that inhibits fast time-averaging, spatial multiplexing in imaging-FCS provides an eightyfold time savings to reach the same S/N ratio as multiple (time-averaged) measurements from a single spot.

  17. Phase-resolved x-ray ferromagnetic resonance measurements in fluorescence yield

    SciTech Connect

    Marcham, M. K.; Keatley, P. S.; Neudert, A.; Hicken, R. J.; Cavill, S. A.; Shelford, L. R.; van der Laan, G.; Telling, N. D.; Childress, J. R.; Katine, J. A.; Shafer, P.; Arenholz, E.

    2010-10-14

    Phase-resolved x-ray ferromagnetic resonance (XFMR) has been measured in fluorescence yield, extending the application of XFMR to opaque samples on opaque substrates. Magnetization dynamics were excited in a Co{sub 50}Fe{sub 50}(0.7)/Ni{sub 90}Fe{sub 10}(5) bilayer by means of a continuous wave microwave excitation, while x-ray magnetic circular dichroism (XMCD) spectra were measured stroboscopically at different points in the precession cycle. By tuning the x-ray energy to the L{sub 3} edges of Ni and Fe, the dependence of the real and imaginary components of the element specific magnetic susceptibility on the strength of an externally applied static bias field was determined. First results from measurements on a Co{sub 50}Fe{sub 50}(0.7)/Ni{sub 90}Fe{sub 10}(5)/Dy(1) sample confirm that enhanced damping results from the addition of the Dy cap.

  18. Fluorescence (TALIF) measurement of atomic hydrogen concentration in a coplanar surface dielectric barrier discharge

    NASA Astrophysics Data System (ADS)

    Mrkvičková, M.; Ráheľ, J.; Dvořák, P.; Trunec, D.; Morávek, T.

    2016-10-01

    Spatially and temporally resolved measurements of atomic hydrogen concentration above the dielectric of coplanar barrier discharge are presented for atmospheric pressure in 2.2% H2/Ar. The measurements were carried out in the afterglow phase by means of two-photon absorption laser-induced fluorescence (TALIF). The difficulties of employing the TALIF technique in close proximity to the dielectric surface wall were successfully addressed by taking measurements on a suitable convexly curved dielectric barrier, and by proper mathematical treatment of parasitic signals from laser-surface interactions. It was found that the maximum atomic hydrogen concentration is situated closest to the dielectric wall from which it gradually decays. The maximum absolute concentration was more than 1022 m-3. In the afterglow phase, the concentration of atomic hydrogen above the dielectric surface stays constant for a considerable time (10 μs-1 ms), with longer times for areas situated farther from the dielectric surface. The existence of such a temporal plateau was explained by the presented 1D model: the recombination losses of atomic hydrogen farther from the dielectric surface are compensated by the diffusion of atomic hydrogen from regions close to the dielectric surface. The fact that a temporal plateau exists even closest to the dielectric surface suggests that the dielectric surface acts as a source of atomic hydrogen in the afterglow phase.

  19. Measurement of the hydrodynamic radius of quantum dots by fluorescence correlation spectroscopy excluding blinking.

    PubMed

    de Thomaz, A A; Almeida, D B; Pelegati, V B; Carvalho, H F; Cesar, C L

    2015-03-19

    One of the most important properties of quantum dots (QDs) is their size. Their size will determine optical properties and in a colloidal medium their range of interaction. The most common techniques used to measure QD size are transmission electron microscopy (TEM) and X-ray diffraction. However, these techniques demand the sample to be dried and under a vacuum. This way any hydrodynamic information is excluded and the preparation process may alter even the size of the QDs. Fluorescence correlation spectroscopy (FCS) is an optical technique with single molecule sensitivity capable of extracting the hydrodynamic radius (HR) of the QDs. The main drawback of FCS is the blinking phenomenon that alters the correlation function implicating in a QD apparent size smaller than it really is. In this work, we developed a method to exclude blinking of the FCS and measured the HR of colloidal QDs. We compared our results with TEM images, and the HR obtained by FCS is higher than the radius measured by TEM. We attribute this difference to the cap layer of the QD that cannot be seen in the TEM images.

  20. Handheld X-ray Fluorescence Spectrometers: Radiation Exposure Risks of Matrix-Specific Measurement Scenarios.

    PubMed

    Rouillon, Marek; Kristensen, Louise J; Gore, Damian B

    2015-07-01

    This study investigates X-ray intensity and dispersion around handheld X-ray fluorescence (XRF) instruments during the measurement of a range of sample matrices to establish radiation exposure risk during operation. Four handheld XRF instruments representing three manufacturers were used on four smooth, flat-lying materials of contrasting matrix composition. Dose rates were measured at 10, 20, 30, and 40 cm intervals every 30° around the instrument at 0 and 45° from the horizontal, as well as vertically from the instrument screen. The analysis of polyethylene recorded dose rates 156 times higher (on average) than steel measurements and 34 times higher than both quartz sand and quartz sandstone. A worst-case exposure scenario was assumed where a user analyses a polyethylene material at arms reach for 1 h each working day for one year. This scenario resulted in an effective body dose of 73.5 μSv, equivalent to three to four chest X-rays (20 μSv) a year, 20 times lower than the average annual background radiation exposure in Australia and well below the annual exposure limit of 1 mSv for non-radiation workers. This study finds the advantages of using handheld XRF spectrometers far outweighs the risk of low radiation exposure linked to X-ray scattering from samples.

  1. Source-corrected two-photon excited fluorescence measurements between 700 and 880 nm

    SciTech Connect

    Fisher, W.G.; Wachter, E.A.; Lytle, F.E.; Armas, M.; Seaton, C.

    1998-04-01

    Passively mode-locked titanium:sapphire (Ti:S) lasers are capable of generating a high-frequency train of transform-limited subpico-second pulses, producing peak powers near 10{sup 5}thinspW at moderate average powers. The low energy per pulse ({lt}20 nJ) permits low fluence levels to be maintained in tightly focused beams, reducing the possibility of saturating fluorescence transitions. These properties, combined with a wavelength tunability from approximately 700 nm to 1 {mu}m, provide excellent opportunities for studying simultaneous two-photon excitation (TPE). However, pulse formation is very sensitive to a variety of intracavity parameters, including group velocity dispersion compensation, which leads to wavelength-dependent pulse profiles as the wavelength is scanned. This wavelength dependence can seriously distort band shapes and apparent peak heights during collection of two-photon spectral data. Since two-photon excited fluorescence is proportional to the product of the peak and average powers, it is not possible to obtain source-independent spectra by using average power correction schemes alone. Continuous-wave, single-mode lasers can be used to generate source-independent two-photon data, but these sources are four to five orders of magnitude less efficient than the mode-locked Ti:S laser and are not practical for general two-photon measurements. Hence, a continuous-wave, single-mode Ti:S laser has been used to collect a source-independent excitation spectrum for the laser dye Coumarin 480. This spectrum may be used to correct data collected with multimode sources; this possibility is demonstrated by using a simple ratiometric method to collect accurate TPE spectra with the mode-locked Ti:S laser. An approximate value of the two-photon cross section for Coumarin 480 is also given. {copyright} {ital 1998} {ital Society for Applied Spectroscopy}

  2. Electronically excited dipole moment of 4-aminobenzonitrile from thermochromic absorption and fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Kawski, A.; Kukliński, B.; Bojarski, P.

    2006-07-01

    The effect of temperature on absorption and fluorescence spectra of 4-aminobenzonitrile (ABN) in 1,2-dichloroethane is studied for temperature ranging from 296 K to 343 K. The analysis of absorption and fluorescence band shift on the basis of Bilot and Kawski theory [L. Bilot, A. Kawski, Z. Naturforsch. 17a (1962) 621], for the known dipole moment in the ground state μg = 5.92 D, and α/ a3 = 0.5 ( α is the polarizability and a is the Onsager interaction radius of the solute) yields for ABN: (1) the empirical Onsager interaction radius a = 3.3 Å, (2) the dipole moment in the excited S 1 state μe = 7.14 D which agrees very well with the value of μe = 7.20 D obtained by Borst et al. [D.R. Borst, T.M. Korter, D.W. Pratt, Chem. Phys. Lett. 350 (2001) 485] from Stark effect studies. Both values of μe concern free ABN molecule and differ significantly from the values of μg (8.0 D, 8.5 D and 8.3 D in cyclohexane, benzene and 1,4-dioxane, respectively) obtained by Schuddeboom et al. [W. Schuddeboom, S.A. Jonker, J.M. Warman, U. Leinhos, W. Kühnle, K.A. Zachariasse, J. Phys. Chem. 96 (1992) 10809] from the time-resolved microwave conductivity measurements which are solvent-dependent. The group moment additivity law in the case of ABN molecule is approximately applicable, both in the ground and in the excited electronic state.

  3. The Measurement of Radiative Lifetimes Using Laser-Induced Fluorescence: Experimental Review and Astrophysical Application

    NASA Astrophysics Data System (ADS)

    Den Hartog, E. A.; Lawler, J. E.; Sneden, C.

    2005-01-01

    One of the standard methods for determining atomic transition probabilities is to combine branching fractions measured using Fourier-transform spectrometry with radiative lifetimes measurements using laser-induced fluorescence (LIF). This combination of techniques provides an efficient method for measuring large sets of accurate, absolute transition probabilities. The radiative lifetimes, which provide the overall scaling for the transition probabilities, can be measured routinely to ± 5% accuracy using time-resolved LIF. Although the time-resolved LIF technique we use does not achieve the accuracy of fast-beam LIF, the time-resolved technique does enable us to make measurements at a far greater rate (hundreds of level lifetimes per year). Care must be taken, however, to understand and control the systematic effects in time-resolved LIF measurements to maintain ± 5% accuracy over a wide dynamic range and hundreds of lifetime measurements. Over the last 25 years, we have measured lifetimes for 47 spectra using time-resolved LIF. Our atomic beam source can produce a slow beam of neutral and singly ionized atoms of nearly any element. Lifetimes from 2 ns to ~2 µs can be measured for energy levels ranging from 15,000 to ~60,000 cm-1. In this review we will describe our method of measuring radiative lifetimes with an emphasis on possible errors and techniques used for controlling them. The electronic bandwidth, linearity, and overall fidelity of the fast photomultiplier, cable connections, and transient waveform digitizer are concerns. Possible errors from atomic collisions, radiation trapping, Zeeman quantum beats, hyperfine quantum beats, atoms/ions escaping from the observation region before radiating, and from radiative cascade through lower levels must be understood and controlled. We will then present a recent example of the application of our transition probability data to abundance determinations in the sun and in metal-poor halo stars (Den Hartog E A et al

  4. A Dual Functional Electroactive and Fluorescent Probe for Coupled Measurements of Vesicular Exocytosis with High Spatial and Temporal Resolution.

    PubMed

    Liu, Xiaoqing; Savy, Alexandra; Maurin, Sylvie; Grimaud, Laurence; Darchen, François; Quinton, Damien; Labbé, Eric; Buriez, Olivier; Delacotte, Jérôme; Lemaître, Frédéric; Guille-Collignon, Manon

    2017-02-20

    In this work, Fluorescent False Neurotransmitter 102 (FFN102), a synthesized analogue of biogenic neurotransmitters, was demonstrated to show both pH-dependent fluorescence and electroactivity. To study secretory behaviors at the single-vesicle level, FFN102 was employed as a new fluorescent/electroactive dual probe in a coupled technique (amperometry and total internal reflection fluorescence microscopy (TIRFM)). We used N13 cells, a stable clone of BON cells, to specifically accumulate FFN102 into their secretory vesicles, and then optical and electrochemical measurements of vesicular exocytosis were experimentally achieved by using indium tin oxide (ITO) transparent electrodes. Upon stimulation, FFN102 started to diffuse out from the acidic intravesicular microenvironment to the neutral extracellular space, leading to fluorescent emissions and to the electrochemical oxidation signals that were simultaneously collected from the ITO electrode surface. The correlation of fluorescence and amperometric signals resulting from the FFN102 probe allows real-time monitoring of single exocytotic events with both high spatial and temporal resolution. This work opens new possibilities in the investigation of exocytotic mechanisms.

  5. Formaldehyde preparation methods for pressure and temperature dependent laser-induced fluorescence measurements.

    PubMed

    Burkert, A; Müller, D; Rieger, S; Schmidl, G; Triebel, W; Paa, W

    2015-12-01

    Formaldehyde is an excellent tracer for the early phase of ignition of hydrocarbon fuels and can be used, e.g., for characterization of single droplet ignition. However, due to its fast thermal decomposition at elevated temperatures and pressures, the determination of concentration fields from laser-induced fluorescence (LIF) measurements is difficult. In this paper, we address LIF measurements of this important combustion intermediate using a calibration cell. Here, formaldehyde is created from evaporation of paraformaldehyde. We discuss three setups for preparation of formaldehyde/air mixtures with respect to their usability for well-defined heating of formaldehyde/air mixtures. The "basic setup" uses a resist heater around the measurement cell for investigation of formaldehyde near vacuum conditions or formaldehyde/air samples after sequential admixing of air. The second setup, described for the first time in detail here, takes advantage of a constant flow formaldehyde/air regime which uses preheated air to reduce the necessary time for gas heating. We used the constant flow system to measure new pressure dependent LIF excitation spectra in the 343 nm spectral region (41 (4) absorption band of formaldehyde). The third setup, based on a novel concept for fast gas heating via excitation of SF6 (chemically inert gas) using a TEA (transverse excitation at atmospheric pressure) CO2 laser, allows to further minimize both gas heating time and thermal decomposition. Here, an admixture of CO2 is served for real time temperature measurement based on Raman scattering. The applicability of the fast laser heating system has been demonstrated with gas mixtures of SF6 + air, SF6 + N2, as well as SF6 + N2 + CO2 at 1 bar total pressure.

  6. Formaldehyde preparation methods for pressure and temperature dependent laser-induced fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Burkert, A.; Müller, D.; Rieger, S.; Schmidl, G.; Triebel, W.; Paa, W.

    2015-12-01

    Formaldehyde is an excellent tracer for the early phase of ignition of hydrocarbon fuels and can be used, e.g., for characterization of single droplet ignition. However, due to its fast thermal decomposition at elevated temperatures and pressures, the determination of concentration fields from laser-induced fluorescence (LIF) measurements is difficult. In this paper, we address LIF measurements of this important combustion intermediate using a calibration cell. Here, formaldehyde is created from evaporation of paraformaldehyde. We discuss three setups for preparation of formaldehyde/air mixtures with respect to their usability for well-defined heating of formaldehyde/air mixtures. The "basic setup" uses a resist heater around the measurement cell for investigation of formaldehyde near vacuum conditions or formaldehyde/air samples after sequential admixing of air. The second setup, described for the first time in detail here, takes advantage of a constant flow formaldehyde/air regime which uses preheated air to reduce the necessary time for gas heating. We used the constant flow system to measure new pressure dependent LIF excitation spectra in the 343 nm spectral region (414 absorption band of formaldehyde). The third setup, based on a novel concept for fast gas heating via excitation of SF6 (chemically inert gas) using a TEA (transverse excitation at atmospheric pressure) CO2 laser, allows to further minimize both gas heating time and thermal decomposition. Here, an admixture of CO2 is served for real time temperature measurement based on Raman scattering. The applicability of the fast laser heating system has been demonstrated with gas mixtures of SF6 + air, SF6 + N2, as well as SF6 + N2 + CO2 at 1 bar total pressure.

  7. The Pathophysiological Mechanism Underlying Brugada Syndrome. Depolarization versus Repolarization

    PubMed Central

    Wilde, Arthur A.M.; Postema, Pieter G.; Di Diego, José M.; Viskin, Sami; Morita, Hiroshi; Fish, Jeffrey M.; Antzelevitch, Charles

    2010-01-01

    This Point/Counterpoint presents a scholarly debate of the mechanisms underlying the electrocardiographic and arrhythmic manifestations of Brugada syndrome (BrS), exploring in detail the available evidence in support of the repolarization vs. depolarization hypothesis. PMID:20659475

  8. Depolarizing collisions with hydrogen: Neutral and singly ionized alkaline earths

    SciTech Connect

    Manso Sainz, Rafael; Ramos, Andrés Asensio; Bueno, Javier Trujillo; Aguado, Alfredo

    2014-06-20

    Depolarizing collisions are elastic or quasielastic collisions that equalize the populations and destroy the coherence between the magnetic sublevels of atomic levels. In astrophysical plasmas, the main depolarizing collider is neutral hydrogen. We consider depolarizing rates on the lowest levels of neutral and singly ionized alkali earths Mg I, Sr I, Ba I, Mg II, Ca II, and Ba II, due to collisions with H°. We compute ab initio potential curves of the atom-H° system and solve the quantum mechanical dynamics. From the scattering amplitudes, we calculate the depolarizing rates for Maxwellian distributions of colliders at temperatures T ≤ 10,000 K. A comparative analysis of our results and previous calculations in the literature is completed. We discuss the effect of these rates on the formation of scattering polarization patterns of resonant lines of alkali earths in the solar atmosphere, and their effect on Hanle effect diagnostics of solar magnetic fields.

  9. Measurements of cell wall mechanical properties using optically trapped fluorescent microspheres

    NASA Astrophysics Data System (ADS)

    Ermilov, Sergey; Qian, Feng; Murdock, David; Brownell, William E.; Anvari, Bahman

    2004-10-01

    Information on plasma membrane (PM) and cell wall mechanical properties is important for many biophysical applications, especially for those, which involve cells, undergoing significant mechanical stress (red blood cells, outer hair cells, fibrocytes, etc.). Optical tweezers is frequently used to study PM mechanics, particularly by pulling long PM tethers. One of the limitations on using optical tweezers to study cell wall mechanics is associated with transillumination technique of the trapped object position sensing, which prevents accurate mechanical testing in the proximity to the cell. In this work we use an optical tweezers in conjunction with a position-sensing system, which spectrally separates signals from the trapped fluorescent microsphere and imaging background. We have used this setup to study mechanics of the cell wall and PM separated from the underlying cytoskeleton on human embryonic kidney cells. We measured the force exerted by the cell on the trapped microsphere as a function of the cell wall displacement during the process of tether formation, and as a function of time during the process of tether growth and relaxation. Tethering force - cell wall displacement profiles have shown a behavior, implying that tether formation process starts with elastic deformation of the intact cell wall, followed by the plastic deformations and sliding of the PM over the underlying cytoskeleton, and ends with the local separation of a PM. Tethering force - cell wall displacement profiles have been used to estimate tether formation force, stiffness parameter of the cell wall and the works of tether formation, elastic and plastic deformations of the cell wall, related to the mechanical properties of a composite cell wall and cell wall - plasma membrane association strength. Temporal steady-state and relaxation tethering force profiles have been similar to the ones measured using transillumination position sensing, however average force values have been smaller in

  10. Mapping cropland GPP in the north temperate region with space measurements of chlorophyll fluorescence

    NASA Astrophysics Data System (ADS)

    Guanter, L.; Zhang, Y.; Jung, M.; Joiner, J.; Voigt, M.; Huete, A. R.; Zarco-Tejada, P.; Frankenberg, C.; Lee, J.; Berry, J. A.; Moran, S. M.; Ponce-Campos, G.; Beer, C.; Camps-Valls, G.; Buchmann, N. C.; Gianelle, D.; Klumpp, K.; Cescatti, A.; Baker, J. M.; Griffis, T.

    2013-12-01

    Monitoring agricultural productivity is important for optimizing management practices in a world under a continuous increase of food and biofuel demand. We used new space measurements of sun-induced chlorophyll fluorescence (SIF), a vegetation parameter intrinsically linked to photosynthesis, to capture photosynthetic uptake of the crop belts in the north temperate region. The following data streams and procedures have been used in this analysis: (1) SIF retrievals have been derived from measurements of the MetOp-A / GOME-2 instrument in the 2007-2011 time period; (2) ensembles of process-based and data-driven biogeochemistry models have been analyzed in order to assess the capability of global models to represent crop gross primary production (GPP); (3) flux tower-based GPP estimates covering the 2007-2011 time period have been extracted over 18 cropland and grassland sites in the Midwest US and Western Europe from the Ameriflux and the European Fluxes Database networks; (4) large-scale NPP estimates have been derived by the agricultural inventory data sets developed by USDA-NASS and Monfreda et al. The strong linear correlation between the SIF space retrievals and the flux tower-based GPP, found to be significantly higher than that between reflectance-based vegetation indices (EVI, NDVI and MTCI) and GPP, has enabled the direct upscaling of SIF to cropland GPP maps at the synoptic scale. The new crop GPP estimates we derive from the scaling of SIF space retrievals are consistent with both flux tower GPP estimates and agricultural inventory data. These new GPP estimates show that crop productivity in the US Western Corn Belt, and most likely also in the rice production areas in the Indo-Gangetic plain and China, is up to 50-75% higher than estimates by state-of-the-art data-driven and process-oriented biogeochemistry models. From our analysis we conclude that current carbon models have difficulties in reproducing the special conditions of those highly productive

  11. Correlates of spreading depolarization in human scalp electroencephalography.

    PubMed

    Drenckhahn, Christoph; Winkler, Maren K L; Major, Sebastian; Scheel, Michael; Kang, Eun-Jeung; Pinczolits, Alexandra; Grozea, Cristian; Hartings, Jed A; Woitzik, Johannes; Dreier, Jens P

    2012-03-01

    It has been known for decades that suppression of spontaneous scalp electroencephalographic activity occurs during ischaemia. Trend analysis for such suppression was found useful for intraoperative monitoring during carotid endarterectomy, or as a screening tool to detect delayed cerebral ischaemia after aneurismal subarachnoid haemorrhage. Nevertheless, pathogenesis of such suppression of activity has remained unclear. In five patients with aneurismal subarachnoid haemorrhage and four patients with decompressive hemicraniectomy after malignant hemispheric stroke due to middle cerebral artery occlusion, we here performed simultaneously full-band direct and alternating current electroencephalography at the scalp and direct and alternating current electrocorticography at the cortical surface. After subarachnoid haemorrhage, 275 slow potential changes, identifying spreading depolarizations, were recorded electrocorticographically over 694 h. Visual inspection of time-compressed scalp electroencephalography identified 193 (70.2%) slow potential changes [amplitude: -272 (-174, -375) µV (median quartiles), duration: 5.4 (4.0, 7.1) min, electrocorticography-electroencephalography delay: 1.8 (0.8, 3.5) min]. Intervals between successive spreading depolarizations were significantly shorter for depolarizations with electroencephalographically identified slow potential change [33.0 (27.0, 76.5) versus 53.0 (28.0, 130.5) min, P = 0.009]. Electroencephalography was thus more likely to display slow potential changes of clustered than isolated spreading depolarizations. In contrast to electrocorticography, no spread of electroencephalographic slow potential changes was seen, presumably due to superposition of volume-conducted electroencephalographic signals from widespread cortical generators. In two of five patients with subarachnoid haemorrhage, serial magnetic resonance imaging revealed large delayed infarcts at the recording site, while electrocorticography showed clusters

  12. Scattering and Depolarization of Electromagnetic Waves--Full Wave Solutions.

    DTIC Science & Technology

    1984-01-01

    Analysis," Proceedings of the International Union of Radio Science URSI Conference at Ciudad Universitaria , Madrid, August 1983, in press. . . 13...rough land and seat3 J. The full wave approach was also used to determine the scattering and depolarization of radio waves in irregular spheroidal struc...Full Wave Solutions," Radio Science, Vol. 17, No. 5, September-October 1982, pp. 1055-1066. 4. "Scattering and Depolarization by Rough Surfaces: Full

  13. Catecholamine secretion, calcium levels and calcium influx in response to membrane depolarization in bovine chromaffin cells.

    PubMed

    Calvo, S; Granja, R; González-García, C; Ceña, V

    1995-09-01

    In this paper, we show that exposure of chromaffin cells to high K+ (75 mM) for 5 min releases about 15% of total norepinephrine and 8% of total epinephrine contained in chromaffin cells. The measured resting membrane potential of these cells was -55 mV. Long (10 s) depolarizing electrical pulses applied from a holding potential of -55 mV to 5 mV, that would produce a depolarization similar to exposure to high K+ (75 mM), induced an inward Ca2+ current that inactivated with a time constant of about 0.8 s and promoted the influx of about 1 fmol of Ca2+ into the cell. Both high K+ and electrically-induced depolarization increased intracellular Ca2+ levels to a similar value (about 350 nM). Extrapolation would indicate that total Ca2+ influx in high K+ (75 mM)-stimulated 10(6) chromaffin cells would amount to 1 nmol which would promote the secretion of about 4.9 nmol of norepinephrine and 3.5 nmol of epinephrine from 10(6) chromaffin cells. The results indicate that Ca2+ influx in response to depolarization is short-lived, likely due to Ca(2+)-dependent inactivation of voltage-dependent Ca2+ channels. However, intracellular Ca2+ levels remain high as long as depolarization is present and long after Ca2+ influx has ceased. This would suggest that some processes related to either Ca2+ buffering or extrusion from the cell may be voltage dependent.

  14. Contractile responses to rat urotensin II in resting and depolarized basilar arteries.

    PubMed

    Porras-González, Cristina; Ureña, Juan; Egea-Guerrero, Juan José; Gordillo-Escobar, Elena; Murillo-Cabezas, Francisco; González-Montelongo, María del Carmen; Muñoz-Sánchez, María Angeles

    2014-03-01

    The effects of human urotensin II (hUII) on the vascular tone of different animal species has been studied extensively. However, little has been reported on the vasoactive effects of rat urotensin (rUII) in murine models. The aim of the present study was to investigate the effects of rUII on vasoreactivity in rat basilar arteries. Basilar arteries from adult male Wistar rats (300-350 g) were isolated, cut in rings, and mounted on a small vessel myograph to measure isometric tension. rUII concentrations were studied in both resting and depolarized state. To remove endothelial nitric oxide effects from the rUII response, we treated selected arterial rings with Nω-nitro-L-arginine methyl ester (L-NAME). 10 μM rUII produced a potent vasoconstrictor response in rat basilar arteries with intact endothelium, while isometric forces remained unaffected in arterial rings treated with lower rUII concentrations. Although L-NAME did not have a significant effect on 10 μM rUII-evoked contraction, it slightly increased arterial ring contraction elicited by 1 μM rUII. In depolarized arteries, dose-dependent rUII increased depolarization-induced contractions. This effect was suppressed by L-NAME. Our results show that the rat basilar artery has a vasoconstrictor response to rUII. The most potent vasoconstrictor effect was produced by lower doses of rUII (0.1 and 1 μM) in depolarized arteries with intact endothelium. This effect could facilitate arterial vasospasm in vascular pathophysiological processes such as subarachnoid hemorrhage and hypertension, when sustained depolarization and L-type Ca(2+) channel activation are present.

  15. Application of ultrafast gold luminescence to measuring the instrument response function for multispectral multiphoton fluorescence lifetime imaging

    NASA Astrophysics Data System (ADS)

    Talbot, Clifford B.; Patalay, Rakesh; Munro, Ian; Warren, Sean; Ratto, Fulvio; Matteini, Paolo; Pini, Roberto; Breunig, H. Georg; König, Karsten; Chu, Antony C.; Stamp, Gordon W.; Neil, Mark A. A.; French, Paul M. W.; Dunsby, Chris

    2011-07-01

    When performing multiphoton fluorescence lifetime imaging in multiple spectral emission channels, an instrument response function must be acquired in each channel if accurate measurements of complex fluorescence decays are to be performed. Although this can be achieved using the reference reconvolution technique, it is difficult to identify suitable fluorophores with a mono-exponential fluorescence decay across a broad emission spectrum. We present a solution to this problem by measuring the IRF using the ultrafast luminescence from gold nanorods. We show that ultrafast gold nanorod luminescence allows the IRF to be directly obtained in multiple spectral channels simultaneously across a wide spectral range. We validate this approach by presenting an analysis of multispectral autofluorescence FLIM data obtained from human skin ex vivo.

  16. Spin depolarization effect induced by charge state conversion of nitrogen vacancy center in diamond

    NASA Astrophysics Data System (ADS)

    Chen, Xiang-Dong; Zhou, Lei-Ming; Zou, Chang-Ling; Li, Cong-Cong; Dong, Yang; Sun, Fang-Wen; Guo, Guang-Can

    2015-09-01

    The electron spin of the negatively charged the nitrogen vacancy center (NV- ) in diamond can be optically polarized through intersystem crossing, which enables the defect to be used for quantum computation and metrology. In this work, we studied the electron spin depolarization effect of the NV center induced by charge state conversion, which was proven to be a spin-independent process. The spin-state initialization fidelity was largely affected by the charge state conversion process. As a result, the optical polarization of the electron spin decreased about 14 %(31 % ) with a high-power continuous-wave (pulsed) green laser. Moreover, the undefined fluorescence anomalous saturation effect of the NV center was analyzed and explained in detail based on the spin depolarization. The results demonstrated that a weak laser should be used for initialization of the NV center. In addition, the power and polarization of a laser for NV spin detection should be carefully adjusted to obtain the highest fluorescence signal. Our work also provided information that can increase the understanding of the charge state conversion and spin polarization processes of the NV center for quantum information and sensing.

  17. Nuclear Resonance Fluorescence Measurements on ^237Np for Security and Safeguards Applications

    NASA Astrophysics Data System (ADS)

    Angell, C. T.; Joshi, T.; Yee, Ryan; Norman, E. B.; Kulp, W. D.; Warren, G. A.; Korbly, S.; Klimenko, A.; Wilson, C.; Copping, R.; Shuh, D. K.

    2009-10-01

    The smuggling of nuclear material and the diversion of fissile material for covert weapon programs both present grave risks to world security. Methods are needed to detect nuclear material smuggled in cargo, and for proper material accountability in civilian fuel re-processing facilities. Nuclear resonance fluorescence (NRF) is a technique that can address both needs. It is a non-destructive active interrogation method that provides isotope-specific information. It works by using a γ-ray beam to resonantly excite levels in a nucleus and observing the γ-rays emitted whose energy and intensity are characteristic of that isotope. ^237Np presents significant safeguard challenges; it is fissile yet currently has fewer safeguard restrictions. NRF measurements on ^237Np will expand the nuclear database and will permit designing interrogation and assay systems. Measurements were made using the bremsstrahlung beam at the HVRL at MIT on a 7 g target of ^237Np with two incident electron energies of 2.8 and 3.1 MeV. Results will be presented with discussion of the relevant nuclear structure necessary to predict levels in other actinides.

  18. Computation of two-dimensional electric field from the ion laser induced fluorescence measurements

    SciTech Connect

    Spektor, Rostislav

    2010-09-15

    This paper presents a method of computing two-dimensional electric field from ion laser induced fluorescence (LIF) measurements in a plasma flow. The expression for the field is derived by taking velocity moments of the Boltzmann equation for ions. It was found that the pressure tensor, related to the width of the ion velocity distribution, plays a critical role in the computation of the electric field. Even with the assumption of cold ion flow, the pressure tensor contribution may be significant when velocity spread is caused by other forces. Such a situation occurs in the flow of a Hall thruster, where velocity spread is caused by the ions born at different potentials. LIF measurements of the cylindrical hall thruster plume were used to demonstrate practical application of the derived method. Whenever the pressure tensor components are small as compared to the mean ion drift velocity, the electric field calculations reduce to a simple expression given in terms of mean ion drift velocity and its divergence.

  19. Direct measurement of efflux in Pseudomonas aeruginosa using an environment-sensitive fluorescent dye.

    PubMed

    Iyer, Ramkumar; Erwin, Alice L

    2015-01-01

    Resistance-Nodulation-Division (RND) family pumps AcrB and MexB are the major efflux routes in Escherichia coli and Pseudomonas aeruginosa respectively. Fluorescent environment-sensitive dyes provide a means to study efflux pump function in live bacterial cells in real-time. Recently, we demonstrated the utility of this approach using the dye Nile Red to quantify AcrB-mediated efflux and measured the ability of antibiotics and other efflux pump substrates to compete with efflux of Nile Red, independent of antibacterial activity. Here, we extend this method to P. aeruginosa and describe a novel application that permits the comparison and rank-ordering of bacterial strains by their inherent efflux potential. We show that glucose and l-malate re-energize Nile Red efflux in P. aeruginosa, and we highlight differences in the glucose dependence and kinetics of efflux between P. aeruginosa and E. coli. We quantify the differences in efflux among a set of P. aeruginosa laboratory strains, which include PAO1, the hyper-sensitive strain ATCC 35151 and its parent, ATCC 12055. Efflux of Nile Red in P. aeruginosa is mediated by MexAB-OprM and is slower than in E. coli. In conclusion, we describe an efflux measurement tool for use in antibacterial drug discovery and basic research on P. aeruginosa efflux pumps.

  20. Flow Property Measurement Using Laser-Induced Fluorescence in the NASA Ames Interaction Heating Facility

    NASA Technical Reports Server (NTRS)

    Grinstead, Jay Henderson; Porter, Barry J.; Carballo, Julio Enrique

    2011-01-01

    The spectroscopic diagnostic technique of two photon absorption laser-induced fluorescence (TALIF) of atomic species has been applied to single-point measurements of velocity and static temperature in the NASA Ames Interaction Heating Facility (IHF) arc jet. Excitation spectra of atomic oxygen and nitrogen were recorded while scanning a tunable dye laser over the absorption feature. Thirty excitation spectra were acquired during 8 arc jet runs at two facility operating conditions; the number of scans per run varied between 2 and 6. Curve fits to the spectra were analyzed to recover their Doppler shifts and widths, from which the flow velocities and static temperatures, respectively, were determined. An increase in the number of independent flow property pairs from each as-measured scan was obtained by extracting multiple lower-resolution scans. The larger population sample size enabled the mean property values and their uncertainties for each run to be characterized with greater confidence. The average plus or minus 2 sigma uncertainties in the mean velocities and temperatures for all 8 runs were plus or minus 1.4% and plus or minus 11%, respectively.

  1. Fluorescence of prostate-specific antigen as measured with a portable 1D scanner

    NASA Astrophysics Data System (ADS)

    Kim, Byeong C.; Jeong, Jin H.; Jeong, Dong S.; Kim, Young M.; Oh, Sang W.; Choi, Eui Y.; Kim, Jae H.; Nahm, Kie B.

    2005-01-01

    Prostate-specific antigen (PSA) is an androgen-dependent glycoprotein protease (M.W. 33 kDa) and a member of kallikrein super-family of serine protease, and has chymotrypsin-like enzymatic activity. It is synthesized by the prostate epithelial cells and found in the prostate gland and seminal plasma as a major protein. It is widely used as a clinical marker for diagnosis, screening, monitoring and prognosis of prostate cancer. In normal male adults, the concentration of PSA in the blood is below 4 ng/ml and this value increases in patients with the prostate cancer or the benign prostatic hyperplasia (BPH) due to its leakage into the circulatory system. As such, systematic monitoring of the PSA level in the blood can provide critical information about the progress of the prostatic disease. We have developed a compact integral system that can quantitatively measure the concentration of total PSA in human blood. This system utilizes the fluorescence emitted from the dye molecules attached to PSA molecules after appropriate immunoassay-based processing. Developed for the purpose of providing an affordable means of fast point-of-care testing of the prostate cancer, this system proved to be able to detect the presence of the PSA at the level of 0.18 ng/ml in less than 12 minutes, with the actual measurement taking less than 2 minutes. The design concept for this system is presented together with the result for a few representative samples.

  2. A new total antioxidant potential measurements using RP-HPLC assay with fluorescence detection.

    PubMed

    Głód, Bronisław K; Piszcz, Paweł; Czajka, Katarzyna; Zarzycki, Paweł K

    2011-05-01

    In this paper, an improved total antioxidant potential (TAP) estimation using high-performance liquid chromatographic (HPLC) assay with fluorometric detection has been described. The principle of this method is based on the hydroxyl radicals generated in the Fenton-like reaction and subsequently detected using hydroxyterephthalic acid (HTPA), which is a reaction product of hydroxyl radicals and terephthalic acid (TPA), working as a sensing compound. HTPA quantity in the samples was measured by fluorescence detector working at excitation and emission wavelengths equal to 312 and 428 nm, respectively. A number of key experimental conditions including the influence of the reaction (incubation) time on the surface areas of HTPA peaks, concentration of Fe(II) ions as well as the influence of concentration of TPA on the surface area of the chromatographic peak of HTPA were optimized to the characteristic feature of TAP measurements. The elaborated assay has been used to evaluate TAP values of selected low-molecular mass compounds like pyrogallol, tryptamine, and n-alcohols (methanol, ethanol, and n-propanol) as well as chlorogenic and ascorbic acids and benzoic acid derivatives, which are commonly present in the food samples.

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

    PubMed Central

    Shlezinger, Neta; Eizner, Elad; Dubinchik, Stas; Minz-Dub, Anna; Tetroashvili, Rachel; Reider, Adi; Sharon, Amir

    2014-01-01

    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. PMID:28357220

  4. Unmixing of fluorescence spectra to resolve quantitative time-series measurements of gene expression in plate readers

    PubMed Central

    2014-01-01

    Background To connect gene expression with cellular physiology, we need to follow levels of proteins over time. Experiments typically use variants of Green Fluorescent Protein (GFP), and time-series measurements require specialist expertise if single cells are to be followed. Fluorescence plate readers, however, a standard in many laboratories, can in principle provide similar data, albeit at a mean, population level. Nevertheless, extracting the average fluorescence per cell is challenging because autofluorescence can be substantial. Results Here we propose a general method for correcting plate reader measurements of fluorescent proteins that uses spectral unmixing and determines both the fluorescence per cell and the errors on that fluorescence. Combined with strain collections, such as the GFP fusion collection for budding yeast, our methodology allows quantitative measurements of protein levels of up to hundreds of genes and therefore provides complementary data to high throughput studies of transcription. We illustrate the method by following the induction of the GAL genes in Saccharomyces cerevisiae for over 20 hours in different sugars and argue that the order of appearance of the Leloir enzymes may be to reduce build-up of the toxic intermediate galactose-1-phosphate. Further, we quantify protein levels of over 40 genes, again over 20 hours, after cells experience a change in carbon source (from glycerol to glucose). Conclusions Our methodology is sensitive, scalable, and should be applicable to other organisms. By allowing quantitative measurements on a per cell basis over tens of hours and over hundreds of genes, it should increase our understanding of the dynamic changes that drive cellular behaviour. PMID:24495318

  5. I Situ Laser Interferometry and Fluorescence Quenching Measurements of Poly(methyl Methacrylate) Thin Film Dissolution.

    NASA Astrophysics Data System (ADS)

    Wang, Fei

    The dissolution mechanisms of poly(methyl methacrylate) (PMMA) thin films in selected organic solvents was investigated. The dissolution was monitored using an in situ laser interferometry and fluorescence quenching (LIFQ) technique. Phenanthrene -labeled PMMA (Phe-PMMA) was used as a probe. Solutions of PMMA in toluene were spin-coated onto sapphire substrate to form films approximately 1 μm thick. The LIFQ results show that for PMMA film dissolution the transition layer thickness increases until the dissolution reaches its steady state. Then this final transition layer thickness (FTL) does not change until solvent vanguard molecules reach the surface of the substrate. Thermal history effects on PMMA film dissolution were examined. The dissolution rate decreases with increasing baking temperature and reaches a constant value for annealing at 150^circC. The results show that the thermal history has negligible effect on the factor of reduction f obtained from interferometry measurements. Fluorescence quenching measurements, by contrast, suggest that transition layer thickness decreases with increasing baking temperature. This suggests that the fluorescence quenching part of the LIFQ experiment is sensitive to the Fickian precursor portion of the solvent concentration profile in the film. The dissolution of PMMA films in acetone, 2-butanone, and 2-pentanone was studied. The results show that the dissolution rate decreases significantly with increasing solvent molecular size. Significant differences are found for FTL values calculated from LIFQ experiments and those calculated from f obtained by laser interferometry. Values of f are essentially identical in three solvents used. The effect of non-solvent on PMMA dissolution was studied by using 2-propanol and 2-butanone mixtures as solvents. The dissolution rate decreases with increasing non-solvent content. This indicates a strong thermodynamic effect, especially at high concentration of non-solvent. Molecular weight

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

    PubMed

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

    2014-09-30

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

  7. Measurement of intracellular pH in pancreatic duct cells: a new method for calibrating the fluorescence data.

    PubMed

    Hegyi, Péter; Rakonczay, Zoltán; Gray, Mike A; Argent, Barry E

    2004-05-01

    Pancreatic duct cells secrete the bicarbonate ions found in pancreatic juice. Impairment of ductal bicarbonate secretion, as occurs in cystic fibrosis, has serious consequences for pancreatic function and for the structural integrity of the gland. As bicarbonate is a buffer ion, the accurate measurement of intracellular pH (pHi) in duct cells is an important technique for studying the mechanisms of bicarbonate transport. Commonly, pHi is measured using the fluorescent dye biscarboxyethylcarboxyfluorescein (BCECF). The purpose of this study was to develop a new technique for the accurate calibration of BCECF fluorescent signals. Our results indicate that BCECF fluorescence is not only dependent on pHi but also on the total fluorescence intensity of the detected area (which may be influenced by dye loading, dye leakage, and the shutter size on the photomultiplier). The outcome is that one calibration curve is not sufficient for accurate determination of pHi. In fact, an appropriate calibration curve must be selected for each individual experiment. Moreover, the calibration plot is only linear over a narrow range of pHi values. In conclusion, we have developed a new technique that should be applicable to all cell types for the accurate calibration of fluorescent signals from the pH-sensitive dye BCECF.

  8. Pulsing Depolarization: New Method of In-Vivo Blood Glucometry

    NASA Astrophysics Data System (ADS)

    Romanov, Dmitri; Shvartsman, Leonid; Fine, Ilya

    2002-03-01

    We propose a new system of non-invasive measurements enabling one to monitor in-vivo the glucose content of the blood. The proposed method is based on the transmission oximetry technique [1] and on the model of red blood cell aggregation [2]. We utilize the known birefregence of glucose solutions (in this case, the blood plasma) and the absence of this optical activity on the part of the red cells. Thus, we model the blood as a birefrigent host medium filled with optically non-active inclusions of spheroidal shape, arbitrarily oriented and having one semiaxis growing with time during the cell aggregation. We trace the evolution of initially linearly polarized laser beam in this system, and calculate the ellipticity and the azimuthal angle of the resulting partially depolarized light. The azimuthal angle is obtained as time-independent quantity being a function of the hematocrit, glucose content and thickness of the host medium layer, while the ellipticity depends also on the red cell aggregation, and thus oscillates with the heartbeats. Measurements of both the azimuthal angle and the ellipticity provide us with sufficient data to determine the glucose content of the blood. [1] I. Fine, A. Weinreb, Med. and Biol. Eng. and Comput., 31, 516 (1993). [2] L.D. Shvartsman, I. Fine, SPIE Proc., 4162, 120 (2000).