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Sample records for fluorescence depolarization measurements

  1. Fluorescence depolarization measurements on oriented membranes.

    PubMed Central

    Adler, M; Tritton, T R

    1988-01-01

    We describe the theory and experimental application of fluorescence depolarization measurements on small molecules bound to oriented phospholipid bilayers. The results yield insight into both the orientation and the rotational motion of fluorophores in a membrane environment. To accomplish this the angular distribution of polarized fluorescence intensities is measured on a membrane preparation consisting of stacked phospholipid bilayers oriented in a known coordinate system. Considerably more information is available from this data than in comparable solution phase measurements. Three parameters are derived from the data: the rate of rotational diffusion and the second and fourth degree order parameters. These latter two parameters provide an assessment of the average distribution of fluorophore orientation in the membrane bilayer. The data have been carefully examined for systematic experimental artifacts and new protocols are presented which help to eliminate errors that have not been amply treated in the past. We present data for two types of fluorescent molecules: (a) conventional membrane probes like diphenylhexatriene, perylene and anthroyloxy fatty acids; and (b) the anticancer agent adriamycin and several congeneric anthracycline antibiotics. The results show that the hydrocarbon core of membranes is more rigid than previously thought, particularly above the thermal phase transition temperature. We also show that the orientation of small molecules is sensitive to both the phospholipid composition and to the interaction of specific functional groups with the lipid bilayer. The results are discussed in terms of energetic models describing the general patterns for the binding of small molecules to biological membranes. Images FIGURE 1 PMID:3165033

  2. Fluorescence depolarization in the presence of excitation energy migration in partly ordered polymer films

    NASA Astrophysics Data System (ADS)

    Synak, A.; Gondek, G.; Bojarski, P.; Kułak, L.; Kubicki, A.; Szabelski, M.; Kwiek, P.

    2004-11-01

    Excitation energy migration is studied for 3,3'-diethylthiacyanine iodide in uniaxially stretched and unstretched poly(vinyl alcohol) films. Time-resolved and steady-state measurements of fluorescence depolarization yield quite different results for disordered and partly ordered systems. Contrary to disordered systems, for which strong concentration depolarization was observed, partly ordered systems exhibit much weaker depolarization as a result of preferential angular distribution of transition dipole moments of identical fluorophores participating in energy migration. Difference of the same origin was found for time-resolved emission anisotropy measurements. These results are in agreement with those obtained from the Monte-Carlo simulation.

  3. Analyzing fluorophore electronic structure and depolarization by fluorescence polarizing angle spectrum

    SciTech Connect

    Mu, Taotao; Chen, Siying Zhang, Yinchao; Chen, He; Guo, Pan

    2014-07-21

    In this Letter, a method, based on stokes parameters, is developed to observe the angular displacement between the excitation and emission moments. Experiments demonstrate that when combined with degree of polarization spectrums, we can acquire the depolarization caused by angular displacement or energy migration. The method presented in this Letter can be easily realized with the existing fluorescence measuring system and may potentially make it convenient to study the fluorophore electronic structure or the mechanism of fluorescence anisotropy.

  4. EPR and fluorescence depolarization studies on bovine cardiac myosin.

    PubMed

    Stone, D B; Mendelson, R A; Botts, J; Cheung, P H

    1981-09-01

    To test for possible differences in local conformation and S1 flexibility, bovine cardiac and rabbit skeletal myosins were labeled with a fluorophore (1,5-IAEDANS) and a spin label having iodoacetamide reactivity. The marked activation of the Ca2+-ATPase (6- to 8-fold) and inhibition of the K+ (EDTA)-ATPase (80-90%) by both labels indicated specific labeling of the fast-reacting thiols (SH1) of both myosins. Fluorescence depolarization studies of 1,5-IAEDANS-labeled cardiac myosin indicated that, like skeletal myosin, the SI moieties of cardiac myosin exhibit considerable segmental flexibility with respect to the rod portion of the molecule. This indicates that segmental flexibility may be a property of all myosins. Cardiac and skeletal myosins immobilized spin labels to approximately the same extent, indicating a similarity in steric restraints around the SH1 thiol of the two myosins. The magnitude of the changes in spin label mobility accompanying binding of MgADP and hydrolysis of MgATP was reduced in cardiac myosin relative to skeletal myosin. This suggests that the lower catalytic center activity of cardiac myosin is associated with more restricted conformational changes accompanying formation of M.ADP and M.ADP.Pi. From measurements of spin label mobility, the affinity of cardiac and skeletal myosin for ADP were similar: Kd (ADP) = 7 microM, n = 1.6. The EPR spectrum of spin labels attached to cardiac and skeletal myosin showed similar saturation effects upon actin binding indicating immobilization of myosin heads occurs with both proteins.

  5. Reorientational motion of a cross-link junction in a poly(dimethylsiloxane) network measured by time-resolved fluorescence depolarization

    SciTech Connect

    Stein, A.D. ); Hoffman, D.A. ); Frank, C.W. ); Fayer, M.D. )

    1992-02-15

    The reorientational dynamics of a cross-link junction in poly(dimethylsiloxane) networks, measured by the fluorescence anisotropy decay of a chromophore tagged to the cross-link, have been investigated over a range of temperatures from {ital T}{sub {ital g}}+75 to {ital T}{sub {ital g}}+150. The probe chromophore, 1-dimethylamino-5-sulfonylnaphthalene amide (dansyl amide), is pendant to a trifunctional silane that acts as a cross-linking molecule. In cyclohexanol, the fluorescence anisotropy decay is in agreement with Debye--Stokes--Einstein hydrodynamic theory (rotational diffusion) demonstrating that the cross-linker can be used as a probe of orientational relaxation. The fluorescence anisotropy decays at a rapid rate in an end-linked poly(dimethyl siloxane) network reflecting fast reorientational motion of the cross-link junction. This reorientation appears diffusive and has a temperature dependence in accord with the Williams--Landel--Ferry equation. A model is proposed that suggests that reorientation and translational motion of the cross-link occur simultaneously and are both coupled to fluctuations of the polymer chain ends.

  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. The depolarization - attenuated backscatter relation: CALIPSO lidar measurements vs. theory.

    PubMed

    Hu, Yongxiang; Vaughan, Mark; Liu, Zhaoyan; Lin, Bing; Yang, Ping; Flittner, David; Hunt, Bill; Kuehn, Ralph; Huang, Jiangping; Wu, Dong; Rodier, Sharon; Powell, Kathy; Trepte, Charles; Winker, David

    2007-04-30

    Using measurements obtained by the Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observations (CALIPSO) satellite, relationships between layer-integrated depolarization ratio (delta) and layer-integrated attenuated backscatter (gamma) are established for moderately thick clouds of both ice and water. A new and simple form of the delta-gamma relation for spherical particles, developed from Monte Carlo simulations and suitable for both water clouds and spherical aerosol particles, is found to agree well with the observations. A high-backscatter, low-depolarization delta-gamma relationship observed for some ice clouds is shown to result primarily from horizontally oriented plates and implies a preferential lidar ratio - depolarization ratio relation in nature for ice cloud particles containing plates.

  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. Far wing depolarization of light - Generalized absorption profiles. [in laser fluorescence spectroscopy of Sr vapor

    NASA Technical Reports Server (NTRS)

    Thomann, P.; Burnett, K.; Cooper, J.

    1981-01-01

    An absorption (and/or emission) event which takes place during a strong collision is called a 'correlated event'. It is discussed how correlated events affect the far red wing depolarization of fluorescence. Attention is given to an atomic vapor which is irradiated by linearly polarized light of a frequency on the red side of the resonance line. Two limiting cases are considered, corresponding to excitation in the impact region and in the quasi-static wing. In the quasi-static wing, absorption of a photon followed by fluorescence (rather than Rayleigh scattering), occurs mostly during a collision. Correlated events dominate the scattering process. Expressions derived for the polarization of the fluorescent light are applied to far red wing depolarization. It is found that the polarization of the fluorescent light does not go to zero in the far wing, but depends crucially on the detailed nature of the anisotropy in the long-range part of the interatomic potential.

  10. 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.

  11. Fluorescence depolarization of rhodamine 6G in glycerol: a photon-counting test of three-dimensional excitation transport theory

    SciTech Connect

    Anfinrud, P.A.; Hart, D.E.; Hedstrom, J.F.; Struve, W.S.

    1986-05-22

    Time-correlated photon counting has been used to measure fluorescence concentration depolarization for rhodamine 6G in glycerol. The excitation transport theory developed by Gochanour, Andersen, and Fayer yields good approximations to the experimental decay profiles over the concentration range 1.7 x 10/sup -4/ to 2.4 x 10/sup -3/ M. Although the differences between optimized theoretical and experimental profiles are fractionally small, they are readily characterized under present counting statistics. They prove to be dominated by experimental artifacts, arising from excitation trapping by rhodamine 6G aggregates and from self-absorption in solution cells thicker than approx. 10 ..mu..m.

  12. Experimental verification of depolarization effects in bioelectrical impedance measurement.

    PubMed

    Chen, Xiaoyan; Lv, Xinqiang; Du, Meng

    2014-01-01

    The electrode polarization effects on bioelectrical impedance measurement at low-frequency cannot be ignored. In this paper, the bioelectrical data of mice livers are measured to specify the polarization effects on the bio-impedance measurement data. We firstly introduce the measurement system and methodology. Using the depolarization method, the corrected results are obtained. Besides, the specific effects of electrode polarization on bio-impedance measurement results are investigated using comparative analysis of the previous and posterior correction results from dielectric spectroscopy, Cole-Cole plot, conductivity and spectroscopy of dissipation tangent. Experimental results show that electrode polarization has a significant influence on the characteristic parameters of mouse liver tissues. To be specific, we see a low-frequency limit resistance R0 increase by 19.29%, a reactance peak XP increase by 8.50%, a low-frequency limit conductivity Kl decrease by 17.65% and a dissipation peak tangent decrease by 160%.

  13. Determining the orientation of transition moments and depolarization by fluorescence polarizing angle spectrum.

    PubMed

    Mu, Taotao; Chen, Siying; Zhang, Yinchao; Guo, Pan; Chen, He

    2015-05-01

    In this paper, fluorescence polarizing angle spectrum, combined with degree of polarization(DOP), is proposed to determine the spatial orientation of transition dipole moments (TDMs) and depolarization of chlorophyll in solution. It is found that, due to the oriented TDMs under polarized laser excitation, the projections of angle of polarization(AOP) and DOP on the three orthogonal planes are different from each other. Experiments demonstrate that we can acquire the spatial orientation by detecting the projections of AOP on two orthogonal planes (xOz and yOz). Meanwhile, The depolarization can also be determined by the DOP spectrums. The validity of this method has been verified by another projection on the xOy plane.

  14. Polar stratospheric cloud measurements by means of depolarization lidar in the Antarctic

    NASA Astrophysics Data System (ADS)

    Stefanutti, L.

    1991-02-01

    POLE (Polar Ozone Lidar Experiment) is a cooperative project between the French and Italian services. It was started with the implementation of a first depolarization backscattering lidar for measurements both of background stratospheric aerosols and Polar Stratospheric Clouds (PSCs). A complex Ozone lidar was also installed which will replace the backscattering system and extend its measurements to tropospheric and stratospheric Ozone and to stratospheric and mesospheric temperatures. This new system allows also the measurement of the backscattered and depolarized signal produced by PSCs and background aerosols. The depolarization technique seemed to be quite efficient in the detection of different types of PSCs.

  15. Polar stratospheric cloud measurements by means of depolarization lidar in the Antarctic

    NASA Technical Reports Server (NTRS)

    Stefanutti, L.

    1991-01-01

    POLE (Polar Ozone Lidar Experiment) is a cooperative project between the French and Italian services. It was started with the implementation of a first depolarization backscattering lidar for measurements both of background stratospheric aerosols and Polar Stratospheric Clouds (PSCs). A complex Ozone lidar was also installed which will replace the backscattering system and extend its measurements to tropospheric and stratospheric Ozone and to stratospheric and mesospheric temperatures. This new system allows also the measurement of the backscattered and depolarized signal produced by PSCs and background aerosols. The depolarization technique seemed to be quite efficient in the detection of different types of PSCs.

  16. Independent flexible motion of submolecular domains of the Ca2+,Mg2+-ATPase of sarcoplasmic reticulum measured by time-resolved fluorescence depolarization of site-specifically attached probes.

    PubMed

    Suzuki, S; Kawato, S; Kouyama, T; Kinosita, K; Ikegami, A; Kawakita, M

    1989-09-19

    The Ca2+-transporting ATPase of rabbit skeletal muscle sarcoplasmic reticulum was site-specifically labeled with either N-(1-anilinonaphth-4-yl)maleimide (ANM) or 5-[[(iodoacetamido)-ethyl]amino]naphthalene-1-sulfonate (IAEDANS), and the segmental motion of submolecular domains of the ATPase molecule was examined by means of time-resolved and steady-state fluorescence anisotropy measurements. The ANM-binding domain showed wobbling with a rotational relaxation time phi = 69 ns in the absence of free Ca2+ without any independent wobbling of the ANM moiety. The IAEDANS-binding domain showed a significantly slower wobbling with phi = 190 ns in the absence of Ca2+. The present results demonstrated for the first time that the ATPase molecule is composed of distinct domains whose mobilities are considerably different from each other. The binding of Ca2+ to the transport site increased the segmental motion of ANM-labeled domain, leading to a phi value of 65 ns. Solubilization of the ANM-labeled SR membranes by deoxycholate led to a further increase in the segmental flexibility (phi = 48 ns in the absence of free Ca2+), indicating that the mobility of the ANM-binding domain was considerably restricted through interaction with the membrane. The mobility of the ANM-binding domain of solubilized ATPase was also increased to some extent upon binding of Ca2+.

  17. 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.

  18. Excimer fluorescence compared to depolarization in the flow cytometric characterization of lateral membrane mobility in platelets

    NASA Astrophysics Data System (ADS)

    Rothe, Gregor; Schaefer, Buerk; Wimmer, Martin S.; Schmitz, Gerd

    1998-04-01

    An altered cellular membrane fluidity secondary to changes of cholesterol metabolism is a potentially important mechanism in the pathogenesis of atherosclerosis. Especially in blood platelets an increased sensitivity for stimulation dependent aggregation which is a risk factor for thrombosis has been experimentally linked to disorders of lipid and lipoprotein metabolism. The goal of this study was the development of a flow cytometric assay for the direct analysis of cellular membrane microviscosity in correlation to activation associated phenotypic changes of platelets in vitro. The analysis of fluorescence polarization following the staining of hydrophobic lipid regions of cell membranes with the fluorescent dye 1,6-diphenyl-1,3,5-hexatriene (DPH) is a well established method for the analysis of membrane fluidity. The extent of fluorescence anisotropy dependent on the rotational mobility of this fluorochrome is indirectly proportional to the microviscosity of the stained membrane subcompartment. In this study, an alternative and more simple method based on the diffusion dependent excimer formation of pyrenedecanoic acid (PDA) (J. Immunol. Methods 96:225-31, 1987) was characterized in comparison to the DPH method as a reference. Human platelets showed a rapid uptake of both DPH and PDA resulting in the staining primarily of the plasma membrane after up to 30 min of incubation. Staining analyzed at 351 nm excitation resulted in a saturation of the depolarization coefficient of DPH at 20 (mu) M but an increase of the excimer to monomer ratio of PDA with increasing dye concentration. A 'membrane fluidity coefficient' which saturated at 5 (mu) M PDA was calculated as the excimer fluorescence divided through the square of monomer fluorescence thereby correcting for the influence of dye concentration on excimer formation. The temperature dependent changes of membrane viscosity were further used as a model for the comparison of both methods. Cells analyzed at temperatures

  19. 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.

  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. Practical depolarization-ratio-based inversion procedure: lidar measurements of the Eyjafjallajökull ash cloud over the Netherlands.

    PubMed

    Donovan, David Patrick; Apituley, Arnoud

    2013-04-10

    In this paper we present a technique for estimating optical backscatter and extinction profiles using lidar, which exploits the difference between the observed linear volume depolarization ratio at 355 nm and the corresponding expected aerosol-only depolarization ratio. The technique is specific to situations where a single strongly depolarizing species is present and the associated linear particulate depolarization ratio may be presumed to be known to within a reasonable degree of accuracy (on the order of 10%). The basic principle of the technique is extended to deal with situations where a depolarizing fraction is mixed with nondepolarizing aerosol. In general, since the relative depolarization interchannel calibration is much more stable than the absolute system calibration, the depolarization-based technique is easier to implement than conventional techniques that require a profile-by-profile calibration or, equivalently, an identification of aerosol-free altitude intervals. This in particular allows for unattended data analysis and makes the technique well-suited to be part of a broader (volcanic ash) surveillance system. The technique is demonstrated by applying it to the analysis of aerosol layers resulting from the 2010 eruptions of the Eyjafjallajökull volcano in Iceland. The measurements were made at the Cabauw remote-sensing site in the central Netherlands. By comparing the results of the depolarization-based inversion with a more conventional manual inversion procedure as well as Raman lidar results, it is demonstrated that the technique can be successfully applied to the particular case of 355 nm depolarization lidar volcanic ash soundings, including cases in which the ash is mixed with nondepolarizing aerosol. PMID:23670771

  2. Depolarization Ratio of Clouds Measured by Multiple-Field of view Multiple Scattering Polarization Lidar

    NASA Astrophysics Data System (ADS)

    Okamoto, Hajime; Sato, Kaori; Makino, Toshiyuki; Nishizawa, Tomoaki; Sugimoto, Nobuo; Jin, Yoshitaka; Shimizu, Atsushi

    2016-06-01

    We have developed the Multiple Field of view Multiple Scattering Polarization Lidar (MFMSPL) system for the study of optically thick low-level clouds. It has 8 telescopes; 4 telescopes for parallel channels and another 4 for perpendicular channels. The MFMSPL is the first lidar system that can measure depolarization ratio for optically thick clouds where multiple scattering is dominant. Field of view of each channel was 10mrad and was mounted with different angles ranging from 0 mrad (vertical) to 30mrad. And footprint size from the total FOV was achieved to be close to that of Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observation (CALIPSO) lidar at the altitude of 1km in order to reproduce similar degree of multiple scattering effects as observed from space. The MFMSPL has started observations since June 2014 and has been continuously operated at National Institute for Environmental Studies (NIES) in Tsukuba, Japan. Observations proved expected performance such that measured depolarization ratio was comparable to the one observed by CALIPSO lidar.

  3. Measurement of the Linear Depolarization Ratio of Aged Dust at Three Wavelengths (355, 532 and 1064 nm) Simultaneously over Barbados

    NASA Astrophysics Data System (ADS)

    Haarig, Moritz; Althausen, Dietrich; Ansmann, Albert; Klepel, André; Baars, Holger; Engelmann, Ronny; Groß, Silke; Freudenthaler, Volker

    2016-06-01

    A ground-based polarization Raman lidar is presented, that is able to measure the depolarization ratio at three wavelengths (355, 532 and 1064 nm) simultaneously. This new feature is implemented for the first time in a Raman lidar. It provides a full dataset of 3 backscatter coefficients, two extinction coefficients and 3 depolarization ratios (3+2+3 lidar system). To ensure the data quality, it has been compared to the well characterized two-wavelength polarization lidar POLIS. Measurements of long-range transported dust have been performed in the framework of the Saharan Aerosol Long-Range Transport and Aerosol-Cloud-Interaction Experiment (SALTRACE) in the Caribbean.

  4. Modification of DNA dynamics by platinum drug binding: a time-dependent fluorescence depolarization study of the interaction of cis- and trans-diamminedichloroplatinum(II) with DNA.

    PubMed

    Millar, D P; Ho, K M; Aroney, M J

    1988-11-15

    The interaction of calf thymus DNA with the antitumor drug cis-diamminedichloroplatinum(II), and with the clinically ineffective trans isomer, is studied by time-dependent fluorescence depolarization spectroscopy of intercalated ethidium. The effect of the platinum compounds on the rapid torsional motions of DNA in solution is observed via depolarization of the ethidium fluorescence. The depolarization data are successfully analyzed with an elastic model of DNA dynamics and yield a value for the product of the torsional rigidity of the DNA and the friction factor for DNA twisting. The dependence of this quantity on the degree of platination of the DNA is determined for each isomer. At low levels of platination, the cis isomer increases the solute-solvent friction acting on the DNA torsional motions, which we attribute to local kinking of the helix axis at the sites of platination. At high levels of platination, the cis isomer decreases the torsional rigidity of the DNA, indicating that disruption of DNA duplex structure occurs under these conditions. The binding of the trans isomer to DNA has no effect on the torsional rigidity or the friction. The present results are compared with other findings on the interaction of these platinum compounds with DNA.

  5. Benefit of depolarization ratio at λ = 1064 nm for the retrieval of the aerosol microphysics from lidar measurements

    NASA Astrophysics Data System (ADS)

    Gasteiger, J.; Freudenthaler, V.

    2014-11-01

    A better quantification of aerosol properties is required for improving the modelling of aerosol effects on weather and climate. This task is methodologically demanding due to the diversity of the microphysical properties of aerosols and the complex relation between their microphysical and optical properties. Advanced lidar systems provide spatially and temporally resolved information on the aerosol optical properties that is sufficient for the retrieval of important aerosol microphysical properties. Recently, the mass concentration of transported volcanic ash, which is relevant for the flight safety of aeroplanes, was retrieved from measurements of such lidar systems in southern Germany. The relative uncertainty of the retrieved mass concentration was on the order of ±50%. The present study investigates improvements of the retrieval accuracy when the capability of measuring the linear depolarization ratio at 1064 nm is added to the lidar setup. The lidar setups under investigation are based on those of MULIS and POLIS of the Ludwig-Maximilians-Universität in Munich (Germany) which measure the linear depolarization ratio at 355 and 532 nm with high accuracy. The improvements are determined by comparing uncertainties from retrievals applied to simulated measurements of this lidar setup with uncertainties obtained when the depolarization at 1064 nm is added to this setup. The simulated measurements are based on real lidar measurements of transported Eyjafjallajökull volcano ash. It is found that additional 1064 nm depolarization measurements significantly reduce the uncertainty of the retrieved mass concentration and effective particle size. This significant improvement in accuracy is the result of the increased sensitivity of the lidar setup to larger particles. The size dependence of the depolarization does not vary strongly with refractive index, thus we expect similar benefits for the retrieval in case of measurements of other volcanic ash compositions and

  6. Polar winter cloud depolarization measurements with the CANDAC Rayleigh-Mie-Raman Lidar

    NASA Astrophysics Data System (ADS)

    McCullough, E. M.; Nott, G. J.; Duck, T. J.; Sica, R. J.; Doyle, J. G.; Pike-thackray, C.; Drummond, J. R.

    2011-12-01

    Clouds introduce a significant positive forcing to the Arctic radiation budget and this is strongest during the polar winter when shortwave radiation is absent (Intrieri et al., 2002). The amount of forcing depends on the occurrence probability and optical depth of the clouds as well as the cloud particle phase (Ebert and Curry 1992). Mixed-phase clouds are particularly complex as they involve interactions between three phases of water (vapour, liquid and ice) coexisting in the same cloud. Although significant progress has been made in characterizing wintertime Arctic clouds (de Boer et al., 2009 and 2011), there is considerable variability in the relative abundance of particles of each phase, in the morphology of solid particles, and in precipitation rates depending on the meteorology at the time. The Canadian Network for the Detection of Atmospheric Change (CANDAC) Rayleigh-Mie-Raman Lidar (CRL) was installed in the Canadian High Arctic at Eureka, Nunavut (80°N, 86°W) in 2008-2009. The remotely-operated system began with measurement capabilities for multi-wavelength aerosol extinction, water vapour mixing ratio, and tropospheric temperature profiles, as well as backscatter cross section coefficient and colour ratio. In 2010, a new depolarization channel was added. The capability to measure the polarization state of the return signal allows the characterization of the cloud in terms of liquid and ice water content, enabling the lidar to probe all three phases of water in these clouds. Lidar depolarization results from 2010 and 2011 winter clouds at Eureka will be presented, with a focus on differences in downwelling radiation between mixed phase clouds and ice clouds. de Boer, G., E.W. Eloranta, and M.D. Shupe (2009), Arctic mixed-phase stratiform cloud properties from multiple years of surface-based measurements at two high-latitude locations, Journal of Atmospheric Sciences, 66 (9), 2874-2887. de Boer, G., H. Morrison, M. D. Shupe, and R. Hildner (2011

  7. 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…

  8. 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.

  9. 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. PMID:19844319

  10. 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

  11. 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

  12. [Fluorescent probes for intracellular Mg2+ measurement].

    PubMed

    Komatsu, Hirokazu; Suzuki, Yoshio; Suzuki, Koji

    2004-08-01

    Recently, fluorescent probes are widely used as tools for dynamical measurement of ion distributions and concentrations in cells. They are highly sensitive, and offer imaging by the use of fluorescent microscopy in easily and less cell damaging way. This paper discusses the selectivity and optical character of the three novel Mg(2+) fluorescent probes. KMG-20AM offers ratiometric quantative measurement of Mg(2+), KMG-104 provides high-sensitive qualitative analysis and 3-D measurement. With those improved Mg(2+) fluorescent probes, the physiological and pathological role of Mg(2+) are going to be more and more clear. PMID:15577092

  13. Depolarization by high-aperture focusing

    NASA Astrophysics Data System (ADS)

    Bahlmann, Karsten; Hell, Stefan W.

    2002-05-01

    We propose and demonstrate a method employing ferroelectric monomolecular layers, by which it is possible to precisely measure the planar light field polarization in the focus of a lens. This method allowed us to establish for the first time to our knowledge, the perpendicularly oriented field that is anticipated at high apertures. For a numerical aperture 1.4 oil immersion lens illuminated with linearly polarized plane waves, the integral of the modulus square of the perpendicular component amounts to (1.51r0.2) % of that of the initial polarization. It is experimentally proven that depolarization decreases with decreasing aperture angle and increases when using annular apertures. Annuli formed by a central obstruction with a diameter of 89 % of that of the entrance pupil raise the integral to 5.5 %. This compares well with the value of 5.8% predicted by electromagnetic focusing theory; however, the depolarization is also due to imperfections connected with focusing by refraction. Besides fluorescence microscopy and single molecule spectroscopy, the measured intensity of the depolarized component in the focal plane is relevant to all forms of light spectroscopy combining strong focusing with polarization analysis.

  14. Depolarized FRET (depolFRET) on the cell surface: FRET control by photoselection.

    PubMed

    Bene, László; Gogolák, Péter; Ungvári, Tamás; Bagdány, Miklós; Nagy, István; Damjanovich, László

    2016-02-01

    Sensitivity of FRET in hetero- and homo-FRET systems on the photoselected orientation distribution of donors has been proven by using polarized and depolarized light for excitation. FRET as well as donor and acceptor anisotropies have been simultaneously measured in a dual emission-polarization scheme realized in a conventional flow cytometer by using single laser excitation and applying fluorophore-conjugated mAbs against the MHCI and MHCII cell surface receptors. Depolarization of the originally polarized light have been achieved by using crystal depolarizers based on Cornu's principle, a quarter-wave plate for circular polarization, and a parallel beam splitter acting as a diagonal-polarizer for dual-polarization excitation. Simultaneous analysis of intensity-based FRET efficiency and acceptor depolarization equivocally report that depolarization of light may increase FRET in an amount depending on the acceptor-to-donor concentration ratio. Acceptor depolarization turned to be more sensitive to FRET than donor hyper-polarization and even than intensity-based FRET efficiency. It can be used as a sensitive tool for monitoring changes in the dynamics of the donor-acceptor pairs. The basic observations of FRET enhancement and increased acceptor depolarization obtained for hetero-FRET are paralleled by analog observations of homo-FRET enhancements under depolarized excitation. In terms of the orientation factor for FRET, the FRET enhancements on depolarization in the condition of the macroscopically isotropic orientation distributions such as those of the cell surface bound fluorophores report on the presence of local orientation mismatches of the donor and acceptor preventing the optimal FRET in the polarized case, which may be eliminated by the excitation depolarization. A theory of fluorescence anisotropy for depolarized excitation is also presented. PMID:26657258

  15. Remote measurement of ocean temperature from depolarization in Raman scattering. [using laser beam

    NASA Technical Reports Server (NTRS)

    Chang, C. H.; Young, L. A.

    1975-01-01

    Ocean temperatures may be mapped in three dimensions from an aircraft down to depths of 2 to 4 attenuation lengths by monitoring Raman radiation backscattered from a laser beam. This paper describes laboratory experiments on the temperature dependence of Raman spectra of saline solutions and calculations of the expected performance of a field system. The Raman technique is of greatest potential utility in coastal and estuarine waters where gradients are relatively high. It is estimated that seawater temperature may be measured to a statistical precision of 0.5 C at depths to four attenuation lengths in two meter water, using one Joule of transmitted laser energy.

  16. 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.

  17. Quartz crystal microbalance measurements of mitochondrial depolarization predicting chemically induced toxicity of vascular cells and macrophages.

    PubMed

    Dewilde, Abiche H; Wang, Gang; Zhang, Jianping; Marx, Kenneth A; Therrien, Joel M; Braunhut, Susan J

    2013-08-01

    Quartz crystal microbalances (QCMs) measure mass on the nanogram (ng) scale. We built novel QCMs as toxicity biosensors incorporating living cells. Human endothelial cells or canine macrophages were equilibrated on QCM crystal surfaces until stable oscillation frequencies occurred. Vehicle or sodium azide (NaN3) (25-100 mM) was added to these QCMs while continuously collecting crystal oscillation frequency data. At these doses, NaN3 alters mitochondrial membrane permeability and causes mitochondrial swelling and intrinsic apoptosis. Our studies demonstrated no frequency change in QCMs with untreated cells or without cells but NaN3. If NaN3 was added to either cell type within QCMs, 5 to 8 min later increases in oscillation frequency (Δf) occurred (400-1600 Hz) that correlated with dose. All frequency changes reverted to baseline by 15 min. In parallel, during the first 30 min, no change in cell or nuclear areas, or in actin or microtubule distributions, was detected. Yet, mitochondrial size and membrane permeability increased significantly during, but not after, 5 to 8 min. Viability studies confirmed dose-dependent toxicity that was predicted and proportionate to the 5- to 8-min Δf. These studies confirm that cell-based QCMs can detect early events in intrinsic apoptosis and reveal unique kinetic information about events occurring within subcellular structures in response to toxins.

  18. Fluorescence lifetime measurements in flow cytometry

    NASA Astrophysics Data System (ADS)

    Beisker, Wolfgang; Klocke, Axel

    1997-05-01

    Fluorescence lifetime measurements provide insights int eh dynamic and structural properties of dyes and their micro- environment. The implementation of fluorescence lifetime measurements in flow cytometric systems allows to monitor large cell and particle populations with high statistical significance. In our system, a modulated laser beam is used for excitation and the phase shift of the fluorescence signal recorded with a fast computer controlled digital oscilloscope is processed digitally to determine the phase shift with respect to a reference beam by fast fourier transform. Total fluorescence intensity as well as other parameters can be determined simultaneously from the same fluorescence signal. We use the epi-illumination design to allow the use of high numerical apertures to collect as much light as possible to ensure detection of even weak fluorescence. Data storage and processing is done comparable to slit-scan flow cytometric data using data analysis system. The results are stored, displayed, combined with other parameters and analyzed as normal listmode data. In our report we discuss carefully the signal to noise ratio for analog and digital processed lifetime signals to evaluate the theoretical minimum fluorescence intensity for lifetime measurements. Applications to be presented include DNA staining, parameters of cell functions as well as different applications in non-mammalian cells such as algae.

  19. Fluorescence lifetime measurements in heterogeneous scattering medium

    NASA Astrophysics Data System (ADS)

    Nishimura, Goro; Awasthi, Kamlesh; Furukawa, Daisuke

    2016-07-01

    Fluorescence lifetime in heterogeneous multiple light scattering systems is analyzed by an algorithm without solving the diffusion or radiative transfer equations. The algorithm assumes that the optical properties of medium are constant in the excitation and emission wavelength regions. If the assumption is correct and the fluorophore is a single species, the fluorescence lifetime can be determined by a set of measurements of temporal point-spread function of the excitation light and fluorescence at two different concentrations of the fluorophore. This method is not dependent on the heterogeneity of the optical properties of the medium as well as the geometry of the excitation-detection on an arbitrary shape of the sample. The algorithm was validated by an indocyanine green fluorescence in phantom measurements and demonstrated by an in vivo measurement.

  20. Depolarization of cell membrane is associated with an increase in ciliary beat frequency (CBF).

    PubMed

    Mao, H; Wong, L B

    1995-10-24

    We hypothesize that activation of muscarinic cholinergic receptors depolarizes the cell membrane of the mammalian ciliated cells which in turn causes an increase of CBF. To test this hypothesis, a di-8-ANEPPS fluorescence photon counting and nonstationary heterodyne laser light scattering system was developed to measure cell membrane potential (psi) and CBF in cultured ovine tracheal ciliated cells simultaneously. Carbachol dose dependently depolarized the cell membrane with a corresponding stimulation of CBF. The carbachol induced depolarization of cell membrane and increases of CBF were inhibited by prior application of either atropine or verapamil or amiloride. These novel data suggest that depolarization of the cell membrane and the corresponding stimulation of CBF caused by the activation of muscarinic receptors of the mammalian ciliated cells are dependent on the influx of either extracellular Ca2+ or Na+. PMID:7488025

  1. Fluorescence anisotropy measurements under shock compression

    NASA Astrophysics Data System (ADS)

    Wang, Jue; Bassett, Will; Banishev, Alexandr; Dlott, Dana

    2015-06-01

    Fluorescence anisotropy measurements, where the parallel and perpendicular polarized emissions from probe molecules are acquired simultaneously, provide direct measurement of molecular rotational dynamics. In our experiments, the fluorescence from rhodamine 6G dye in various materials under GPa shocks produced by laser-driven flyer plates is collected, separated into two orthogonally-polarized beams using a Wollaston prism and detected with a streak camera. In liquids, the molecular rotations result from rotational diffusion and in solids from shear flow. The rotation rates can be used to determine the viscosity of the shocked medium.

  2. Depolarization remote sensing by orthogonality breaking.

    PubMed

    Fade, Julien; Alouini, Mehdi

    2012-07-27

    A new concept devoted to sensing the depolarization strength of materials from a single measurement is proposed and successfully validated on a variety of samples. It relies on the measurement of the orthogonality breaking between two orthogonal states of polarization after interaction with the material to be characterized. Due to orthogonality preservation between the two states after propagation in birefringent media, this measurement concept is shown to be perfectly suited to depolarization remote sensing through fibers, opening the way to real-time depolarization endoscopy.

  3. 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.

  4. 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.

  5. Ice clouds and Asian dust studied with lidar measurements of particle extinction-to-backscatter ratio, particle depolarization, and water-vapor mixing ratio over Tsukuba.

    PubMed

    Sakai, Tetsu; Nagai, Tomohiro; Nakazato, Masahisa; Mano, Yuzo; Matsumura, Takatsugu

    2003-12-20

    The tropospheric particle extinction-to-backscatter ratio, the depolarization ratio, and the water-vapor mixing ratio were measured by use of a Raman lidar and a polarization lidar during the Asian dust seasons in 2001 and 2002 in Tsukuba, Japan. The apparent (not corrected for multiple-scattering effects) extinction-to-backscatter ratios (Sp) showed a dependence on the relative humidity with respect to ice (RHice) obtained from the lidar-derived water-vapor mixing ratio and radiosonde-derived temperature; they were mostly higher than 30 sr in dry air (RHice < 50%), whereas they were mostly lower than 30 sr in ice-supersaturated air (RHice > or = 100%), where the apparent extinction coefficients were larger than 0.036 km(-1). Both regions showed mean particle depolarization ratios of 20%-22%. Comparisons with theoretical calculations and the previous experiments suggest that the observed dependence of Sp on RHice is attributed to the difference in the predominant particles: nonspherical aerosols (mainly the Asian dust) in dry air and cloud particles in ice-supersaturated air. PMID:14717284

  6. Ice clouds and Asian dust studied with lidar measurements of particle extinction-to-backscatter ratio, particle depolarization, and water-vapor mixing ratio over Tsukuba.

    PubMed

    Sakai, Tetsu; Nagai, Tomohiro; Nakazato, Masahisa; Mano, Yuzo; Matsumura, Takatsugu

    2003-12-20

    The tropospheric particle extinction-to-backscatter ratio, the depolarization ratio, and the water-vapor mixing ratio were measured by use of a Raman lidar and a polarization lidar during the Asian dust seasons in 2001 and 2002 in Tsukuba, Japan. The apparent (not corrected for multiple-scattering effects) extinction-to-backscatter ratios (Sp) showed a dependence on the relative humidity with respect to ice (RHice) obtained from the lidar-derived water-vapor mixing ratio and radiosonde-derived temperature; they were mostly higher than 30 sr in dry air (RHice < 50%), whereas they were mostly lower than 30 sr in ice-supersaturated air (RHice > or = 100%), where the apparent extinction coefficients were larger than 0.036 km(-1). Both regions showed mean particle depolarization ratios of 20%-22%. Comparisons with theoretical calculations and the previous experiments suggest that the observed dependence of Sp on RHice is attributed to the difference in the predominant particles: nonspherical aerosols (mainly the Asian dust) in dry air and cloud particles in ice-supersaturated air.

  7. 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.

  8. Depolarization measurements on the ATS-6 20 GHz downlink A description of the VPI&SU experiment and some initial results. [meteorological precipitation effects

    NASA Technical Reports Server (NTRS)

    Bostian, C. W.; Stutzman, W. L.; Manus, E. A.; Wiley, P. H.; Marshall, R. E.

    1975-01-01

    The experiment considered is mainly concerned with the depolarizing effects of precipitation at millimeter wavelengths. Excessive depolarization introduces cross talk into communication systems which employ orthogonal polarization for frequency reuse. An understanding of atmospheric depolarization phenomena is, therefore, required for the design of future earth-satellite communications systems. Attenuation and cross polarization ratio data obtained under various meteorological conditions, including rain and a snowstorm, are presented.

  9. 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. PMID:21725434

  10. Fluorescent nanosensors for intracellular measurements: synthesis, characterization, calibration, and measurement

    PubMed Central

    Desai, Arpan S.; Chauhan, Veeren M.; Johnston, Angus P. R.; Esler, Tim; Aylott, Jonathan W.

    2013-01-01

    Measurement of intracellular acidification is important for understanding fundamental biological pathways as well as developing effective therapeutic strategies. Fluorescent pH nanosensors are an enabling technology for real-time monitoring of intracellular acidification. The physicochemical characteristics of nanosensors can be engineered to target specific cellular compartments and respond to external stimuli. Therefore, nanosensors represent a versatile approach for probing biological pathways inside cells. The fundamental components of nanosensors comprise a pH-sensitive fluorophore (signal transducer) and a pH-insensitive reference fluorophore (internal standard) immobilized in an inert non-toxic matrix. The inert matrix prevents interference of cellular components with the sensing elements as well as minimizing potentially harmful effects of some fluorophores on cell function. Fluorescent nanosensors are synthesized using standard laboratory equipment and are detectable by non-invasive widely accessible imaging techniques. The outcomes of studies employing this technology are dependent on reliable methodology for performing measurements. In particular, special consideration must be given to conditions for sensor calibration, uptake conditions and parameters for image analysis. We describe procedures for: (1) synthesis and characterization of polyacrylamide and silica based nanosensors, (2) nanosensor calibration and (3) performing measurements using fluorescence microscopy. PMID:24474936

  11. 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.

  12. 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.

  13. 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.

  14. Dissolved-oxygen quenching of in-situ fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Chudyk, Wayne; Tonaszuck, David; Pohlig, Kenneth

    1993-04-01

    In-situ fluorescence measurements of aromatic organic ground water contaminants do not always agree with gas chromatographic methods. Dissolved oxygen quenching of fluorescence may be an interferant in field measurements. Two standard fluorescent aromatics, quinine sulfate and naphthalene, were evaluated in this study. Over the range of dissolved oxygen concentrations expected to be encountered in the field, no effects of oxygen quenching on fluorescence of these compounds was observed. Quenching of quinine sulfate fluorescence by sodium chloride was observed using this system. Sodium chloride quenching was shown to follow the Stern-Volmer relation.

  15. 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.

  16. 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.

  17. 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. PMID:23566827

  18. 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.

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

    PubMed

    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

  20. 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

  1. Depolarization of polarized light caused by high altitude clouds. 2: Depolarization of lidar induced by water clouds.

    PubMed

    Sun, Y Y; Li, Z P

    1989-09-01

    A model for calculation of the depolarization of a laser beam propagating through a water cloud is described, in which multiple scattering up to sixth order is included using a Monte Carlo technique. The influence of beam geometry, drop size distribution, and cloud extinction coefficient on the depolarization is discussed. Good agreement between calculated and measured profiles of the depolarization observed in water clouds has been obtained.

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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

    2015-10-01

    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.

  7. 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

    2015-10-01

    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. PMID:26397162

  8. Measuring initiator caspase activation by bimolecular fluorescence complementation.

    PubMed

    Parsons, Melissa J; Bouchier-Hayes, Lisa

    2015-01-01

    Initiator caspases, including caspase-2, -8, and -9, are activated by the proximity-driven dimerization that occurs after their recruitment to activation platforms. Here we describe the use of caspase bimolecular fluorescence complementation (caspase BiFC) to measure this induced proximity. BiFC assays rely on the use of a split fluorescent protein to identify protein-protein interactions in cells. When fused to interacting proteins, the fragments of the split fluorescent protein (which do not fluoresce on their own) can associate and fluoresce. In this protocol, we use the fluorescent protein Venus, a brighter and more photostable variant of yellow fluorescent protein (YFP), to detect the induced proximity of caspase-2. Plasmids encoding two fusion products (caspase-2 fused to either the amino- or carboxy-terminal halves of Venus) are transfected into cells. The cells are then treated with an activating (death) stimulus. The induced proximity (and subsequent activation) of caspase-2 in the cells is visualized as Venus fluorescence. The proportion of Venus-positive cells at a single time point can be determined using fluorescence microscopy. Alternatively, the increase in fluorescence intensity over time can be evaluated by time-lapse confocal microscopy. The caspase BiFC strategy described here should also work for other initiator caspases, such as caspase-8 or -9, as long as the correct controls are used. PMID:25561623

  9. 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.

  10. Membrane depolarization in PC-12 cells during hypoxia is regulated by an O2-sensitive K+ current.

    PubMed

    Zhu, W H; Conforti, L; Czyzyk-Krzeska, M F; Millhorn, D E

    1996-08-01

    The effects of hypoxia on K+ current (IK), resting membrane potential, and cytosolic free Ca2+ in rat pheochromocytoma (PC-12) cells were studied. Whole cell voltage- and current-clamp experiments were performed to measure IK and membrane potential, respectively. Cytosolic free Ca2+ level was measured using the Ca(2+)-sensitive fluorescent dye fura 2. Depolarizing voltage steps to +50 mV from a holding potential of -90 mV elicited a slowly inactivating, tetraethylammonium chloride-sensitive, and Ca(2+)-insensitive IK that was reversibly inhibited by reduced O2 tension. Graded reduction in PO2 (from 150 to 0 mmHg) induced a graded inhibition of O2-sensitive IK [IK(O2)] up to 46% at 0 mmHg. Moreover, hypoxia induced a 19-mV membrane depolarization and a twofold increase in cytosolic free Ca2+. In Ca(2+)-free condition, inhibition of IK(O2) induced an 8-mV depolarization, suggesting that inhibition of IK(O2) was responsible for initiating depolarization. The effect of reduced PO2 on the current-voltage relationship showed a reduction of outward current and a 14-mV shift in the reversal potential comparable with the amount of depolarization measured in current clamp experiments. Neither Ca(2+)-activated IK nor inwardly rectifying IK are responsible for the hypoxia-induced depolarization. In conclusion, PC-12 cells express an IK(O2), inhibition of which leads to membrane depolarization and increased intracellular Ca2+, making the PC-12 clonal cell line a useful model for studying the molecular and biophysical mechanisms that mediate O2 chemosensitivity. PMID:8770007

  11. Measurement of Fluorescence Spectra from Ambient Aerosol Particles Using Laser-induced Fluorescence Technique

    NASA Astrophysics Data System (ADS)

    Taketani, F.; Kanaya, Y.; Nakamura, T.; Moteki, N.; Takegawa, N.

    2011-12-01

    To obtain the information of composition of organic aerosol particles in atmosphere, we developed an instrument using laser-induced fluorescence (LIF) technique. To measure the fluorescence from a particle, we employed two lasers. Scattering light signal derived from a single particle upon crossing the 635nm-CW laser triggers the 266nm-pulsed laser to excite the particle. Fluorescence from the particle in the wavelength range 300-600nm is spectrally dispersed by a grating spectrometer and then detected by a 32-Ch photo-multiplier tube(PMT). The aerosol stream is surrounded by a coaxial sheath air flow and delivered to the optical chamber at atmospheric pressure. Using PSL particles with known sizes, we made a calibration curve to estimate particle size from scattering light intensity. With the current setup of the instrument we are able to detect both scattering and fluorescence from particles whose diameters are larger than 0.5um. Our system was able to differentiate particles composed of mono-aromatic species (e.g. Tryptophan) from those of Riboflavin, by their different fluorescence wavelengths. Also, measurements of fluorescence spectra of ambient particles were demonstrated in our campus in Yokosuka city, facing Tokyo bay in Japan. We obtained several types of florescence spectra in the 8 hours. Classification of the measured fluorescence spectra will be discussed in the presentation.

  12. Characterization of long-range transported Saharan dust by means of ground-based Raman and depolarization lidar measurements at Barbados

    NASA Astrophysics Data System (ADS)

    Gross, S.; Freudenthaler, V.; Schäfler, A.; Toledano, C.; Wiegner, M.; Weinzierl, B.; Ehret, G.

    2014-12-01

    In June and July 2013 the Saharan Aerosol Long-range Transport and Aerosol-Cloud-Interaction Experiment (SALTRACE) took place at Barbados to investigate long-range transported dust after its transport over the Atlantic Ocean. Ground-based lidar measurements were performed to characterize the optical properties of long-range transported dust, and to investigate possible changes of the optical properties during transport. Measurements of the particle linear depolarization ratio (PLDR) at 355 nm and 532 nm and of the vibrational Raman signals at 387 nm and 607 nm were conducted with the small portable lidar system POLIS of the Meteorological Institute (MIM) of the Ludwig-Maximilians-Universität München in cooperation with the Institute of Atmospheric Physics (IPA) of the German Aerospace Centre (DLR). The measurements were performed in a routine manner every day for 3 hours starting at sunrise and another 3 hours starting at sunset (combined Raman and PLDR). During heavy dust events or during the formation of tropical storm Chantal measurements were performed continuously up to 48 hours. During SALTRACE the vertical aerosol distribution was dominated by a three layer structure. The boundary layer was dominated by marine air masses in most of the cases. Above the boundary layer a mixing layer with very variable structure was found up to heights of 1.5 km to 2 km. This layer was often affected by cumulus clouds. The Saharan dust layer was observed in a height range of about 2 km to about 3.5 km. In some cases the upper boundary was even 5 km. In our presentation we will give an overview over the general measurement situation and the POLIS measurements. We will present first results of the PLDR and the lidar ratio of long-range transported dust, as well as of the marine dominated air masses in the boundary layer. Furthermore we will compare our findings with results found for fresh Saharan dust and dust at the beginning of the long-range transport investigated during

  13. Fluorescence Measurement of Burned Skin Tissues

    NASA Astrophysics Data System (ADS)

    de Pedro, Hector Michael; Chang, Chuan-I.; Nguyen, Hue; Malko, Anton; Zarnani, Faranak; Glosser, Robert; Maas, D.; Idris, A.

    2011-03-01

    Early removal of affected tissues from burn patients can significantly increase the success of their recovery, since burns continue to spread and damage surrounding tissues after hours of injury. The rationale behind this procedure is that burns trigger the body's immune system to overreact, causing additional damage. Therefore it is important to locate and identify the burn (area and thickness) so that it can be removed as quickly as possible. Our project explores the use of autofluorescence as a tool to identify the burned tissues from healthy ones. Here we present that our fluorescence results show differences between burned and normal skin in both its spectra and lifetime.

  14. System and method for measuring fluorescence of a sample

    SciTech Connect

    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.

  15. 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.

  16. 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.

  17. Optimization of variable fluorescence measurements of phytoplankton communities with cyanobacteria.

    PubMed

    Simis, Stefan G H; Huot, Yannick; Babin, Marcel; Seppälä, Jukka; Metsamaa, Liisa

    2012-04-01

    Excitation-emission fluorescence matrices of phytoplankton communities were simulated from laboratory-grown algae and cyanobacteria cultures, to define the optical configurations of theoretical fluorometers that either minimize or maximize the representation of these phytoplankton groups in community variable fluorescence measurements. Excitation sources that match the photosystem II (PSII) action spectrum of cyanobacteria do not necessarily lead to equal representation of cyanobacteria in community fluorescence. In communities with an equal share of algae and cyanobacteria, inducible PSII fluorescence in algae can be retrieved from community fluorescence under blue excitation (450-470 nm) with high accuracy (R (2) = 1.00). The highest correlation between community and cyanobacterial variable fluorescence is obtained under orange-red excitation in the 590-650 nm range (R (2) = 0.54). Gaussian band decomposition reveals that in the presence of cyanobacteria, the emission detection slit must be narrow (up to 10 nm) and centred on PSII chlorophyll-a emission (~683 nm) to avoid severe dampening of the signal by weakly variable phycobilisomal fluorescence and non-variable photosystem I fluorescence. When these optimizations of the optical configuration of the fluorometer are followed, both cyanobacterial and algal cultures in nutrient replete exponential growth exhibit values of the maximum quantum yield of charge separation in PSII in the range of 0.65-0.7.

  18. 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.

  19. Depolarization synthesis: understanding the optics of Mueller matrix depolarization.

    PubMed

    Cloude, Shane R

    2013-04-01

    In this paper, we consider the forward problem in depolarization by optical systems. That is, we seek a compact parameterization that allows us to take an arbitrary "pure" optical system (namely one defined by a single Mueller-Jones matrix) and model all possible ways in which that system can depolarize light. We model this structure using compound unitary transformations and illustrate physical interpretation of the parameters involved by considering four examples, the family of depolarizers generated by scattering by random nonspherical particle clouds. We then turn attention to circular polarizers before considering all ways in which mirror reflection can cause depolarization. Finally, we consider a numerical example applied to a published Mueller matrix for backscatter from chiral turbid media.

  20. Measuring and Sorting Cell Populations Expressing Isospectral Fluorescent Proteins with Different Fluorescence Lifetimes

    PubMed Central

    Naivar, Mark; Houston, Jessica P.; Brent, Roger

    2014-01-01

    Study of signal transduction in live cells benefits from the ability to visualize and quantify light emitted by fluorescent proteins (XFPs) fused to different signaling proteins. However, because cell signaling proteins are often present in small numbers, and because the XFPs themselves are poor fluorophores, the amount of emitted light, and the observable signal in these studies, is often small. An XFP's fluorescence lifetime contains additional information about the immediate environment of the fluorophore that can augment the information from its weak light signal. Here, we constructed and expressed in Saccharomyces cerevisiae variants of Teal Fluorescent Protein (TFP) and Citrine that were isospectral but had shorter fluorescence lifetimes, ∼1.5 ns vs ∼3 ns. We modified microscopic and flow cytometric instruments to measure fluorescence lifetimes in live cells. We developed digital hardware and a measure of lifetime called a “pseudophasor” that we could compute quickly enough to permit sorting by lifetime in flow. We used these abilities to sort mixtures of cells expressing TFP and the short-lifetime TFP variant into subpopulations that were respectively 97% and 94% pure. This work demonstrates the feasibility of using information about fluorescence lifetime to help quantify cell signaling in living cells at the high throughput provided by flow cytometry. Moreover, it demonstrates the feasibility of isolating and recovering subpopulations of cells with different XFP lifetimes for subsequent experimentation. PMID:25302964

  1. 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.

  2. Fluorescence resonance energy transfers measurements on cell surfaces via fluorescence polarization.

    PubMed Central

    Cohen-Kashi, Meir; Moshkov, Sergey; Zurgil, Naomi; Deutsch, Mordechai

    2002-01-01

    A method has been developed for the determination of the efficiency of fluorescence resonance energy transfer efficiency between moieties located on cell surfaces by performing individual cell fluorescence polarization (FP) measurements. The absolute value of energy transfer efficiency (E) is calculated on an individual cell basis. The examination of this methodology was carried out using model experiments on human T lymphocyte cells. The cells were labeled with fluorescein-conjugated Concanavalin A (ConA) as donor, or rhodamine-conjugated ConA as acceptor. The experiments and results clearly indicate that determination of E via FP measurements is possible, efficient, and more convenient than other methods. PMID:12202365

  3. 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.

  4. 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.

  5. 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. PMID:18855420

  6. 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.

  7. Molecular mechanism for depolarization-induced modulation of Kv channel closure.

    PubMed

    Labro, Alain J; Lacroix, Jerome J; Villalba-Galea, Carlos A; Snyders, Dirk J; Bezanilla, Francisco

    2012-11-01

    Voltage-dependent potassium (Kv) channels provide the repolarizing power that shapes the action potential duration and helps control the firing frequency of neurons. The K(+) permeation through the channel pore is controlled by an intracellularly located bundle-crossing (BC) gate that communicates with the voltage-sensing domains (VSDs). During prolonged membrane depolarizations, most Kv channels display C-type inactivation that halts K(+) conduction through constriction of the K(+) selectivity filter. Besides triggering C-type inactivation, we show that in Shaker and Kv1.2 channels (expressed in Xenopus laevis oocytes), prolonged membrane depolarizations also slow down the kinetics of VSD deactivation and BC gate closure during the subsequent membrane repolarization. Measurements of deactivating gating currents (reporting VSD movement) and ionic currents (BC gate status) showed that the kinetics of both slowed down in two distinct phases with increasing duration of the depolarizing prepulse. The biphasic slowing in VSD deactivation and BC gate closure was strongly correlated in time and magnitude. Simultaneous recordings of ionic currents and fluorescence from a probe tracking VSD movement in Shaker directly demonstrated that both processes were synchronized. Whereas the first slowing originates from a stabilization imposed by BC gate opening, the subsequent slowing reflects the rearrangement of the VSD toward its relaxed state (relaxation). The VSD relaxation was observed in the Ciona intestinalis voltage-sensitive phosphatase and in its isolated VSD. Collectively, our results show that the VSD relaxation is not kinetically related to C-type inactivation and is an intrinsic property of the VSD. We propose VSD relaxation as a general mechanism for depolarization-induced slowing of BC gate closure that may enable Kv1.2 channels to modulate the firing frequency of neurons based on the depolarization history. PMID:23071266

  8. 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

  9. Biochip Image Grid Normalization Absolute Signal Fluorescence Measurement Using

    SciTech Connect

    Alferov, Oleg

    2001-04-17

    This software was developed to measure absolute fluorescent intensities of gel pads on a microchip in units defined by a standard fluorescent slide. It can accomodate varying measurement conditions (e.g. exposure time, sensitivity of detector, resolution of detector, etc.) as well as fluorescent microscopes with non-uniform sensitivity across their field of view allowing the user to compare measurements done on different detectors with varying exposure times, sensitivities, and resolutions. The software is designed both to operate Roper Scientific, Inc. cameras and to use image files produced by the program supplied with that equipment for its calculations. the intensity of the gel pad signal is computed so as to reduce background influence.

  10. Biochip Image Grid Normalization Absolute Signal Fluorescence Measurement Using

    2001-04-17

    This software was developed to measure absolute fluorescent intensities of gel pads on a microchip in units defined by a standard fluorescent slide. It can accomodate varying measurement conditions (e.g. exposure time, sensitivity of detector, resolution of detector, etc.) as well as fluorescent microscopes with non-uniform sensitivity across their field of view allowing the user to compare measurements done on different detectors with varying exposure times, sensitivities, and resolutions. The software is designed both tomore » operate Roper Scientific, Inc. cameras and to use image files produced by the program supplied with that equipment for its calculations. the intensity of the gel pad signal is computed so as to reduce background influence.« less

  11. Particle shape as revealed by spectral depolarization

    NASA Astrophysics Data System (ADS)

    Duncan, Donald D.; Thomas, Michael E.

    2008-02-01

    Remote detection of airborne biological agents is a problem of contemporary interest. At issue is the discrimination of scatter from such harmful species and the scatter from naturally occurring particles such as hydrosols, pollen, and dust. The feature that our detection scheme attempts to exploit is that the species of interest typically have a characteristic size and shape that produce a unique signature in the depolarization of the scattered light. Through the use of T-matrix calculations we demonstrate the effect and show processing algorithms for quantifying and interpreting the measurements. Simulations suggest that it is possible to retrieve an estimate of the aspect ratio from a measurement of the spectral behavior of the linear depolarization in either the forward-scatter or back-scatter regime.

  12. Fluorescence molecular tomographic image reconstruction based on reduced measurement data

    NASA Astrophysics Data System (ADS)

    Zou, Wei; Wang, Jiajun; Feng, David Dagan; Fang, Erxi

    2015-07-01

    The analysis of fluorescence molecular tomography is important for medical diagnosis and treatment. Although the quality of reconstructed results can be improved with the increasing number of measurement data, the scale of the matrices involved in the reconstruction of fluorescence molecular tomography will also become larger, which may slow down the reconstruction process. A new method is proposed where measurement data are reduced according to the rows of the Jacobian matrix and the projection residual error. To further accelerate the reconstruction process, the global inverse problem is solved with level-by-level Schur complement decomposition. Simulation results demonstrate that the speed of the reconstruction process can be improved with the proposed algorithm.

  13. 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.

  14. Measurements of extrinsic fluorescence in Intralipid and polystyrene microspheres

    PubMed Central

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

    2014-01-01

    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. PMID:25136497

  15. 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.

  16. Fluorescence measurements for chemical optical fiber sensor of cobalt

    NASA Astrophysics Data System (ADS)

    Mazikowski, Adam; Kaczmarek, Emil

    2006-10-01

    Opto-chemical sensors are sensors of quantities (pH level, heavy metal ions concentration), detection of which can be performed optically. These sensors utilize various optical phenomena such as changes of fluorescence in the presence of a certain agent. Many substances available and interesting from the sensor point of view exhibit different properties in solution and after physical and/or chemical mounting on glass slide or optical fiber. Because of this it is necessary to investigate application possibilities of a certain substance in well defined metrological environment. In this paper we described system for measuring fluorescence of sensing materials. We proposed system utilizing emission and absorption spectra separation and phase-sensitive detection. As an example of such system a fluorescence sensor of cobalt was of our interest. We described sample preparation process and measured some properties of chosen chemical substances. Achieved results are the basis for further research.

  17. Ice-cloud depolarization of backscatter for CO(2) and other infrared lidars.

    PubMed

    Eberhard, W L

    1992-10-20

    The depolarization of backscatter from ice particles at the CO(2) lidar wavelength of 10.59 microm was investigated through field measurements, with simultanous depolarization measurements taken at 0.6943 microm for comparison. The depolarization ratio at the infrared wavelength was usually at or below the lidar's sensitivity limit of 0.01, which is dramatically smaller than the typical 0.5 linear depolarization ratio for short-wave lidars. This behavior is explained by the strong absorption of ice at the infrared wavelength. Depolarization measurements at a 10.59-microm wavelength cannot discriminate between ice and water clouds in the manner of short-wave lidars. A possibility exists for more prominent depolarization at shorter CO(2) lidar wavelengths (e.g., 9.115 microm), but additional research is required. Depolarization at the 2.09-microm wavelength is predicted to be substantial and useful for hydrometeor observations.

  18. 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.

  19. 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…

  20. Measurement of in vitro microtubule polymerization by turbidity and fluorescence.

    PubMed

    Mirigian, Matthew; Mukherjee, Kamalika; Bane, Susan L; Sackett, Dan L

    2013-01-01

    Tubulin polymerization may be conveniently monitored by the increase in turbidity (optical density, or OD) or by the increase in fluorescence intensity of diamidino-phenylindole. The resulting data can be a quantitative measure of microtubule (MT) assembly, but some care is needed in interpretation, especially of OD data. Buffer formulations used for the assembly reaction significantly influence the polymerization, both by altering the critical concentration for polymerization and by altering the exact polymer produced-for example, by increasing the production of sheet polymers in addition to MT. Both the turbidity and the fluorescence methods are useful for demonstrating the effect of MT-stabilizing or -destabilizing additives.

  1. 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.

  2. 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.

  3. Laboratory investigations of mineral dust near-backscattering depolarization ratios

    NASA Astrophysics Data System (ADS)

    Järvinen, E.; Kemppinen, O.; Nousiainen, T.; Kociok, T.; Möhler, O.; Leisner, T.; Schnaiter, M.

    2016-07-01

    Recently, there has been increasing interest to derive the fractions of fine- and coarse-mode dust particles from polarization lidar measurements. For this, assumptions of the backscattering properties of the complex dust particles have to be made either by using empirical data or particle models. Laboratory measurements of dust backscattering properties are important to validate the assumptions made in the lidar retrievals and to estimate their uncertainties. Here, we present laboratory measurements of linear and circular near-backscattering (178°) depolarization ratios of over 200 dust samples measured at 488 and 552 nm wavelengths. The measured linear depolarization ratios ranged from 0.03 to 0.36 and were strongly dependent on the particle size. The strongest size-dependence was observed for fine-mode particles as their depolarization ratios increased almost linearly with particle median diameter from 0.03 to 0.3, whereas the coarse-mode particle depolarization values stayed rather constant with a mean linear depolarization ratio of 0.27. The depolarization ratios were found to be insensitive to the dust source region or thin coating of the particles or to changes in relative humidity. We compared the measurements with results of three different scattering models. With certain assumptions for model particle shape, all the models were capable of correctly describing the size-dependence of the measured dust particle, albeit the model particles significantly differed in composition, shape and degree of complexity. Our results show potential for distinguishing the dust fine- and coarse-mode distributions based on their depolarization properties and, thus, can serve the lidar community as an empirical reference.

  4. 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.

  5. Depolarization in the SLC Collider Arcs

    SciTech Connect

    Emma, P.; Limberg, T.; Rossmanith, R.

    1994-06-01

    In the 1993 running cycle of the Stanford Linear Collider, electron spin polarization measurements with a Moller polarimeter at the end of the linac and a Compton polarimeter near the interaction point (IP) indicated a relative polarization loss of up to 20% across the arc. The authors present calculations of the depolarizing effects where variations in energy, energy spread and transverse emittance as well as changes in orbit and initial spin orientation are taken into account. They compare their results with measurements and conclude that, in standard operating conditions, the relative polarization loss is only 3{+-}2%.

  6. 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.

  7. Near-infrared spark source excitation for fluorescence lifetime measurements

    NASA Astrophysics Data System (ADS)

    Birch, D. J. S.; Hungerford, G.; Imhof, R. E.

    1991-10-01

    We have extended the range of excitation wavelengths from spark sources used in single photon timing fluorometry into the near infrared by means of the all-metal coaxial flashlamp filled with an argon-hydrogen gas mixture. At 750 nm this mixture gives ˜15 times the intensity available from pure hydrogen for a comparable pulse duration. Measurements are demonstrated by using the laser dye IR-140 in acetone, for which a fluorescence lifetime of 1.20 ns is recorded.

  8. 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.

  9. Depolarization of light by rough surface of scattering phantoms

    NASA Astrophysics Data System (ADS)

    Tchvialeva, Lioudmila; Markhvida, Igor; Lee, Tim K.; Doronin, Alexander; Meglinski, Igor

    2013-02-01

    The growing interest in biomedical optics to the polarimetric methods push researchers to better understand of light depolarization during scattering in and on the surface of biological tissues. Here we study the depolarization of light propagated in silicone phantoms. The phantoms with variety of surface roughness and bulk optical properties are designed to imitate human skin. Free-space speckle patterns in parallel (III) and perpendicular (I⊥) direction in respect to incident polarization are used to get the depolarization ratio of backscattered light DR = (III - I⊥)/( III + I⊥). The Monte Carlo model developed in house is also applied to compare simulated DR with experimentally measured. DR dependence on roughness, concentration and size of scattering particles is analysed. A weak depolarization and negligible response to scattering of the medium are observed for phantoms with smooth surfaces, whereas for the surface roughness in order to the mean free path the depolarization ratio decreases and reveals dependence on the bulk scattering coefficient. In is shown that the surface roughness could be a key factor triggering the ability of tissues' characterization by depolarization ratio.

  10. Laser-induced fluorescence measurement of combustion chemistry intermediates

    NASA Technical Reports Server (NTRS)

    Crosley, David R.

    1986-01-01

    Laser-induced fluorescence (LIF) can measure the trace (often free radical) species encountered as intermediates in combustion chemistry; OH, CS, NH, NS, and NCO are typical of the species detected in flames by LIF. Attention is given to illustrative experiments designed to accumulate a quantitative data base for LIF detection in low pressure flow systems and flames, as well as to flame measurements conducted with a view to the detection of new chemical intermediaries that may deepen insight into the chemistry of combustion.

  11. 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.

  12. 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

  13. Intracellular pH measurements made simple by fluorescent protein probes and the phasor approach to fluorescence lifetime imaging.

    PubMed

    Battisti, Antonella; Digman, Michelle A; Gratton, Enrico; Storti, Barbara; Beltram, Fabio; Bizzarri, Ranieri

    2012-05-25

    A versatile pH-dependent fluorescent protein was applied to intracellular pH measurements by means of the phasor approach to fluorescence lifetime imaging. By this fit-less method we obtain intracellular pH maps under resting or altered physiological conditions by single-photon confocal or two-photon microscopy.

  14. Lidar for multiple backscattering and depolarization observations.

    PubMed

    Allen, R J; Platt, C M

    1977-12-01

    A lidar system that can measure multiple scattering and depolarization in the atmosphere is being used to study the climatic effects of cirrus clouds and to perform other investigations. The lidar system and its novel aspects are described in this paper. The influence of multiple scattering on noise, signal, and SNR is considered. Special receiver field stops incorporated for multiple scattering measurements, use of low voltage to control the photomultiplier tube gain, and a precision power/energy monitor are described. A technique for aligning transmitter and receiver axes and measuring transmitter beamwidth is presented. The multiple-scattered components of backscattered light are determined by inserting a center-blocked field stop to restrict the receiver field of view to the region outside of the diverging transmitted beam. Typical returns with and without the opaque field stop indicate the amplitude of multiple scattering from cirrus clouds and prove the feasibility of this technique. The depolarization ratio delta and backscatter coefficients from an altostratus cloud illustrate the potential of these quantities for the study of cloud structure and phase.

  15. Measuring Phagosome pH by Ratiometric Fluorescence Microscopy.

    PubMed

    Nunes, Paula; Guido, Daniele; Demaurex, Nicolas

    2015-01-01

    Phagocytosis is a fundamental process through which innate immune cells engulf bacteria, apoptotic cells or other foreign particles in order to kill or neutralize the ingested material, or to present it as antigens and initiate adaptive immune responses. The pH of phagosomes is a critical parameter regulating fission or fusion with endomembranes and activation of proteolytic enzymes, events that allow the phagocytic vacuole to mature into a degradative organelle. In addition, translocation of H(+) is required for the production of high levels of reactive oxygen species (ROS), which are essential for efficient killing and signaling to other host tissues. Many intracellular pathogens subvert phagocytic killing by limiting phagosomal acidification, highlighting the importance of pH in phagosome biology. Here we describe a ratiometric method for measuring phagosomal pH in neutrophils using fluorescein isothiocyanate (FITC)-labeled zymosan as phagocytic targets, and live-cell imaging. The assay is based on the fluorescence properties of FITC, which is quenched by acidic pH when excited at 490 nm but not when excited at 440 nm, allowing quantification of a pH-dependent ratio, rather than absolute fluorescence, of a single dye. A detailed protocol for performing in situ dye calibration and conversion of ratio to real pH values is also provided. Single-dye ratiometric methods are generally considered superior to single wavelength or dual-dye pseudo-ratiometric protocols, as they are less sensitive to perturbations such as bleaching, focus changes, laser variations, and uneven labeling, which distort the measured signal. This method can be easily modified to measure pH in other phagocytic cell types, and zymosan can be replaced by any other amine-containing particle, from inert beads to living microorganisms. Finally, this method can be adapted to make use of other fluorescent probes sensitive to different pH ranges or other phagosomal activities, making it a generalized

  16. Chloride transport across placental microvillous membranes measured by fluorescence

    SciTech Connect

    Illsley, N.P.; Glaubensklee, C.; Davis, B.; Verkman, A.S. )

    1988-12-01

    Chloride transport across human placental microvillous vesicle membrane was investigated using the fluorescent probe SPQ (6-methoxy-N(3-sulfopropyl)quinolinium). Chloride influx (J{sub Cl}) was calculated from the initial rate of quenching of intravesicular SPQ fluorescence by chloride. J{sub Cl} measured by SPQ fluorescence was not significantly different from J{sub Cl} measured by uptake of {sup 36}Cl; SPQ did not affect measurements of J{sub Cl}. J{sub Cl} was increased 51% by a 58-mV membrane potential. Voltage-stimulated J{sub Cl} showed a saturable dependence on chloride concentration with a dissociation constant (K{sub d}) of 18 {plus minus} 5 mM and was inhibited by diphenylamine-2-carboxylate with an apparent inhibitory constant of 0.13 {plus minus} 0.03 mM. The activation energy calculated for voltage-stimulated J{sub Cl} was 4.6 {plus minus} 0.6 kcal/mol. J{sub Cl} was also stimulated by a reduction in the external pH from 7.0 to 5.5 (internal pH = 70). pH-stimulated chloride influx was increased by trans-HCO{sub 3} and was inhibited by dihydro-4,4{prime}-diisothiocyano-2,2{prime}-disulfonic stilbene. Uptake of {sup 36}Cl into microvillous vesicles was stimulated by trans-Cl. pH-stimulated J{sub Cl} showed a saturable dependence on chloride with a K{sub d} of 38 {plus minus} 6 mM but was not affected by membrane potential. No evidence was found for Na- or K-coupled chloride cotransport. These findings demonstrate the presence of a saturable chloride conductance and an electroneutral chloride-bicarbonate exchanger in the placental microvillous membrane.

  17. Measuring Phagosome pH by Ratiometric Fluorescence Microscopy.

    PubMed

    Nunes, Paula; Guido, Daniele; Demaurex, Nicolas

    2015-12-07

    Phagocytosis is a fundamental process through which innate immune cells engulf bacteria, apoptotic cells or other foreign particles in order to kill or neutralize the ingested material, or to present it as antigens and initiate adaptive immune responses. The pH of phagosomes is a critical parameter regulating fission or fusion with endomembranes and activation of proteolytic enzymes, events that allow the phagocytic vacuole to mature into a degradative organelle. In addition, translocation of H(+) is required for the production of high levels of reactive oxygen species (ROS), which are essential for efficient killing and signaling to other host tissues. Many intracellular pathogens subvert phagocytic killing by limiting phagosomal acidification, highlighting the importance of pH in phagosome biology. Here we describe a ratiometric method for measuring phagosomal pH in neutrophils using fluorescein isothiocyanate (FITC)-labeled zymosan as phagocytic targets, and live-cell imaging. The assay is based on the fluorescence properties of FITC, which is quenched by acidic pH when excited at 490 nm but not when excited at 440 nm, allowing quantification of a pH-dependent ratio, rather than absolute fluorescence, of a single dye. A detailed protocol for performing in situ dye calibration and conversion of ratio to real pH values is also provided. Single-dye ratiometric methods are generally considered superior to single wavelength or dual-dye pseudo-ratiometric protocols, as they are less sensitive to perturbations such as bleaching, focus changes, laser variations, and uneven labeling, which distort the measured signal. This method can be easily modified to measure pH in other phagocytic cell types, and zymosan can be replaced by any other amine-containing particle, from inert beads to living microorganisms. Finally, this method can be adapted to make use of other fluorescent probes sensitive to different pH ranges or other phagosomal activities, making it a generalized

  18. 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.

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

    PubMed

    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.

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. Characterization of Geiger mode avalanche photodiodes for fluorescence decay measurements

    NASA Astrophysics Data System (ADS)

    Jackson, John C.; Phelan, Don; Morrison, Alan P.; Redfern, R. Michael; Mathewson, Alan

    2002-05-01

    Geiger mode avalanche photodiodes (APD) can be biased above the breakdown voltage to allow detection of single photons. Because of the increase in quantum efficiency, magnetic field immunity, robustness, longer operating lifetime and reduction in costs, solid-state detectors capable of operating at non-cryogenic temperatures and providing single photon detection capabilities provide attractive alternatives to the photomultiplier tube (PMT). Shallow junction Geiger mode APD detectors provide the ability to manufacture photon detectors and detector arrays with CMOS compatible processing steps and allows the use of novel Silicon-on-Insulator(SoI) technology to provide future integrated sensing solutions. Previous work on Geiger mode APD detectors has focused on increasing the active area of the detector to make it more PMT like, easing the integration of discrete reaction, detection and signal processing into laboratory experimental systems. This discrete model for single photon detection works well for laboratory sized test and measurement equipment, however the move towards microfluidics and systems on a chip requires integrated sensing solutions. As we move towards providing integrated functionality of increasingly nanoscopic sized emissions, small area detectors and detector arrays that can be easily integrated into marketable systems, with sensitive small area single photon counting detectors will be needed. This paper will demonstrate the 2-dimensional and 3-dimensional simulation of optical coupling that occurs in Geiger mode APDs. Fabricated Geiger mode APD detectors optimized for fluorescence decay measurements were characterized and preliminary results show excellent results for their integration into fluorescence decay measurement systems.

  5. Telescope Array measurement of UHECR composition from stereoscopic fluorescence detection

    NASA Astrophysics Data System (ADS)

    Stroman, Thomas; Bergman, Douglas; Abu Zayyad, Tareq

    2014-03-01

    The chemical composition of ultra-high-energy cosmic rays (UHECRs) is an important constraint on models of UHECR production and propagation, and must be determined experimentally. A UHECR-induced extensive air shower's longitudinal development is dictated by the energy per nucleon of the primary particle. The observed distribution of atmospheric slant depths (Xmax) is therefore sensitive to the composition, facilitating measurement of the relative abundances of ``light'' (proton-like) and ``heavy'' (iron-like) primary UHECR particles. The Telescope Array (TA) experiment, the northern hemisphere's largest UHECR detector, includes three fluorescence detector (FD) stations that record the longitudinal development of the extensive air showers produced by UHECR arrivals. ``Stereo'' observation of individual showers by multiple FDs tightly constrains the trajectory reconstruction, allowing a precise measurement of Xmax as well as energy. We will present the stereo TA data from six years of operation and progress toward a measurement of chemical composition.

  6. Comparison of plate reader-based methods with fluorescence microscopy for measurements of intracellular calcium levels for the assessment of in vitro neurotoxicity.

    PubMed

    Meijer, Marieke; Hendriks, Hester S; Heusinkveld, Harm J; Langeveld, Wendy T; Westerink, Remco H S

    2014-12-01

    The intracellular calcium concentration ([Ca(2+)]i) is an important readout for in vitro neurotoxicity since calcium is critically involved in many essential neurobiological processes, including neurotransmission, neurodegeneration and neurodevelopment. [Ca(2+)]i is often measured with considerable throughput at the level of cell populations with plate reader-based assays or with lower throughput at the level of individual cells with fluorescence microscopy. However, these methodologies yield different quantitative and qualitative results. In recent years, we demonstrated that the resolution and sensitivity of fluorescence microscopy is superior compared to plate reader-based assays. However, it is currently unclear if the use of plate reader-based assays results in more 'false negatives' or 'false positives' in neurotoxicity screening studies. In the present study, we therefore compared a plate reader-based assay with fluorescence microscopy using a small test set of environmental pollutants consisting of dieldrin, lindane, polychlorinated biphenyl 53 (PCB53) and tetrabromobisphenol-A (TBBPA). Using single-cell fluorescence microscopy, we demonstrate that all test chemicals reduce the depolarization-evoked increase in [Ca(2+)]i, whereas lindane, PCB53 and TBBPA also increase basal [Ca(2+)]i, though via different mechanisms. Importantly, none of these effects were confirmed with the plate reader-based assay. We therefore conclude that standard plate reader-based methods are not sufficiently sensitive and reliable to measure the highly dynamic and transient changes in [Ca(2+)]i that occur during chemical exposure.

  7. 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.

  8. Strain measurements in thermally grown alumina scales using ruby fluorescence

    SciTech Connect

    Veal, B.W.; Natesan, K.; Koshelev, I.; Grimsditch, M.; Renusch, D. Hou, P.Y.

    1996-12-31

    We have measured strains in alumina scales thermally grown on Fe-Cr- Al alloys by exploiting the strain dependence of the ruby luminescence line. Measurements were done on Fe-5Cr-28Al and Fe-18Cr-10Al (at.%, bal. Fe) oxidized between 300-1300 C with periodic cycling to room temperature. Significantly different levels of strain buildup were observed in scales on these alloys. Results on similar alloys containing a dilute reactive element (Zr or Hf) are also presented. We observe that scales on alloys containing a reactive element (RE) can support higher strains than scales on RE-free alloys. With the luminescence technique, strain relief associated with spallation thresholds is readily observed. In early stage oxidation, the evolution of transition phases is monitored using Raman and fluorescence spectroscopies. The fluorescence technique also provides a sensitive probe of early stage formation of {alpha}-Al{sub 2}O{sub 3}. It appears that, in presence of Cr{sub 2}O{sub 3} or Fe{sub 2}O{sub 3}, the {alpha}-alumina phase can form at anomalously low temperatures.

  9. Rapid fluorescence-based measurement of toxicity in anaerobic digestion.

    PubMed

    Chen, Jian Lin; Ortiz, Raphael; Xiao, Yeyuan; Steele, Terry W J; Stuckey, David C

    2015-05-15

    A rapid fluorescence measurement based on resazurin reduction was developed and applied for the detection of toxicants/inhibitors to anaerobic digestion metabolism. By initially using a pure facultative anaerobic strain, Enterococcus faecalis as a model organism, this technique proved to be fast and sensitive when detecting the model toxicant, pentachlorophenol (PCP). The technique revealed significant metabolic changes in Enterococcus faecalis with a PCP spike ranging from 0.05 to 100 mg/L, and could detect PCP's toxicity to E. faecalis at a concentration of only 0.05 mg/L in 8 min. Furthermore, by extending this technique to a mixed anaerobic sludge, not only could the effect of 0.05-100 mg/L PCP be determined on anaerobic digestion metabolism within 10 min, but also its rate of biogas production. These results suggest that a resazurin-based fluorescence measurement can potentially be incorporated into a microfluidic system to develop a biosensor for the real-time monitoring, control and early warning of toxicant/inhibitor loads in the influent to an anaerobic digestion system.

  10. 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.

  11. 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.

  12. Probing intrinsic anisotropies of fluorescence: Mueller matrix approach.

    PubMed

    Saha, Sudipta; Soni, Jalpa; Chandel, Shubham; Kumar, Uday; Ghosh, Nirmalya

    2015-08-01

    We demonstrate that information on “intrinsic” anisotropies of fluorescence originating from preferential orientation/organization of fluorophore molecules can be probed using a Mueller matrix of fluorescence. For this purpose, we have developed a simplified model to decouple and separately quantify the depolarization property and the intrinsic anisotropy properties of fluorescence from the experimentally measured fluorescence Mueller matrix. Unlike the traditionally defined fluorescence anisotropy parameter, the Mueller matrix-derived fluorescence polarization metrics, namely, fluorescence diattenuation and polarizance parameters, exclusively deal with the intrinsic anisotropies of fluorescence. The utility of these newly derived fluorescence polarimetry parameters is demonstrated on model systems exhibiting multiple polarimetry effects, and an interesting example is illustrated on biomedically important fluorophores, collagen. PMID:26301796

  13. 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

  14. Quantitative fluorescent speckle microscopy (QFSM) to measure actin dynamics.

    PubMed

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

    2012-10-01

    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 meiotic/mitotic spindle. Here, focus is 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 the microinjection of fluorophore-labeled actin into cells, which generate a low ratio of fluorescently labeled to endogenously unlabeled actin monomers. Spinning disk confocal or wide-field imaging then visualizes fluorophore clusters (two to eight 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

  15. Prolonged irradiation of enhanced cyan fluorescent protein or Cerulean can invalidate Forster resonance energy transfer measurements.

    PubMed

    Hoffmann, Birgit; Zimmer, Thomas; Klöcker, Nikolaj; Kelbauskas, Laimonas; König, Karsten; Benndorf, Klaus; Biskup, Christoph

    2008-01-01

    Since its discovery, green fluorescent protein (GFP) and its variants have proven to be a good and convenient fluorescent label for proteins: GFP and other visible fluorescent proteins (VFPs) can be fused selectively to the protein of interest by simple cloning techniques and develop fluorescence without additional cofactors. Among the steadily growing collection of VFPs, several pairs can be chosen that can serve as donor and acceptor fluorophores in Forster resonance energy transfer (FRET) experiments. Among them, the cyan fluorescent proteins (ECFP/Cerulean) and the enhanced yellow fluorescent protein (EYFP) are most commonly used. We show that ECFP and Cerulean have some disadvantages despite their common use: Upon irradiation with light intensities that are commonly used for intensity- and lifetime-based FRET measurements, both the fluorescence intensity and the fluorescence lifetime of ECFP and Cerulean decrease. This can hamper both intensity- and lifetime-based FRET measurements and emphasizes the need for control measurements to exclude these artifacts. PMID:18601529

  16. 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.

  17. A continuous fluorescent assay for protein prenyltransferases measuring diphosphate release.

    PubMed

    Pais, June E; Bowers, Katherine E; Stoddard, Andrea K; Fierke, Carol A

    2005-10-15

    Protein farnesyltransferase and protein geranylgeranyltransferase type I catalyze the transfer of a 15- and a 20-carbon prenyl group, respectively, from a prenyl diphosphate to a cysteine residue at the carboxyl terminus of target proteins, with the concomitant release of diphosphate. Common substrates include oncogenic Ras proteins, which are implicated in up to 30% of all human cancers, making prenyltransferases a viable target for chemotherapeutic drugs. A coupled assay has been developed to measure the rate constant of diphosphate (PPi) dissociation during the prenyltransferase reaction under both single and multiple turnover conditions. In this assay, the PPi group produced in the prenyltransferase reaction is rapidly cleaved by inorganic pyrophosphatase to form phosphate (Pi), which is then bound by a coumarin-labeled phosphate binding protein from Escherichia coli, resulting in a fluorescence increase. The observed rate constant for PPi release is equal to the rate constant of prenylation of the peptide, as measured by other assays, so that this nonradioactive assay can be used to measure prenyltransferase activity under either single or multiple turnover conditions. This assay can be adapted for high-throughput screening for potential prenyltransferase substrates and inhibitors.

  18. 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.

  19. 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

  20. 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.

  1. Lysophosphatidylcholine cell depolarization: increased membrane permeability for use in the determination of cell membrane potentials

    SciTech Connect

    Gallo, R.L.; Wersto, R.P.; Notter, R.H.; Finkelstein, J.N.

    1984-12-01

    Current techniques for the determination of cellular membrane potentials based on the uptake of a radiolabeled lipophilic cation, (3H)triphenylmethylphosphonium, and the cyanine dye, DiOC5(3), were analyzed in terms of the proportions of these probes which are accumulated due to potential-dependent and potential-independent forces. Measurements were made of probe uptake in two model systems: rabbit type II pneumocytes and human promyelocytic HL60 cells. For both cell types, the membrane potential-independent component of triphenylmethylphosphonium uptake was found to be a function of several variables, including the length of exposure of the cells to the transport facilitator tetraphenylboron, the concentration of tetraphenylboron, and the integrity of the cell membrane. To accurately determine the magnitude of the potential-independent component of probe uptake by type II and HL60 cells, the cell-permeabilizing agent lysophosphatidylcholine was used. The ability of lysophosphatidylcholine to depolarize cell membranes and accurately predict membrane potential-independent accumulation was found to be equal to or superior to several other techniques commonly used to achieve membrane depolarization (e.g. gramicidin, valinomycin plus high external potassium). Lysophosphatidylcholine cell treatment was found to be a simple, rapid, and accurate technique to increase cell membrane permeability and allow equilibration of intra- and extracellular ions. The method is shown to be useful for determining membrane potential-independent accumulation of both radiolabeled and fluorescent probes of membrane potential.

  2. 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

  3. 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%.

  4. Electron beam dispersion measurements in nitrogen using two-dimensional imaging of N2(+) fluorescence

    NASA Astrophysics Data System (ADS)

    Clapp, L. H.; Twiss, R. G.; Cattolica, R. J.

    Experimental results are presented related to the radial spread of fluorescence excited by 10 and 20 KeV electron beams passing through nonflowing rarefied nitrogen at 293 K. An imaging technique for obtaining species distributions from measured beam-excited fluorescence is described, based on a signal inversion scheme mathematically equivalent to the inversion of the Abel integral equation. From fluorescence image data, measurements of beam radius, integrated signal intensity, and spatially resolved distributions of N2(+) first-negative-band fluorescence-emitting species have been made. Data are compared with earlier measurements and with an heuristic beam spread model.

  5. Depolarized light source for fiber optic sensors

    NASA Astrophysics Data System (ADS)

    Burns, W. K.; Moeller, R. P.

    1991-12-01

    An apparatus comprised of a depolarized light source for fiber optic sensors is disclosed. In a preferred embodiment, the depolarized light source of the apparatus comprises: a first laser for generating a first beam at a first frequency with the first beam having a linear polarization state; a second laser for generating a second beam at a second frequency with the second beam having a linear polarization state; a means for rotating the polarization state of the second beam so that the first and second beams have orthogonal linear polarization states with respect to each other; and a means for combining the first beam with the polarization-rotated second beam to obtain a composite beam which is depolarized. In a system operation, the apparatus comprises a source of depolarized light; an integrated optic modulator means for receiving a modulator drive signal; a first low-birefringence fiber for conveying the depolarized light beam from the depolarized light source to the integrated optic modulator means (the integrated optic modulator mean modulating only one linear polarization state in the depolarized light beam as a function of the modulator drive signal to produce a modulated beam); a photodetector; and a second low-birefringence fiber for conveying the modulated light beam to the photodetector with the photodetector being responsive to the modulated light beam for developing an electrical signal proportional to the modulator drive signal.

  6. 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

  7. Ice depolarization on low-angle 2 GHz satellite downlinks

    NASA Technical Reports Server (NTRS)

    Stutzman, W. L.; Bostian, C. W.; Tsolakis, A.; Pratt, T.

    1984-01-01

    The impact of ice depolarization on the statistical performance of satellite downlinks were investigated. Propagation data recorded during 1979 and 1980 to see what impact of ice depolarization on link performance were analyzed. The effects on the cross polarization discrimination (XPD) statistics amounted to at most a 2 to 4 dB reduction in the XPD values which rain would have produced for a given percentage of time. Ice depolarization had no effect on the statistics of XPD values below the 0.01% level. Most of the severe ice depolarization events were associated with drops in barometric pressure and the passage of intense cold fronts through our area. Ice contents as the product of three individually undetermined quantities were defined: ice particle density, ice cloud thickness, and the average volume of the ice crystals. It is indicated that populations of ice particle with ice contents on the order of 0.002 m4/m3 are probably responsible for the lower values of measured XPD.

  8. 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

  9. 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.

  10. 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.

  11. Atmospheric Depolarization Lidar Experimental Receiver: A Space Shuttle Hitchhiker Payload

    NASA Technical Reports Server (NTRS)

    McGill, Matthew J.; Scott, V. Stanley; Spinhirne, James D.

    1998-01-01

    Development work is underway at the Goddard Space Flight Center to construct a depolarization measuring atmospheric lidar receiver. The Atmospheric Lidar (AL) is tentatively scheduled to fly on the Space Shuttle in a late-1999 time frame. The AL will fly in conjunction with the Shuttle Laser Altimeter (SLA) and the Infrared Spectral Imaging Radiometer (ISIR) to provide a comprehensive package of atmospheric aerosol and cloud information. The AL operates in conjunction with the SLA laser transmitter and measures profiles of atmospheric backscatter at 532 nm. The receiver system discriminates between the parallel and perpendicular polarizations of the backscattered signal, thus providing depolarization ratios for scattering from clouds. The lidar receiver also provides cloud height and thickness measurements to complement the brightness temperature measurements generated by the ISIR thermal imager. The function of the AL is twofold. The primary function is to provide range-resolved measurements of atmospheric aerosol backscatter and depolarization ratio at 532 nm with 75 m vertical resolution. The scientific purpose of these measurements is to determine composition of clouds based on the depolarization ratio (i.e cloud content is water or ice), to determine cloud height and thickness, and to gain further understanding of the global distribution of aerosols. This information, when coupled with the cloud brightness measured by the ISIR thermal imager will provide a significant amount of information on cloud composition and radiative effects, particularly for cirrus and sub-visual cirrus clouds. A secondary function of the AL is to serve as an in-space test bed for lidar technology advancements, including a fully fiber-coupled receiver and photon counting from space. In addition, the data obtained by the AL will be used to develop software for the Geoscience Laser Altimeter System (GLAS) flight mission.

  12. 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.

  13. Measurement of exhaust gas recirculation rate by laser-induced fluorescence in engine

    NASA Astrophysics Data System (ADS)

    Morin, C.; Modica, V.; Guibert, P.

    2008-10-01

    The objective of this study is to measure by planar laser-induced fluorescence the exhaust gas recirculation (EGR) rate in the combustion chamber of an optical engine to quantify the stratification phenomena used in the new combustion strategy. From the results obtained in a high pressure-high temperature (HP-HT) facility, the tracer chosen for this aim is 3-pentanone. This paper presents a quantitative measurement of the EGR rate in the engine and a post-processing model with a correction and calibration procedure by considering the influence of temperature and pressure on the absorption cross-section and the 3-pentanone fluorescence quantum yield from the results established in the HP-HT facility. The stratification phenomena are quantified by using 3-pentanone fluorescence for two different configurations of EGR introduction in the engine. The local fluorescence measurements in the HP-HT facility are also compared with planar fluorescence measurements in the optical engine.

  14. Measurement of protein-like fluorescence in river and waste water using a handheld spectrophotometer.

    PubMed

    Baker, Andy; Ward, David; Lieten, Shakti H; Periera, Ryan; Simpson, Ellie C; Slater, Malcolm

    2004-07-01

    Protein-like fluorescence intensity in rivers increases with increasing anthropogenic DOM inputs from sewerage and farm wastes. Here, a portable luminescence spectrophotometer was used to investigate if this technology could be used to provide both field scientists with a rapid pollution monitoring tool and process control engineers with a portable waste water monitoring device, through the measurement of river and waste water tryptophan-like fluorescence from a range of rivers in NE England and from effluents from within two waste water treatment plants. The portable spectrophotometer determined that waste waters and sewerage effluents had the highest tryptophan-like fluorescence intensity, urban streams had an intermediate tryptophan-like fluorescence intensity, and the upstream river samples of good water quality the lowest tryptophan-like fluorescence intensity. Replicate samples demonstrated that fluorescence intensity is reproducible to +/- 20% for low fluorescence, 'clean' river water samples and +/- 5% for urban water and waste waters. Correlations between fluorescence measured by the portable spectrophotometer with a conventional bench machine were 0.91; (Spearman's rho, n = 143), demonstrating that the portable spectrophotometer does correlate with tryptophan-like fluorescence intensity measured using the bench spectrophotometer.

  15. 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

  16. 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.

  17. 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

  18. 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.

  19. 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.

  20. Red and far red Sun-induced chlorophyll fluorescence as a measure of plant photosynthesis

    NASA Astrophysics Data System (ADS)

    Rossini, M.; Nedbal, L.; Guanter, L.; Ač, A.; Alonso, L.; Burkart, A.; Cogliati, S.; Colombo, R.; Damm, A.; Drusch, M.; Hanus, J.; Janoutova, R.; Julitta, T.; Kokkalis, P.; Moreno, J.; Novotny, J.; Panigada, C.; Pinto, F.; Schickling, A.; Schüttemeyer, D.; Zemek, F.; Rascher, U.

    2015-03-01

    Remote estimation of Sun-induced chlorophyll fluorescence emitted by terrestrial vegetation can provide an unparalleled opportunity to track spatiotemporal variations of photosynthetic efficiency. Here we provide the first direct experimental evidence that the two peaks of the chlorophyll fluorescence spectrum can be accurately mapped from high-resolution radiance spectra and that the signal is linked to variations in actual photosynthetic efficiency. Red and far red fluorescence measured using a novel airborne imaging spectrometer over a grass carpet treated with an herbicide known to inhibit photosynthesis was significantly higher than the corresponding signal from an equivalent untreated grass carpet. The reflectance signal of the two grass carpets was indistinguishable, confirming that the fast dynamic changes in fluorescence emission were related to variations in the functional status of actual photosynthesis induced by herbicide application. Our results from a controlled experiment at the local scale illustrate the potential for the global mapping of terrestrial photosynthesis through space-borne measurements of chlorophyll fluorescence.

  1. 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.

  2. 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.

  3. 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.

  4. Micro-light guides: a new method for measuring tissue fluorescence and reflectance.

    PubMed

    Ji, S; Chance, B; Nishiki, K; Smith, T; Rich, T

    1979-03-01

    Three-way light guides containing one or more strands of 25-micron or 80-micron diameter optical fibers in each channel have been constructed and used to measure the NADH fluorescence and UV reflectance from mitochondrial suspensions, the perfused, hemoglobin-free rat liver, and the perfused beating interventricular septum of the rabbit. The optical changes measured with these so-called micro-light guides, which have channels containing one or several strands of optical fibers less than 100 micron, are comparable in magnitude with those measured using much larger conventional light guides. The effect of light scattering on the fluorescence channel has been determined and an empirical equation for correcting the fluorescence channel for light scattering has been obtained for mitochondrial suspensions. A mathematical equation characterizing the optical behavior of a two-way micro-light guide has been derived and has been shown to account satisfactorily for reflectance and fluorescence measurements of a mat surface in air.

  5. 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.

  6. 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.

  7. Biliary fluorescent aromatic compounds (FACs) measured by fixed wavelength fluorescence (FF) in several marine fish species from the NW Mediterranean.

    PubMed

    Insausti, David; Carrasson, Maite; Maynou, Francesc; Cartes, Joan E; Solé, Montserrat

    2009-11-01

    The fixed wavelength fluoresce (FF) method was used to estimate the levels of fluorescent aromatic compounds (FACs) in the bile of fourteen fish species of commercial and/or ecological interest. Sampling was carried out in the NW Mediterranean at depths ranging from 50 to 1000 m during four seasonal cruises. During the summer sampling period, some species were also collected from another site (Vilanova fishing grounds) for comparison. Baseline levels of the polycyclic aromatic hydrocarbons (PAHs) naphthalene, phenanthrene, pyrene and benzo[a]pyrene were measured. Some seasonality was observed, with reduced FF levels in summer and no differences among sites, consistent with sediment PAH levels. We discuss our results in relation to fish phylogeny, season, depth, diet, trophic level and swimming capacity. Overall FF levels indicated differences among species; the suprabenthic feeders from shallow and deep communities, and Mullus barbatus in particular, displayed elevated FF values and are potential candidates for additional monitoring studies.

  8. Comparison of cone and cone shell configuration for depth sensitive fluorescence measurements in turbid media

    NASA Astrophysics Data System (ADS)

    Ong, Yi Hong; Liu, Quan

    2014-03-01

    We have developed a novel cone shell illumination and detection configurations using combination of axicon lenses for depth sensitive fluorescence spectroscopy. The probe was demonstrated experimentally to be able to selectively detecting fluorescence from different depths from a two-layered turbid agar phantom. In addition to enhanced contrast of subsurface fluorescence measurement as compared to a conventional cone configuration implemented by a microscope objective lens, the axicon lenses based setup eliminated the need of moving the objective lens up or down to achieve depth sensitive measurements, which effectively improves the consistency of optical coupling thus would be preferred in a clinical setting.

  9. 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

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

    SciTech Connect

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

    2010-03-15

    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 {mu}m{sup 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.

  11. 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.

  12. 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.

  13. 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. PMID:19568951

  14. Fluorescence spectroscopy of kerosene vapour at high temperatures and pressures: potential for gas turbines measurements

    NASA Astrophysics Data System (ADS)

    Orain, M.; Baranger, P.; Ledier, C.; Apeloig, J.; Grisch, F.

    2014-09-01

    Laser-induced fluorescence spectroscopy of kerosene vapour was performed in a heated test cell operating between 450 and 900 K, at pressure from 0.1 to 3.0 MPa, for oxygen molar fraction between 0 and 21 %, with different laser excitation wavelengths (248, 266, 282 and 308 nm). Results show that, depending on the laser excitation scheme, kerosene fluorescence spectrum exhibits one or two fluorescence bands in the UV-visible range (attributed to aromatics naturally present in kerosene fuel). Fluorescence intensity of these bands decreases with increasing temperature, pressure and oxygen molar fraction. Different imaging strategies were derived from spectroscopic findings to simultaneously measure temperature and equivalence ratio fields in kerosene/air sprays, or flame structure and fuel spatial distribution in kerosene/air aeronautical combustors, by means of planar laser-induced fluorescence on kerosene vapour (K-PLIF).

  15. Bright Fraction of Single-Walled Carbon Nanotubes through Correlated Fluorescence and Topography Measurements.

    PubMed

    Nogaj, Lisa J; Smyder, Julie A; Leach, Kathryn E; Tu, Xiaomin; Zheng, Ming; Krauss, Todd D

    2015-07-16

    Correlated measurements of fluorescence and topography were performed for individual single-walled carbon nanotubes (SWNTs) on quartz using epifluorescence confocal microscopy and atomic force microscopy (AFM). Surprisingly, only ~11% of all SWNTs in DNA-wrapped samples were found to be highly emissive on quartz, suggesting that the ensemble fluorescence quantum yield is low because only a small population of SWNTs fluoresces strongly. Qualitatively similar conclusions were obtained from control studies using a sodium cholate surfactant system. To accommodate AFM measurements, excess surfactant was removed from the substrate. Though individual SWNTs on nonrinsed and rinsed surfaces displayed differences in fluorescence intensities and line widths, arising from the influence of the local environment on individual SWNT optical measurements, photoluminescence data from both samples displayed consistent trends.

  16. 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.

  17. 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.

  18. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2016-02-01

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

  20. 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.

  1. 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.

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

    PubMed

    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, ϵ , is around 5 in lipid droplets and 25<ϵ<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.

  3. 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...

  4. 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.

  5. τFCS: Multi-Method Global Analysis Enhances Resolution and Sensitivity in Fluorescence Fluctuation Measurements

    PubMed Central

    Anthony, Neil R.; Berland, Keith M.

    2014-01-01

    Fluorescence fluctuation methods have become invaluable research tools for characterizing the molecular-level physical and chemical properties of complex systems, such as molecular concentrations, dynamics, and the stoichiometry of molecular interactions. However, information recovery via curve fitting analysis of fluctuation data is complicated by limited resolution and challenges associated with identifying accurate fit models. We introduce a new approach to fluorescence fluctuation spectroscopy that couples multi-modal fluorescence measurements with multi-modal global curve fitting analysis. This approach yields dramatically enhanced resolution and fitting model discrimination capabilities in fluctuation measurements. The resolution enhancement allows the concentration of a secondary species to be accurately measured even when it constitutes only a few percent of the molecules within a sample mixture, an important new capability that will allow accurate measurements of molecular concentrations and interaction stoichiometry of minor sample species that can be functionally important but difficult to measure experimentally. We demonstrate this capability using τFCS, a new fluctuation method which uses simultaneous global analysis of fluorescence correlation spectroscopy and fluorescence lifetime data, and show that τFCS can accurately recover the concentrations, diffusion coefficients, lifetimes, and molecular brightness values for a two component mixture over a wide range of relative concentrations. PMID:24587370

  6. Quantitative liquid and vapor distribution measurements in evaporating fuel sprays using laser-induced exciplex fluorescence

    NASA Astrophysics Data System (ADS)

    Fansler, Todd D.; Drake, Michael C.; Gajdeczko, Boguslaw; Düwel, Isabell; Koban, Wieland; Zimmermann, Frank P.; Schulz, Christof

    2009-12-01

    Fully quantitative two-dimensional measurements of liquid- and vapor-phase fuel distributions (mass per unit volume) from high-pressure direct-injection gasoline injectors are reported for conditions of both slow and rapid vaporization in a heated, high-pressure spray chamber. The measurements employ the coevaporative gasoline-like fluorobenzene (FB)/diethylmethylamine (DEMA)/hexane exciplex tracer/fuel system. In contrast to most previous laser-induced exciplex-fluorescence (LIEF) experiments, the quantitative results here include regions in which liquid and vapor fuel coexist (e.g. near the injector exit). A unique aspect is evaluation of both vapor- and liquid-phase distributions at varying temperature and pressure using only in situ vapor-phase fluorescence calibration measurements at room temperature and atmospheric pressure. This approach draws on recent extensive measurements of the temperature-dependent spectroscopic properties of the FB-DEMA exciplex system, in particular on knowledge of the quantum efficiencies of the vapor-phase and liquid-phase (exciplex) fluorescence. In addition to procedures necessary for quantitative measurements, we discuss corrections for liquid-vapor crosstalk (liquid fluorescence that overlaps the vapor-fluorescence bandpass), the unknown local temperature due to vaporization-induced cooling, and laser-sheet attenuation by scattering and absorption.

  7. 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.

  8. 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.

  9. Effect of biofilm on fluorescence measurements derived from fast repetition rate fluorometers.

    PubMed

    Patil, Jagadish S; Saino, Toshiro

    2015-01-01

    This study evaluates, for the first time, the influence of biofilms on single and double optical window (SOW and DOW, respectively) fast repetition rate fluorometer (FRRF) measurements of microalgal photosystem-II initial fluorescence (F0), maximum fluorescence (Fm), variable fluorescence (Fv = Fm - F0), quantum yield (Fv/Fm) and functional absorption cross section (σPSII)]. Biofilms with chlorophyll > 0.1 μg cm(-2) and > 0.3 μgcm(-2) on SOW and DOW, respectively, produced a substantial increase in fluorescence. However, the relative magnitude of biofouling effects depended on sample chlorophyll concentrations, being more critical at concentrations < 1 mg m(-3). In DOW-FRRF, biofilms affected F0 (increased) and Fv/Fm (decreased) but not Fv and σPSII, whereas in SOW-FRRF, biofilms increased fluorescence and showed a variable effect on Fv/Fm and σPSII, because only biofilms on SOW attained actual Fm. As a result, the biofilm effect was substantial on SOW-FRRF measurements. On the other hand, the neutral-density filters (representing non-chlorophyll containing biofilms) with different transmission levels reduced the fluorescence signal. Correction procedures for the above photosystem-II parameters are proposed here.

  10. Simultaneous measurement of oscillations in oxygen evolution and chlorophyll a fluorescence in leaf pieces.

    PubMed

    Walker, D A; Sivak, M N; Prinsley, R T; Cheesbrough, J K

    1983-11-01

    In spinach (Spinacia oleracea) and barley (Hordeum vulgare) leaves, chlorophyll a fluorescence and O(2) evolution have been measured simultaneously following re-illumination after a dark interval or when steady state photosynthesis has been perturbed by changes in the gas phase. In high CO(2) concentrations, both O(2) and fluorescence can display marked dampening oscillations that are antiparallel but slightly out of phase (a rise or fall in fluorescence anticipating a corresponding fall or rise in O(2) by about 10 to 15 seconds). Infrared gas analysis measurements showed that CO(2) uptake behaved like O(2) evolution both in the period of oscillation (about 1 minute) and in its relation to fluorescence. In the steady state, oscillations were initiated by increases in CO(2) or by increases or decreases in O(2). Oscillations in O(2) or CO(2) did not occur without associated oscillations in fluorescence and the latter were a sensitive indicator of the former. The relationship between such oscillations in photosynthetic carbon assimilation and chlorophyl a fluorescence is discussed in the context of the effect of ATP or NADPH consumption on known quenching mechanisms. PMID:16663255

  11. 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.

  12. 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

  13. Fluorescence spectroscopy: a rapid, noninvasive method for measurement of skin surface thickness of topical agents.

    PubMed

    Rhodes, L E; Diffey, B L

    1997-01-01

    We report the quantification of skin surface thickness of topical agents by in vivo fluorescence spectroscopy, and demonstrate its potential uses for assessment of application technique and substantivity. A series of studies were performed on forearm skin of eight normal subjects using three creams which have intrinsic fluorescence: a sunscreen (Neutrogena SPF15 waterproof cream), an antiseptic (Hewlett's cream) and a steroid (Trimovate (clobetasone butyrate) cream). Initially, the dose-response relationship was established for each agent by applying a series of five doses (0.5-8 microliters/cm2) and measuring cream fluorescence using appropriate excitation and emission wavelengths. Next, the influence of application technique was examined by comparing light application of cream with firm rubbing. Substantivity of the three creams was assessed on dry skin by taking fluorescence measurements over 8 h. Finally, water resistance of 2 microliters/cm2 of sunscreen and antiseptic cream were compared by measuring fluorescence after each of four water immersions. The fluorescence intensity was strongly correlated with the logarithm of surface density. r = 1.0, 0.92 and 0.98 for sunscreen, antiseptic and steroid creams, respectively, allowing derivation of a simple expression for equivalent thickness. Surface thickness of each cream was lower following firm rubbing compared with light application (P < 0.01). The rate constants for reduction of surface density of the three creams with time on dry skin were not significantly different. However, on washed skin, the rate constant was higher for Hewlett's than Neutrogena cream (0.503 and 0.243 h. respectively, P = 0.02), with a higher rate for each cream on wet compared with dry skin (P < 0.001). Hence, fluorescence spectroscopy is a simple, rapid method for measurement of cream thickness in vivo. The many potential applications in dermatology include quantitative assessment of application technique and substantivity of topical

  14. Laser induced fluorescence measurements of dissolved oxygen concentration fields near air bubble surfaces

    NASA Astrophysics Data System (ADS)

    Roy, Sabita; Duke, Steve R.

    2000-09-01

    This article describes a laser-induced fluorescence (LIF) technique for measuring dissolved oxygen concentration gradients in water near the surface of an air bubble. Air bubbles are created at the tip of a needle in a rectangular bubble column filled with water that contains pyrenebutyric acid (PBA). The fluorescence of the PBA is induced by a planar pulse of nitrogen laser light. Oxygen transferring from the air bubble to the deoxygenated water quenches the fluorescence of the PBA. Images of the instantaneous and two-dimensional fluorescence field are obtained by a UV-intensified charge-coupled device (CCD) camera. Quenching of fluorescence intensity is determined at each pixel in the CCD image to measure dissolved oxygen concentration. Two-dimensional concentration fields are presented for a series of measurements of oxygen transfer from 1.6 mm bubbles suspended on the tip of a needle in a quiescent fluid. The images show the spatially varying concentration profiles, gradients, and boundary layer thicknesses at positions around the bubble surfaces. These direct and local measurements of concentration behavior within the mass transfer boundary layer show the potential of this LIF technique for the development of general and mechanistic models for oxygen transport across the air-water interface.

  15. 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.

  16. 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.

  17. Quantitative fluorescence measurements of the OH radical in high pressure methane flames

    SciTech Connect

    Battles, B.E.; Hanson, R.K. )

    1992-07-01

    A method for quantifying laser-induced fluorescence signals from the OH radical in high-pressure flames is presented. The fluorescence signal per unit OH mole fraction is modeled as a function of temperature, pressure, and overall flame stoichiometry. Known values of the collisional quenching cross sections as a function of temperature are used to model the electronic quench rate. The reverse A - X (1.0) Q15 transition is used with broadband collection to measure single-point fluorescence produced by a pulsed Nd:YAG-pumped, frequency-doubled dye laser. Laser absorption and thermocouples are used to measure absolute OH concentration and temperature, respectively, which are used to confirm the validity of the model. Measurements are made in CH4/O2/N2 flames up to 10 atm. 13 refs.

  18. Quantitative fluorescence measurements of the OH radical in high pressure methane flames

    NASA Technical Reports Server (NTRS)

    Battles, B. E.; Hanson, R. K.

    1992-01-01

    A method for quantifying laser-induced fluorescence signals from the OH radical in high-pressure flames is presented. The fluorescence signal per unit OH mole fraction is modeled as a function of temperature, pressure, and overall flame stoichiometry. Known values of the collisional quenching cross sections as a function of temperature are used to model the electronic quench rate. The reverse A - X (1.0) Q15 transition is used with broadband collection to measure single-point fluorescence produced by a pulsed Nd:YAG-pumped, frequency-doubled dye laser. Laser absorption and thermocouples are used to measure absolute OH concentration and temperature, respectively, which are used to confirm the validity of the model. Measurements are made in CH4/O2/N2 flames up to 10 atm.

  19. 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…

  20. Experimental Test of New Technique to Overcome Spin Depolarizing Resonances

    SciTech Connect

    Raymond, R. S.; Chao, A. W.; Krisch, A. D.; Leonova, M. A.; Morozov, V. S.; Sivers, D. W.; Wong, V. K.; Ganshvili, A.; Gebel, R.; Lehrach, A.; Lorentz, B.; Maier, R.; Prasuhn, D.; Stockhorst, H.; Welsch, D.; Hinterberger, F.; Kondratenko, A. M.

    2009-08-04

    We recently tested a new spin resonance crossing technique, Kondratenko Crossing (KC) by sweeping an rf solenoid's frequency through an rf-induced spin resonance with both the KC an traditional Fast Crossing (FC) patterns. Using both rf bunched and unbunched 1.85 GeV/c polarized deuterons stored in COSY, we varied the parameters of both crossing patterns. Compared to FC with the same crossing speed, KC reduced the depolarization by measured factors of 4.7+-0.3 and 19+-{sub 5}{sup 12} for unbunched and bunched beams, respectively. This clearly showed the large potential benefit of Kondratenko Crossing over Fast Crossing.

  1. Calibration of the Pierre Auger Observatory fluorescence detectors and the effect on measurements

    NASA Astrophysics Data System (ADS)

    Gookin, Ben

    The Pierre Auger Observatory is a high-energy cosmic ray observatory located in Malargue, Mendoza, Argentina. It is used to probe the highest energy particles in the Universe, with energies greater than 1018 eV, which strike the Earth constantly. The observatory uses two techniques to observe the air shower initiated by a cosmic ray: a surface detector composed of an array of more than 1600 water Cherenkov tanks covering 3000 km2, and 27 nitrogen fluorescence telescopes overlooking this array. The Cherenkov detectors run all the time and therefore have high statistics on the air showers. The fluorescence detectors run only on clear moonless nights, but observe the longitudinal development of the air shower and make a calorimetric measure of its energy. The energy measurement from the the fluorescence detectors is used to cross calibrate the surface detectors, and makes the measurements made by the Auger Observatory surface detector highly model-independent. The calibration of the fluorescence detectors is then of the utmost importance to the measurements of the Observatory. Described here are the methods of the absolute and multi-wavelength calibration of the fluorescence detectors, and improvements in each leading to a reduction in calibration uncertainties to 4% and 3.5%, respectively. Also presented here are the effects of introducing a new, and more detailed, multi-wavelength calibration on the fluorescence detector energy estimation and the depth of the air shower maximum measurement, leading to a change of 1+-0.03% in the absolute energy scale at 1018 eV, and a negligible change in the measurement on shower maximum.

  2. 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

  3. Exploring Neuronal Bistability at the Depolarization Block

    PubMed Central

    Dovzhenok, Andrey; Kuznetsov, Alexey S.

    2012-01-01

    Many neurons display bistability–coexistence of two firing modes such as bursting and tonic spiking or tonic spiking and silence. Bistability has been proposed to endow neurons with richer forms of information processing in general and to be involved in short-term memory in particular by allowing a brief signal to elicit long-lasting changes in firing. In this paper, we focus on bistability that allows for a choice between tonic spiking and depolarization block in a wide range of the depolarization levels. We consider the spike-producing currents in two neurons, models of which differ by the parameter values. Our dopaminergic neuron model displays bistability in a wide range of applied currents at the depolarization block. The Hodgkin-Huxley model of the squid giant axon shows no bistability. We varied parameter values for the model to analyze transitions between the two parameter sets. We show that bistability primarily characterizes the inactivation of the Na+ current. Our study suggests a connection between the amount of the Na+ window current and the length of the bistability range. For the dopaminergic neuron we hypothesize that bistability can be linked to a prolonged action of antipsychotic drugs. PMID:22900051

  4. 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.

  5. 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.

  6. 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.

  7. Direct measurement of off-state trapping rate fluctuations in single quantum dot fluorescence.

    PubMed

    Cordones, Amy A; Bixby, Teresa J; Leone, Stephen R

    2011-08-10

    Fluorescence decay times measured during the off-state of single CdSe/ZnS quantum dot blinking are found to decrease with increasing off-state duration, contradicting the charging model widely considered to explain the blinking phenomenon. The change in the nonradiative process of a short off-state duration compared to a long one is investigated here through simultaneous measurement of fluorescence decay and blinking behavior. The results are investigated in the framework of two models based on fluctuating trapping rates.

  8. Measurement of partial L fluorescence yields of bismuth using synchrotron radiation.

    PubMed

    Ménesguen, Yves; Boyer, Bruno; Rodrigues, Matias; Lépy, Marie-Christine

    2016-03-01

    Tunable monochromatic photon radiation was used to measure transmission of a bismuth target in the energy range from 7keV to 20keV. Partial L fluorescence yields of bismuth were obtained by combining measurement of the fluorescence induced by photoionization of the bismuth target and X-rays from the radioactive decay of (210)Pb. Several photon energies have been used to successively ionize the L subshells, which allowed detailed analysis of the rearrangement spectra and determination of the X-ray relative intensities of the L1, L2 and L3 series. PMID:26651165

  9. 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.

  10. 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.

  11. A novel fluorescent sensor for measurement of CFTR function by flow cytometry.

    PubMed

    Vijftigschild, Lodewijk A W; van der Ent, Cornelis K; Beekman, Jeffrey M

    2013-06-01

    Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene cause cystic fibrosis. CFTR-dependent iodide transport measured by fluorescent quenching of ectopically expressed halide-sensitive yellow fluorescent protein (YFP) is widely being used to study CFTR function by microscopy or plate readers. Since YFP fluorescence in these systems is dependent on YFP expression levels and iodide concentration, differences in sensor expression level between experimental units are normalized at the start of each experiment. To allow accurate measurement of CFTR function by flow cytometry, we reasoned that co-expression of an iodide insensitive fluorescent protein would allow for normalization of sensor expression levels and more accurate quantification of CFTR function. Our data indicated that dsRed and mKate fluorescence are iodide insensitive, and we determined an optimal format for co-expression of these fluorescent proteins with halide-sensitive YFP. We showed using microscopy that ratiometric measurement (YFP/mKate) corrects for differences in sensor expression levels. Ratiometric measurements were essential to accurately measure CFTR function by flow cytometry that we here describe for the first time. Mixing of wild type or mutant CFTR expressing cells indicated that addition of approximately 10% of wild type CFTR expressing cells could be distinguished by ratiometric YFP quenching. Flow cytometric ratiometric YFP quenching also allowed us to study CFTR mutants associated with differential residual function upon ectopic expression. Compared with conventional plate-bound CFTR function assays, the flow cytometric approach described here can be used to study CFTR function in suspension cells. It may be further adapted to study CFTR function in heterologous cell populations using cell surface markers and selection of cells that display high CFTR function by cell sorting.

  12. Algal fluorescence: impact and potential for retrieval from measurements of the underwater degree of polarization

    NASA Astrophysics Data System (ADS)

    Ahmed, S.; Tonizzo, A.; Ibrahim, A.; Gilerson, A.; Gross, B.; Moshary, F.

    2012-09-01

    Algorithms for retrieving inherent optical properties (IOPs) in coastal waters from remote sensing of water leaving reflectance spectra, are increasingly focused on red and near infrared (NIR) spectral bands, since the simple blue - green ratio approaches, valid in open oceans, fail when in coastal waters with strongly scattering inorganic particles and colored dissolved organic matter (CDOM). NIR spectra can however be significantly impacted by overlapping chlorophyll a fluorescence, and considerable progress has been made to quantify its contribution, and hence achieve more accurate [Chl] retrievals. Recently we have been studying multiangular hyperspectral polarization characteristics of underwater scattered light, using our recently developed Stokes vector polarimeter to fully measure Stokes parameters. From these studies, information on IOPs, in particular the characteristics of non - algal particles (NAP), which are the primary source of underwater polarized elastic scattering, can be obtained. Multiangular hyperspectral polarization measurements, combined with those of IOPs collected in eutrophic waters of Chesapeake/Virginia and New York Harbor/Hudson River areas, showed that chlorophyll a fluorescence markedly impacts (reduces) the underwater degree of polarization (DOP) in the 650 - 700 nm spectral region. By noting the unpolarized nature of algal fluorescence and the partially polarized properties of elastic scattering, we are able to separate the chlorophyll a fluorescence signal from the total reflectance. The analysis is based on comparisons of experimental measurements with vector/scalar radiative transfer computations using measured IOPs as inputs. Relationships between change in observed DOP and fluorescence contributions are examined, and the possibility of using DOP measurements for underwater fluorescence retrieval is evaluated for different scattering geometries.

  13. A portable time-domain LED fluorimeter for nanosecond fluorescence lifetime measurements

    NASA Astrophysics Data System (ADS)

    Wang, Hongtao; Qi, Ying; Mountziaris, T. J.; Salthouse, Christopher D.

    2014-05-01

    Fluorescence lifetime measurements are becoming increasingly important in chemical and biological research. Time-domain lifetime measurements offer fluorescence multiplexing and improved handling of interferers compared with the frequency-domain technique. In this paper, an all solid-state, filterless, and highly portable light-emitting-diode based time-domain fluorimeter (LED TDF) is reported for the measurement of nanosecond fluorescence lifetimes. LED based excitation provides more wavelengths options compared to laser diode based excitation, but the excitation is less effective due to the uncollimated beam, less optical power, and longer latency in state transition. Pulse triggering and pre-bias techniques were implemented in our LED TDF to improve the peak optical power to over 100 mW. The proposed pulsing circuit achieved an excitation light fall time of less than 2 ns. Electrical resetting technique realized a time-gated photo-detector to remove the interference of the excitation light with fluorescence. These techniques allow the LED fluorimeter to accurately measure the fluorescence lifetime of fluorescein down to concentration of 0.5 μM. In addition, all filters required in traditional instruments are eliminated for the non-attenuated excitation/emission light power. These achievements make the reported device attractive to biochemical laboratories seeking for highly portable lifetime detection devices for developing sensors based on fluorescence lifetime changes. The device was initially validated by measuring the lifetimes of three commercial fluorophores and comparing them with reported lifetime data. It was subsequently used to characterize a ZnSe quantum dot based DNA sensor.

  14. A portable time-domain LED fluorimeter for nanosecond fluorescence lifetime measurements

    SciTech Connect

    Wang, Hongtao; Salthouse, Christopher D.; Qi, Ying; Mountziaris, T. J.

    2014-05-15

    Fluorescence lifetime measurements are becoming increasingly important in chemical and biological research. Time-domain lifetime measurements offer fluorescence multiplexing and improved handling of interferers compared with the frequency-domain technique. In this paper, an all solid-state, filterless, and highly portable light-emitting-diode based time-domain fluorimeter (LED TDF) is reported for the measurement of nanosecond fluorescence lifetimes. LED based excitation provides more wavelengths options compared to laser diode based excitation, but the excitation is less effective due to the uncollimated beam, less optical power, and longer latency in state transition. Pulse triggering and pre-bias techniques were implemented in our LED TDF to improve the peak optical power to over 100 mW. The proposed pulsing circuit achieved an excitation light fall time of less than 2 ns. Electrical resetting technique realized a time-gated photo-detector to remove the interference of the excitation light with fluorescence. These techniques allow the LED fluorimeter to accurately measure the fluorescence lifetime of fluorescein down to concentration of 0.5 μM. In addition, all filters required in traditional instruments are eliminated for the non-attenuated excitation/emission light power. These achievements make the reported device attractive to biochemical laboratories seeking for highly portable lifetime detection devices for developing sensors based on fluorescence lifetime changes. The device was initially validated by measuring the lifetimes of three commercial fluorophores and comparing them with reported lifetime data. It was subsequently used to characterize a ZnSe quantum dot based DNA sensor.

  15. A portable time-domain LED fluorimeter for nanosecond fluorescence lifetime measurements.

    PubMed

    Wang, Hongtao; Qi, Ying; Mountziaris, T J; Salthouse, Christopher D

    2014-05-01

    Fluorescence lifetime measurements are becoming increasingly important in chemical and biological research. Time-domain lifetime measurements offer fluorescence multiplexing and improved handling of interferers compared with the frequency-domain technique. In this paper, an all solid-state, filterless, and highly portable light-emitting-diode based time-domain fluorimeter (LED TDF) is reported for the measurement of nanosecond fluorescence lifetimes. LED based excitation provides more wavelengths options compared to laser diode based excitation, but the excitation is less effective due to the uncollimated beam, less optical power, and longer latency in state transition. Pulse triggering and pre-bias techniques were implemented in our LED TDF to improve the peak optical power to over 100 mW. The proposed pulsing circuit achieved an excitation light fall time of less than 2 ns. Electrical resetting technique realized a time-gated photo-detector to remove the interference of the excitation light with fluorescence. These techniques allow the LED fluorimeter to accurately measure the fluorescence lifetime of fluorescein down to concentration of 0.5 μM. In addition, all filters required in traditional instruments are eliminated for the non-attenuated excitation/emission light power. These achievements make the reported device attractive to biochemical laboratories seeking for highly portable lifetime detection devices for developing sensors based on fluorescence lifetime changes. The device was initially validated by measuring the lifetimes of three commercial fluorophores and comparing them with reported lifetime data. It was subsequently used to characterize a ZnSe quantum dot based DNA sensor.

  16. Evaluation of slide based cytometry (SBC) for concentration measurements of fluorescent dyes in solution

    NASA Astrophysics Data System (ADS)

    Pierzchalski, Arkadiusz; Marecka, Monika; Müller, Hans-Willy; Bocsi, József; Tárnok, Attila

    2009-02-01

    Flow cytometers (FCM) are built for particle measurements. In principle, concentration measurement of a homogeneous solution is not possible with FCM due to the lack of a trigger signal. In contrast to FCM slide based cytometry systems could act as tools for the measurement of concentrations using volume defined cell counting chambers. These chambers enable to analyze a well defined volume. Sensovation AG (Stockach, Germany) introduced an automated imaging system that combines imaging with cytometric features analysis. Aim of this study was to apply this imaging system to quantify the fluorescent molecule concentrations. The Lumisens (Sensovation AG) slide-based technology based on fluorescence digital imaging microscopy was used. The instrument is equipped with an inverted microscope, blue and red LEDs, double band-pass filters and a high-resolution cooled 16-bit digital camera. The instrument was focussed on the bottom of 400μm deep 6 chamber slides (IBIDI GmbH, Martinsried, Germany) or flat bottom 96 well plates (Greiner Bio One GmbH, Frickenhausen, Germany). Fluorescent solutions were imaged under 90% pixel saturation in a broad concentration range (FITC: 0.0002-250 μg/ml, methylene blue (MethB): 0.0002-250 μg/ml). Exposition times were recorded. Images were analysed by the iCys (CompuCyte Corp., Cambridge, MA, USA) image analysis software with the phantom contour function. Relative fluorescence intensities were calculated from mean fluorescence intensities per phantom contours divided by the exposition time. Solution concentrations could be distinguished over a broad dynamic range of 3.5 to 5.5 decades log (range FITC: 0.0002-31.25μg/ml, MethB: 0.0076-31.25μg/ml) with a good linear relationship between dye concentration and relative fluorescence intensity. The minimal number of fluorescent molecules per pixel as determined by the mean fluorescence intensity and the molecular weight of the fluorochrome were about 800 molecules FITC and ~2.000 MethB. The

  17. 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.

  18. Ischemia-induced spreading depolarization in the retina.

    PubMed

    Srienc, Anja I; Biesecker, Kyle R; Shimoda, Angela M; Kur, Joanna; Newman, Eric A

    2016-09-01

    Cortical spreading depolarization is a metabolically costly phenomenon that affects the brain in both health and disease. Following severe stroke, subarachnoid hemorrhage, or traumatic brain injury, cortical spreading depolarization exacerbates tissue damage and enlarges infarct volumes. It is not known, however, whether spreading depolarization also occurs in the retina in vivo. We report now that spreading depolarization episodes are generated in the in vivo rat retina following retinal vessel occlusion produced by photothrombosis. The properties of retinal spreading depolarization are similar to those of cortical spreading depolarization. Retinal spreading depolarization waves propagate at a velocity of 3.0 ± 0.1 mm/min and are associated with a negative shift in direct current potential, a transient cessation of neuronal spiking, arteriole constriction, and a decrease in tissue O2 tension. The frequency of retinal spreading depolarization generation in vivo is reduced by administration of the NMDA antagonist MK-801 and the 5-HT(1D) agonist sumatriptan. Branch retinal vein occlusion is a leading cause of vision loss from vascular disease. Our results suggest that retinal spreading depolarization could contribute to retinal damage in acute retinal ischemia and demonstrate that pharmacological agents can reduce retinal spreading depolarization frequency after retinal vessel occlusion. Blocking retinal spreading depolarization generation may represent a therapeutic strategy for preserving vision in branch retinal vein occlusion patients.

  19. Ischemia-induced spreading depolarization in the retina.

    PubMed

    Srienc, Anja I; Biesecker, Kyle R; Shimoda, Angela M; Kur, Joanna; Newman, Eric A

    2016-09-01

    Cortical spreading depolarization is a metabolically costly phenomenon that affects the brain in both health and disease. Following severe stroke, subarachnoid hemorrhage, or traumatic brain injury, cortical spreading depolarization exacerbates tissue damage and enlarges infarct volumes. It is not known, however, whether spreading depolarization also occurs in the retina in vivo. We report now that spreading depolarization episodes are generated in the in vivo rat retina following retinal vessel occlusion produced by photothrombosis. The properties of retinal spreading depolarization are similar to those of cortical spreading depolarization. Retinal spreading depolarization waves propagate at a velocity of 3.0 ± 0.1 mm/min and are associated with a negative shift in direct current potential, a transient cessation of neuronal spiking, arteriole constriction, and a decrease in tissue O2 tension. The frequency of retinal spreading depolarization generation in vivo is reduced by administration of the NMDA antagonist MK-801 and the 5-HT(1D) agonist sumatriptan. Branch retinal vein occlusion is a leading cause of vision loss from vascular disease. Our results suggest that retinal spreading depolarization could contribute to retinal damage in acute retinal ischemia and demonstrate that pharmacological agents can reduce retinal spreading depolarization frequency after retinal vessel occlusion. Blocking retinal spreading depolarization generation may represent a therapeutic strategy for preserving vision in branch retinal vein occlusion patients. PMID:27389181

  20. Direct spectrometry: a new alternative for measuring the fluorescence of composite resins and dental tissues.

    PubMed

    da Silva, Tm; de Oliveira, Hpm; Severino, D; Balducci, I; Huhtala, Mfrl; Gonçalves, Sep

    2014-01-01

    The aim of this study was to evaluate the fluorescence intensity of different composite resins and compare those values with the fluorescence intensity of dental tissues. Different composite resins were used to make 10 discs (2 mm in depth and 4 mm in diameter) of each brand, divided into groups: 1) Z (Filtek Z350, 3M ESPE), 2) ES (Esthet-X, Dentsply), 3) A (Amelogen Plus, Ultradent), 4) DVS (Durafill-VS, Heraeus Kulzer) with 2 mm composite resin for enamel (A2), 5) OES ([Esthet-X] opaque-OA [1 mm] + enamel-A2 [1 mm]); 6) ODVSI ([Charisma-Opal/Durafill-VSI], opaque-OM (1 mm) + translucent [1mm]), and 7) DVSI ([Durafill- VSI] translucent [2 mm]). Dental tissue specimens were obtained from human anterior teeth cut in a mesiodistal direction to obtain enamel, dentin, and enamel/dentin samples (2 mm). The fluorescence intensity of specimens was directly measured using an optic fiber associated with a spectrometer (Ocean Optics USB 4000) and recorded in graphic form (Origin 8.0 program). Data were submitted to statistical analysis using Dunnet, Tukey, and Kruskall-Wallis tests. Light absorption of the composite resins was obtained in a spectral range from 250 to 450 nm, and that of dental tissues was between 250 and 300 nm. All composite resins were excited at 398 nm and exhibited maximum emissions of around 485 nm. Fluorescence intensity values for all of the resins showed statistically significant differences (measured in arbitrary units [AUs]), with the exception of groups Z and DVS. Group DVSI had the highest fluorescence intensity values (13539 AU), followed by ODVS (10440 AU), DVS (10146 AU), ES (3946 AU), OES (3841 AU), A (3540 AU), and Z (1146 AU). The fluorescence intensity values for the composite resins differed statistically from those of dental tissues (E=1380 AU; D=6262 AU; E/D=3251 AU). The opacity interfered with fluorescence intensity, and group Z demonstrated fluorescence intensity values closest to that of tooth enamel. It is concluded that the

  1. Evidence that plasma membrane electrical potential is required for vesicular stomatitis virus infection of MDCK cells: a study using fluorescence measurements through polycarbonate supports.

    PubMed

    Akeson, M; Scharff, J; Sharp, C M; Neville, D M

    1992-01-01

    We used fluorescence microscopy of Madin-Darby Canine Kidney (MDCK) cells grown on polycarbonate filters to study a possible link between plasma membrane electrical potential (delta psi pm) and infectivity of vesicular stomatitis virus (VSV). Complete substitution of K+ for extracellular Na+ blocks VSV infection of MDCK cells as well as baby hamster kidney (BHK) cells. When we independently perfused the apical and basal-lateral surfaces of high resistance monolayers, high K+ inhibited VSV infection of MDCK cells only when applied to the basal-lateral side; high K+ applied apically had no effect on VSV infection. This morphological specificity correlates with a large decrease in delta psi pm of MDCK cells when high K+ buffer is perfused across the basal-lateral surface. Depolarization of the plasma membrane by 130 mM basal K+ causes a sustained increase of cytosol pH in MDCK cells from 7.3 to 7.5 as reported by the fluorescent dye BCECF. Depolarization also causes a transient increase of cytosol Ca2+ from 70 to 300 nM as reported by the dye Fura-2. Neither increase could explain the block of VSV infectivity by plasma membrane depolarization. One alternative hypothesis is that delta psi pm facilitates membrane translocation of viral macromolecules as previously described for colicins, mitochondrial import proteins, and proteins secreted by Escherichia coli.

  2. Time resolved laser induced fluorescence measurements: Considerations when using Nd:YAG based system

    NASA Astrophysics Data System (ADS)

    Rabasovic, Maja S.; Sevic, Dragutin; Terzic, Mira; Marinkovic, Bratislav P.

    2012-05-01

    Time-resolved laser-induced fluorescence (TR-LIF) and the laser induced breakdown spectroscopy (LIBS) have been shown to be methods which are fast and sensitive to provide information about the constituents in analyzed samples. TR-LIF and LIBS have similar hardware requirements. In this paper, we analyze some characteristics of TR-LIF/LIBS system implemented in our laboratory, considering the fact that the excitation part of the system is based on Nd:YAG laser and Optical Parametric Oscillator (OPO). The laser is more than powerful enough (365 mJ at 1064 nm, variable OPO output >5 mJ) for LIBS, but somehow slow (the length of fundamental laser harmonic output pulse is about 5 ns) for fluorescence measurements in our present area of interest, namely plants and food products. Fortunately, the pulse length of tunable OPO output (320-475 nm) is less then 1 ns, so by means of a correct deconvolution procedure it is possible to measure the fluorescence lifetimes in the range as small as a few nanoseconds. The fluorescence detection part of our system is based on picosecond streak camera. Using the fluorescent dyes (Rhodamine B and Fluorescein) ethanol solutions we verified the analyzing capabilities of our TR-LIF system.

  3. Measurement of chlorophyll a fluorescence with an airborne fluorosensor

    NASA Technical Reports Server (NTRS)

    Jarrett, O., Jr.; Brown, C. A., Jr.; Campbell, J. W.; Houghton, W. M.; Poole, L. R.

    1979-01-01

    Phytoplankton biomass and diversity among various algal species are important for marine productivity assessments. The spatial heterogeneity of phytoplankton in coastal and estuarine environments complicates estimates of total biomass using conventional surface sampling techniques. Since synoptic or near-synoptic data can be quite useful in these studies, this area is a natural focal point for development of remote sensors. However, it is very difficult to sense phytoplankton density and diversity with spacecraft-borne passive sensors primarily because modulation in the signal due to phytoplankton is of the same order as that of atmospheric effects. The same sensors mounted on aircraft may be able to detect and quantify high concentrations of phytoplankton (blooms), but the current lack of knowledge about the spectral reflectance signatures of the major phytoplankton color groups rules out any diversity measurements by this type of sensor. An active fluorosensor mounted on a low-flying aircraft or helicopter is not limited by any of these constraints. A brief survey of the four currently active systems is presented.

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

    PubMed

    Qian, Kun; Yu, Na; Tucker, Kristal R; Levitan, Edwin S; Canavier, Carmen C

    2014-12-01

    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.

  5. 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.

  6. 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.

  7. Probe pressure effects on human skin diffuse reflectance and fluorescence spectroscopy measurements.

    PubMed

    Lim, Liang; Nichols, Brandon; Rajaram, Narasimhan; Tunnell, James W

    2011-01-01

    Diffuse reflectance and fluorescence spectroscopy are popular research techniques for noninvasive disease diagnostics. Most systems include an optical fiber probe that transmits and collects optical spectra in contact with the suspected lesion. The purpose of this study is to investigate probe pressure effects on human skin spectroscopic measurements. We conduct an in-vivo experiment on human skin tissue to study the short-term (<2 s) and long-term (>30 s) effects of probe pressure on diffuse reflectance and fluorescence measurements. Short-term light probe pressure (P0<9 mN∕mm2) effects are within 0 ± 10% on all physiological properties extracted from diffuse reflectance and fluorescence measurements, and less than 0±5% for diagnostically significant physiological properties. Absorption decreases with site-specific variations due to blood being compressed out of the sampled volume. Reduced scattering coefficient variation is site specific. Intrinsic fluorescence shows a large standard error, although no specific pressure-related trend is observed. Differences in tissue structure and morphology contribute to site-specific probe pressure effects. Therefore, the effects of pressure can be minimized when the pressure is small and applied for a short amount of time; however, long-term and large pressures induce significant distortions in measured spectra. PMID:21280899

  8. Probe pressure effects on human skin diffuse reflectance and fluorescence spectroscopy measurements

    PubMed Central

    Lim, Liang; Nichols, Brandon; Rajaram, Narasimhan; Tunnell, James W.

    2011-01-01

    Diffuse reflectance and fluorescence spectroscopy are popular research techniques for noninvasive disease diagnostics. Most systems include an optical fiber probe that transmits and collects optical spectra in contact with the suspected lesion. The purpose of this study is to investigate probe pressure effects on human skin spectroscopic measurements. We conduct an in-vivo experiment on human skin tissue to study the short-term (<2 s) and long-term (>30 s) effects of probe pressure on diffuse reflectance and fluorescence measurements. Short-term light probe pressure (P0 < 9 mN∕mm2) effects are within 0 ± 10% on all physiological properties extracted from diffuse reflectance and fluorescence measurements, and less than 0 ± 5% for diagnostically significant physiological properties. Absorption decreases with site-specific variations due to blood being compressed out of the sampled volume. Reduced scattering coefficient variation is site specific. Intrinsic fluorescence shows a large standard error, although no specific pressure-related trend is observed. Differences in tissue structure and morphology contribute to site-specific probe pressure effects. Therefore, the effects of pressure can be minimized when the pressure is small and applied for a short amount of time; however, long-term and large pressures induce significant distortions in measured spectra. PMID:21280899

  9. Measurement of air and nitrogen fluorescence light yields induced by electron beam for UHECR experiments

    NASA Astrophysics Data System (ADS)

    Colin, P.; Chukanov, A.; Grebenyuk, V.; Naumov, D.; Nédélec, P.; Nefedov, Y.; Onofre, A.; Porokhovoi, S.; Sabirov, B.; Tkatchev, L.; Macfly Collaboration

    2007-06-01

    Most of the Ultra High Energy Cosmic Ray (UHECR) experiments and projects (HiRes, AUGER, TA, EUSO, TUS, etc.) use air fluorescence to detect and measure extensive air showers (EAS). The precise knowledge of the Fluorescence Light Yield (FLY) is of paramount importance for the reconstruction of UHECR. The MACFLY—Measurement of Air Cherenkov and Fluorescence Light Yield—experiment has been designed to perform such FLY measurements. In this paper we will present the results of FLY in the 290-440 nm wavelength range for dry air and pure nitrogen, both excited by electrons with energy of 1.5 MeV, 20 GeV and 50 GeV. The experiment uses a 90Sr radioactive source for low energy measurement and a CERN SPS e - beam for high energy. We find that the FLY is proportional to the deposited energy ( Ed) in the gas and we show that the air fluorescence properties remain constant independently of the electron energy. At the reference point: atmospheric dry air at 1013 hPa and 23 °C, the ratio FLY/ Ed = 17.6 photon/MeV with a systematic error of 13.2%.

  10. 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.

  11. Quantitative in vivo imaging of the lung using time-domain fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Ma, Guobin; Jean-Jacques, Muriel; Melanson-Drapeau, Lysanne; Khayat, Mario

    2009-02-01

    In this paper, nebulized or intravenous cetuximab (also known as Erbitux) labeled with NIR dyes is administered in the lungs of the mouse and imaged using a time-domain fluorescence imaging system (Optix(R)). Time resolved measurements provide lifetime of the fluorescent probes. In addition, through time-of-flight information contained in the data, one can also assess probe localization and concentration distribution quantitatively. Results shown include suppression of tissue autofluorescence by lifetime gating and recovery of targeted and non-targeted distributions of cetuximab labeled with the NIR fluorophores.

  12. 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.

  13. Remotely Measured Terrestrial Chlorophyll Fluorescence Using Airborne G-LiHT and APFS Sensors

    NASA Astrophysics Data System (ADS)

    Cook, W. B.; Yee, J. H.; Corp, L. A.; Cook, B. D.; Huemmrich, K. F.

    2014-12-01

    In September 2014 the Goddard Lidar, Hyperspectral and Thermal (G-LiHT) and the APL/JHU Airborne Plant Fluorescence Sensor (APFS) were flown together on a NASA Langley King Air over vegetated targets in North Carolina and Virginia. The instruments provided high spatial and spectral resolution data in the visible and near infrared, down-welling irradiance, elevation maps, and thermal imagery. Ground validation data was also collected concurrently. Here we report the results of these measurements and show the feasibility of using these types of instruments for collection the fluorescence and other information essential for ecological and carbon cycle studies.

  14. 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.

  15. Measuring surface dynamics of biomolecules by total internal reflection fluorescence with photobleaching recovery or correlation spectroscopy.

    PubMed

    Thompson, N L; Burghardt, T P; Axelrod, D

    1981-03-01

    The theoretical basis of a new technique for measuring equilibrium adsorption/desorption kinetics and surface diffusion of fluorescent-labeled solute molecules at solid surfaces has been developed. The technique combines total internal reflection fluorescence (TIR) with either fluorescence photobleaching recovery (FPR) or fluorescence correlation spectroscopy (FCS). A laser beam totally internally reflects at a solid/liquid interface; the shallow evanescent field in the liquid excites the fluorescence of surface adsorbed molecules. In TIR/FPR, adsorbed molecules are bleaching by a flash of the focused laser beam; subsequent fluorescence recovery is monitored as bleached molecules exchange with unbleached ones from the solution or surrounding nonilluminated regions of the surface. In TIR/FCS, spontaneous fluorescence fluctuations due to individual molecules entering and leaving a well-defined portion of the evanescent field are autocorrelated. Under appropriate experimental conditions, the rate constants and surface diffusion coefficient can be readily obtained from the TIR/FPR and TIR/FCS curves. In general, the shape of the theoretical TIR/FPR and TIR/FCS curves depends in a complex manner upon the bulk and surface diffusion coefficients, the size of the iluminated or observed region, and the adsorption/desorption/kinetic rate constants. The theory can be applied both to specific binding between immobilized receptors and soluble ligands, and to nonspecific adsorption processes. A discussion of experimental considerations and the application of this technique to the adsorption of serum proteins on quartz may be found in the accompanying paper (Burghardt and Axelrod. 1981. Biophys. J. 33:455). PMID:7225515

  16. 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.

  17. Fluorescence intensity changes associated with contractile activation in frog muscle stained with Nile Blue A.

    PubMed Central

    Bezanilla, F; Horowicz, P

    1975-01-01

    1. Extrinsic fluorescence intensity changes were studied in frog semitendinosus muscles stained with Nile Blue A in response to electrical stimulation. Muscles were stretched and put into hypertonic solutions to prevent movement. The muscles were illuminated at 90 degrees to their long axis with a narrow beam of light at a central wave-length of 6250 . Fluorescence emission was measured at 90 degrees to the exciting light using a filter which absorbed light of wave-lengths shorter than 6400 . 2. In response to a single stimulus the fluorescence intensity increases briefly. The fluorescence response is propagated at a constant velocity of about 1.5 m/sec. The average ratio of the maximum fluorescence intensity change to the resting fluorescence is 4.5 times 10-3 for supramaximal shocks. The fluorescence intensity change starts early in the falling phase of the action potential. 3. The fluorescence intensity change increases when nitrate replaces chloride and decreases when D2O replaces H2O. The rates of rise and fall of the fluorescence change was unaffected by nitrate replacement of chloride but are slowed where D2O replaces H2O. The rates of rise and fall of the fluorescence change increase with increasing temperature for all solutions used. The peak fluorescence intensity change, however, goes through a maximum at about 17 degrees C for aqueous chloride and nitrate solutions in the range of 10-25 degrees C. With D2O solutions, the peak fluorescence intensity increases monotonically in this range of temperatures. 4. The fluorescence intensity change in response to trains of action potentials are not additive. 5. Depolarization of muscles treated with tetrodotoxin using triangular-shaped fluid electrodes produces an increase in fluorescence at about the same threshold values required to elicit tension in preparations that are not fully stretched. The fluorescence intensity change precedes in time tension development. Near threshold depolarizations, the delay in

  18. Patterning pallet arrays for cell selection based on high-resolution measurements of fluorescent biosensors

    PubMed Central

    Shadpour, Hamed; Zawistowski, Jon S.; Herman, Annadele; Hahn, Klaus; Allbritton, Nancy L.

    2011-01-01

    Pallet arrays enable cells to be separated while they remain adherent to a surface and provide a much greater range of cell selection criteria relative to that of current technologies. However there remains a need to further broaden cell selection criteria to include dynamic intracellular signaling events. To demonstrate the feasibility of measuring cellular protein behavior on the arrays using high resolution microscopy, the surfaces of individual pallets were modified to minimize the impact of scattered light at the pallet edges. The surfaces of the three-dimensional pallets on an array were patterned with a coating such as fibronectin using a customized stamping tool. Micropatterns of varying shape and size were printed in designated regions on the pallets in single or multiple steps to demonstrate the reliability and precision of patterning molecules on the pallet surface. Use of a fibronectin matrix stamped at the center of each pallet permitted the localization of H1299 and mouse embryonic fibroblast (MEF) cells to the pallet centers and away from the edges. Compared to pallet arrays with fibronection coating the entire top surface, arrays with a central fibronectin pattern increased the percentage of cells localized to the pallet center by 3-4 fold. Localization of cells to the pallet center also enabled the physical separation of cells from optical artifacts created by the rough pallet side walls. To demonstrate the measurement of dynamic intracellular signaling on the arrays, fluorescence measurements of high spatial resolution were performed using a RhoA GTPase biosensor. This biosensor utilized fluorescence resonance energy transfer (FRET) between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) to measure localized RhoA activity in cellular ruffles at the cell periphery. These results demonstrated the ability to perform spatially resolved measurements of fluorescence-based sensors on the pallet arrays. Thus, the patterned pallet

  19. Patterning pallet arrays for cell selection based on high-resolution measurements of fluorescent biosensors.

    PubMed

    Shadpour, Hamed; Zawistowski, Jon S; Herman, Annadele; Hahn, Klaus; Allbritton, Nancy L

    2011-06-24

    Pallet arrays enable cells to be separated while they remain adherent to a surface and provide a much greater range of cell selection criteria relative to that of current technologies. However there remains a need to further broaden cell selection criteria to include dynamic intracellular signaling events. To demonstrate the feasibility of measuring cellular protein behavior on the arrays using high resolution microscopy, the surfaces of individual pallets were modified to minimize the impact of scattered light at the pallet edges. The surfaces of the three-dimensional pallets on an array were patterned with a coating such as fibronectin using a customized stamping tool. Micropatterns of varying shape and size were printed in designated regions on the pallets in single or multiple steps to demonstrate the reliability and precision of patterning molecules on the pallet surface. Use of a fibronectin matrix stamped at the center of each pallet permitted the localization of H1299 and mouse embryonic fibroblast (MEF) cells to the pallet centers and away from the edges. Compared to pallet arrays with fibronectin coating the entire top surface, arrays with a central fibronectin pattern increased the percentage of cells localized to the pallet center by 3-4-fold. Localization of cells to the pallet center also enabled the physical separation of cells from optical artifacts created by the rough pallet side walls. To demonstrate the measurement of dynamic intracellular signaling on the arrays, fluorescence measurements of high spatial resolution were performed using a RhoA GTPase biosensor. This biosensor utilized fluorescence resonance energy transfer (FRET) between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) to measure localized RhoA activity in cellular ruffles at the cell periphery. These results demonstrated the ability to perform spatially resolved measurements of fluorescence-based sensors on the pallet arrays. Thus, the patterned pallet arrays

  20. 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

    2015-11-01

    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. PMID:26426293

  1. 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

    2015-11-01

    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.

  2. 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.

  3. 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.

  4. 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.

  5. Spectroscopic fluorescence measurements of lamb and human heart tissue in vitro

    NASA Astrophysics Data System (ADS)

    Filippidis, George; Zacharakis, Giannis; Kochiadakis, G. E.; Chrysostomakis, S. I.; Vardas, P. E.; Fotakis, Costas; Papazoglou, Theodore G.

    2003-10-01

    Laser-induced fluorescence spectra were obtained during the exposure of lamb heart (n=20) tissue to Argon-ion radiation (457.9nm). Fluorescence spectra from different heart compartments (the left and right atria and ventricles, the myocardium, the epicardium, and the aorta) were recorded. Simple algebraic algorithms based on the spectral intensity variation were constructed in order to detect spectral features and characterize the different cardiac compartments. Additionally, it was investigated whether each chamber exhibited constant spectral response. After the end of each experiment the lamb hearts were stored in formalin (10%). The samples were irradiated again after forty eight (48) hours in order to investigate the spectral differences that appear due to formalin conservation. Similar fluorescence measurements were taken from a limited number of human heart tissues (n=2) ex vivo.

  6. 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.

  7. Dynamic depolarization in plasmonic metal nanoparticles

    NASA Astrophysics Data System (ADS)

    Apell, S. Peter; Zorić, Igor; Langhammer, Christoph

    2016-08-01

    At very low photon energies most metals have a very large and negative dielectric function. For the response of a metal nanoparticle to an external field in this limit, this means that the particular choice of metal does not matter and the localized surface plasmon energy mainly depends on the shape and size of the particle. Here, we present a theoretical framework to describe this situation and unearth the interplay between the depolarization factor of the problem at hand and the dielectric function of the particle. Available experimental results compare favorably with our theoretical framework.

  8. 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. PMID:27409221

  9. 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.

  10. 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.

  11. 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.

  12. 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 ).

  13. Characterization of depolarizing optical media by means of the entropy factor: application to biological tissues

    NASA Astrophysics Data System (ADS)

    Pereda Cubián, David; Arce Diego, José Luis; Rentmeesters, Raf

    2005-01-01

    Polarized light imaging is a potential tool to obtain an adequate description of the properties of depolarizing media such as biological tissues. In many biomedical applications, for instance, dermatology, ophthalmology, or urology, imaging polarimetry provides a noninvasive diagnosis of a wide range of disease states, and, likewise, it could be applied to the study of internal tissues though the use of endoscopes that use optical fibers. We introduce an algebraic method, based on the Mueller-coherence matrix, for a clearer analysis of the polarization characteristics of depolarizing media via the entropy factor. First-order errors introduced by the measurement system are corrected. Entropy defines three kinds of media according to their depolarizing behavior, and several examples corresponding to each region are shown. The calculation of this factor provides clearer information than that provided by the traditional Mueller matrix in the analysis of biological tissue properties by polarization measurement techniques.

  14. 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.

  15. 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.

  16. Fluorescence anisotropy imaging microscopy for homo-FRET in living cells.

    PubMed

    Tramier, Marc; Coppey-Moisan, Maïté

    2008-01-01

    In this chapter, we present the basic physical principles of the fluorescence anisotropy imaging microscopy (FAIM) and its application to study FP-tagged protein dynamics and interaction in live cells. The Förster mechanism of electronic energy transfer can occur between like chromophores (homo-fluorescence resonance energy transfer, homo-FRET) inducing fluorescence depolarization and can be monitored by fluorescence anisotropy. The energy transfer rate is fast compared to the rotational time of proteins, and therefore its detection as a fast depolarization process in the fluorescence anisotropy can be easily discriminated from rotational motion. Quantitative analysis of fluorescence anisotropy decays provides information on structural parameters: distance between the two interacting chromophores and spatial orientation between the chromophores within dimeric proteins. Fluorescence anisotropy decay is not easy to measure in living cells under the microscope and the instrumentations are necessarily sophisticated. In contrast, any type of microscope can be used to measure the steady-state anisotropy. Interestingly, two-photon excitation steady-state FAIM is a powerful tool for qualitative analysis of macromolecule interactions in living cells and can be used easily for time-lapse homo-FRET.

  17. 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.

  18. Comparison of Air Fluorescence and Ionization Measurements of E.M. Shower Depth Profiles: Test of a UHECR Detector Technique

    SciTech Connect

    Belz, J.; Cao, Z.; Huentemeyer, P.; Jui, C.C.H.; Martens, K.; Matthews, J.; Maestas, M.; Smith, J.; Sokolsky, P.; Springer, R.W.; Thomas, J.; Thomas, S.; Chen, P.; Field, Clive; Hast, C.; Iverson, R.; Ng, J.S.T.; Odian, A.; Reil, K.; Vincke, H.; Walz, D.; /SLAC /Montana U. /Rutgers U., Piscataway /Taiwan, Natl. Taiwan U.

    2005-10-07

    Measurements are reported on the fluorescence of air as a function of depth in electromagnetic showers initiated by bunches of 28.5 GeV electrons. The light yield is compared with the expected and observed depth profiles of ionization in the showers. It validates the use of atmospheric fluorescence profiles in measuring ultra high energy cosmic rays.

  19. New portable photoacoustic and fluorescence photometer for field measurement of photosynthesis

    NASA Astrophysics Data System (ADS)

    Bélanger, Raymond; Paquette, André; N'soukpoé-Kossi, Christophe N.; Leblanc, Roger M.

    1993-05-01

    A new portable photoacoustic and fluorescence photometer has been built. The instrument is especially made for field measurements but can also be used indoors. This new instrument has many advantages. It can measure the photosynthetic O2 evolution and energy storage, and the vitality index in the same sample. The system is very compact, which makes it easy to transport. A small electrical generator satisfies the 110 V power requirement for field applications. All manipulations are computer controlled including the data acquisition and treatment. The photoacoustic signal-to-noise ratio for carbon black is the same under field conditions as in the laboratory (˜2×104) at 130 Hz. Results obtained on declining sugar maple trees in the field are presented. The combination of photoacoustic and fluorescence measurements in one instrument represents a very powerful tool in photosynthesis research.

  20. 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.

  1. Real-time focus and overlay measurement by the use of fluorescent markers

    NASA Astrophysics Data System (ADS)

    Maas, Diederik; van Zwet, Erwin

    2014-04-01

    In lithography, overlay control is getting increasingly complex. Advanced Process Control (APC) is introduced to minimize excursions from the process window for the present exposure. APC uses metrology data of previously exposed wafers, hence, there is always a delay of tens of minutes before the required information is available. This paper proposes the combination of a patterned expose beam and a patterned fluorescent marker on a wafer to generate a fluorescent signal that carries real-time information of the focus and/or position error of the expose pattern with the pattern on the wafer. A practical realization requires some changes to the exposure process, stepper design and reticle lay-out. Firstly, a matched pair of markers on the wafer and reticle is required. Secondly, the generated fluorescent signal must be measured, for example with a (spectrally filtered) photon counter close to the expose area of the wafer. At last, the markers from the previous lithography step shall, after development, be filled with fluorescent material. This deposition requires an additional process step. Photon budget calculations suggest an overlay measurement accuracy of less than a tenth of a nm (real-time).

  2. 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.

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

    NASA Astrophysics Data System (ADS)

    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.

    2016-09-01

    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 obtained spatially resolved measurements of Ti K-α emission. Density profiles were measured from K-α intensity. Ti ionization state distributions and electron temperatures were inferred by fitting K-α spectra to spectra from CRETIN simulations. This work 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.

  4. Impact of Emission Anisotropy on Fluorescence Spectroscopy and FRET Distance Measurements

    PubMed Central

    Ivanov, Vassili; Li, Min; Mizuuchi, Kiyoshi

    2009-01-01

    Abstract The objective of this report is to provide a practical and improved method for estimating Förster resonance energy transfer distance measurement error due to unknown angles in the dipole orientation factor based on emission anisotropy measurements. We improve on the method of Dale et al. (1979), which has minor mistakes and is frequently interpreted in overly optimistic ways in the literature. To facilitate proper fluorescence intensity measurements, we also evaluated instrument parameters that could impact the measurement. The apparent fluorescence intensity of isotropic samples depends on the sample emission anisotropy, fluorometer geometry, and optical apertures. We separate parameters of the sample, and those of the cylindrically symmetric illumination source and detector in the equations describing results of unpolarized and polarized fluorescence intensity measurements. This approach greatly simplifies calculations compared with the more universal method of Axelrod (1989). We provide a full computational method for calculating the Förster resonance energy transfer distance error and present a graph describing distance error in the simplest case. PMID:19651051

  5. 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

  6. Measurements of Ambient OH and HO2 by Laser-Induced Fluorescence Using FAGE

    NASA Astrophysics Data System (ADS)

    Dusanter, S.; Vimal, D.; Stevens, P.

    2005-12-01

    Measurements of OH (hydroxyl) and HO2 (hydroperoxyl) radicals provide a critical test of our understanding of the fast photochemistry of the atmosphere, but are challenging because of their short lifetimes and low concentrations. Several instruments developed during the last decade have successfully made measurements of these important radicals. However, these measurements have shown that there are still gaps in our understanding of OH and HO2 radical chemistry in the atmosphere. Additional measurements of OH and HO2 are needed to constrain and test current models of atmospheric chemistry. We will present a detailed description of our new Fluorescence Assay by Gas Expansion (FAGE) system and our ongoing work toward an automated field instrument, focusing on its characteristics in terms of sensitivity, limit of detection, selectivity, temporal resolution, stability and calibration. In this technique, ambient air is expanded through a pinhole into a low pressure cell. The OH radicals are then electronically excited using a transition in the (0, 0) band of the A-X system near 308 nm. The resulting fluorescence, which is proportional to the OH concentration, is collected and quantified. HO2 is converted into OH by adding a small flow of NO inside the fluorescence cell. Finally, we will present measurements of OH and HO2 concentrations on the Indiana University, Bloomington campus.

  7. 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.

  8. On the collective network of ionic liquid/water mixtures. IV. Kinetic and rotational depolarization.

    PubMed

    Schröder, Christian; Sega, Marcello; Schmollngruber, Michael; Gailberger, Elias; Braun, Daniel; Steinhauser, Othmar

    2014-05-28

    Dielectric spectroscopy is a measure of the collective Coulomb interaction in liquid systems. Adding ionic liquids to an aqueous solution results in a decrease of the static value of the generalized dielectric constant which cannot be attributed to kinetic depolarization models characterized by the static conductivity and rotational relaxation constant. However, a dipolar Poisson-Boltzmann model computing the water depolarization in the proximity of ions is not only successful for simple electrolytes but also in case of molecular ionic liquids. Moreover, our simple geometric hydration model is also capable to explain the dielectric depolarization. Both models compute the dielectric constant of water and obtain the overall dielectric constant by averaging the values of its components, water and the ionic liquid, weighted by their volume occupancies. In this sense, aqueous ionic liquid mixtures seem to behave like polar mixtures.

  9. Tissue distribution and real-time fluorescence measurement of a tumor-targeted nanodevice by a two photon optical fiber fluorescence probe

    NASA Astrophysics Data System (ADS)

    Thomas, Thommey P.; Ye, Jing Yong; Yang, Chu-Sheng; Myaing, Monthiri; Majoros, Istvan J.; Kotlyar, Alina; Cao, Zhengyi; Norris, Theodore B.; Baker, James R., Jr.

    2006-02-01

    Real-time fluorescence measurement in deep tumors in live animals (or humans) by conventional methods has significant challenges. We have developed a two-photon optical fiber fluorescence (TPOFF) probe as a minimally invasive technique for quantifying fluorescence in solid tumors in live mice. Here we demonstrate TPOFF for real-time measurements of targeted drug delivery dynamics to tumors in live mice. 50-femtosecond laser pulses at 800 nm were coupled into a single mode optical fiber and delivered into the tumor through a 27-gauge needle. Fluorescence was collected back through the same fiber, filtered, and detected with photon counting. Biocompatible dendrimer-based nanoparticles were used for targeted delivery of fluorescent materials into tumors. Dendrimers with targeting agent folic acid and fluorescent reporter 6-TAMRA (G5-6T-FA) were synthesized. KB cell tumors expressing high levels of FA receptors were developed in SCID mice. We initially demonstrated the specific uptake of the targeted conjugates into tumor, kidney and liver, using the TPOFF probe. The tumor fluorescence was then taken in live mice at 30 min, 2 h and 24 h with the TPOFF probe. G5-6T-FA accumulated in the tumor with maximum mean levels reaching 673 +/- 67 nM at the 2 h time point. In contrast, the levels of a control, non-targeted conjugate (G5-6T) at 2 h reached a level of only 136 +/- 28 nM in tumors, and decrease quickly. This indicates that the TPOFF probe can be used as a minimally invasive detection system for quantifying the specific targeting of a fluorescent nanodevice on a real-time basis.

  10. Observations of the spectral dependence of particle depolarization ratio of aerosols using NASA Langley airborne High Spectral Resolution Lidar

    NASA Astrophysics Data System (ADS)

    Burton, S. P.; Hair, J. W.; Kahnert, M.; Ferrare, R. A.; Hostetler, C. A.; Cook, A. L.; Harper, D. B.; Berkoff, T. A.; Seaman, S. T.; Collins, J. E.; Fenn, M. A.; Rogers, R. R.

    2015-09-01

    Particle depolarization ratio is presented for three case studies from the NASA Langley airborne High Spectral Resolution Lidar-2 (HSRL-2). Particle depolarization ratio from lidar is an indicator of non-spherical particles and is sensitive to the fraction of non-spherical particles and their size. The HSRL-2 instrument measures depolarization at three wavelengths: 355, 532, and 1064 nm. The three measurement cases presented here include two cases of dust aerosol and one case of smoke aerosol. These cases have partial analogs in earlier HSRL-1 depolarization measurements at 532 and 1064 nm and in literature, but the availability of three wavelengths gives additional insight into different scenarios for non-spherical particles in the atmosphere. A case of transported Saharan dust has a spectral dependence with a peak of 0.30 at 532 nm with smaller particle depolarization ratios of 0.27 and 0.25 at 1064 and 355 nm, respectively. A case of locally generated wind-blown North American dust has a maximum of 0.38 at 1064 nm, decreasing to 0.37 and 0.24 at 532 and 355 nm, respectively. The cause of the maximum at 1064 nm is inferred to be very large particles that have not settled out of the dust layer. The smoke layer has the opposite spectral dependence, with the peak of 0.24 at 355 nm, decreasing to 0.09 and 0.02 at 532 and 1064 nm. The depolarization in the smoke case is inferred to be due to the presence of coated soot aggregates. We also point out implications for the upcoming EarthCARE satellite, which will measure particle depolarization ratio only at 355 nm. At 355 nm, the particle depolarization ratios for all three of our case studies are very similar, indicating that smoke and dust may be more difficult to separate with EarthCARE measurements than heretofore supposed.

  11. A 20 Ghz Depolarization Experiment Using the ATS-6 Satellite

    NASA Technical Reports Server (NTRS)

    Bostian, C. W.; Manus, E. A.; Marshall, R. E.; Pendrak, H. N.; Stutzman, W. L.; Wiley, P. H.; Kauffman, S. R.

    1975-01-01

    A depolarization experiment using the 20 GHz downlink from the ATS-6 satellite was described. The following subjects were covered: (1) an operational summary of the experiment, (2) a description of the equipment used with emphasis on improvements made to the signal processing receiver used with the ATS-5 satellite, (3) data on depolarization and attenuation in one snow storm and two rain storms at 45 deg elevation, (4) data on low angle propagation, (5) conclusions about depolarization on satellite paths, and (6) recommendations for the depolarization portion of the CTS experiment.

  12. 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.

  13. 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

  14. Fluorescence measurements of anion transport by the GABA receptor in reconstituted membrane preparations

    SciTech Connect

    Dunn, S.M.J.; Shelman, R.A.; Agey, M.W. )

    1989-03-21

    A fluorescence assay for measuring the functional properties of the GABA{sub A} receptor in reconstituted membrane vesicles is described. This assay is based on a method previously described to measure monovalent cation transport mediated by the nicotinic acetylcholine receptor in membranes from Torpedo electric organ. The GABA{sub A} receptor has been solubilized from bovine brain membranes and reconstituted into phospholipid vesicles. Influx of chloride or iodide into the vesicles has been measured in stopped-flow experiments by monitoring the fluorescence quench of an anion-sensitive fluorophore trapped within the vesicles. Muscimol, a GABA{sub A} receptor agonist, stimulated a rapid uptake of either chloride or iodide. Stimulation of chloride influx was dependent on the concentration of muscimol, and the midpoint of the dose-response curve occurred at approximately 0.3 {mu}M. Agonist-stimulated uptake was enhanced by diazepam and blocked by desensitization and by the antagonists bicuculline and picrotoxin. These receptor-mediated effects are shown to be qualitatively similar to measurements of {sup 36}Cl{sup {minus}} and {sup 125}I{sup {minus}} efflux using synaptoneurosomes prepared from rat cerebral cortex. The advantages of the fluorescence method in terms of its improved time resolution, sensitivity, and suitability for quantitating GABA{sub A} receptor function are discussed.

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

    PubMed

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

    2015-11-01

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

  16. 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.

  17. The reproducibility of 109Cd-based X-ray fluorescence measurements of bone lead.

    PubMed Central

    Gordon, C L; Webber, C E; Chettle, D R

    1994-01-01

    We assessed the reproducibility of X-ray fluorescence-based lead measurements from multiple measurements made on a low-concentration plaster of paris phantom and in five subjects measured five times on two occasions. Over a 6-month period, 220 measurements of the same phantom were obtained and showed a standard deviation of 1.29 micrograms Pb (g plaster of paris)-1. The two sets of in vivo measurements were made 10 months apart and revealed a mean standard deviation of 3.4 micrograms Pb (g bone mineral)-1 and 5.1 micrograms Pb (g bone mineral)-1 for males and females, respectively. Our measured standard deviation exceeded by 20-30% the calculated standard deviation associated with a single measurement both in the phantom and in subjects. This indicates that some variance is introduced during the measurement process. Operator learning and consistency significantly minimized this increased variability. Measured lead concentrations of the left and right tibia in 14 subjects showed no significant differences between legs. As a result, either tibia can be sampled and compared over time. The levels of reproducibility we report here mean that X-ray fluorescence-based determinations of bone lead concentrations are reliable both over the short and long term. Thus, reasonably sized confidence intervals can be placed on detected changes in concentration and should permit acquisition of longitudinal data within a reasonable length of time. Images Figure 1. PMID:7895710

  18. Monitoring sperm mitochondrial respiration response in a laser trap using ratiometric fluorescence

    NASA Astrophysics Data System (ADS)

    Mei, Adrian; Botvinick, Elliot; Berns, Michael

    2005-08-01

    Sperm motility is an important area in understanding male infertility. Various techniques, such as the Computer Assisted Sperm Analysis (CASA), have been used to understand sperm motility. Sperm motility is related to the energy (ATP) production of sperm. ATP is produced by the depolarization of the membrane potential of the inner membrane of the mitochondria. In this study, a mitochondrial dye, JC-1, has been used to monitor the energetics of the mitochondria. This fluorescent dye can emit at two different wavelengths, depending on the membrane potential of the mitochondria. It can fluoresce green at low membrane potential and red at high membrane potential. The ratio of the two colors (red/green) allows for an accurate measurement of the change of membrane potential. Various experiments were conducted to quantify the behavior of the dye within the sperm and the reaction of the sperm to trap. Sperm were trapped using laser tweezers. Results have shown that the ratio drops dramatically when sperm are trapped, indicating a depolarization of the membrane. The physiological response to this depolarization is yet to be determined, but the studies indicate that the sperm could have been slightly damaged by the laser. However, knowing that sperm depolarizes their membrane when trapped can help understand how sperm react to their environment and consequently help treat male infertility.

  19. Post-depolarization potentiation of GABAA receptors: A novel mechanism regulating tonic conductance in hippocampal neurons

    PubMed Central

    Ransom, Christopher B.; Wu, Yuanming; Richerson, George B.

    2010-01-01

    Ambient GABA in the brain activates GABAA receptors to produce tonic inhibition. Membrane potential influences both GABA transport and GABAA receptors and could thereby regulate tonic inhibition. We investigated the voltage-dependence of tonic currents in cultured rat hippocampal neurons using patch clamp techniques. Tonic GABAA conductance increased with depolarization from 15±3 pS/pF at −80 mV to 29±5 pS/pF at −40 mV. Inhibition of vesicular or nonvesicular GABA release did not prevent voltage-dependent increases of tonic conductance. Currents evoked with exogenous GABA (1 µM) were outwardly-rectifying, similar to tonic currents due to endogenous GABA. These results indicate that the voltage-dependent increase of tonic conductance was due to intrinsic GABAA receptor properties rather than an elevation of ambient GABA. Following transient depolarization to +40 mV, endogenous tonic currents measured at −60 mV were increased by 75±17%. This novel form of tonic current modulation, termed post-depolarization potentiation (PDP), recovered with a time constant of 63 s, was increased by exogenous GABA, and inhibited by GABAA receptor antagonists. Measurements of EGABA showed PDP was due to increased conductance and not a change in the anion gradient. To assess the functional significance of PDP, we used voltage-clamp waveforms that replicated epileptiform activity. PDP was produced by this pathophysiologic depolarization. These data show that depolarization produces prolonged potentiation of tonic conductance due to voltage-dependent properties of GABAA receptors. These properties are well suited to limit excitability during pathophysiologic depolarization accompanied by rises in ambient GABA, such as occur during seizures and ischemia. PMID:20519542

  20. Technique for measurement of fluorescence lifetime by use of stroboscopic excitation and continuous-wave detection.

    PubMed

    Matthews, D R; Summers, H D; Njoh, K; Errington, R J; Smith, P J; Barber, P; Ameer-Beg, S; Vojnovic, B

    2006-03-20

    A study of the practicality a simple technique for obtaining time-domain information that uses continuous wave detection of fluorescence is presented. We show that this technique has potential for use in assays for which a change in the lifetime of an indicator occurs in reaction to an analyte, in fluorescence resonance energy transfer, for example, and could be particularly important when one is carrying out such measurements in the scaled-down environment of a lab on a chip (biochip). A rate-equation model is presented that allows an objective analysis to be made of the relative importance of the key measurement parameters: optical saturation of the fluorophore and period of the excitation pulse. An experimental demonstration of the technique that uses a cuvette-based analysis of a carbocyanine dye and for which the excitation source is a 650 nm wavelength, self-pulsing AlGaInP laser diode is compared with the model.

  1. Measuring evaporation rates of laser-trapped droplets by use of fluorescent morphology-dependent resonances.

    PubMed

    Pastel, R; Struthers, A

    2001-05-20

    Morphology-dependent resonances (MDRs) are used to measure accurately the evaporation rates of laser-trapped 1- to 2-mum droplets of ethylene glycol. Droplets containing 3 x 10(-5) M Rhodamine-590 laser dye are optically trapped in a 20-mum hollow fiber by two counterpropagating 150-mW, 800-nm laser beams. A weaker 532-nm laser excites the dye, and fluorescence emission is observed near 560 nm as the droplet evaporates. A complete series of first-order TE and TM MDRs dominates the fluorescent output. MDR mode identification sizes the droplets and provides accurate evaporation rates. We verify the automated MDR mode identification by counting fringes in a videotape of the experiment. The longitudinal spring constant of the trap, measured by analysis of the videotaped motion of droplets perturbed from the trap center, provides independent verification of the laser's intensity within the trap.

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

    PubMed

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

    2016-08-01

    Confocal endomicroscopy is a powerful tool for in vivo real-time imaging at cellular resolution inside a living body without tissue resection. Microscopic fluorescence lifetime measurement can provide information about localized biochemical conditions such as pH and the concentrations of oxygen and calcium. We hypothesized that combining these techniques could assist accurate cancer discrimination by providing both biochemical and morphological information. We designed a dual-mode experimental setup for confocal endomicroscopic imaging and fluorescence lifetime measurement and applied it to a mouse xenograft model of activated human pancreatic cancer generated by subcutaneous injection of AsPC-1 tumor cells. Using this method with pH-sensitive sodium fluorescein injection, we demonstrated discrimination between normal and cancerous tissues in a living mouse. With further development, this method may be useful for clinical cancer detection. PMID:27547835

  3. [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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. Precise Measurement of the Absolute Yield of Fluorescence Photons in Atmospheric Gases

    SciTech Connect

    Ave, M.; Bohacova, M.; Daumiller, K.; Di Carlo, P.; Di Giulio, C.; Luis, P.Facal San; Gonzales, D.; Hojvat, C.; Horandel, J.R.; Hrabovsky, M.; Iarlori, M.; /INFN, Aquila /Karlsruhe, Inst. Technol.

    2011-01-01

    We have performed a measurement of the absolute yield of fluorescence photons at the Fermilab Test Beam. A systematic uncertainty at 5% level was achieved by the use of Cherenkov radiation as a reference calibration light source. A cross-check was performed by an independent calibration using a laser light source. A significant improvement on the energy scale uncertainty of Ultra-High Energy Cosmic Rays is expected.

  11. Measurement of optical trapping forces by use of the two-photon-excited fluorescence of microspheres.

    PubMed

    Kachynski, A V; Kuzmin, A N; Pudavar, H E; Kaputa, D S; Cartwright, A N; Prasad, P N

    2003-12-01

    A novel technique for the calibration of laser trapping systems that utilizes two-photon-excited fluorescence of commercial dye-stained microspheres has been demonstrated. The trapping forces as well as the trapping efficiency have been measured for various liquid environments and trapping depths. The trapping efficiency in water was found to decrease with an increase of trapping depths because of the enlargement of the trapping beam waist caused by aberrations of the optical system.

  12. Valley depolarization in monolayer WSe2.

    PubMed

    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

  13. 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.

  14. Observations of the spectral dependence of linear particle depolarization ratio of aerosols using NASA Langley airborne High Spectral Resolution Lidar

    NASA Astrophysics Data System (ADS)

    Burton, S. P.; Hair, J. W.; Kahnert, M.; Ferrare, R. A.; Hostetler, C. A.; Cook, A. L.; Harper, D. B.; Berkoff, T. A.; Seaman, S. T.; Collins, J. E.; Fenn, M. A.; Rogers, R. R.

    2015-12-01

    Linear particle depolarization ratio is presented for three case studies from the NASA Langley airborne High Spectral Resolution Lidar-2 HSRL-2). Particle depolarization ratio from lidar is an indicator of non-spherical particles and is sensitive to the fraction of non-spherical particles and their size. The HSRL-2 instrument measures depolarization at three wavelengths: 355, 532, and 1064 nm. The three measurement cases presented here include two cases of dust-dominated aerosol and one case of smoke aerosol. These cases have partial analogs in earlier HSRL-1 depolarization measurements at 532 and 1064 nm and in literature, but the availability of three wavelengths gives additional insight into different scenarios for non-spherical particles in the atmosphere. A case of transported Saharan dust has a spectral dependence with a peak of 0.30 at 532 nm with smaller particle depolarization ratios of 0.27 and 0.25 at 1064 and 355 nm, respectively. A case of aerosol containing locally generated wind-blown North American dust has a maximum of 0.38 at 1064 nm, decreasing to 0.37 and 0.24 at 532 and 355 nm, respectively. The cause of the maximum at 1064 nm is inferred to be very large particles that have not settled out of the dust layer. The smoke layer has the opposite spectral dependence, with the peak of 0.24 at 355 nm, decreasing to 0.09 and 0.02 at 532 and 1064 nm, respectively. The depolarization in the smoke case may be explained by the presence of coated soot aggregates. We note that in these specific case studies, the linear particle depolarization ratio for smoke and dust-dominated aerosol are more similar at 355 nm than at 532 nm, having possible implications for using the particle depolarization ratio at a single wavelength for aerosol typing.

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

    PubMed

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

    2015-12-15

    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.

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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

    2015-01-01

    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

  1. Depolarization of decaying counterflow turbulence in He II.

    PubMed

    Barenghi, C F; Gordeev, A V; Skrbek, L

    2006-08-01

    We present experimental evidence backed up by numerical simulations that the steady-state vortex tangle created in He II by heat-transfer counterflow is strongly polarized. When the heater that generates the counterflow turbulence is switched off, the vortex tangle decays, the vortex lines randomize their spatial orientation and the tangle's polarization decreases. The process of depolarization slows down the recovery of the transverse second sound signal which measures the vortex line density; at some values of parameters it even leads to a net decrease of the amplitude of the transverse second sound prior to reaching the universal -32 power temporal law decay typical of classical homogeneous isotropic turbulence in a finite-sized channel. PMID:17025541

  2. Depolarization of decaying counterflow turbulence in He II

    SciTech Connect

    Barenghi, C. F.; Gordeev, A. V.; Skrbek, L.

    2006-08-15

    We present experimental evidence backed up by numerical simulations that the steady-state vortex tangle created in He II by heat-transfer counterflow is strongly polarized. When the heater that generates the counterflow turbulence is switched off, the vortex tangle decays, the vortex lines randomize their spatial orientation and the tangle's polarization decreases. The process of depolarization slows down the recovery of the transverse second sound signal which measures the vortex line density; at some values of parameters it even leads to a net decrease of the amplitude of the transverse second sound prior to reaching the universal -3/2 power temporal law decay typical of classical homogeneous isotropic turbulence in a finite-sized channel.

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

    SciTech Connect

    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.

  4. "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…

  5. 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.

  6. [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.

  7. [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. PMID:23928069

  8. 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.

  9. Experimental observation of universality in depolarized light scattering.

    PubMed

    Puentes, Graciana; Voigt, Dirk; Aiello, Andrea; Woerdman, J P

    2005-12-01

    Experimental results on light depolarization due to multimode scattering are reported. By means of polarization tomography, we characterize the depolarizing power and the polarization entropy of a broad class of optically scattering media and confirm the recently predicted universal behavior of these two quantities [Phys. Rev. Lett. 94, 090406 (2005)].

  10. Depolarization of ultracold neutrons during their storage in material bottles

    NASA Astrophysics Data System (ADS)

    Serebrov, A. P.; Lasakov, M. S.; Vassiljev, A. V.; Krasnoschekova, I. A.; Rudnev, Yu. P.; Fomin, A. K.; Varlamov, V. E.; Geltenbort, P.; Butterworth, J.; Young, A. R.; Pesavento, U.

    2003-07-01

    The depolarization of ultracold neutrons (UCN) during their storage in traps has been investigated. The neutron spin-flip probability for the materials studied amounts to ∼(1-2)×10 -5 per collision and does not depend on the temperature. The possible connection between the phenomenon of UCN depolarization and that of anomalous losses is discussed.

  11. 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).

  12. 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.

  13. 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).

  14. Spot measurements of intermediate frequency electric fields in the vicinity of compact fluorescent lamps.

    PubMed

    Bakos, József; Nagy, Noémi; Juhász, Péter; Thuróczy, György

    2010-12-01

    Starting in 2009, certain types of incandescent light bulbs will be withdrawn from the market in the European Union and elsewhere. However, compact fluorescent lamps that are among the candidates to replace them produce intermediate frequency electric fields (EFs) much higher than any other device or appliance previously available to the general public. Measurement results of these EFs showed that the maximum recorded EF strength in the 1.2-100 kHz frequency range in close proximity to the lamps was > 42 V m(-1) for all tested lamps. In nine cases, the field strength exceeded 87 V m(-1) and the highest measured value was 216 V m(-1).

  15. 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.

  16. Stability of some Cactaceae proteins based on fluorescence, circular dichroism, and differential scanning calorimetry measurements.

    PubMed

    Gorinstein, S; Zemser, M; Vargas-Albores, F; Ochoa, J L; Paredes-Lopez, O; Scheler, C; Aksu, S; Salnikow, J

    1999-02-01

    Characterization of three cactus proteins (native and denatured) from Machaerocereus gummosus (Pitahaya agria), Lophocereu schottii (Garambullo), and Cholla opuntia (Cholla), was based on electrophoretic, fluorescence, CD (circular dichroism), DSC (differential scanning calorimetry), and FT-IR (Fourier transform infrared) measurements. The obtained results of intrinsic fluorescence, DSC, and CD were dissimilar for the three species of cactus, providing evidence of differences in secondary and tertiary structures. Cactus proteins may be situated in the following order corresponding to their relative stability: Machaerocereus gummosus (Pitahaya agria) > Cholla opuntia (Cholla) > Lophocereu schottii (Garambullo). Thermodynamic properties of proteins and their changes upon denaturation (temperature of denaturation, enthalphy, and the number of ruptured hydrogen bonds) were correlated with the secondary structure of proteins and disappearance of alpha-helix.

  17. Stability of some Cactaceae proteins based on fluorescence, circular dichroism, and differential scanning calorimetry measurements.

    PubMed

    Gorinstein, S; Zemser, M; Vargas-Albores, F; Ochoa, J L; Paredes-Lopez, O; Scheler, C; Aksu, S; Salnikow, J

    1999-02-01

    Characterization of three cactus proteins (native and denatured) from Machaerocereus gummosus (Pitahaya agria), Lophocereu schottii (Garambullo), and Cholla opuntia (Cholla), was based on electrophoretic, fluorescence, CD (circular dichroism), DSC (differential scanning calorimetry), and FT-IR (Fourier transform infrared) measurements. The obtained results of intrinsic fluorescence, DSC, and CD were dissimilar for the three species of cactus, providing evidence of differences in secondary and tertiary structures. Cactus proteins may be situated in the following order corresponding to their relative stability: Machaerocereus gummosus (Pitahaya agria) > Cholla opuntia (Cholla) > Lophocereu schottii (Garambullo). Thermodynamic properties of proteins and their changes upon denaturation (temperature of denaturation, enthalphy, and the number of ruptured hydrogen bonds) were correlated with the secondary structure of proteins and disappearance of alpha-helix. PMID:10333299

  18. 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

  19. Measuring protein interactions using Förster resonance energy transfer and fluorescence lifetime imaging microscopy

    PubMed Central

    Day, Richard N.

    2013-01-01

    The method of fluorescence lifetime imaging microscopy (FLIM) is a quantitative approach that can be used to detect Förster Resonance Energy Transfer (FRET). The use of FLIM to measure the FRET that results from the interactions between proteins labeled with fluorescent proteins (FPs) inside living cells provides a non-invasive method for mapping interactomes. Here, the use of the phasor plot method to analyze frequency domain (FD) FLIM measurements is described, and measurements obtained from cells producing the 'FRET standard' fusion proteins are used to validate the FLIM system for FRET measurements. The FLIM FRET approach is then used to measure both homologous and heterologous protein-protein interactions (PPI) involving the CCAAT/enhancer-binding protein alpha (C/EBPα). C/EBPα is a transcription factor that controls cell differentiation, and localizes to heterochromatin where it interacts with the heterochromatin protein 1 alpha (HP1α). The FLIM-FRET method is used to quantify the homologous interactions between the FP-labeled basic leucine zipper (BZip) domain of C/EBPα. Then the heterologous interactions between the C/EBPa BZip domain and HP1a are quantified using the FRET-FLIM method. The results demonstrate that the basic region and leucine zipper (BZip) domain of C/EBPα is sufficient for the interaction with HP1α in regions of heterochromatin. PMID:23806643

  20. 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.

  1. Automatic microfluidic fluorescence-array measurement system for detecting organic phosphate.

    PubMed

    Chang, Hsing-Cheng; Lin, Jung-Chin; Lin, Shyan-Lung; Chang, I-Nan; Lin, Chern-Sheng; Chen, Shi-Yao

    2015-01-01

    In this study, an automatic microfluidic fluorescence-array measurement system is developed to detect the concentration of organic phosphate based on the luminol-hydrogen peroxide catalytic fluorescent mechanism. Not only sample quantity and cost can be reduced, but also detection time, accuracy and precision can be improved in the system. The system is composed of a CCD image module, a stepper motor with driver, a microfluidic fluorescence array, a background light elimination module, and a dynamic image-analyzed interface. The pesticides of chlorpyrifos and fenitrothion of organic phosphate are chosen as experimental samples. Only a 2.5 μ l quantity of sample is required to have a fast response time of 1.4 second. Experimental results show that the sensitivities of chlorpyrifos and fenitrothion are 1.88 V/ppm in the range of 0.166 ∼ 10 ppm with averaged error of 1.66% and 0.32 V/ppm in the range of 0.03 ∼ 10 ppm with averaged error of 1.68% respectively. The organophosphorus effective detection range of the developed system covers the legal prescription for pesticide residues.

  2. 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.

  3. A Practical Solution for 77 K Fluorescence Measurements Based on LED Excitation and CCD Array Detector

    PubMed Central

    Lamb, Jacob; Forfang, Kristin; Hohmann-Marriott, Martin

    2015-01-01

    The fluorescence emission spectrum of photosynthetic microorganisms at liquid nitrogen temperature (77 K) provides important insights into the organization of the photosynthetic machinery of bacteria and eukaryotes, which cannot be observed at room temperature. Conventionally, to obtain such spectra, a large and costly table-top fluorometer is required. Recently portable, reliable, and largely maintenance-free instruments have become available that can be utilized to accomplish a wide variety of spectroscopy-based measurements in photosynthesis research. In this report, we show how to build such an instrument in order to record 77K fluorescence spectra. This instrument consists of a low power monochromatic light-emitting diode (LED), and a portable CCD array based spectrometer. The optical components are coupled together using a fiber optic cable, and a custom made housing that also supports a dewar flask. We demonstrate that this instrument facilitates the reliable determination of chlorophyll fluorescence emission spectra for the cyanobacterium Synechocystis sp. PCC 6803, and the green alga Chlamydomonas reinhardtii. PMID:26177548

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. Satellite ultraviolet measurements of nitric oxide fluorescence with a diffusive transport model.

    NASA Technical Reports Server (NTRS)

    Rusch, D. W.

    1973-01-01

    Twilight measurements of fluorescence in the (1, 0) gamma band of nitric oxide were made from June 1967 to January 1969 by an ultraviolet scanning spectrometer on board the polar orbiting satellite Ogo 4. Nitric oxide vertical column emission rates were measured between solar zenith angles of 93 and 98 deg. Seasonal and latitudinal variations were found to be less than a factor of 1.3, the scatter and uncertainty in the data prohibiting more precise determinations from being made. Time independent chemical diffusion models for the vertical distribution of nitric oxide agree well with profiles measured from sounding rockets. The column emission rates calculated from the theoretical models are larger than the satellite measurements by a factor of 3.

  12. 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.

  13. 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.

  14. 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.

  15. 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

  16. High-Resolution, Noninvasive, Two-Photon Fluorescence Measurement of Molecular Concentrations in Corneal Tissue

    PubMed Central

    Cui, Liping; Huxlin, Krystel R.; Xu, Lisen; MacRae, Scott

    2011-01-01

    Purpose. To perform high-resolution, noninvasive, calibrated measurements of the concentrations and diffusion profiles of fluorescent molecules in the live cornea after topical application to the ocular surface. Methods. An 800-nm femtosecond laser was used to perform two-photon fluorescence (TPF) axial scanning measurements. Calibration solutions consisting of sodium fluorescein (Na-Fl; concentration range, 0.01%–2.5%) and riboflavin (concentration range, 0.0125%–0.1%) were tested in well slides, and TPF signals were assessed. Excised feline eyeballs preserved in corneal storage medium and with either intact or removed corneal epithelia were then treated with Na-Fl, riboflavin, or fluorescein dextran (Fl-d) of different molecular weight (MW) for 30 minutes. Calibrated TPF was then used immediately to measure the concentration of these molecules across the central corneal depth. Results. The axial resolution of our TPF system was 6 μm, and a linear relationship was observed between TPF signal and low concentrations of most fluorophores. Intact corneas treated with Na-Fl or riboflavin exhibited a detectable penetration depth of only approximately 20 μm, compared with approximately 400 to 600 μm when the epithelium was removed before fluorophore application. Peak concentrations for intact corneas were half those attained with epithelial removal. Debrided corneas treated with 2,000,000 MW Fl-d showed a half-maximum penetration depth of 156.7 μm compared with 384 μm for the 3,000 MW dextran. The peak concentration of the high MW dextran was one quarter that of the lower MW dextran. Conclusions. TPF is an effective, high-resolution, noninvasive method of quantifying the diffusion and concentration of fluorescent molecules across the cornea. PMID:21228379

  17. Fast implementation for fluorescence tomography based on coordinate descent with limited measurements

    NASA Astrophysics Data System (ADS)

    Xue, Zhenwen; Qin, Chenghu; Wu, Ping; Yang, Xin; Tian, Jie

    2012-03-01

    Fluorescence molecular tomography (FMT) can three-dimensionally resolve molecular activities in in vivo small animal through the reconstruction of the distribution of fluorescent probes. Due to large number of unknowns and limited measurements from the surfaces of small animals, the FMT problem is often ill-posed and ill-conditioned. Though various L2-norm regularizations can make the solution stable, they usually make the solution over-smoothed. During the early stages of tumor detection, fluorescent sources that indicate the distribution of tumors are usually small and sparse, which can be regarded as a type of a priori information. L1-norm regularizations have been incorporated to promote the sparsity of optical tomographic problems. In this paper, an efficient method with the L1-norm regularization based on coordinate descent is proposed to solve the FMT problem with extremely limited measurements. The proposed method minimizes the objective by solving a sequence of scalar minimization subproblems in multi-variable minimization. Each subproblem improves the estimate of the solution via minimizing along a determined coordinate with all other coordinates fixed. This algorithm first updates the coordinate that makes the energy decrease the most. Non-existence of matrix-vector multiplication in the iteration process makes the proposed algorithm time-efficient. To evaluate this method, we compare it to the iterated-shrinkage-based algorithm with L1-norm regularization in numerical experiments. The proposed algorithm is able to obtain satisfactory reconstruction results even when the measurements are very limited. Besides, the proposed algorithm is about two orders of magnitude faster than the iterated-shrinkage-based algorithm, which enables the proposed algorithm into practical applications.

  18. 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.

  19. 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

  20. Recent Developments in Fluorescence Correlation Spectroscopy for Diffusion Measurements in Planar Lipid Membranes

    PubMed Central

    Macháň, Radek; Hof, Martin

    2010-01-01

    Fluorescence correlation spectroscopy (FCS) is a single molecule technique used mainly for determination of mobility and local concentration of molecules. This review describes the specific problems of FCS in planar systems and reviews the state of the art experimental approaches such as 2-focus, Z-scan or scanning FCS, which overcome most of the artefacts and limitations of standard FCS. We focus on diffusion measurements of lipids and proteins in planar lipid membranes and review the contributions of FCS to elucidating membrane dynamics and the factors influencing it, such as membrane composition, ionic strength, presence of membrane proteins or frictional coupling with solid support. PMID:20386647

  1. Glass transition of associated solvents studied by fluorescence measurement of doped chromophores

    NASA Astrophysics Data System (ADS)

    Ye, Jing Yong; Hattori, Toshiaki; Inouye, Hideyuki; Ueta, Hiroshi; Nakatsuka, Hiroki; Maruyama, Yoshihiro; Ishikawa, Mitsuru

    1996-04-01

    The fluorescence lifetime of a triphenylmethane dye, malachite green, doped in three glass-forming associated solvents, 1-propanol, propylene glycol, and glycerol, was measured in a wide temperature range. For each sample three temperature regimes were found in the temperature dependence of the nonradiative relaxation time of malachite green. The lower crossover temperature corresponds to the calorimetric glass transition temperature, and the higher one, 30-50 K above the lower one, is attributed to the critical temperature that is predicted by mode-coupling theory.

  2. Microscopic Study of Glass Transition: Time-Resolved Fluorescence Measurements of Doped Dye Molecules

    NASA Astrophysics Data System (ADS)

    Nakatsuka, H.; Ye, J. Y.; Hattori, T.; Maruyama, Y.; Ishikawa, M.

    The microscopic dynamics of several monomeric and polymeric glass formers has been investigated by the time-resolved fluorescence measurement of doped malachite green molecules in a wide temperature range. For monomers and a polymer without side chains, beside a kink around the calorimetric glass transition temperature Tg, another crossover at Tc about 30 - 50 K above Tg has been clearly observed, which is in agreement with the prediction of the mode-coupling theory. On the other hand, for the complex polymers with side chains, although we could not distinguish any singularities above Tg, we observed another kink below Tg, which can be attributed to the side-chain motions.

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

    NASA Astrophysics Data System (ADS)

    Goeckner, M. J.; Malik, Shamim 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+2 [the P12 component of the X 2Σ+g(ν=0)→B 2Σ+u(ν=0) band] in a N2 plasma. The strength of the laser-induced fluorescence was used as a measure of the temporally and spatially varying ion density. The expanding sheath was produced by applying a -5 kV pulse to a polished planar electrode in the plasma source ion implantation device [J. R. Conrad et al., J. Vac. Sci. Technol. A 8, 3146 (1990)]. The laser beam was aligned normal to the surface and was reflected off the center of the electrode. The LIF diagnostic used here is nonperturbing whereas previous researchers have used Langmuir probes, which perturb the plasma, to make their measurements. As such, the data reported here represent a benchmark measurement of pulsed sheaths and allow a better comparison between experimental measurements and theoretical predictions. It has been found that the sheath edge moves approximately 16 times faster than the ion-acoustic velocity during the early part of the pulse, t<1 μs, and then slows to approximately the ion-acoustic velocity after 6 μs. In addition to the LIF measurements, a biased probe was used far from the cathode to determine the sheath edge location. Good agreement is found when the LIF and probe data are compared. The LIF data also are compared to the predictions of a simulation that is based on a time-varying two-fluid model of the sheath [G. A. Emmert and M. A. Henry, J. Appl. Phys. 71, 113 (1992)]. While the predictions of the model show moderate agreement with the data, substantial discrepancies are observed. These discrepancies are attributed to a number of physical phenomena that are not included in the present model.

  4. First principles calculations of air fluorescence efficiencies with comparisons to measurements

    SciTech Connect

    Colman, J. J.

    2004-01-01

    The fluorescence efficiencies used in calculating the optical emissions produced by energetic particles and penetrating radiation in air are derived for the most part from the measurements, of Davidson and O'neil, Mitchell and Hartman. These efficiencies were obtained from experiments conducted at various air pressures and in the absence of an applied electric field. In this paper we describe detailed 4.5 dimensional (two spatial dimensions and 2.5 phase space dimensions) kinetic calculations for the electron distribution function resulting from the injection of energetic electrons into air at various pressures. We choose beam parameters and dimensions that are directly relevant to the original Davidson and O'Neil experiments. From the electron distribution function and measured excitation cross-sections we then compute the optical efficiencies for a large number of nitrogen and oxygen lines across the electromagnetic spectrum from 320.0 nm to 800.0 nm. A comparison with various measurements is presented. We also present results from simulations with an applied electric field. The computed fluorescence efficiencies can be used to determine the optical emissions associated with high-altitude discharges driven by runaway air breakdown and results are discussed in a separate poster. We have also recalculated optical emission rates that are applicable to discharges dominated by. conventional breakdown for comparison with Taranenko et al. These rates are also used in our self-consistent sprite simulations.

  5. The Monte Carlo modelling of in vivo x-ray fluorescence measurement of lead in tissue.

    PubMed

    Wallace, J D

    1994-10-01

    A Monte Carlo model has been developed, using the EGS4 code, to model the in vivo x-ray fluorescence (XRF) measurement of Pb in non-superficial bone/tissue. Unlike previous work in this field the current model incorporates a correction for Doppler broadening of the Compton scatter peak due to the electron momentum distribution of the medium (tissue/water) in which the photons are Compton scattered by convolving the Compton peak of the Monte Carlo generated spectrum with a modified Compton profile for water. This correction improves the agreement between the measured spectral shape obtained using an experimental in vivo x-ray fluorescence Pb analyser with a 109Cd/180 degrees source/geometry combination, measuring a bone phantom at depth in water and the generated spectral shape obtained from the equivalent Monte Carlo model. The model enables improved estimates to be made of the spectral background beneath the Pb Kalpha1 and Kalpha2 x-ray peaks compared with estimates based on simpler models that assume that Compton interactions are with 'free' electrons and hence permits better optimization of in vivo analyser system design. PMID:15551542

  6. X-ray fluorescence analysis (XFA) of thyroidal iodine content (TIC) with an improved measuring system.

    PubMed

    Reiners, C; Hänscheid, H; Lassmann, M; Tiemann, M; Kreissl, M; Rendl, J; Bier, D

    1998-01-01

    X-ray fluorescence analysis is based on the principal that the electron structure of stable iodine in the thyroid is excited by Americium-241 gamma rays to emit a characteristic fluorescence radiation which is proportional to the amount of iodine present in the gland. A stationary measuring system consisting of a 11.1 GBq Am-241 source and a high-purity Germanium detector with spectrum analyser has been improved by a PC guided method for sonographic definition of the measuring volume. The lower limit of detectibility of the system corresponds to 0.01 mg of Iodine per ml of thyroid volume; the in vivo precision given as coefficient of variation amounts to 15%. The thyroid is exposed with a radiation dose of 6 microSv per measurement. First studies with this improved system carried out in 50 female volunteers between 20 and 40 years of age with normal thyroid volumes resulted in a mean iodine concentration of the thyroid of 0.665 +/- 0.304 mg/ml. The mean iodine excretion in urine was normal with 10.8 +/- 10.4 microg/dl. PMID:9865551

  7. Measurement of air-fluorescence-light yield induced by an electromagnetic shower

    NASA Astrophysics Data System (ADS)

    MACFLY Collaboration; Colin, P.; Chukanov, A.; Grebenyuk, V.; Naumov, D.; Nédélec, P.; Nefedov, Yu.; Onofre, A.; Porokhovoi, S.; Sabirov, B.; Tkatchev, L.

    2009-01-01

    For most of the ultra-high-energy cosmic ray (UHECR) experiments and projects (HiRes, AUGER, TA, JEM-EUSO, TUS, …), the detection technique of extensive air showers is based, at least, on the measurement of the air-fluorescence-induced signal. The knowledge of the fluorescence-light yield (FLY) is of paramount importance for the UHECR energy reconstruction. The MACFLY experiment was designed to perform absolute measurements of the air FLY and to study its properties. Here, we report the result of measurement of dry-air FLY induced by 50 GeV electromagnetic showers as a function of the shower age and as a function of the pressure. The experiment was performed at CERN using a SPS-electron-test-beam line. The result shows the air FLY is proportional to the energy deposited in air (Ed). The ratio FLY/Ed and its pressure dependence remain constant independently of shower age, and more generally, independently of the excitation source used (single-electron track or air shower).

  8. 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.

  9. Rapid parallel measurements of macroautophagy and mitophagy in mammalian cells using a single fluorescent biosensor

    PubMed Central

    Sargsyan, A.; Cai, J.; Fandino, L. B.; Labasky, M. E.; Forostyan, T.; Colosimo, L. K.; Thompson, S. J.; Graham, T. E.

    2015-01-01

    Mitochondrial dysfunction is implicated in many human diseases and occurs in normal aging. Mitochondrial health is maintained through organelle biogenesis and repair or turnover of existing mitochondria. Mitochondrial turnover is principally mediated by mitophagy, the trafficking of damaged mitochondria to lysosomes via macroautophagy (autophagy). Mitophagy requires autophagy, but is itself a selective process that relies on specific autophagy-targeting mechanisms, and thus can be dissociated from autophagy under certain circumstances. Therefore, it is important to assess autophagy and mitophagy together and separately. We sought to develop a robust, high-throughput, quantitative method for monitoring both processes in parallel. Here we report a flow cytometry-based assay capable of rapid parallel measurements of mitophagy and autophagy in mammalian cells using a single fluorescent protein biosensor. We demonstrate the ability of the assay to quantify Parkin-dependent selective mitophagy in CCCP-treated HeLa cells. In addition, we show the utility of the assay for measuring mitophagy in other cell lines, as well as for Parkin-independent mitophagy stimulated by deferiprone. The assay makes rapid measurements (10,000 cells per 6 seconds) and can be combined with other fluorescent indicators to monitor distinct cell populations, enabling design of high-throughput screening experiments to identify novel regulators of mitophagy in mammalian cells. PMID:26215030

  10. Extracellular Space Volume Measured by Two-Color Pulsed Dye Infusion with Microfiberoptic Fluorescence Photodetection

    PubMed Central

    Magzoub, Mazin; Zhang, Hua; Dix, James A.; Verkman, A.S.

    2009-01-01

    The extracellular space (ECS) is the aqueous matrix surrounding cells in solid tissues. The only method to measure ECS volume fraction (α) in vivo has been tetramethylammonium iontophoresis, a technically challenging method developed more than 25 years ago. We report a simple, quantitative method to measure α by microfiberoptic fluorescence detection of a self-quenched green dye, calcein, and a reference red dye, sulforhodamine 101, after pulsed iontophoretic infusion. The idea is that the maximum increase in calcein fluorescence after iontophoresis is proportional to the aqueous volume into which the dye is deposited. We validated the method theoretically, and experimentally, using cell-embedded gels with specified α and ECS viscosity. Measurements in living mice gave α of 0.20 ± 0.01 in brain, 0.13 ± 0.02 in kidney and 0.074 ± 0.01 in skeletal muscle. The technical simplicity of the ”pulsed-infusion microfiberoptic photodetection” method developed here should allow elucidation of the relatively understudied biological roles of the ECS. PMID:19289063

  11. 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.

  12. Airborne laser-induced oceanic chlorophyll fluorescence: solar-induced quenching corrections by use of concurrent downwelling irradiance measurements.

    PubMed

    Hoge, F E; Wright, C W; Swift, R N; Yungel, J K

    1998-05-20

    Airborne laser-induced (and water Raman-normalized) spectral fluorescence emissions from oceanic chlorophyll were obtained during variable downwelling irradiance conditions induced by diurnal variability and patchy clouds. Chlorophyll fluorescence profiles along geographically repeated inbound and outbound flight track lines, separated in time by approximately 3-6 h and subject to overlying cloud movement, were found to be identical after corrections made with concurrent downwelling irradiance measurements. The corrections were accomplished by a mathematical model containing an exponential of the ratio of the instantaneous-to-average downwelling irradiance. Concurrent laser-induced phycoerythrin fluorescence and chromophoric dissolved organic matter fluorescence were found to be invariant to downwelling irradiance and thus, along with sea-surface temperature, established the near constancy of the oceanic surface layer during the experiment and validated the need for chlorophyll fluorescence quenching corrections over wide areas of the ocean.

  13. General Principles for Measuring Resting Membrane Potential and Ion Concentration Using Fluorescent Bioelectricity Reporters

    PubMed Central

    Adams, Dany Spencer; Levin, Michael

    2014-01-01

    This overview provides the basic information needed to understand, choose, and use fluorescent bioelectricity reporters (FBRs), where bioelectricity is defined as cell processes that involve ions or ion flux. While traditional methods of measuring these characteristics are still valid and necessary, the utility of FBRs has facilitated measurement of these properties under circumstances that are not possible with microelectrodes. Specifically, these dyes can be used to achieve subcellular resolution, to measure many cells simultaneously in vivo, and to track bioelectric gradients over long time periods despite cell movements and divisions. This article covers the basic principles underlying the interpretation of the dye signals, describes essential steps for troubleshooting, optimizing data collection, analysis, and presentation, and provides compilations of information that are useful for choosing FBRs for particular projects. PMID:22474653

  14. A cryogenically cooled photofragment fluorescence instrument for measuring stratospheric water vapor

    NASA Technical Reports Server (NTRS)

    Weinstock, Elliot M.; Schwab, James J.; Nee, Jan Bai; Schwab, M. J.; Anderson, James G.

    1990-01-01

    An instrument developed for high-resolution daytime measurements of water vapor in the stratosphere using the technique of photofragment fluorescence is examined. A detailed description of all aspects of the instrument, as well as the results of its first two flights, are presented. The main areas of concern were optical baffling, cryogen transfer, water vapor measurement without contamination, and a dual path absorption measurement. Results of the second flight test indicate that the problems of instrument and gondola contamination, identified in the first flight test, were solved. A signal-to-noise ratio of about 50:1 for 10 sec of averaging throughout the stratosphere is achieved, as well as an altitude resolution of better than 100 m.

  15. 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.

  16. Spreading convulsions, spreading depolarization and epileptogenesis in human cerebral cortex

    PubMed Central

    Major, Sebastian; Pannek, Heinz-Wolfgang; Woitzik, Johannes; Scheel, Michael; Wiesenthal, Dirk; Martus, Peter; Winkler, Maren K.L.; Hartings, Jed A.; Fabricius, Martin; Speckmann, Erwin-Josef; Gorji, Ali

    2012-01-01

    Spreading depolarization of cells in cerebral grey matter is characterized by massive ion translocation, neuronal swelling and large changes in direct current-coupled voltage recording. The near-complete sustained depolarization above the inactivation threshold for action potential generating channels initiates spreading depression of brain activity. In contrast, epileptic seizures show modest ion translocation and sustained depolarization below the inactivation threshold for action potential generating channels. Such modest sustained depolarization allows synchronous, highly frequent neuronal firing; ictal epileptic field potentials being its electrocorticographic and epileptic seizure its clinical correlate. Nevertheless, Leão in 1944 and Van Harreveld and Stamm in 1953 described in animals that silencing of brain activity induced by spreading depolarization changed during minimal electrical stimulations. Eventually, epileptic field potentials were recorded during the period that had originally seen spreading depression of activity. Such spreading convulsions are characterized by epileptic field potentials on the final shoulder of the large slow potential change of spreading depolarization. We here report on such spreading convulsions in monopolar subdural recordings in 2 of 25 consecutive aneurismal subarachnoid haemorrhage patients in vivo and neocortical slices from 12 patients with intractable temporal lobe epilepsy in vitro. The in vitro results suggest that γ-aminobutyric acid-mediated inhibition protects from spreading convulsions. Moreover, we describe arterial pulse artefacts mimicking epileptic field potentials in three patients with subarachnoid haemorrhage that ride on the slow potential peak. Twenty-one of the 25 subarachnoid haemorrhage patients (84%) had 656 spreading depolarizations in contrast to only three patients (12%) with 55 ictal epileptic events isolated from spreading depolarizations. Spreading depolarization frequency and depression

  17. Spreading convulsions, spreading depolarization and epileptogenesis in human cerebral cortex.

    PubMed

    Dreier, Jens P; Major, Sebastian; Pannek, Heinz-Wolfgang; Woitzik, Johannes; Scheel, Michael; Wiesenthal, Dirk; Martus, Peter; Winkler, Maren K L; Hartings, Jed A; Fabricius, Martin; Speckmann, Erwin-Josef; Gorji, Ali

    2012-01-01

    Spreading depolarization of cells in cerebral grey matter is characterized by massive ion translocation, neuronal swelling and large changes in direct current-coupled voltage recording. The near-complete sustained depolarization above the inactivation threshold for action potential generating channels initiates spreading depression of brain activity. In contrast, epileptic seizures show modest ion translocation and sustained depolarization below the inactivation threshold for action potential generating channels. Such modest sustained depolarization allows synchronous, highly frequent neuronal firing; ictal epileptic field potentials being its electrocorticographic and epileptic seizure its clinical correlate. Nevertheless, Leão in 1944 and Van Harreveld and Stamm in 1953 described in animals that silencing of brain activity induced by spreading depolarization changed during minimal electrical stimulations. Eventually, epileptic field potentials were recorded during the period that had originally seen spreading depression of activity. Such spreading convulsions are characterized by epileptic field potentials on the final shoulder of the large slow potential change of spreading depolarization. We here report on such spreading convulsions in monopolar subdural recordings in 2 of 25 consecutive aneurismal subarachnoid haemorrhage patients in vivo and neocortical slices from 12 patients with intractable temporal lobe epilepsy in vitro. The in vitro results suggest that γ-aminobutyric acid-mediated inhibition protects from spreading convulsions. Moreover, we describe arterial pulse artefacts mimicking epileptic field potentials in three patients with subarachnoid haemorrhage that ride on the slow potential peak. Twenty-one of the 25 subarachnoid haemorrhage patients (84%) had 656 spreading depolarizations in contrast to only three patients (12%) with 55 ictal epileptic events isolated from spreading depolarizations. Spreading depolarization frequency and depression

  18. 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.

  19. 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.

  20. The Identification of Mitogen Responding Subpopulations of Human Lymphocytes by Flow Polarimeter Fluorescence Measurements.

    NASA Astrophysics Data System (ADS)

    Chan, Sandra Lynn

    I have developed a method to identify the mitogen responding subpopulation of human peripheral blood lymphocytes. This method employs a flow polarimeter to measure the distribution of the intensity and the polarization of intracellular fluorescein fluorescence in suspensions of mononuclear cells isolated on density gradients from the peripheral blood of donors. I have used the change in the fluorescence of cells exposed to the mitogens PHA and Con A to identify the responding cells and to quantitate this number. I have found that for most donors, the responding cells constitute about 20-40% of the lymphocyte population. The percent of responding cells decreases to zero in patients with acute lymphocytic leukemia (2 patients) and chronic lymphocyte leukemia (10 patients). For a variety of patients with other types of cancer, the responding fraction was not significantly different from healthy controls. Moreover, the number of responding cells does not appear to be age dependent in the age range of 20-80 years. I also found that the change in fluorescence polarization correlated strongly with changes in fluorescence intensity induced by mitogens--the number of responding cells, therefore can be estimated either from the intensity or polarization distributions. The shapes of fluorescence distributions depend strongly on a number of variables including the composition and density of the lymphocyte isolating medium, the mitogen and dye concentrations, the length of incubation with mitogen or dye, and the potassium, calcium, and magnesium concentrations in the medium. In the case of fluorescein, I have worked out a methodology that allows a consistent estimate of the responding lymphocyte number. I have also investigated the use of the dye carbocyanine for the same purpose. This dye presumably identifies the mitogen responding lymphocytes on the basis of changes in membrane potential. The results with carbocyanine were found to depend on a number of variables and I could

  1. 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

  2. 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

  3. Simple relation between lidar multiple scattering and depolarization for water clouds.

    PubMed

    Hu, Yongxiang; Liu, Zhaoyan; Winker, David; Vaughan, Mark; Noel, Vincent; Bissonnette, Luc; Roy, Gilles; McGill, Matthew

    2006-06-15

    An empirical relationship is derived between the multiple-scattering fraction and the linear depolarization ratio by using Monte Carlo simulations of water clouds measured by backscatter lidar. This relationship is shown to hold for clouds having a wide range of extinction coefficients, mean droplet sizes, and droplet size distribution widths. The relationship is also shown to persist for various instrument fields of view and for measurements made within broken cloud fields. The results obtained from the Monte Carlo simulations are verified by using multiple-field-of-view lidar measurements. For space-based lidars equipped to measure linear depolarization ratios, this new relationship can be used to accurately assess signal perturbations due to multiple scattering within nonprecipitating water clouds.

  4. 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

  5. A Simple and Rapid Protocol for Measuring Neutral Lipids in Algal Cells Using Fluorescence

    PubMed Central

    Storms, Zachary J.; Cameron, Elliot; de la Hoz Siegler, Hector; McCaffrey, William C.

    2014-01-01

    Algae are considered excellent candidates for renewable fuel sources due to their natural lipid storage capabilities. Robust monitoring of algal fermentation processes and screening for new oil-rich strains requires a fast and reliable protocol for determination of intracellular lipid content. Current practices rely largely on gravimetric methods to determine oil content, techniques developed decades ago that are time consuming and require large sample volumes. In this paper, Nile Red, a fluorescent dye that has been used to identify the presence of lipid bodies in numerous types of organisms, is incorporated into a simple, fast, and reliable protocol for measuring the neutral lipid content of Auxenochlorella protothecoides, a green alga. The method uses ethanol, a relatively mild solvent, to permeabilize the cell membrane before staining and a 96 well micro-plate to increase sample capacity during fluorescence intensity measurements. It has been designed with the specific application of monitoring bioprocess performance. Previously dried samples or live samples from a growing culture can be used in the assay. PMID:24961928

  6. Atmospheric ammonia measurement using a VUV/photo-fragmentation laser-induced fluorescence technique.

    PubMed

    Schendel, J S; Stickel, R E; Vandijk, C A; Sandholm, S T; Davis, D D; Bradshaw, J D

    1990-11-20

    Vacuum ultraviolet/photofragmentation laser-induced fluorescence has been demonstrated to be a highly specific and sensitive method for the quantitative measurement of atmospheric ammonia (NH(3)). The fluorescence detected in this approach results from the two 193-nm photon photofragmentation step NH(3)?NH(2)? NH(b(1)Sigma(+)) followed by the excitation of the NH(b(1)Sigma(+)) NH(c(1)Pi) transition via a 450-nm photon with final emission being observed from the NH(c(1) Pi) NH(a(1)Delta) transition at 325 nm. Limits of detection for the instrumentpresented here are < 10 pptv and < 4 pptv for 1- and 5-min integration periods, respectively, in ambient sampling conditions. The technique is free from interferences and system performance does not significantly degrade in adverse sampling conditions (i.e., rain, fog, clouds, haze, etc.). Spectroscopic selectivity in the NH(b(1)Sigma(+))?NH(c(1)Pi) transition is sufficient to resolve (15)NH(3) and (14)NH(3) contributions for use in atmospheric tracer studies. Average ammonia measurements at Stone Mountain, GA, ranged from approximately 110 pptv for air temperatures <5 degrees C to approximately 240 pptv for air temperatures >/=<5 degrees C over the period from Dec. 1987 to the end of Apr. 1988.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. Measurement of glycated haemoglobin in whole blood by a novel fluorescence quenching assay.

    PubMed

    Blincko, S; Anzetse, J; Edwards, R

    2000-07-01

    We describe a method for the specific measurement of glycated Hb (GHb) by fluorescence quenching. Whole blood is added to lysing solution, then the lysate is mixed with eosin-boronic acid solution and reacted for at least 5 min at room temperature. The quenching of the fluorescence of the eosin-boronic acid solution is proportional to the concentration of GHb present. Total Hb concentration was measured by absorbance and the GHb expressed as a percentage of the total Hb. Comparison with a commercial high-performance liquid chromatography (HPLC) system for HbA1c showed: %GHb=1.30 (SD 0.04) %HbA1c + 1.36 (SD 0.30), S(y/x) 0.803, n=95, r=0.965 (SD=standard deviation). Intra-assay coefficients of variation were <2.5% (for GHb concentrations in the range 6-20%) and inter-assay coefficients of variation were <4.1% (10 assays on six samples with GHb concentrations in the range 6-20%). Linearity of response was demonstrated by dilution. The effect of adding exogenous glucose, bilirubin and triglycerides was tested on samples with low, medium and high GHb concentrations. No significant interference was found. Variation of haematocrit over the range 0.4-0.6 also had no significant effect on percentage GHb. Preliminary results with samples containing variant Hb (HbAS and HbAC) indicated good agreement with HPLC for these samples also. PMID:10902866

  12. 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.

  13. 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

  14. 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

  15. 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.

  16. A model for depolarized radar echoes from Mars

    NASA Technical Reports Server (NTRS)

    Thompson, T. W.; Moore, H. J.

    1989-01-01

    The depolarized radar echoes from Mars are modeled using a combination of remote-sensing observations. The model reproduces the variations of the total radar cross-sections with longitude observed by Goldstone (1986) along 7 S, yields larger magnitudes of total radar cross-sections along 22 N than those along 7 S, and produces depolarized echo spectra that broadly match those observed by the Arecibo radar in 1980 and 1982. The model indicates that volcanoes and lava plains of the Tharsis-Alba Patera, Elysium, and Amazonia regions have the strongest depolarized echoes from the entire planet. Rock populations for the moon and Mars are estimated assuming depolarized radar echoes result from rocks with radii between 1.3 and three times the wavelength.

  17. UV Raman and Fluorescence for Multi-Species Measurement in Hydrocarbon-Fueled High-Speed Propulsion

    NASA Technical Reports Server (NTRS)

    Skaggs, Patricia Annette; Nandula, Sastri P.; Pitz, Robert W.

    1999-01-01

    This report documents work performed through the NASA Graduate Student Researchers Program, Grant No. NGT3-52316. Research performed included investigation of two-line fluorescence imaging of OH for temperature measurement and an investigation of negative flame speeds for modeling of premixed turbulent flames. The laboratory work and initial analysis of the fluorescence imaging was performed at NASA Glen Research Center with follow up analysis at Vanderbilt University. The negative flame speed investigation was performed using an opposed jet flow simulation program at Vanderbilt University. The fluorescence imaging work is presented first followed by the negative flame speed investigation.

  18. Nuclear depolarization and absolute sensitivity in magic-angle spinning cross effect dynamic nuclear polarization.

    PubMed

    Mentink-Vigier, Frédéric; Paul, Subhradip; Lee, Daniel; Feintuch, Akiva; Hediger, Sabine; Vega, Shimon; De Paëpe, Gaël

    2015-09-14

    Over the last two decades solid state Nuclear Magnetic Resonance has witnessed a breakthrough in increasing the nuclear polarization, and thus experimental sensitivity, with the advent of Magic Angle Spinning Dynamic Nuclear Polarization (MAS-DNP). To enhance the nuclear polarization of protons, exogenous nitroxide biradicals such as TOTAPOL or AMUPOL are routinely used. Their efficiency is usually assessed as the ratio between the NMR signal intensity in the presence and the absence of microwave irradiation εon/off. While TOTAPOL delivers an enhancement εon/off of about 60 on a model sample, the more recent AMUPOL is more efficient: >200 at 100 K. Such a comparison is valid as long as the signal measured in the absence of microwaves is merely the Boltzmann polarization and is not affected by the spinning of the sample. However, recent MAS-DNP studies at 25 K by Thurber and Tycko (2014) have demonstrated that the presence of nitroxide biradicals combined with sample spinning can lead to a depolarized nuclear state, below the Boltzmann polarization. In this work we demonstrate that TOTAPOL and AMUPOL both lead to observable depolarization at ≈110 K, and that the magnitude of this depolarization is radical dependent. Compared to the static sample, TOTAPOL and AMUPOL lead, respectively, to nuclear polarization losses of up to 20% and 60% at a 10 kHz MAS frequency, while Trityl OX63 does not depolarize at all. This experimental work is analyzed using a theoretical model that explains how the depolarization process works under MAS and gives new insights into the DNP mechanism and into the spin parameters, which are relevant for the efficiency of a biradical. In light of these results, the outstanding performance of AMUPOL must be revised and we propose a new method to assess the polarization gain for future radicals.

  19. 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.

  20. Quantitative measurement of Ca2+ and Zn2+ in mammalian cells using genetically encoded fluorescent biosensors

    PubMed Central

    Park, J. Genevieve; Palmer, Amy E.

    2014-01-01

    Summary Genetically encoded, ratiometric, fluorescent biosensors can be used to quantitatively measure intracellular ion concentrations in living cells. We describe important factors to consider when selecting a Ca2+ or Zn2+ biosensor, such as the sensor’s dissociation constant (Kd’) and its dynamic range. We also discuss the limits of quantitative measurement using these sensors and reasons why a sensor may perform differently in different biological systems or subcellular compartments. We outline protocols for 1) quickly confirming sensor functionality in a new biological system, 2) calibrating a sensor to convert a sensor’s FRET ratio to ion concentration, and 3) titrating a sensor in living cells to obtain its Kd’ under different experimental conditions. PMID:24052378

  1. Rates of benthic protozoan grazing on free and attached sediment bacteria measured with fluorescently stained sediment.

    PubMed

    Starink, M; Krylova, I N; Bär-Gilissen, M J; Bak, R P; Cappenberg, T E

    1994-07-01

    In order to determine the importance of benthic protozoa as consumers of bacteria, grazing rates have been measured by using monodispersed fluorescently labeled bacteria (FLB). However, high percentages of nongrazing benthic protists are reported in the literature. These are related to serious problems of the monodispersed FLB method. We describe a new method using 5-(4,6-dichlorotriazin-2-yl)-aminofluorescein (DTAF)-stained sediment to measure in situ bacterivory by benthic protists. This method is compared with the monodispersed FLB technique. Our estimates of benthic bacterivory range from 61 to 73 bacteria protist h and are about twofold higher than the results of the monodispersed FLB method. The number of nongrazing protists after incubation for 15 min with DTAF-stained sediment is in agreement with theoretical expectation. We also tested the relative affinity for FLB of protists and discuss the results with respect to a grazing model.

  2. Fluorescence Resonance Energy Transfer Microscopy for Measuring Chromatin Complex Structure and Dynamics.

    PubMed

    Cherubini, Alessandro; Zippo, Alessio

    2016-01-01

    The Polycomb group (PcG) proteins form regulatory complexes that modify the chromatin structure and silence their target genes. Recent works have found that the composition of Polycomb complexes is highly dynamic. Defining the different protein components of each complex is fundamental for better understanding their biological functions. Fluorescent resonance energy transfer (FRET) is a powerful tool to measure protein-protein interactions, in nanometer order and in their native cellular environment. Here we describe the preparation and execution of a typical FRET experiment using CFP-tagged protein as donor and YFP-tagged protein as acceptor. We further show that FRET can be used in a competition assay to measure binding affinities of different components of the same chromatin complex. PMID:27659982

  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. Experimental phase diagram of negatively supercoiled DNA measured by magnetic tweezers and fluorescence.

    PubMed

    Vlijm, Rifka; Mashaghi, Alireza; Bernard, Stéphanie; Modesti, Mauro; Dekker, Cees

    2015-02-21

    The most common form of DNA is the well-known B-structure of double-helix DNA. Many processes in the cell, however, exert force and torque, inducing structural changes to the DNA that are vital to biological function. Virtually all DNA in cells is in a state of negative supercoiling, with a DNA structure that is complex. Using magnetic tweezers combined with fluorescence imaging, we here study DNA structure as a function of negative supercoiling at the single-molecule level. We classify DNA phases based on DNA length as a function of supercoiling, down to a very high negative supercoiling density σ of -2.5, and forces up to 4.5 pN. We characterize plectonemes using fluorescence imaging. DNA bubbles are visualized by the binding of fluorescently labelled RPA, a eukaryotic single-strand-binding protein. The presence of Z-DNA, a left-handed form of DNA, is probed by the binding of Zα77, the minimal binding domain of a Z-DNA-binding protein. Without supercoiling, DNA is in the relaxed B-form. Upon going toward negative supercoiling, plectonemic B-DNA is being formed below 0.6 pN. At higher forces and supercoiling densities down to about -1.9, a mixed state occurs with plectonemes, multiple bubbles and left-handed L-DNA. Around σ = -1.9, a buckling transition occurs after which the DNA end-to-end length linearly decreases when applying more negative turns, into a state that we interpret as plectonemic L-DNA. By measuring DNA length, Zα77 binding, plectoneme and ssDNA visualisation, we thus have mapped the co-existence of many DNA structures and experimentally determined the DNA phase diagram at (extreme) negative supercoiling.

  5. 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.

  6. A deterministic method for studying depolarization in turbid media

    NASA Astrophysics Data System (ADS)

    Clark, Julia P.

    2016-05-01

    There are a number of interesting experimental and Monte Carlo results regarding the persistence of polarization in turbid media; however, there is not a good theoretical understanding of this phenomenon. These results include circular polarization memory in strongly scattering anisotropic media and the impact of polydisperse scatterers on the depolarization rate. In this work we use the spectrum of the discretized vector radiative transport equation to investigate to study circular depolarization in strongly scattering media.

  7. 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.

  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. Primary afferent depolarization and frequency processing in auditory afferents.

    PubMed

    Baden, Tom; Hedwig, Berthold

    2010-11-01

    Presynaptic inhibition is a widespread mechanism modulating the efficiency of synaptic transmission and in sensory pathways is coupled to primary afferent depolarizations. Axonal terminals of bush-cricket auditory afferents received 2-5 mV graded depolarizing inputs, which reduced the amplitude of invading spikes and indicated presynaptic inhibition. These inputs were linked to a picrotoxin-sensitive increase of Ca(2+) in the terminals. Electrophysiological recordings and optical imaging showed that in individual afferents the sound frequency tuning based on spike rates was different from the tuning of the graded primary afferent depolarizations. The auditory neuropil of the bush-cricket Mecopoda elongata is tonotopically organized, with low frequencies represented anteriorly and high frequencies represented posteriorly. In contrast graded depolarizing inputs were tuned to high-frequencies anteriorly and to low-frequencies posteriorly. Furthermore anterior and posterior axonal branches of individual afferents received different levels of primary afferent depolarization depending on sound frequency. The presence of primary afferent depolarization in the afferent terminals indicates that presynaptic inhibition may shape the synaptic transmission of frequency-specific activity to auditory interneurons.

  10. 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.

  11. Impaired neurovascular coupling to ictal epileptic activity and spreading depolarization in a patient with subarachnoid hemorrhage: Possible link to blood–brain barrier dysfunction

    PubMed Central

    Winkler, Maren K. L.; Chassidim, Yoash; Lublinsky, Svetlana; Revankar, Gajanan S.; Major, Sebastian; Kang, Eun-Jeung; Oliveira-Ferreira, Ana I.; Woitzik, Johannes; Sandow, Nora; Scheel, Michael; Friedman, Alon; Dreier, Jens P.

    2013-01-01

    SUMMARY Spreading depolarization describes a sustained neuronal and astroglial depolarization with abrupt ion translocation between intraneuronal and extracellular space leading to a cytotoxic edema and silencing of spontaneous activity. Spreading depolarizations occur abundantly in acutely injured human brain and are assumed to facilitate neuronal death through toxic effects, increased metabolic demand, and inverse neurovascular coupling. Inverse coupling describes severe hypoperfusion in response to spreading depolarization. Ictal epileptic events are less frequent than spreading depolarizations in acutely injured human brain but may also contribute to lesion progression through increased metabolic demand. Whether abnormal neurovascular coupling can occur with ictal epileptic events is unknown. Herein we describe a patient with aneurysmal subarachnoid hemorrhage in whom spreading depolarizations and ictal epileptic events were measured using subdural opto-electrodes for direct current electrocorticography and regional cerebral blood flow recordings with laser-Doppler flowmetry. Simultaneously, changes in tissue partial pressure of oxygen were recorded with an intraparenchymal oxygen sensor. Isolated spreading depolarizations and clusters of recurrent spreading depolarizations with persistent depression of spontaneous activity were recorded over several days followed by a status epilepticus. Both spreading depolarizations and ictal epileptic events where accompanied by hyperemic blood flow responses at one optode but mildly hypoemic blood flow responses at another. Of note, quantitative analysis of Gadolinium-diethylene-triamine-pentaacetic acid (DTPA)–enhanced magnetic resonance imaging detected impaired blood–brain barrier integrity in the region where the optode had recorded the mildly hypoemic flow responses. The data suggest that abnormal flow responses to spreading depolarizations and ictal epileptic events, respectively, may be associated with blood

  12. 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

  13. Plant chlorophyll fluorescence: active and passive measurements at canopy and leaf scales with different nitrogen treatments.

    PubMed

    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.

  14. 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.

  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. 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.

  17. 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.

  18. OH-Planar Fluorescence Measurements of Pressurized, Hydrogen Premixed Flames in the SimVal Combustor

    SciTech Connect

    Strakey, P.A.; Woodruff, S.D.; Williams, T.C.; Schefer, R.W.

    2008-07-01

    Planar laser-induced fluorescence measurements of the hydroxyl radical in lean, premixed natural gas flames augmented with hydrogen are presented. The experiments were conducted in the Simulation Validation combustor at the National Energy Technology Laboratory at operating pressures from 1 to 8 atmospheres. The data, which were collected in a combustor with well-controlled boundary conditions, are intended to be used for validating computational fluid dynamics models under conditions directly relevant to land-based gas turbine engines. The images, which show significant effects of hydrogen on local flame quenching, are discussed in terms of a turbulent premixed combustion regime and nondimensional parameters such as Karlovitz number. Pressure was found to thin the OH region, but only had a secondary effect on overall flame shape compared with the effects of hydrogen addition, which was found to decrease local quenching and shorten the turbulent flame brush. A method to process the individual images based on local gradients of fluorescence intensity is proposed, and results are presented. Finally, the results of several large eddy simulations are presented and compared with the experimental data in an effort to understand the issues related to model validation, especially for simulations that do not include OH as an intermediate species.

  19. Trifluoperazine and dibucaine-induced inhibition of glutamate-induced mitochondrial depolarization in rat cultured forebrain neurones.

    PubMed

    Hoyt, K R; Sharma, T A; Reynolds, I J

    1997-11-01

    1. Glutamate receptor activation has been previously shown to result in mitochondrial depolarization and activation of the mitochondrial permeability transition pore in cultured neurones. In this study, we characterized the effects of two putative permeability transition inhibitors, namely trifluoperazine and dibucaine, on mitochondrial depolarization in rat intact, cultured forebrain neurones. 2. Permeability transition was monitored by following mitochondrial depolarization in neurones loaded with the mitochondrial membrane potential-sensitive fluorescent indicator, JC-1. Trifluoperazine (10 20 microM) and dibucaine (50-100 microM) inhibited or delayed the onset of glutamate-induced permeability transition. 3. We also investigated the effects of trifluoperazine and dibucaine on neuronal recovery from glutamate-induced Ca2+ loads. Trifluoperazine affected Ca2+ recovery in a manner similar to the mitochondrial Na+/Ca2+ exchange inhibitor, CGP-37157, while dibucaine had no apparent effect on Ca2+ recovery. Therefore, inhibition of permeability transition does not appear to be involved in Ca2+ recovery from glutamate-induced Ca2+ loads. 4. Trifluoperazine and dibucaine did not inhibit [3H]-dizocilpine binding at the concentrations that prevented mitochondrial depolarization. 5. These studies suggest that trifluoperazine and dibucaine inhibit permeability transition in intact neurones. Trifluoperazine also appears to inhibit mitochondrial Na+/Ca2+ exchange. These drugs should prove to be valuable tools in the further study of the role of mitochondrial permeability transition in glutamate-induced neuronal death.

  20. Trifluoperazine and dibucaine-induced inhibition of glutamate-induced mitochondrial depolarization in rat cultured forebrain neurones

    PubMed Central

    Hoyt, Kari R; Sharma, Terre A; Reynolds, Ian J

    1997-01-01

    Glutamate receptor activation has been previously shown to result in mitochondrial depolarization and activation of the mitochondrial permeability transition pore in cultured neurones. In this study, we characterized the effects of two putative permeability transition inhibitors, namely trifluoperazine and dibucaine, on mitochondrial depolarization in rat intact, cultured forebrain neurones. Permeability transition was monitored by following mitochondrial depolarization in neurones loaded with the mitochondrial membrane potential-sensitive fluorescent indicator, JC-1. Trifluoperazine (10–20 μM) and dibucaine (50–100 μM) inhibited or delayed the onset of glutamate-induced permeability transition. We also investigated the effects of trifluoperazine and dibucaine on neuronal recovery from glutamate-induced Ca2+ loads. Trifluoperazine affected Ca2+ recovery in a manner similar to the mitochondrial Na+/Ca2+ exchange inhibitor, CGP-37157, while dibucaine had no apparent effect on Ca2+ recovery. Therefore, inhibition of permeability transition does not appear to be involved in Ca2+ recovery from glutamate-induced Ca2+ loads. Trifluoperazine and dibucaine did not inhibit [3H]-dizocilpine binding at the concentrations that prevented mitochondrial depolarization. These studies suggest that trifluoperazine and dibucaine inhibit permeability transition in intact neurones. Trifluoperazine also appears to inhibit mitochondrial Na+/Ca2+ exchange. These drugs should prove to be valuable tools in the further study of the role of mitochondrial permeability transition in glutamate-induced neuronal death. PMID:9384493

  1. Time and frequency-domain measurement of ground-state recovery times in red fluorescent proteins.

    PubMed

    Manna, Premashis; Jimenez, Ralph

    2015-04-16

    The field of bioimaging and biosensors has been revolutionized by the discovery of fluorescent proteins (FPs) and their use in live cells. FPs are characterized with rich photodynamics due to the presence of nonfluorescent or dark states which are responsible for fluorescence intermittency or "blinking", which has been exploited in several localization-based super-resolution techniques that surpass the diffraction-limited resolution of conventional microscopy. Molecules that convert to these dark states recover to the ground states either spontaneously or upon absorption of another photon, depending on the particular FP and the structural transition that is involved. In this work, we demonstrate time- and frequency-domain methods for the measurement of the ground-state recovery (GSR) times of FPs both in live cells and in solutions. In the time-domain method, we excited the sample with millisecond pulses at varying dark times to obtain percent-recovery. In the frequency-domain method, dark-state hysteresis was employed to obtain the positive phase shift or "phase advance". We extracted the GSR time constants from our measurements using calculations and simulations based on a three-state model system. The GSR time constants of the red FPs studied in these experiments fall in the range from μs to msec time-scales. We find that the time- and frequency-domain techniques are complementary to each other. While accurate GSR times can be extracted from the time-domain technique, frequency-domain measurements are primarily sensitive to the rates of dark-state conversion (DSC) processes. A correlation between GSR times, DSC, and photobleaching rates for the red FPs mCherry, TagRFP-T, and Kriek were observed. These time- and frequency-domain methods can be used in high-throughput screening and sorting of FPs clones based on GSR time constant and photostability and will therefore be valuable for the development of new photoswitchable or photoactivatable FPs.

  2. 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.

  3. 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.

  4. 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

  5. Fully adaptive FEM based fluorescence optical tomography from time-dependent measurements with area illumination and detection.

    PubMed

    Joshi, Amit; Bangerth, Wolfgang; Hwang, Kildong; Rasmussen, John C; Sevick-Muraca, Eva M

    2006-05-01

    Using an area-illumination and area-detection scheme, we acquire fluorescence frequency domain measurements from a tissue phantom with an embedded fluorescent target and obtain tomographic reconstructions of the interior fluorescence absorption map with an adaptive finite element based scheme. The tissue phantom consisted of a clear acrylic cubic box (512 ml) filled with 1% Liposyn solution, while the fluorescent targets were 5 mm diameter glass bulbs filled with 1 microM Indocyanine Green dye solution in 1% Liposyn. Frequency domain area illumination and detection employed a planar excitation source using an expanded intensity modulated (100 MHz) 785 nm diode laser light and a gain modulated image intensified charge coupled device camera, respectively. The excitation pattern was characterized by isolating the singly scattered component with cross polarizers and was input into a dual adaptive finite element-based scheme for three dimensional reconstructions of fluorescent targets embedded beneath the phantom surface. Adaptive mesh refinement techniques allowed efficient simulation of the incident excitation light and the reconstruction of fluorescent targets buried at the depths of 1 and 2 cm. The results demonstrate the first clinically relevant noncontact fluorescence tomography with adaptive finite element methods.

  6. 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.

  7. Two-Photon Absorption Laser Induced Fluorescence Measurements of Neutral Density in Helicon Plasma

    NASA Astrophysics Data System (ADS)

    Galante, Matthew

    2013-10-01

    Neutral particles play a critical role in nearly all plasmas, from the pedestal region of a tokamak fusion plasma to industrial plasma processing systems. In fusion plasmas, neutrals at the edge serve as both a source of particles and also a sink of momentum and energy. Control of the edge plasma density in tokamaks is critical for the transition to H-mode plasmas and the role of neutrals in modifying the plasma rotation in the edge is an area of active research. However, few methods exist to make localized, direct neutral density measurements. We have developed a new diagnostic based on two-photon absorption laser induced fluorescence (TALIF). We use a high intensity (5 MW/cm2), narrow bandwidth (0.1 cm-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. In this talk I will 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 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 through a single optical port in the vacuum vessel. The wavelength resolution of the diagnostic is sufficient to separate hydrogen and deuterium spectra and I will present measurements from mixed hydrogen and deuterium plasmas that demonstrate isotopic abundance measurements are feasible with the TALIF system. Time and spatially resolved measurements also allow us to explore the effects of wall recycling and pulse repetition rates on the neutral density profile in the plasma source. Work supported in part by US DOE under DE-FC02-04ER54698.

  8. A Possible Origin of Linear Depolarization Observed at Vertical Incidence in Rain

    NASA Technical Reports Server (NTRS)

    Jameson, A. R.; Durden, S. L.

    1996-01-01

    Recent observations by two different nadir-pointing airborne radars with some polarization capabilities have detected surprisingly large linear depolarization ratios at times in convective tropical rain. This depolarization can be explained if the rain is considered to be a mixture of a group of apparent spheres and another group of drops that are distorted in the horizontal plane perpendicular to the direction of propagation of the incident wave. If confirmed in future observations, this suggests that at times the larger raindrops are oscillating, in part, because of collisions with smaller drops. Since many of the interpretations of radar polarization measurements in rain by ground-based radars presume that the raindrop shapes correspond to those of the well-known "equilibrium" drops, the present observations may require adjustments to some radar polarization algorithms for estimating rainfall rate, for example, if the shape perturbations observed at nadir also apply to measurements along other axes as well.

  9. Measuring lead, mercury, and uranium by in vivo X-ray fluorescence

    NASA Astrophysics Data System (ADS)

    O'Meara, Joanne Michelle

    X-ray fluorescence (XRF) has been demonstrated to be a useful technique for measuring trace quantities of heavy metals in various tissues within the body. This thesis investigates a means of improving the measurement of lead in bone, as well as increasing the existing sensitivity of measuring kidney mercury content. The XRF measurement of uranium is also explored. This work assesses the feasibility of a normalisation method for the 57Co/90° system, in relating detected signal to the lead content of the sample. The feasibility of normalisation has been shown, which reduces subject dose and improves system transportability, as well as removes subjectivity, by eliminating the need for acquiring planar x-ray images of the measurement site. In the measurement of renal mercury concentrations, a gain in sensitivity increasing the x-ray tube operating voltage of the current system is investigated. It found that 250 kV, rather than 175 kV, and a titanium rather than uranium filter, results in a 2.5 +/- 0.2 times gain in sensitivity. This potential improvement could have profound clinical implications for the accuracy of occupational monitoring, and for assessing whether there is a quantitative relationship between biological fluid levels and mercury content in this critical organ. The XRF measurement of bone uranium content is also explored. Both source-excited and polarised systems have been developed, however, the sensitivity is currently beyond that which is useful for occupational monitoring of exposure to this toxin. The particular case of measuring uranium in survivors of "Friendly Fire" incidents (from Operation Desert Storm) is investigated, and the first detectable quantity of uranium has been observed in a member of this cohort, with the XRF system designed and built during the course of this work.

  10. Two photon absorption laser induced fluorescence measurements of neutral density in a helicon plasmaa)

    NASA Astrophysics Data System (ADS)

    Galante, M. E.; Magee, R. M.; Scime, E. E.

    2014-05-01

    We have developed a new diagnostic based on two-photon absorption laser induced fluorescence (TALIF). We use a high intensity (5 MW/cm2), narrow bandwidth (0.1 cm-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.

  11. 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.

  12. 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.

  13. Measurement of the UHECR Energy Spectrum by the Telescope Array Fluorescence Detectors

    NASA Astrophysics Data System (ADS)

    Stroman, Thomas; Bergman, Douglas

    2013-04-01

    Ultra-high-energy cosmic rays (UHECRs), subatomic charged particles of extraterrestrial origin and with kinetic energies near or exceeding 10^18 eV, are very rare. The Telescope Array (TA) experiment in western Utah is the northern hemisphere's largest UHECR detector, and consists of three atmospheric fluorescence detectors (FDs) and a ground array of 507 scintillator detectors. In stand-alone ``monocular'' operation, the FDs can observe the widest range in primary UHECR energies. One FD employs refurbished hardware from the High-Resolution Fly's Eye experiment; the remaining two FDs were designed for TA and employ new hardware and analysis. We will present the UHECR energy spectrum measured by the FDs in monocular mode using data collected during the first four years of operation.

  14. 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.

  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. LABORATORY MEASUREMENTS OF NiH BY FOURIER TRANSFORM DISPERSED FLUORESCENCE

    SciTech Connect

    Vallon, Raphael; Richard, Cyril; Crozet, Patrick; Wannous, Ghassan; Ross, Amanda

    2009-05-01

    Red and orange bands of laser-induced fluorescence in NiH have been recorded on a Fourier transform interferometer at Doppler resolution. The spectra show strong transitions to low-lying vibronic states which are not thermally populated in a laboratory source, and therefore do not appear in laser excitation spectra, but which would be expected to contribute significantly to any stellar spectrum. The strongest bands belong to the G[{omega}' 5/2]-X {sub 2} {sup 2}{delta}{sub 3/2}, I[{omega}' 3/2]-X {sub 2}, and {sup 2}{delta}{sub 3/2} I[{omega}' 3/2]-W {sub 1} {sup 2}{pi}{sub 3/2} systems. Measurements are reported for {sup 58}NiH, {sup 60}NiH, and {sup 62}NiH.

  17. 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

  18. Measurement of gas density and temperature distributions in strongly rotating UF/sub 6/ using laser-induced fluorescence

    SciTech Connect

    Gentry, R.A.; Caldwell, S.E.; White, R.W.

    1981-01-01

    A new technique for using Laser Induced Fluorescence (LIF) signals to measure the distribution of gas density and temperature in strongly rotating UF/sub 6/ gas is presented. An external pulsed laser is used to excite the rotating UF/sub 6/ gas, producing an exponentially decaying fluorescence signal. A multi-channel fiber optics system simultaneously collects the fluorescence signals emanating from a number of points in the gas. The signals from each optical channel are digitized and processed to determine the fluorescence signal intensity and decay lifetime at each of the points of observation by means of a least squares fitting process. Gas densities and temperatures are then determined from the intensity and lifetime data. A recently constructed LIF probe system is described and an analysis of the unfolding techniques necessary to process the signal data is presented. Preliminary data, obtained in tests of the probe system in a laboratory rotor, are presented.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. Classification of particle effective shape ratios in cirrus clouds based on the lidar depolarization ratio.

    PubMed

    Noel, Vincent; Chepfer, Helene; Ledanois, Guy; Delaval, Arnaud; Flamant, Pierre H

    2002-07-20

    A shape classification technique for cirrus clouds that could be applied to future spaceborne lidars is presented. A ray-tracing code has been developed to simulate backscattered and depolarized lidar signals from cirrus clouds made of hexagonal-based crystals with various compositions and optical depth, taking into account multiple scattering. This code was used first to study the sensitivity of the linear depolarization rate to cloud optical and microphysical properties, then to classify particle shapes in cirrus clouds based on depolarization ratio measurements. As an example this technique has been applied to lidar measurements from 15 mid-latitude cirrus cloud cases taken in Palaiseau, France. Results show a majority of near-unity shape ratios as well as a strong correlation between shape ratios and temperature: The lowest temperatures lead to high shape ratios. The application of this technique to space-borne measurements would allow a large-scale classification of shape ratios in cirrus clouds, leading to better knowledge of the vertical variability of shapes, their dependence on temperature, and the formation processes of clouds.

  5. Faraday signature of magnetic helicity from reduced depolarization

    SciTech Connect

    Brandenburg, Axel; Stepanov, Rodion

    2014-05-10

    Using one-dimensional models, we show that a helical magnetic field with an appropriate sign of helicity can compensate the Faraday depolarization resulting from the superposition of Faraday-rotated polarization planes from a spatially extended source. For radio emission from a helical magnetic field, the polarization as a function of the square of the wavelength becomes asymmetric with respect to zero. Mathematically speaking, the resulting emission occurs then either at observable or at unobservable (imaginary) wavelengths. We demonstrate that rotation measure (RM) synthesis allows for the reconstruction of the underlying Faraday dispersion function in the former case, but not in the latter. The presence of positive magnetic helicity can thus be detected by observing positive RM in highly polarized regions in the sky and negative RM in weakly polarized regions. Conversely, negative magnetic helicity can be detected by observing negative RM in highly polarized regions and positive RM in weakly polarized regions. The simultaneous presence of two magnetic constituents with opposite signs of helicity is shown to possess signatures that can be quantified through polarization peaks at specific wavelengths and the gradient of the phase of the Faraday dispersion function. Similar polarization peaks can tentatively also be identified for the bi-helical magnetic fields that are generated self-consistently by a dynamo from helically forced turbulence, even though the magnetic energy spectrum is then continuous. Finally, we discuss the possibility of detecting magnetic fields with helical and non-helical properties in external galaxies using the Square Kilometre Array.

  6. 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

  7. 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. PMID:26037330

  8. 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.

  9. Single-shot volumetric laser induced fluorescence (VLIF) measurements in turbulent flows seeded with iodine.

    PubMed

    Wu, Yue; Xu, Wenjiang; Lei, Qingchun; Ma, Lin

    2015-12-28

    This work reports the experimental demonstration of single-shot visualization of turbulent flows in all three spatial dimensions (3D) based on volumetric laser induced fluorescence (VLIF). The measurements were performed based on the LIF signal of iodine (I2) vapor seeded in the flow. In contrast to established planar LIF (PLIF) technique, the VLIF technique excited the seeded I2 vapor volumetrically by a thick laser slab. The volumetric LIF signals emitted were then simultaneously collected by a total of five cameras from five different orientations, based on which a 3D tomographic reconstruction was performed to obtain the 3D distribution of the I2 vapor in the target flow. Single-shot measurements (with a measurement duration of a few ns) were demonstrated in a 50 mm × 50 mm × 50 mm volume with a nominal spatial resolution of 0.42 mm and an actual resolution of ~0.71 mm in all three dimensions (corresponding to a total of 120 × 120 × 120 voxels). PMID:26832005

  10. Measurement Of Atmospheric Peroxy Radicals By Chemical Conversion And Laser-induced Fluorescence Technique

    NASA Astrophysics Data System (ADS)

    Ren, X.; Naik, C.; Mao, J.; Harder, H.; Martinez, M.; Lesher, R.; Brune, W. H.

    2005-12-01

    A new method for measuring atmospheric peroxy radicals is described based on chemical conversion and laser-induced fluorescence (LIF) technique. Peroxy radicals are quantitatively converted into hydroperoxyl radicals (HO2) by the reactions with NO in a low-pressure reactor. The produced HO2 is then detected with an LIF instrument. The characterization and response of this instrument has been evaluated through the laboratory experiments as well as numeric simulations. Relative responses of different organic groups of peroxy radicals to HO2 were measured and the conversion coefficients agree generally well with the model calculations. The dependence of conversion coefficients on different experiment conditions was investigated. For HO2, the LIF signal is calibrated with an HO2 source produced by the photolysis of H2O via a low-pressure mercury lamp. Field measurements of peroxy radicals using this method were conducted at a rural site and preliminary results are presented. The estimated accuracy of the derived HOxROx concentrations is about 40% with a 2σ confidence level. Typical detection limit is about 0.2 pptv for 1-minute averaging times.

  11. 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.

  12. 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}

  13. 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.

  14. Research in Depolarization and Extinction Coefficient of Particles in Tibetan Plateau by Lidar

    NASA Astrophysics Data System (ADS)

    Dai, Guangyao; Song, Xiaoquan; Zhai, Xiaochun; Wu, Songhua

    2016-06-01

    Vertical profiles of the depolarization ratio and the extinction coefficient of atmospheric particles in Tibetan Plateau were measured with the OUC Water Vapor, Cloud and Aerosol Lidar during the 3rd Tibetan Plateau Atmospheric Expedition Experiment Campaign in 2013 and 2014. The cloud types and phases, the spatial temporal distribution of the aerosols and the boundary layer height in the Tibetan Plateau were obtained using polarization lidar technique. In this paper, the depolarization ratio was validated with CALIOP polarization simultaneous data, and the extinction coefficient was retrieved by the Fernald method. The result implied that the atmosphere in the Tibetan Plateau was quite clean with low aerosol load and serious pollution. The ice-water mixed cumulus, water cumulus or stratus clouds in Litang and Nagqu were occurred and classified, respectively. The boundary layer height in Nagqu at average altitude over 4600 m was obtained at around 200 m-300 m, which was commonly lower than that in other observed sites.

  15. Generation of vector beams using a double-wedge depolarizer: Non-quantum entanglement

    NASA Astrophysics Data System (ADS)

    Samlan, C. T.; Viswanathan, Nirmal K.

    2016-07-01

    Propagation of horizontally polarized Gaussian beam through a double-wedge depolarizer generates vector beams with spatially varying state of polarization. Jones calculus is used to show that such beams are maximally nonseparable on the basis of even (Gaussian)-odd (Hermite-Gaussian) mode parity and horizontal-vertical polarization state. The maximum nonseparability in the two degrees of freedom of the vector beam at the double wedge depolarizer output is verified experimentally using a modified Sagnac interferometer and linear analyser projected interferograms to measure the concurrence 0.94±0.002 and violation of Clauser-Horne-Shimony-Holt form of Bell-like inequality 2.704±0.024. The investigation is carried out in the context of the use of vector beams for metrological applications.

  16. Diurnal feeding rhythms in north sea copepods measured by Gut fluorescence, digestive enzyme activity and grazing on labelled food

    NASA Astrophysics Data System (ADS)

    Baars, M. A.; Oosterhuis, S. S.

    Results obtained with three methods for measuring feeding rhythms of copepods were different. Gut fluorescence showed clear day-night variation during 2 out of 3 cruises at the Oyster Ground in the North Sea. The species studied ( Pseudocalanus, Temora, Centropages, Calanus) had highest gut fluorescence during the night in May and September, the larger species demonstrating the largest difference. Gut fluorescence was positively correlated with ambient chlorophyll concentrations. Gut clearance rate was not dependent on temperature but on gut fullness. Gut passage times at high gut fluorescence levels were ˜25 minutes, at low levels 2 hours. In grazing experiments with 14C labelled food, filtering rates declined after 5 to 15 minutes, presumably before the first defecation of radioactive material. Filtering rates in Temora were higher at night than by day during May and July, but not in Pseudocalanus and Calanus during September. No diurnal pattern of amylase and tryptic activity was found, except in July for amylase but then probably due to vertical migration. The activity of these digestive enzymes appeared to be least and gut fluorescence most suitable for the detection of grazing rhythms. The occurrence of high fluorescence levels at night in all species studied suggests that intermittent feeding by copepods on phytoplankton is a general phenomenon from spring to autumn. The increase in foraging activity appeared to start well before complete darkness, during May and July even one hour or more before sunset.

  17. 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.

  18. 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. PMID:26724008

  19. Can the light scattering depolarization ratio of small particles be greater than 1/3?

    PubMed

    Khlebtsov, Nikolai G; Melnikov, Andrei G; Bogatyrev, Vladimir A; Dykman, Lev A; Alekseeva, Anna V; Trachuk, Lyubov A; Khlebtsov, Boris N

    2005-07-21

    According to the theory of light scattering by small randomly oriented particles, the depolarized ratio of the scattered intensities, I(vh)/I(vv), cannot exceed 1/3. Here we show that this conclusion does not hold for nonspherical plasmon resonant metal particles. Our analysis is based on the Rayleigh approximation and the exact T-matrix method as applied to spheroids and circular cylinders with semispherical ends. For small particles, the condition I(vh)/I(vv) >1/3 can be satisfied within the upper left quadrant of the complex relative dielectric permeability Real(eps) < -2 (rods) and within the upper unit semicircle centered at Real(eps) = -1 (disks). For gold nanorods with the axis ratio exceeding 2, the maximal theoretical values I(vh)/I(vv) lie between 1/3 and 3/4 at wavelengths of 550-650 nm. The extinction and static light scattering spectra (450-850 nm, at 90 degrees degrees) as well as the depolarized ratio of He-Ne laser light scattering were measured with gold nanospheres (the average diameters of 21, 29, and 46 nm) and nanorods (the longitudinal plasmon resonance peak positions at 655, 692, and 900 nm). The measured depolarization ratios of nanospheres (0.07-0.16) and nanorods (0.3-0.48) are in good agreement with theoretical calculations based on estimations of the average particle size and shape. PMID:16852700

  20. Depolarization-contraction coupling in short frog muscle fibers. A voltage clamp study

    PubMed Central

    1984-01-01

    Short muscle fibers (1.5 mm) were dissected from hindlimb muscles of frogs and voltage clamped with two microelectrodes to study phenomena related to depolarization-contraction coupling. Isometric myograms obtained in response to depolarizing pulses of durations between 10 and 500 ms and amplitudes up to 140 mV had the following properties. For suprathreshold pulses of fixed duration (in the range of 20-100 ms), the peak tension achieved, the time to peak tension, and contraction duration increased as the internal potential was made progressively more positive. Peak tension eventually saturates with increasing internal potentials. For pulse durations of greater than or equal to 50 ms, the rate of tension development becomes constant for increasing internal potentials when peak tensions become greater than one-third of the maximum tension possible. Both threshold and maximum steepness of the relation between internal potential and peak tension depend on pulse duration. The relation between the tension-time integral and the stimulus amplitude-duration product was examined. The utility of this relation for excitation-contraction studies is based on the observation that once a depolarizing pulse configuration has elicited maximum tension, further increases in either stimulus duration or amplitude only prolong the contractile response, while the major portion of the relaxation phase after the end of a pulse is exponential, with a time constant that is not significantly affected by either the amplitude or the duration of the pulse. Hence, the area under the tension-response curve provides a measure of the availability to troponin of the calcium released from the sarcoplasmic reticulum in response to membrane depolarization. The results from this work complement those obtained in experiments in which intramembrane charge movements related to contractile activation were studied and those in which intracellular Ca++ transients were measured. PMID:6611386

  1. 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

  2. Steady-State Acceptor Fluorescence Anisotropy Imaging under Evanescent Excitation for Visualisation of FRET at the Plasma Membrane

    PubMed Central

    Devauges, Viviane; Matthews, Daniel R.; Aluko, Justin; Nedbal, Jakub; Levitt, James A.; Poland, Simon P.; Coban, Oana; Weitsman, Gregory; Monypenny, James; Ng, Tony; Ameer-Beg, Simon M.

    2014-01-01

    We present a novel imaging system combining total internal reflection fluorescence (TIRF) microscopy with measurement of steady-state acceptor fluorescence anisotropy in order to perform live cell Förster Resonance Energy Transfer (FRET) imaging at the plasma membrane. We compare directly the imaging performance of fluorescence anisotropy resolved TIRF with epifluorescence illumination. The use of high numerical aperture objective for TIRF required correction for induced depolarization factors. This arrangement enabled visualisation of conformational changes of a Raichu-Cdc42 FRET biosensor by measurement of intramolecular FRET between eGFP and mRFP1. Higher activity of the probe was found at the cell plasma membrane compared to intracellularly. Imaging fluorescence anisotropy in TIRF allowed clear differentiation of the Raichu-Cdc42 biosensor from negative control mutants. Finally, inhibition of Cdc42 was imaged dynamically in live cells, where we show temporal changes of the activity of the Raichu-Cdc42 biosensor. PMID:25360776

  3. Simultaneous measurement of water volume and pH in single cells using BCECF and fluorescence imaging microscopy.

    PubMed

    Alvarez-Leefmans, Francisco J; Herrera-Pérez, José J; Márquez, Martín S; Blanco, Víctor M

    2006-01-15

    Regulation and maintenance of cell water volume and intracellular pH (pHi) are vital functions that are interdependent; cell volume regulation affects, and is in turn affected by, changes in pHi. Disruption of either function underlies various pathologies. To study the interaction and kinetics of these two mechanisms, we developed and validated a quantitative fluorescence imaging microscopy method to measure simultaneous changes in pHi and volume in single cells loaded with the fluorescent probe BCECF. CWV is measured at the excitation isosbestic wavelength, whereas pHi is determined ratiometrically. The method has a time resolution of <1 s and sensitivity to osmotic changes of approximately 1%. It can be applied in real time to virtually any cell type attached to a coverslip, independently of cellular shape and geometry. Calibration procedures and algorithms developed to transform fluorescence signals into changes in cell water volume (CWV) and examples of applications are presented. PMID:16258035

  4. Consideration for the dynamic depolarization in the effective-medium model for description of optical properties for anisotropic nanostructured semiconductors

    SciTech Connect

    Golovan, L. A.; Zabotnov, S. V. Timoshenko, V. Yu.; Kashkarov, P. K.

    2009-02-15

    The effective-medium model has been generalized within the dipole approximation, with allowance for the shape anisotropy and dynamic depolarization of semiconductor nanoparticles. The calculations revealed nonmonotonic dependences for the birefringence and dichroism on the nanoparticle size. Comparison of the measured and calculated refractive index dispersion of birefringent porous silicon layers in the near-IR region indicates that consideration for the dynamic depolarization gives a better description of the optical properties for this material in comparison with the generally used effective-medium electrostatic approximation.

  5. 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.

  6. 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.

  7. 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.

  8. Water vapor content in the polar atmosphere measured by Lyman-alpha/OH fluorescence method

    NASA Technical Reports Server (NTRS)

    Iwasaka, Y.; Saitoh, S.; Ono, A.

    1985-01-01

    The water vapor of the polar stratosphere possibly plays an important role in various aeronomical processes; for example, OH radical formation through photodissociation of H2O, formation of water cluster ions, radiative energy transfer in the lower stratosphere, condensation onto particulate matter, and so on. In addition to these, it has been speculated, from the viewpoint of global transport and/or budget of water vapor, that the polar stratosphere functions as an active sink. STANFORD (1973) emphasized the existence of the stratospheric Cist cloud in the polar stratosphere which brought a large loss rate of stratospheric water vapor through a so-called freeze-out of cloud particles from the stratosphere into the troposphere. However, these geophysically interesting problems unfortunately remain to be solved, owing to the lack of measurements on water vapor distribution and its temporal variation in the polar stratosphere. The water vapor content measured at Syowa Station (69.00 deg S, 39.35 deg E), Antarctica using a balloon-borne hygrometer (Lyman - alpha/OH fluorescence type) is discussed.

  9. 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.

  10. Fluorescence competition assay measurements of free energy changes for RNA pseudoknots.

    PubMed

    Liu, Biao; Shankar, Neelaabh; Turner, Douglas H

    2010-01-26

    RNA pseudoknots have important functions, and thermodynamic stability is a key to predicting pseudoknots in RNA sequences and to understanding their functions. Traditional methods, such as UV melting and differential scanning calorimetry, for measuring RNA thermodynamics are restricted to temperature ranges around the melting temperature for a pseudoknot. Here, we report RNA pseudoknot free energy changes at 37 degrees C measured by fluorescence competition assays. Sequence-dependent studies for the loop 1-stem 2 region reveal (1) the individual nearest-neighbor hydrogen bonding (INN-HB) model provides a reasonable estimate for the free energy change when a Watson-Crick base pair in stem 2 is changed, (2) the loop entropy can be estimated by a statistical polymer model, although some penalty for certain loop sequences is necessary, and (3) tertiary interactions can significantly stabilize pseudoknots and extending the length of stem 2 may alter tertiary interactions such that the INN-HB model does not predict the net effect of adding a base pair. The results can inform writing of algorithms for predicting and/or designing RNA secondary structures.

  11. Measurements of fluorescence yield of electrons in air under atmospheric conditions: A key parameter for energy of cosmic rays

    NASA Astrophysics Data System (ADS)

    Monnier Ragaigne, D.; Gorodetzky, P.; Blacksley, C.; Wicek, F.; Monard, H.; Dagoret-Campagne, S.

    2012-12-01

    The measurement of the fluorescence yield and its dependence on atmospheric properties such as pressure, temperature or pollutants, are essential to obtain a reliable measurement of the primary energy of cosmic rays. A new type of absolute measurement of the nitrogen fluorescence yield in the air will be performed at LAL using 3 items which will yield an unprecedented precision in all conditions of pressure, temperature, and pollutants. A 5 MeV electron beam will be provided by the new electron accelerator PHIL at LAL(Laboratoire de l'Accélérateur Linéaire, Univ Paris-Sud, CNRS/IN2P3, Orsay). This source will induce florescence yield inside an integrating sphere. The sphere will be surrounded by a spherical envelope to create a temperature controlled chamber (a Dewar). With this setup it will be possible to vary the temperature from -60 C to +40 C and the pressure from 1 to 0.01 atm. An output device on this sphere will be equipped with a set of optical fibers driving the fluorescence light to a Jobin-Yvon spectrometer equipped with an LN_{2} cooled CCD. The fluorescence spectrum in the 300-430 nm range will be accurately measured in steps of 0.1 nm resolution. A PMT equipped with a BG3 filter (the same as on JEM-EUSO) will be set on the sphere to measure the integrated yield. The expected precision of the yield should be better than 5%.

  12. Susceptibility of Primary Sensory Cortex to Spreading Depolarizations

    PubMed Central

    Bogdanov, Volodymyr B.; Middleton, Natalie A.; Theriot, Jeremy J.; Parker, Patrick D.; Abdullah, Osama M.; Ju, Y. Sungtaek; Hartings, Jed A.

    2016-01-01

    Spreading depolarizations (SDs) are recognized as actors in neurological disorders as diverse as migraine and traumatic brain injury (TBI). Migraine aura involves sensory percepts, suggesting that sensory cortices might be intrinsically susceptible to SDs. We used optical imaging, MRI, and field potential and potassium electrode recordings in mice and electrocorticographic recordings in humans to determine the susceptibility of different brain regions to SDs. Optical imaging experiments in mice under isoflurane anesthesia showed that both cortical spreading depression and terminal anoxic depolarization arose preferentially in the whisker barrel region of parietal sensory cortex. MRI recordings under isoflurane, ketamine/xylazine, ketamine/isoflurane, and urethane anesthesia demonstrated that the depolarizations did not propagate from a subcortical source. Potassium concentrations showed larger increases in sensory cortex, suggesting a mechanism of susceptibility. Sensory stimulation biased the timing but not the location of depolarization onset. In humans with TBI, there was a trend toward increased incidence of SDs in parietal/temporal sensory cortex compared with other regions. In conclusion, SDs are inducible preferentially in primary sensory cortex in mice and most likely in humans. This tropism can explain the predominant sensory phenomenology of migraine aura. It also demonstrates that sensory cortices are vulnerable in brain injury. SIGNIFICANCE STATEMENT Spreading depolarizations (SDs) are involved in neurologic disorders as diverse as migraine and traumatic brain injury. In migraine, the nature of aura symptoms suggests that sensory cortex may be preferentially susceptible. In brain injury, SDs occur at a vulnerable time, during which the issue of sensory stimulation is much debated. We show, in mouse and human, that sensory cortex is more susceptible to SDs. We find that sensory stimulation biases the timing but not the location of the depolarizations

  13. Differential gene expression in skeletal muscle cells after membrane depolarization.

    PubMed

    Juretić, Nevenka; Urzúa, Ulises; Munroe, David J; Jaimovich, Enrique; Riveros, Nora

    2007-03-01

    Skeletal muscle is a highly plastic tissue with a remarkable capacity to adapt itself to challenges imposed by contractile activity. Adaptive response, that include hypertrophy and activation of oxidative mechanisms have been associated with transient changes in transcriptional activity of specific genes. To define the set of genes regulated by a depolarizing stimulus, we used 22 K mouse oligonucleotide microarrays. Total RNA from C2C12 myotubes was obtained at 2, 4, 18, and 24 h after high K+ stimulation. cDNA from control and depolarized samples was labeled with cyanine 3 or 5 dyes prior to microarray hybridization. Loess normalization followed by statistical analysis resulted in 423 differentially expressed genes using an unadjusted P-value < or = 0.01 as cut off. Depolarization affects transcriptional activity of a limited number of genes, mainly associated with metabolism, cell communication and response to stress. A number of genes related to Ca2+ signaling pathways are induced at 4 h, reinforcing the potential role of Ca2+ in early steps of signal transduction that leads to gene expression. Significant changes in the expression of molecules involved in muscle cell structure were observed; K+-depolarization increased Tnni1 and Acta1 mRNA levels in both differentiated C2C12 and rat skeletal muscle cells in primary culture. Of these two, depolarization induced slow Ca2+ transients appear to have a role only in the regulation of Tnni1 transcriptional activity. We suggest that depolarization induced expression of a small set of genes may underlie Ca2+ dependent plasticity of skeletal muscle cells. PMID:17146758

  14. Velocity Field Measurements in Rarefied, Hypersonic Flows of Nitrogen Using Laser-Induced Fluorescence of Iodine

    NASA Astrophysics Data System (ADS)

    Cecil, Eric

    Velocity fields are measured in the shock layer and boundary layer on a plate with a cylindrical fin immersed in a hypersonic, free jet of nitrogen, using laser-induced fluorescence (LIF) of iodine. A sheet beam from a single-mode argon laser at 514 nm is used to excite hyperfine components of the P(13), R(15) and P(48), P(103) blended rotational-vibrational lines in the B-X electronic transition for iodine seeded in the flow. The Doppler broadening and shift of these lines, and the relative rotational line strengths are determined for excitation spectra recorded in a planar grid. Using this measurement technique, estimates for iodine of the mass velocity component and kinetic temperature of translation in the direction of laser propagation, rotational temperature, and relative number density are determined at each point. Sectional planes of the flow over the body are investigated at a spatial resolution on the scale of the molecular mean-free-path in the free jet near the plate leading edge. Two directions within each plane are examined, to determine the velocity vector and to investigate translational non-equilibrium. Predictions from two direct simulation Monte Carlo computations of the flow are compared with the measurements. Large values of slip velocity and temperature jump at the plate surface are observed for iodine. Measurements and DSMC predictions indicate strong translational non-equilibrium effects for the iodine in the shock wave and the thick boundary layer on the plate, and are qualitatively consistent with a bimodal velocity distribution function. As a consequence of the ratio of molecular masses, the translational non-equilibrium of iodine is much greater than for nitrogen.

  15. Measuring plant available phosphorus using diffusive gradients in thin films and x-ray fluorescence spectrometry

    NASA Astrophysics Data System (ADS)

    Rothwell, Shane; Surridge, Ben; Dodd, Ian; Quinton, John; Zhang, Hao

    2015-04-01

    Global concerns of phosphorus (P) deficiency limiting crop yields, and finite supplies of mineral P reserves, suggest a need to maximise P use efficiency in agriculture. To accurately predict the availability of soil P to crops, and subsequent P fertiliser recommendations, soil P tests must determine only the P that will be accessed and utilised by a crop. However, there is growing doubt regarding the ability of current extraction techniques (water, bicarbonate, resin) to accurately determine plant-available P across a range of soils. Indeed, the most widely-used test (Olsen P) across all soil types was only designed for alkaline soils and therefore it is inappropriate as a national standard soil test. Thus, there is an urgent need to develop a standard approach to measuring P availability applicable across a range of soil types. Diffusive Gradients in Thin Films (DGT) may be a more accurate technique for measuring the P available to plants than P measured using current extraction techniques because the measurement responds to both soil solution P and the P rapidly resupplied from the solid phase. However, elution by acid extraction of P retained within the resin gel of a DGT device, followed by analysis via inductively coupled plasma-based techniques, typically results in a delay of several days between DGT deployment and reporting of P concentrations. This is currently a significant constraint on the adoption of DGT to determine plant-available P in agricultural soils. Our research seeks to develop a novel combination of two existing techniques, DGT with portable x-ray fluorescence spectrometry (pXRF) to achieve rapid, non-destructive analysis of P within a DGT device, thus significantly reducing the length of time between DGT deployment and the final determination of plant-available P in agricultural soils. We aim to develop DGT-pXRF as a robust routine analytical procedure suitable for analysis of plant available P in a wide range of agricultural soil types.

  16. 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

  17. Are Fluorescence Quantum Yields So Tricky to Measure? A Demonstration Using Familiar Stationery Products

    NASA Astrophysics Data System (ADS)

    Fery-Forgues*, Suzanne; Lavabre, Dominique

    1999-09-01

    Fluorescence quantum yields are used to quantify the efficiency of the emission process. In spite of the importance of these data, experimental directions for their acquisition are rarely given. A general procedure for determining the relative fluorescence quantum yield of solutions is described here, drawing attention to the many pitfalls that students may encounter. Starting materials are common yellow and pink highlighter pens.

  18. PHYTOPLANKTON NUTRIENT STATUS AND VARIABLE FLUORESCENCE MEASUREMENTS IN A GULF COAST ESTUARY

    EPA Science Inventory

    Changes in variable fluorescence parameters such as the maximum quantum yield of fluorescence (a.k.a. photosynthetic efficiency and Fv/Fm) have been related to nutrient status in single-species cultures. To test if changes in Fv/Fm of mixed natural assemblages were related to nut...

  19. Dark-field circular depolarization optical coherence microscopy

    PubMed Central

    Mehta, Kalpesh; Zhang, Pengfei; Yeo, Eugenia Li Ling; Kah, James Chen Yong; Chen, Nanguang

    2013-01-01

    Optical coherence microscopy (OCM) is a widely used structural imaging modality. To extend its application in molecular imaging, gold nanorods are widely used as contrast agents for OCM. However, they very often offer limited sensitivity as a result of poor signal to background ratio. Here we experimentally demonstrate that a novel OCM implementation based on dark-field circular depolarization detection can efficiently detect circularly depolarized signal from gold nanorods and at the same time efficiently suppress the background signals. This results into a significant improvement in signal to background ratio. PMID:24049689

  20. Fluorescence Techniques for Measuring Kinetics of Specific Binding of Hormone to Cell Surface Receptors.

    NASA Astrophysics Data System (ADS)

    Hellen, Edward Herbert

    This thesis presents theoretical calculations and technical advances relevant to total internal reflection/ fluorescence photobleaching recovery (tir/fpr), and results from experiments using tir/fpr to measure the dissociation rate constant of epidermal growth factor (egf) hormone interacting with its receptor molecule on A431 cells. The classical electromagnetic calculations describe fluorescence emission from fluorophores near an interface (possibly metal coated). It is well known that an interface alters the emission properties of nearby fluorophores. Most previous classical calculations model the fluorophore as a fixed-amplitude dipole oscillator. However, for fluorophores under steady illumination, a fixed-power dipole is more appropriate. This modification corresponds to normalizing the fixed-amplitude dipole's intensity by its total dissipated power. The results for the fixed-power model differ nontrivially from the fixed-amplitude model. The observation-angle -dependent intensity as a function of the fluorophore's orientation and distance from the surface is calculated. General expressions are derived for the emission power as observed through a circular-aperture collection system located on either side of the interface. A system for maintaining long-term focus of samples under high-magnification quantitative observation in an epi-illumination optical microscope is described. Focus -dependent changes in the backreflection of an off-axis HeNe laser generate negative feedback signals which drive a dc motor coupled to the fine-focus knob of the microscope. This system has several advantages: (1) it is compatible and nonobstructive with concurrent data acqusition of sample intensities; (2) it requires no alteration of the sample, stage, or objective; (3) it monitors the position of sample areas very near to those under observation; (4) it is inexpensive. The system can hold a glass coverslip sample to within 0.5 μm of its preset focus position. Prismless tir

  1. Speed of disentanglement in multiqubit systems under a depolarizing channel

    SciTech Connect

    Zhang, Fu-Lin Jiang, Yue; Liang, Mai-Lin

    2013-06-15

    We investigate the speed of disentanglement in the multiqubit systems under the local depolarizing channel, in which each qubit is independently coupled to the environment. We focus on the bipartition entanglement between one qubit and the remaining qubits constituting the system, which is measured by the negativity. For the two-qubit system, the speed for the pure state completely depends on its entanglement. The upper and lower bounds of the speed for arbitrary two-qubit states, and the necessary conditions for a state achieving them, are obtained. For the three-qubit system, we study the speed for pure states, whose entanglement properties can be completely described by five local-unitary-transformation invariants. An analytical expression of the relation between the speed and the invariants is derived. The speed is enhanced by the three-tangle which is the entanglement among the three qubits, but reduced by the two-qubit correlations outside the concurrence. The decay of the negativity can be restrained by the other two negativity with the coequal sense. The unbalance between two qubits can reduce the speed of disentanglement of the remaining qubit in the system, and even can retrieve the entanglement partially. For the k-qubit systems in an arbitrary superposition of Greenberger–Horne–Zeilinger state and W state, the speed depends almost entirely on the amount of the negativity when k increases to five or six. An alternative quantitative definition for the robustness of entanglement is presented based on the speed of disentanglement, with comparison to the widely studied robustness measured by the critical amount of noise parameter where the entanglement vanishes. In the limit of large number of particles, the alternative robustness of the Greenberger–Horne–Zeilinger-type states is inversely proportional to k, and the one of the W states approaches 1/√(k)

  2. Measuring Norfloxacin Binding to Trypsin Using a Fluorescence Quenching Assay in an Upper-Division, Integrated Laboratory Course

    ERIC Educational Resources Information Center

    Hicks, Katherine A.

    2016-01-01

    Fluorescence quenching assays are often used to measure dissociation constants that quantify the binding affinity between small molecules and proteins. In an upper-division undergraduate laboratory course, where students work on projects using a guided inquiry-based approach, a binding titration experiment at physiological pH is performed to…

  3. Design and investigation of a series of rhodamine-based fluorescent probes for optical measurements of pH.

    PubMed

    Best, Quinn A; Xu, Ruisong; McCarroll, Matthew E; Wang, Lichang; Dyer, Daniel J

    2010-07-16

    A series of structurally similar fluorescent probes (1-4), synthesized from rhodamine B, were designed to optically measure pH. Each probe had a unique "off-on" response as the solution went from basic to acidic. Probes 1-3 exhibited a spirocyclic quenching of the pyronin B fluorophore, whereas probe 4 was quenched by PET from the amine moiety.

  4. Measurement of Bluetongue Virus Binding to a Mammalian Cell Surface Receptor by an In Situ Immune Fluorescent Staining Technique

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A quantifiable in situ immune fluorescent assay (IFA) was developed to measure bluetongue virus (BTV) binding to mammalian cells. The utility of the assay was demonstrated with both Chinese hamster ovary (CHO) and bovine pulmonary artery endothelial (CPAE) cells. Since heparin sulfate (HS) has been ...

  5. Experimental Assessment and Enhancement of Planar Laser-Induced Fluorescence Measurements of Nitric Oxide in an Inverse Diffusion Flame

    NASA Technical Reports Server (NTRS)

    Partridge, William P.; Laurendeau, Normand M.

    1997-01-01

    We have experimentally assessed the quantitative nature of planar laser-induced fluorescence (PLIF) measurements of NO concentration in a unique atmospheric pressure, laminar, axial inverse diffusion flame (IDF). The PLIF measurements were assessed relative to a two-dimensional array of separate laser saturated fluorescence (LSF) measurements. We demonstrated and evaluated several experimentally-based procedures for enhancing the quantitative nature of PLIF concentration images. Because these experimentally-based PLIF correction schemes require only the ability to make PLIF and LSF measurements, they produce a more broadly applicable PLIF diagnostic compared to numerically-based correction schemes. We experimentally assessed the influence of interferences on both narrow-band and broad-band fluorescence measurements at atmospheric and high pressures. Optimum excitation and detection schemes were determined for the LSF and PLIF measurements. Single-input and multiple-input, experimentally-based PLIF enhancement procedures were developed for application in test environments with both negligible and significant quench-dependent error gradients. Each experimentally-based procedure provides an enhancement of approximately 50% in the quantitative nature of the PLIF measurements, and results in concentration images nominally as quantitative as LSF point measurements. These correction procedures can be applied to other species, including radicals, for which no experimental data are available from which to implement numerically-based PLIF enhancement procedures.

  6. Mueller-Stokes characterization and optimization of a liquid crystal on silicon display showing depolarization.

    PubMed

    Márquez, A; Moreno, I; Iemmi, C; Lizana, A; Campos, J; Yzuel, M J

    2008-02-01

    In this paper we characterize the polarimetric properties of a liquid crystal on silicon display (LCoS), including depolarization and diattenuation which are usually not considered when applying the LCoS in diffractive or adaptive optics. On one hand, we have found that the LCoS generates a certain degree (that can be larger than a 10%) of depolarized light, which depends on the addressed gray level and on the incident state of polarization (SOP), and can not be ignored in the above mentioned applications. The main origin of the depolarized light is related with temporal fluctuations of the SOP of the light reflected by the LCoS. The Mueller matrix of the LCoS is measured as a function of the gray level, which enables for a numerical optimization of the intensity modulation configurations. In particular we look for maximum intensity contrast modulation or for constant intensity modulation. By means of a heuristic approach we show that, using elliptically polarized light, amplitude-mostly or phase-mostly modulation can be obtained at a wavelength of 633 nm.

  7. Large plasma-membrane depolarization precedes rapid blue-light-induced growth inhibition in cucumber

    NASA Technical Reports Server (NTRS)

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

    1989-01-01

    Blue-light (BL)-induced suppression of elongation of etiolated Cucumis sativus L. hypocotyls began after a 30-s lag time, which was halved by increasing the fluence rate from 10 to 100 micromoles m-2 s-1. Prior to the growth suppression, the plasma-membrane of the irradiated cells depolarized by as much as 100 mV, then returned within 2-3 min to near its initial value. The potential difference measured with surface electrodes changed with an identical time course but opposite polarity. The lag time for the change in surface potential showed an inverse dependence on fluence rate, similar to the lag for the growth inhibition. Green light and red light caused neither the electrical response nor the rapid inhibition of growth. The depolarization by BL did not propagate to nonirradiated regions and exhibited a refractory period of about 10 min following a BL pulse. Fluence-response relationships for the electrical and growth responses provide correlational evidence that the plasma-membrane depolarization reflects an event in the transduction chain of this light-growth response.

  8. Local Maps of the Polarization and Depolarization in Organic Ferroelectric Field-Effect Transistors.

    PubMed

    Cai, Ronggang; Jonas, Alain M

    2016-02-24

    We study the local ferroelectric polarization and depolarization of poly(vinylidene fluoride-co-trifluoroethylene) (P(VDF-TrFE)) in p-type ferroelectric field-effect transistors (FeFETs). Piezoresponse force microscopy (PFM) is used to obtain local maps of the polarization on model metal-semiconductor-ferroelectric stacks, and on FeFETs stripped from their top-gate electrode; transfer curves are measured on complete FeFETs. The influence of the semiconductor layer thickness and of the polarity and amplitude of the poling voltage are investigated. In accumulation, the stable "on" state consists of a uniform upward-polarized ferroelectric layer, with compensation holes accumulating at the ferroelectric/semiconducting interface. In depletion, the stable "off" state consists of a depolarized region in the center of the transistor channel, surrounded by partially downward-polarized regions over the source and drain electrodes and neighboring regions. The partial depolarization of these regions is due to the incomplete screening of polarization charges by the charges of the remote electrodes. Therefore, thinner semiconducting layers provide higher downward polarizations, which result in a more depleted transistor channel and a higher charge injection barrier between the electrodes and the semiconductor, leading to lower threshold voltages and higher on/off current values at zero gate bias. Clues for optimization of the devices are finally provided.

  9. Fluorescent probes in biology and medicine: measurement of intracellular pH values in individual cells

    NASA Astrophysics Data System (ADS)

    Slavik, Jan; Cimprich, Petr; Gregor, Martin; Smetana, Karel, Jr.

    1997-12-01

    The application possibilities of fluorescent probes have increased dramatically in the last few years. The main areas are as follows (Slavik, 1994, 1996, 1998). Intracellular ionic cell composition: There are selective ion-sensitive dyes for H+, Ca2+, Mg2+, K+, Na+, Fe3+, Cl-, Zn2+, Cd2+, Hg2+, Pb2+, Ba2+, La3+. Membrane potential: Using the so-called slow (Nernstian dyes) or electrochromic dyes one can assess the value of the transmembrane potential. Membrane fluidity: Fluorescent probes inform about the freedom of rotational and translational movement of membrane proteins and lipids. Selective labeling: Almost any object of interest inside the cell or on its surface can be selectively fluorescently labeled. There are dyes specific for DNA, RNA, oligonucleotides (FISH), Golgi, endoplasmic reticulum, mitochondria, vacuoles, cytoskeleton, etc. Using fluorescent dyes specific receptors may be localized, their conformational changes followed and the polarity of corresponding binding sites accessed. The endocytic pathway may be followed, enzymes and their local enzymatic activity localized. For really selective labeling fluorescent labeled antibodies exist. Imaging: One of the main advantages of fluorescence imaging is its versatility. It allow choice among ratio imaging in excitation, ratio imaging in emission and lifetime imaging. These approaches can be applied to both the classical wide-field fluorescence microscopy and to the laser confocal fluorescence microscopy, one day possibly to the scanning near field optical microscopy. Simultaneous application of several fluorescent dyes: The technical progress in both excitation sources and in detectors allows to extend the excitation deeper in the blue and ultraviolet side and the detection further in the NIR and IR. Consequently, up to 6 peaks in excitation and up to 6 peaks in emission can be followed without any substantial difficulties. Application of dyes such with longer fluorescence lifetimes such as rare earth

  10. Correlates of spreading depolarization in human scalp electroencephalography

    PubMed Central

    Drenckhahn, Christoph; Winkler, Maren K. L.; Major, Sebastian; Scheel, Michael; Kang, Eun-Jeung; Pinczolits, Alexandra; Grozea, Cristian; Hartings, Jed A.; Woitzik, Johannes

    2012-01-01

    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

  11. Ambient measurements of biological aerosol particles near Killarney, Ireland: a comparison between real-time fluorescence and microscopy techniques

    NASA Astrophysics Data System (ADS)

    Healy, D. A.; Huffman, J. A.; O'Connor, D. J.; Pöhlker, C.; Pöschl, U.; Sodeau, J. R.

    2014-02-01

    Primary biological aerosol particles (PBAP) can contribute significantly to the coarse particle burden in many environments, may thus influence climate and precipitation systems as cloud nuclei, and can spread disease to humans, animals, and plants. Measurements of PBAP in natural environments taken at high time- and size- resolution are, however, sparse and so large uncertainties remain in the role that biological particles play in the Earth system. In this study two commercial real-time fluorescence particle sensors and a Sporewatch single-stage particle impactor were operated continuously from 2 August to 2 September 2010 at a rural sampling location in Killarney National Park in south western Ireland. A cascade impactor was operated periodically to collect size-resolved particles during exemplary periods. Here we report the first ambient comparison of the waveband integrated bioaerosol sensor (WIBS-4) with the ultraviolet aerodynamic particle sizer (UV-APS) and also compare these real-time fluorescence techniques with results of fluorescence and optical microscopy of impacted samples. Both real-time instruments showed qualitatively similar behaviour, with increased fluorescent bioparticle concentrations at night when relative humidity was highest and temperature was lowest. The fluorescent particle number from the FL3 channel of the WIBS-4 and from the UV-APS were strongly correlated and dominated by a 3 μm mode in the particle size distribution. The WIBS FL2 channel exhibited particle modes at approx. 1 and 3 μm, and each were correlated with the concentration of fungal spores commonly observed in air samples collected at the site (ascospores, basidiospores, Ganoderma spp.). The WIBS FL1 channel exhibited variable multi-modal distributions turning into a broad featureless single mode after averaging and exhibited poor correlation with fungal spore concentrations, which may be due to the detection of bacterial and non-biological fluorescent particles

  12. Ambient measurements of biological aerosol particles near Killarney, Ireland: a comparison between real-time fluorescence and microscopy techniques

    NASA Astrophysics Data System (ADS)

    Healy, D. A.; Huffman, J. A.; O'Connor, D. J.; Pöhlker, C.; Pöschl, U.; Sodeau, J. R.

    2014-08-01

    Primary biological aerosol particles (PBAPs) can contribute significantly to the coarse particle burden in many environments. PBAPs can thus influence climate and precipitation systems as cloud nuclei and can spread disease to humans, animals, and plants. Measurement data and techniques for PBAPs in natural environments at high time- and size resolution are, however, sparse, and so large uncertainties remain in the role that biological particles play in the Earth system. In this study two commercial real-time fluorescence particle sensors and a Sporewatch single-stage particle impactor were operated continuously from 2 August to 2 September 2010 at a rural sampling location in Killarney National Park in southwestern Ireland. A cascade impactor was operated periodically to collect size-resolved particles during exemplary periods. Here we report the first ambient comparison of a waveband integrated bioaerosol sensor (WIBS-4) with a ultraviolet aerodynamic particle sizer (UV-APS) and also compare these real-time fluorescence techniques with results of fluorescence and optical microscopy of impacted samples. Both real-time instruments showed qualitatively similar behavior, with increased fluorescent bioparticle concentrations at night, when relative humidity was highest and temperature was lowest. The fluorescent particle number from the FL3 channel of the WIBS-4 and from the UV-APS were strongly correlated and dominated by a 3 μm mode in the particle size distribution. The WIBS FL2 channel exhibited particle modes at approx. 1 and 3 μm, and each was correlated with the concentration of fungal spores commonly observed in air samples collected at the site (ascospores, basidiospores, Ganoderma spp.). The WIBS FL1 channel exhibited variable multimodal distributions turning into a broad featureless single mode after averaging, and exhibited poor correlation with fungal spore concentrations, which may be due to the detection of bacterial and non-biological fluorescent

  13. Ultraviolet emission and excitation fluorescence spectroscopic characterization of DMBA-treated Swiss Albino mice skin carcinogenesis for measuring tissue transformation

    NASA Astrophysics Data System (ADS)

    Aruna, Prakasa R.; Hemamalini, Srinivasan; Ebenezar, Jeyasingh; Ganesan, Singaravelu

    2002-05-01

    The ultraviolet fluorescence emission spectra of skin tissues under different pathological conditions were measured at 280nm excitation. At this excitation wavelength, the normal skin showed a primary peak emission at 352nm and this primary peak emission from neoplastic skin shows a blue shift with respect to normal tissue. This blue shift increases as the stage of abnormality increases and it is maximum (19nm) for well-differentiated squamous cell carcinoma. This alteration is further confirmed from fluorescence excitation spectra of the tissues for 340nm emission. The study concludes that the change in the emission of tryptophan around 340nm may be due to partial unfolding of protein.

  14. A step toward standardization: development of accurate measurements of X-ray absorption and fluorescence.

    PubMed

    Chantler, Christopher T; Barnea, Zwi; Tran, Chanh Q; Rae, Nicholas A; de Jonge, Martin D

    2012-11-01

    This paper explains how to take the counting precision available for XAFS (X-ray absorption fine structure) and attenuation measurements, of perhaps one part in 10(6) in special cases, to produce a local variance below 0.01% and an accuracy of attenuation of the order 0.01%, with an XAFS accuracy at a similar level leading to the determination of dynamical bond lengths to an accuracy similar to that obtained by standard and experienced crystallographic measurements. This includes the necessary corrections for the detector response to be linear, including a correction for dark current and air-path energy dependencies; a proper interpretation of the range of sample thicknesses for absorption experiments; developments of methods to measure and correct for harmonic contamination, especially at lower energies without mirrors; the significance of correcting for the actual bandwidth of the beam on target after monochromation, especially for the portability of results and edge structure from one beamline to another; definitions of precision, accuracy and XAFS accuracy suitable for theoretical model analysis; the role of additional and alternative high-accuracy procedures; and discusses some principles regarding data formats for XAFS and for the deposition of data sets with manuscripts or to a database. Increasingly, the insight of X-ray absorption and the standard of accuracy needed requires data with high intrinsic precision and therefore with allowance for a range of small but significant systematic effects. This is always crucial for absolute measurements of absorption, and is of equal importance but traditionally difficult for (usually relative) measurements of fluorescence XAFS or even absorption XAFS. Robust error analysis is crucial so that the significance of conclusions can be tested within the uncertainties of the measurements. Errors should not just include precision uncertainty but should attempt to include estimation of the most significant systematic error

  15. Measurements of NO2 in Maritime Atmosphere in Japan by Laser-Induced Fluorescence Technique

    NASA Astrophysics Data System (ADS)

    Matsumoto, J.; Kajii, Y.

    2001-12-01

    NO2 is one of the most important species in tropospheric photochemistry since it plays a key role as a precursor of ozone. Photostationary-state (PSS) between NO and NO2 is a critical factor for ozone production. It is essential to measure NO2 precisely at the level of pptv in the remote, background region. In this study, a compact and sensitive instrument for direct measurement of NO2 has been developed utilizing laser-induced fluorescence (LIF) technique. For the purpose of simple, compact and stable measurement, the single wavelength excitation by a powerful Nd:YAG laser (532.1 nm, 6500 mW at 10 kHz) is adopted. As a result of improvement, the sensitivity, background signal, dark current and the limit of detection are 0.07 cps ppbv-1 mW-1, 70 cps and 4 pptv (60-s, S/N=1), respectively. These specifications suggest the LIF- NO2 instrument can be utilized to measure NO2 at the level of pptv. Two field observations have been successfully carried out under maritime conditions in Japan. The measurements were conducted in Okinawa Island for 10 days and in Rishiri Island for 18 days. The stability of the instrument was confirmed through these observations. In intercomparison with a chemiluminescence-based detector, excellent agreements between two instruments were shown. Thus, the LIF instrument is confirmed to be reasonable for measuring atmospheric NO2. Finally, PSS of NOx in Rishiri Island is considered. As a result, it is suggested that unidentified species such as halogen oxides can be important in the conversion process of NO to NO2. This additional conversion of NO to NO2 can increase the formation rate of nitric acid. In this case, the increase of formation rate can be estimated as 7 %. Consequently, the high-performance LIF instrument realizes precise consideration about NO2 in PSS of NOx. This compact, simple method is promising to be applied conveniently in remote regions over the world.

  16. Measurement of the fluorescence of crop residues: A tool for controlling soil erosion

    NASA Technical Reports Server (NTRS)

    Daughtry, C. S. T.; Mcmurtrey, J. E., III; Chappelle, E. W.; Hunter, W. J.

    1994-01-01

    Management of crop residues, the portion of a crop left in the field after harvest, is an important conservation practice for minimizing soil erosion and for improving water quality. Quantification of crop residue cover is required to evaluate the effectiveness of conservation tillage practices. Methods are needed to quantify residue cover that are rapid, accurate, and objective. The fluorescence of crop residue was found to be a broadband phenomenon with emission maxima at 420 to 495 nm for excitations of 350 to 420 nm. Soils had low intensity broadband emissions over the 400 to 690 nm region for excitations of 300 to 600 nm. The range of relative fluorescence intensities for the crop residues was much greater than the fluorescence observed of the soils. As the crop residues decompose their blue fluorescence values approach the fluorescence of the soil. Fluorescence techniques are concluded to be less ambiguous and better suited for discriminating crop residues and soils than reflectance methods. If properly implemented, fluorescence techniques can be used to quantify, not only crop residue cover, but also photosynthetic efficiency in the field.

  17. Isolation of Chlamydomonas reinhardtii mutants with altered mitochondrial respiration by chlorophyll fluorescence measurement.

    PubMed

    Massoz, Simon; Larosa, Véronique; Horrion, Bastien; Matagne, René F; Remacle, Claire; Cardol, Pierre

    2015-12-10

    The unicellular green alga Chlamydomonas reinhardtii is a model organism for studying energetic metabolism. Most mitochondrial respiratory-deficient mutants characterized to date have been isolated on the basis of their reduced ability to grow in heterotrophic conditions. Mitochondrial deficiencies are usually partly compensated by adjustment of photosynthetic activity and more particularly by transition to state 2. In this work, we explored the opportunity to select mutants impaired in respiration and/or altered in dark metabolism by measuring maximum photosynthetic efficiency by chlorophyll fluorescence analyses (FV/FM). Out of about 2900 hygromycin-resistant insertional mutants generated from wild type or from a mutant strain deficient in state transitions (stt7 strain), 22 were found to grow slowly in heterotrophic conditions and 8 of them also showed a lower FV/FM value. Several disrupted coding sequences were identified, including genes coding for three different subunits of respiratory-chain complex I (NUO9, NUOA9, NUOP4) or for isocitrate lyase (ICL1). Overall, the comparison of respiratory mutants obtained in wild-type or stt7 genetic backgrounds indicated that the FV/FM value can be used to isolate mutants severely impaired in dark metabolism.

  18. New aspects concerning the energy transfer in carotenoids by measuring intracavity absorption spectra and delayed fluorescence

    NASA Astrophysics Data System (ADS)

    Bettermann, Hans; Bouschen, Werner; Ulrich, Lars; Domnick, Gabriele; Martin, H. D.

    1999-05-01

    The first excited singlet state and the lower energetic triplet states of carotenoids are considered to be involved in the light-harvesting as well as in the photochemical protection of cells, respectively. For this reason, the symmetry-forbidden S 0-S 1 (1 1A g-2 1A g) transitions and the multiplicity-forbidden S 0-T 2 (1 1A g-2 3A g) transition of the model carotenoid 8,13-dimethyl-2,2,19,19-tetramethoxy-icosa-4,6,8,10,12,14,16-heptaene-3,18-dione were investigated by intracavity absorption spectroscopy from low-concentrated ethanolic solutions. Both transitions are shaped by promoting modes caused by Herzberg-Teller coupling and the sequence of these modes allows the precise determination of the non-visible S 0-S 1 (0-0)- and S 0-T 2 (0-0)-transitions. The assignments of the singlet-triplet transitions were additionally supported by measuring delayed fluorescence from crystalline samples by directly exciting vibronic triplet states. The vibronic coupling is promoted by C-H bending vibrations of the chain and mainly by deformation modes of the terminating groups of the carotenoid.

  19. Light emitting diode-based nanosecond ultraviolet light source for fluorescence lifetime measurements

    NASA Astrophysics Data System (ADS)

    Araki, Tsutomu; Misawa, Hiroaki

    1995-12-01

    A compact pulsed-light source is devised from an InGaN/AlGaN double heterostructure light-emitting diode (LED). The LED emits a 450-nm (blue) light under conventional dc operation below 30 mA. When a current larger than 50 mA is applied, the intensity of the 450-nm light saturates, but that of the 380-nm light due to the InGaN component continues to increase. This phenomenon is utilized to realize a nanosecond ultraviolet (UV) light source. Under repetitive, large current pulsing (frequency=10 kHz, pulse width=4 ns, peak current=2 A), the peak LED emission shifts from 450 to 380 nm. Intense light pulses (peak value=40 mW) of 4-ns duration were generated. To evaluate the potential of the pulsed LED as an excitation source, the fluorescence lifetime of a quinine-sulfate solution was measured. The observed lifetime characteristics agreed well with the generally accepted behavior.

  20. A simple approach for measuring FRET in fluorescent biosensors using two-photon microscopy.

    PubMed

    Day, Richard N; Tao, Wen; Dunn, Kenneth W

    2016-11-01

    Genetically encoded fluorescent protein (FP)-based biosensor probes are useful tools for monitoring cellular events in living cells and tissues. Because these probes were developed for one-photon excitation approaches, their broad two-photon excitation (2PE) and poorly understood photobleaching characteristics have made their implementation in studies using two-photon laser-scanning microscopy (TPLSM) challenging. Here we describe a protocol that simplifies the use of Förster resonance energy transfer (FRET)-based biosensors in TPLSM. First, the TPLSM system is evaluated and optimized using FRET standards expressed in living cells, which enables the determination of spectral bleed-through (SBT) and the confirmation of FRET measurements from the known standards. Next, we describe how to apply the approach experimentally using a modified version of the A kinase activity reporter (AKAR) protein kinase A (PKA) biosensor as an example-first in cells in culture and then in hepatocytes in the liver of living mice. The microscopic imaging can be accomplished in a day in laboratories that routinely use TPLSM.

  1. Measurements of population densities of metastable and resonant levels of argon using laser induced fluorescence

    SciTech Connect

    Nikolić, M.; Newton, J.; Sukenik, C. I.; Vušković, L.; Popović, S.

    2015-01-14

    We present a new approach to measure population densities of Ar I metastable and resonant excited states in low temperature Ar plasmas at pressures higher than 1 Torr. This approach combines the time resolved laser induced fluorescence technique with the kinetic model of Ar. The kinetic model of Ar is based on calculating the population rates of metastable and resonant levels by including contributions from the processes that affect population densities of Ar I excited states. In particular, we included collisional quenching processes between atoms in the ground state and excited states, since we are investigating plasma at higher pressures. We also determined time resolved population densities of Ar I 2 p excited states by employing optical emission spectroscopy technique. Time resolved Ar I excited state populations are presented for the case of the post-discharge of the supersonic flowing microwave discharge at pressures of 1.7 and 2.3 Torr. The experimental set-up consists of a pulsed tunable dye laser operating in the near infrared region and a cylindrical resonance cavity operating in TE{sub 111} mode at 2.45 GHz. Results show that time resolved population densities of Ar I metastable and resonant states oscillate with twice the frequency of the discharge.

  2. Organization of FtsZ Filaments in the Bacterial Division Ring Measured from Polarized Fluorescence Microscopy

    PubMed Central

    Si, Fangwei; Busiek, Kimberly; Margolin, William; Sun, Sean X.

    2013-01-01

    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. PMID:24209842

  3. Chemotherapy-Induced Changes in Cardiac Capillary Permeability Measured by Fluorescent Multiple Indicator Dilution

    PubMed Central

    Fernandez-Fernandez, Alicia; Carvajal, Denny A.; Lei, Tingjun; McGoron, Anthony J.

    2014-01-01

    Anthracyclines cause severe irreversible cardiac toxicity. The study of changes in cardiac permeability with chemotherapy could enhance the understanding of mechanisms behind cardiac damage, and provide useful information to evaluate anthracycline cardiotoxicity. Thirty-six rats (12 Sprague-Dawley, 12 Wistar, 12 Fischer-344) were randomly assigned to control (n= 21) or doxorubicin (n = 15), and injected i.p. with a cumulative dose of 18 mg/kg doxorubicin in saline (vehicle) or vehicle over 12 days. Echocardiography was performed at baseline and on day 11. An isolated heart experiment was done on day 12 to obtain perfused heart pressure values, and to measure cardiac capillary permeability using a Texas Red/sodium fluorescein multiple indicator dilution method. Control animals had significantly lower average permeability-surface-area-products (0.035±0.013 cm3/s) than doxorubicin animals (0.066±0.023 cm3/s), PSP±SD, p<0.001. These permeability changes correlated with significant functional changes. There was a significant decline in cardiac function with a deleterious effect of chemotherapy on fractional shortening (p<0.001), left ventricular developed pressure (p<0.001), contractility (p<0.001), and relaxation (p=0.02). Based on our results, cardiac capillary permeability changes can be detected after in vivo chemotherapy treatment using our fluorescent multiple indicator dilution technique, and may provide valuable information in evaluating cardiotoxicity of novel drugs. PMID:25224075

  4. Ion dynamics in a DC magnetron microdischarge measured with laser-induced fluorescence

    NASA Astrophysics Data System (ADS)

    Young, Christopher; Gascon, Nicolas; Lucca Fabris, Andrea; Ito, Tsuyohito; Cappelli, Mark

    2015-11-01

    We present evidence of coherent rotating azimuthal wave structures in a planar DC magnetron microdischarge operated with argon and xenon. The dominant stable mode structure varies with discharge voltage, and high frame rate camera imaging of plasma emission reveals propagating azimuthal waves in the negative E-> × B-> direction. This negative drift direction is attributed to a local field reversal arising from strong density gradients that drive excess ions towards the anode. Observed mode transitions are shown to be consistent with models of gradient drift-wave dispersion in such a field reversal when the fluid representation includes ambipolar diffusion parallel to the magnetic field direction. Time-averaged and time-resolved laser-induced fluorescence measurements interrogate xenon ion dynamics under the action of the field reversal. Time resolution is obtained by synchronizing with the coherent azimuthal wave frequency at fixed mode number. This work is sponsored by the U.S. Air Force Office of Scientific Research with Dr. Mitat Birkan as program manager. C.Y. acknowledges support from the DOE NNSA Stewardship Science Graduate Fellowship under Contract DE-FC52-08NA28752.

  5. A simple approach for measuring FRET in fluorescent biosensors using two-photon microscopy.

    PubMed

    Day, Richard N; Tao, Wen; Dunn, Kenneth W

    2016-11-01

    Genetically encoded fluorescent protein (FP)-based biosensor probes are useful tools for monitoring cellular events in living cells and tissues. Because these probes were developed for one-photon excitation approaches, their broad two-photon excitation (2PE) and poorly understood photobleaching characteristics have made their implementation in studies using two-photon laser-scanning microscopy (TPLSM) challenging. Here we describe a protocol that simplifies the use of Förster resonance energy transfer (FRET)-based biosensors in TPLSM. First, the TPLSM system is evaluated and optimized using FRET standards expressed in living cells, which enables the determination of spectral bleed-through (SBT) and the confirmation of FRET measurements from the known standards. Next, we describe how to apply the approach experimentally using a modified version of the A kinase activity reporter (AKAR) protein kinase A (PKA) biosensor as an example-first in cells in culture and then in hepatocytes in the liver of living mice. The microscopic imaging can be accomplished in a day in laboratories that routinely use TPLSM. PMID:27685098

  6. Noninvasive quantitative measurement of colloid transport in mesoscale porous media using time lapse fluorescence imaging.

    PubMed

    Bridge, Jonathan W; Banwart, Steven A; Heathwaite, A Louise

    2006-10-01

    We demonstrate noninvasive quantitative imaging of colloid and solute transport at millimeter to decimeter (meso-) scale. Ultraviolet (UV) excited fluorescent solute and colloid tracers were independently measured simultaneously during co-advection through saturated quartz sand. Pulse-input experiments were conducted at constant flow rates and ionic strengths 10(-3), 10(-2) and 10(-1) M NaCl. Tracers were 1.9 microm carboxylate latex microspheres and disodium fluorescein. Spatial moments analysis was used to quantify relative changes in mass distribution of the colloid and solute tracers over time. The solute advected through the sand at a constant velocity proportional to flow rate and was described well by a conservative transport model (CXTFIT). In unfavorable deposition conditions increasing ionic strength produced significant reduction in colloid center of mass transport velocity over time. Velocity trends correlated with the increasing fraction of colloid mass retained along the flowpath. Attachment efficiencies (defined by colloid filtration theory) calculated from nondestructive retained mass data were 0.013 +/- 0.03, 0.09 +/- 0.02, and 0.22 +/- 0.05 at 10(-3), 10(-2), and 10(-1) M ionic strength, respectively, which compared well with previously published data from breakthrough curves and destructive sampling. Mesoscale imaging of colloid mass dynamics can quantify key deposition and transport parameters based on noninvasive, nondestructive, spatially high-resolution data.

  7. pH Measurement Using Dual-Wavelength Fluorescent Ratio by Two-Photon Excitation for Mitochondrial Activity

    NASA Astrophysics Data System (ADS)

    Kanazashi, Yasuaki; Li, Yongbo; Onojima, Takumi; Iwami, Kentaro; Ohta, Yoshihiro; Umeda, Norihiro

    2012-11-01

    A mitochondrion has a pH gradient between the two sides of its inner membrane in order to produce adenosine triphosphate (ATP). Because ATP depletion causes numerous diseases, the measurement of the pH value around the mitochondrion is expected to clarify the mechanism of these diseases. In this study, a dual-wavelength pH-sensitive dye was excited by two-photon absorption initiated using a femtosecond pulse laser. In addition, fluorescence from the dye was directly collected from the fluorescent point using the collection-mode probe of a scanning near-field optical microscope. By this proposed method, a pH calibration curve was obtained from the fluorescent intensity ratio of the dye solution, and temporal pH variations with 0.1 s time resolution following the addition of acid were observed. Moreover, mitochondrial activity on the basis of the pH changes was successfully observed in three different mitochondrial densities.

  8. Image processing for non-ratiometric measurement of membrane voltage using fluorescent reporters and pulsed laser illumination.

    PubMed

    Silve, Aude; Rocke, Sarah; Frey, Wolfgang

    2015-06-01

    The measurement of transmembrane voltages induced by pulsed electric field exposure can be achieved by using fluorescent dyes like ANNINE-6. Such approach requires a quantitative determination of the fluorescence intensity along the cell's membrane by image processing. When high temporal resolution is required, the illumination source is frequently a dye-laser which causes high fluctuations in the intensity of illumination which in turn affects the fluorescence intensity and thus the quality of the results. We propose an image processing technique that allows to overcome the fluctuations and to produce quantitative data. It uses the optical background noise as a correcting factor. Standard deviation in the fluctuations is thus efficiently reduced by at least a factor of 2.5. Additionally we draw attention to the fact that the parasitic component of the laser radiation (ASE) can also suppress fluctuations although it deteriorates wavelength precision.

  9. Method for measuring temperatures and densities in hypersonic wind tunnel air flows using laser-induced O2 fluorescence

    NASA Technical Reports Server (NTRS)

    Laufer, Gabriel; Mckenzie, Robert L.; Fletcher, Douglas G.

    1990-01-01

    Laser-induced fluorescence in oxygen, in combination with Raman scattering, is shown to be an accurate means by which temperature, density, and their fluctuations owing to turbulence can be measured in air flows associated with high-speed wind tunnels. For temperatures above 60 K and densities above 0.01 amagat, the uncertainties in the temperature and density measurements can be less than 2 percent, if the signal uncertainties are dominated by photon statistical noise. The measurements are unaffected by collisional quenching and can be achieved with laser fluences for which nonlinear effects are insignificant. Temperature measurements using laser-induced fluorescence alone have been demonstrated at known densities in the range of low temperatures and densities which are expected in a hypersonic wind tunnel.

  10. A method for measuring temperatures and densities in hypersonic wind tunnel air flows using laser-induced O2 fluorescence

    NASA Technical Reports Server (NTRS)

    Laufer, Gabriel; Fletcher, Douglas G.; Mckenzie, Robert L.

    1990-01-01

    Laser-induced fluorescence in oxygen, in combination with Raman scattering, is shown to be an accurate means by which temperature, density, and their fluctuations due to turbulence can be measured in air flows associated with high-speed wind tunnels. For temperatures above 60 K and densities above 0.01 amagat, the uncertainty in the temperature and density measurements can be less than 2 and 3 percent, respectively, if the signal uncertainties are dominated by photon-statistical noise. The measurements are unaffected by collisional quenching and can be achieved with laser fluences for which nonlinear effects are insignificant. Temperature measurements using laser-induced fluorescence alone have been demonstrated at known densities in the range of low temperatures and densities which are expected in a hypersonic wind tunnel.

  11. Method for measuring temperatures and densities in hypersonic wind tunnel air flows using laser-induced O(2) fluorescence.

    PubMed

    Laufer, G; McKenzie, R L; Fletcher, D G

    1990-11-20

    Laser-induced fluorescence in oxygen, in combination with Raman scattering, is shown to be an accurate means by which temperature, density, and their fluctuations owing to turbulence can be measured in air flows associated with high speed wind tunnels. For temperatures above 60 K and densities above 0.01 amagat, the uncertainties in the temperature and density measurements can be <2%, if the signal uncertainties are dominated by photon statistical noise. The measurements are unaffected by collisional quenching and can be achieved with laser fluences for which nonlinear effects are insignificant. Temperature measurements using laser-induced fluorescence alone have been demonstrated at known densities in the range of low temperatures and densities which are expected in a hypersonic wind tunnel.

  12. Phantom validation of Monte Carlo modeling for noncontact depth sensitive fluorescence measurements in an epithelial tissue model.

    PubMed

    Ong, Yi Hong; Zhu, Caigang; Liu, Quan

    2014-08-01

    Experimental investigation and optimization of various optical parameters in the design of depth sensitive optical measurements in layered tissues would require a huge amount of time and resources. A computational method to model light transport in layered tissues using Monte Carlo simulations has been developed for decades to reduce the cost incurred during this process. In this work, we employed the Monte Carlo method to investigate the depth sensitivity achieved by various illumination and detection configurations including both the traditional cone configurations and new cone shell configurations, which are implemented by convex or axicon lenses. Phantom experiments have been carried out to validate the Monte Carlo modeling of fluorescence in a two-layered turbid, epithelial tissue model. The measured fluorescence and depth sensitivity of different illumination–detection configurations were compared with each other. The results indicate excellent agreement between the experimental and simulation results in the trends of fluorescence intensity and depth sensitivity. The findings of this study and the development of the Monte Carlo method for noncontact setups provide useful insight and assistance in the planning and optimization of optical designs for depth sensitive fluorescence measurements.

  13. A dark green fluorescent protein as an acceptor for measurement of Förster resonance energy transfer

    PubMed Central

    Murakoshi, Hideji; Shibata, Akihiro C. E.; Nakahata, Yoshihisa; Nabekura, Junichi

    2015-01-01

    Measurement of Förster resonance energy transfer by fluorescence lifetime imaging microscopy (FLIM-FRET) is a powerful method for visualization of intracellular signaling activities such as protein-protein interactions and conformational changes of proteins. Here, we developed a dark green fluorescent protein (ShadowG) that can serve as an acceptor for FLIM-FRET. ShadowG is spectrally similar to monomeric enhanced green fluorescent protein (mEGFP) and has a 120-fold smaller quantum yield. When FRET from mEGFP to ShadowG was measured using an mEGFP-ShadowG tandem construct with 2-photon FLIM-FRET, we observed a strong FRET signal with low cell-to-cell variability. Furthermore, ShadowG was applied to a single-molecule FRET sensor to monitor a conformational change of CaMKII and of the light oxygen voltage (LOV) domain in HeLa cells. These sensors showed reduced cell-to-cell variability of both the basal fluorescence lifetime and response signal. In contrast to mCherry- or dark-YFP-based sensors, our sensor allowed for precise measurement of individual cell responses. When ShadowG was applied to a separate-type Ras FRET sensor, it showed a greater response signal than did the mCherry-based sensor. Furthermore, Ras activation and translocation of its effector ERK2 into the nucleus could be observed simultaneously. Thus, ShadowG is a promising FLIM-FRET acceptor. PMID:26469148

  14. A dark green fluorescent protein as an acceptor for measurement of Förster resonance energy transfer.

    PubMed

    Murakoshi, Hideji; Shibata, Akihiro C E; Nakahata, Yoshihisa; Nabekura, Junichi

    2015-10-15

    Measurement of Förster resonance energy transfer by fluorescence lifetime imaging microscopy (FLIM-FRET) is a powerful method for visualization of intracellular signaling activities such as protein-protein interactions and conformational changes of proteins. Here, we developed a dark green fluorescent protein (ShadowG) that can serve as an acceptor for FLIM-FRET. ShadowG is spectrally similar to monomeric enhanced green fluorescent protein (mEGFP) and has a 120-fold smaller quantum yield. When FRET from mEGFP to ShadowG was measured using an mEGFP-ShadowG tandem construct with 2-photon FLIM-FRET, we observed a strong FRET signal with low cell-to-cell variability. Furthermore, ShadowG was applied to a single-molecule FRET sensor to monitor a conformational change of CaMKII and of the light oxygen voltage (LOV) domain in HeLa cells. These sensors showed reduced cell-to-cell variability of both the basal fluorescence lifetime and response signal. In contrast to mCherry- or dark-YFP-based sensors, our sensor allowed for precise measurement of individual cell responses. When ShadowG was applied to a separate-type Ras FRET sensor, it showed a greater response signal than did the mCherry-based sensor. Furthermore, Ras activation and translocation of its effector ERK2 into the nucleus could be observed simultaneously. Thus, ShadowG is a promising FLIM-FRET acceptor.

  15. X-Ray Diffraction and Fluorescence Measurements for In Situ Planetary Instruments

    NASA Astrophysics Data System (ADS)

    Hansford, G.; Hill, K. S.; Talboys, D.; Vernon, D.; Ambrosi, R.; Bridges, J.; Hutchinson, I.; Marinangeli, L.

    2011-12-01

    The ESA/NASA ExoMars mission, due for launch in 2018, has a combined X-ray fluorescence/diffraction instrument, Mars-XRD, as part of the onboard analytical laboratory. The results of some XRF (X-ray fluorescence) and XRD (X-ray diffraction) tests using a laboratory chamber with representative performance are reported. A range of standard geological reference materials and analogues were used in these tests. The XRD instruments are core components of the forthcoming NASA Mars Science Laboratory (MSL) and ESA/NASA ExoMars missions and will provide the first demonstrations of the capabilities of combined XRD/XRF instrumentation in situ on an extraterrestrial planetary surface. The University of Leicester team is part of the Italy-UK collaboration that is responsible for building the ExoMars X-ray diffraction instrument, Mars-XRD [1,2]. Mars-XRD incorporates an Fe-55 radioisotope source and three fixed-position charge-coupled devices (CCDs) to simultaneously acquire an X-ray fluorescence spectrum and a diffraction pattern providing a measurement of both elemental and mineralogical composition. The CCDs cover an angular range of 2θ = 6° to 73° enabling the analysis of a wide range of geologically important minerals including phyllosilicates, feldspars, oxides, carbonates and evaporites. The identification of hydrous minerals may help identify past Martian hydrothermal systems capable of preserving traces of life. Here we present some initial findings from XRF and XRD tests carried out at the University of Leicester using an Fe-55 source and X-ray sensitive CCD. The XRF/XRD test system consists of a single CCD on a motorised arm, an Fe-55 X-ray source, a collimator and a sample table which approximately replicate the reflection geometry of the Mars-XRD instrument. It was used to test geological reference standard materials and Martian analogues. This work was funded by the Science and Technology Facilities Council, UK. References [1] Marinangeli, L., Hutchinson, I

  16. In Situ Airborne Measurement of Formaldehyde with a New Laser Induced Fluorescence Instrument

    NASA Astrophysics Data System (ADS)

    Arkinson, H.; Hanisco, T. F.; Cazorla, M.; Fried, A.; Walega, J.

    2012-12-01

    Formaldehyde (HCHO) is a highly reactive and ubiquitous compound in the atmosphere that originates from primary emissions and secondary formation by photochemical oxidation of volatile organic compounds. HCHO is an important precursor to the formation of ozone and an ideal tracer for the transport of boundary layer pollutants to higher altitudes. In situ measurements of HCHO are needed to improve understanding of convective transport mechanisms and the effects of lofted pollutants on ozone production and cloud microphysics in the upper troposphere. The Deep Convective Clouds and Chemistry Project (DC3) field campaign addressed the effects of deep, midlatitude continental convective clouds on the upper troposphere by examining vertical transport of fresh emissions and water aloft and by characterizing subsequent changes in composition and chemistry. Observations targeting convective storms were conducted over Colorado, Alabama, and Texas and Oklahoma. We present measurements of the In Situ Airborne Formaldehyde instrument (ISAF), which uses laser induced fluorescence to achieve the high sensitivity and fast time response required to detect low concentrations in the upper troposphere and capture the fine structure characteristic of convective storm outflow. Preliminary results from DC3 indicate that the ISAF is able to resolve concentrations ranging from under 35 ppt to over 35 ppb, spanning three orders of magnitude, in less than a few minutes. Frequent, abrupt changes in HCHO captured by the ISAF are corroborated by similar patterns observed by simultaneous trace gas and aerosol measurements. Primary HCHO emissions are apparent in cases when the DC-8 flew over combustion sources or biomass burning, and secondary HCHO formation is suggested by observations of enhanced HCHO concurrent with other elevated hydrocarbons. Vertical transport of HCHO is indicated by measurements of over 6 ppb from outflow in the upper troposphere. The DC-8 payload also included the

  17. Electrical perception of the 'death message' in Chara: characterization of K+ -induced depolarization.

    PubMed

    Shimmen, Teruo

    2006-04-01

    When the nodal end of an internodal cell of Chara corallina was subjected to a pulse treatment with artificial cell sap, a depolarization lasted in artificial pond water. This depolarization could also be induced by pulse treatment with KCl solution, indicating that K+ in the artificial cell sap is responsible (K+ -induced depolarization). The depolarization was prolonged in the presence of 2 mM KCl and the prolonged depolarization was terminated by supplementing with either CaCl2, MgCl2 or NaCl. These results supported the hypothesis that K+ released from the killed cell is responsible for generation of the wound-induced membrane depolarization.

  18. Laser-saturated fluorescence of nitric oxide and chemiluminescence measurements in premixed ethanol flames

    SciTech Connect

    Marques, Carla S.T.; Barreta, Luiz G.; Sbampato, Maria E.; dos Santos, Alberto M.

    2010-11-15

    In this study, nitric oxide laser-saturated fluorescence (LSF) measurements were acquired from premixed ethanol flames at atmospheric pressure in a burner. NO-LSF experimental profiles for fuel-rich premixed ethanol flames ({phi} = 1.34 and {phi} = 1.66) were determined through the excitation/detection scheme of the Q{sub 2}(26.5) rotational line in the A{sup 2}{sigma}{sup +} - X{sup 2}{pi} (0,0) vibronic band and {gamma}(0,1) emission band. A calibration procedure by NO doping into the flame was applied to establish the NO concentration profiles in these flames. Chemiluminescent emission measurements in the (0, 0) vibronic emission bands of the OH{sup *} (A{sup 2}{sigma}{sup +} - X{sup 2}{pi}) and CH{sup *}(A{sup 2}{delta} - X{sup 2}{pi}) radicals were also obtained with high spatial and spectral resolution for fuel-rich premixed ethanol flames to correlate them with NO concentrations. Experimental chemiluminescence profiles and the ratios of the integrated areas under emission spectra (A{sub CH*}/A{sub CH*}(max.) and A{sub CH*}/A{sub OH*}) were determined. The relationships between chemiluminescence and NO concentrations were established along the premixed ethanol flames. There was a strong connection between CH{sup *} radical chemiluminescence and NO formation and the prompt-NO was identified as the governing mechanism for NO production. The results suggest the optimum ratio of the chemiluminescence of two radicals (A{sub CH*}/A{sub OH*}) for NO diagnostic purposes. (author)

  19. Rapid virus production and removal as