Stitching Type Large Aperture Depolarizer for Gas Monitoring Imaging Spectrometer

NASA Astrophysics Data System (ADS)

Liu, X.; Li, M.; An, N.; Zhang, T.; Cao, G.; Cheng, S.

2018-04-01

To increase the accuracy of radiation measurement for gas monitoring imaging spectrometer, it is necessary to achieve high levels of depolarization of the incoming beam. The preferred method in space instrument is to introduce the depolarizer into the optical system. It is a combination device of birefringence crystal wedges. Limited to the actual diameter of the crystal, the traditional depolarizer cannot be used in the large aperture imaging spectrometer (greater than 100 mm). In this paper, a stitching type depolarizer is presented. The design theory and numerical calculation model for dual babinet depolarizer were built. As required radiometric accuracies of the imaging spectrometer with 250 mm × 46 mm aperture, a stitching type dual babinet depolarizer was design in detail. Based on designing the optimum structural parmeters the tolerance of wedge angle refractive index, and central thickness were given. The analysis results show that the maximum residual polarization degree of output light from depolarizer is less than 2 %. The design requirements of polarization sensitivity is satisfied.

Mueller matrix signature in advanced fluorescence microscopy imaging

NASA Astrophysics Data System (ADS)

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

2017-02-01

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

Enhancing Continuous Online Microdialysis Using Dexamethasone: Measurement of Dynamic Neurometabolic Changes during Spreading Depolarization.

PubMed

Varner, Erika L; Leong, Chi Leng; Jaquins-Gerstl, Andrea; Nesbitt, Kathryn M; Boutelle, Martyn G; Michael, Adrian C

2017-08-16

Microdialysis is well established in chemical neuroscience as a mainstay technology for real time intracranial chemical monitoring in both animal models and human patients. Evidence shows that microdialysis can be enhanced by mitigating the penetration injury caused during the insertion of microdialysis probes into brain tissue. Herein, we show that retrodialysis of dexamethasone in the rat cortex enhances the microdialysis detection of K + and glucose transients induced by spreading depolarization. Without dexamethasone, quantification of glucose transients was unreliable by 5 days after probe insertion. With dexamethasone, robust K + and glucose transients were readily quantified at 2 h, 5 days, and 10 days after probe insertion. The amplitudes of the K + transients declined day-to-day following probe insertion, and the amplitudes of the glucose transients exhibited a decreasing trend that did not reach statistical significance. Immunohistochemistry and fluorescence microscopy confirm that dexamethasone is highly effective at preserving a healthy probe-brain interface for at least 10 days even though retrodialysis of dexamethasone ceased after 5 days.

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.

[The role of neuroglobin in oxygen-glucose deprivation and reoxygenation-induced mitochondrial depolarization and reactive oxygen species production in SH-SY5Y cells].

PubMed

Deng, S Y; Ai, Y H; Zhang, L N; Wu, L; Chen, C X; Wang, Y M; Liu, Z Y; Huang, L; Peng, Q Y

2017-01-01

Objective: To investigate the role of neuroglobin (NGB) in oxygen-glucose deprivation and reoxygenation (OGD/R) induced mitochondrial depolarization and reactive oxygen species (ROS)production in a human neuroblastoma cell line (SH-SY5Y). Methods: SH-SY5Y cells were transfected with lentivirus to establish a stable cell line of NGB knockdown (KD). After treated with OGD/R, cells were collected at different time points to analyze NGB mRNA and protein levels. Furthermore, cells were stained with JC-1 and DCFH-DA to evaluate mitochondrial depolarization and ROS production by inverted fluorescence microscope. Also, to determine the neurotoxicity, we measured the lactate dehydrogenase(LDH)level in the cell culture medium. Results: After the treatment of OGD/R, the NGB mRNA and protein started to elevate and peak at 4 h and 8 h (2.04±0.35 fold, 1.69±0.18 fold). Compared with the vector group, NGB KD group had much more mitochondrial depolarization [JC-1 red/green (1.10±0.10) vs (1.46±0.11), P <0.05] and ROS production [DCFH-DA fluorescence (36.30±5.32) vs (16.26±2.97), P <0.05]. Furthermore, NGB KD groups had a higher level of LDH release [(63.42±6.14)%vs (49.65±5.09)%, P <0.05]. Conclusions: NGB plays an important role in the homeostasis of mitochondria. Knockdown of NGB results in increased mitochondrial depolarization, ROS production and neurotoxicity under hypoxia circumstances.

Ischemia-induced spreading depolarization in the retina

PubMed Central

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

2016-01-01

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

Depolarization on Earth-space paths

NASA Technical Reports Server (NTRS)

1981-01-01

Sources of depolarization effects on the propagation paths of orthogonally-polarized information channels are considered. The main sources of depolarization at millimeter wave frequencies are hydrometeor absorption and scattering in the troposphere. Terms are defined. Mathematical formulations for the effects of the propagation medium characteristics and antenna performance on signals in dual polarization Earth-space links are presented. Techniques for modeling rain and ice depolarization are discussed.

FM Dye Cycling at the Synapse: Comparing High Potassium Depolarization, Electrical and Channelrhodopsin Stimulation.

PubMed

Kopke, Danielle L; Broadie, Kendal

2018-05-24

FM dyes are used to study the synaptic vesicle (SV) cycle. These amphipathic probes have a hydrophilic head and hydrophobic tail, making them water-soluble with the ability to reversibly enter and exit membrane lipid bilayers. These styryl dyes are relatively non-fluorescent in aqueous medium, but insertion into the outer leaflet of the plasma membrane causes a >40X increase in fluorescence. In neuronal synapses, FM dyes are internalized during SV endocytosis, trafficked both within and between SV pools, and released with SV exocytosis, providing a powerful tool to visualize presynaptic stages of neurotransmission. A primary genetic model of glutamatergic synapse development and function is the Drosophila neuromuscular junction (NMJ), where FM dye imaging has been used extensively to quantify SV dynamics in a wide range of mutant conditions. The NMJ synaptic terminal is easily accessible, with a beautiful array of large synaptic boutons ideal for imaging applications. Here, we compare and contrast the three ways to stimulate the Drosophila NMJ to drive activity-dependent FM1-43 dye uptake/release: 1) bath application of high [K + ] to depolarize neuromuscular tissues, 2) suction electrode motor nerve stimulation to depolarize the presynaptic nerve terminal, and 3) targeted transgenic expression of channelrhodopsin variants for light-stimulated, spatial control of depolarization. Each of these methods has benefits and disadvantages for the study of genetic mutation effects on the SV cycle at the Drosophila NMJ. We will discuss these advantages and disadvantages to assist the selection of the stimulation approach, together with the methodologies specific to each strategy. In addition to fluorescent imaging, FM dyes can be photoconverted to electron-dense signals visualized using transmission electron microscopy (TEM) to study SV cycle mechanisms at an ultrastructural level. We provide the comparisons of confocal and electron microscopy imaging from the different

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.

The elementary events of Ca2+ release elicited by membrane depolarization in mammalian muscle

PubMed Central

Csernoch, L; Zhou, J; Stern, M D; Brum, G; Ríos, E

2004-01-01

Cytosolic [Ca2+] transients elicited by voltage clamp depolarization were examined by confocal line scanning of rat skeletal muscle fibres. Ca2+ sparks were observed in the fibres' membrane-permeabilized ends, but not in responses to voltage in the membrane-intact area. Elementary events of the depolarization-evoked response could be separated either at low voltages (near −50 mV) or at −20mV in partially inactivated cells. These were of lower amplitude, narrower and of much longer duration than sparks, similar to ‘lone embers’ observed in the permeabilized segments. Their average amplitude was 0.19 and spatial half-width 1.3 μm. Other parameters depended on voltage. At −50 mV average duration was 111 ms and latency 185 ms. At −20 mV duration was 203 ms and latency 24 ms. Ca2+ release current, calculated on an average of events, was nearly steady at 0.5–0.6 pA. Accordingly, simulations of the fluorescence event elicited by a subresolution source of 0.5 pA open for 100 ms had morphology similar to the experimental average. Because 0.5 pA is approximately the current measured for single RyR channels in physiological conditions, the elementary fluorescence events in rat muscle probably reflect opening of a single RyR channel. A reconstruction of cell-averaged release flux at −20 mV based on the observed distribution of latencies and calculated elementary release had qualitatively correct but slower kinetics than the release flux in prior whole-cell measurements. The qualitative agreement indicates that global Ca2+ release flux results from summation of these discrete events. The quantitative discrepancies suggest that the partial inactivation strategy may lead to events of greater duration than those occurring physiologically in fully polarized cells. PMID:14990680

The elementary events of Ca2+ release elicited by membrane depolarization in mammalian muscle.

PubMed

Csernoch, L; Zhou, J; Stern, M D; Brum, G; Ríos, E

2004-05-15

Cytosolic [Ca(2+)] transients elicited by voltage clamp depolarization were examined by confocal line scanning of rat skeletal muscle fibres. Ca(2+) sparks were observed in the fibres' membrane-permeabilized ends, but not in responses to voltage in the membrane-intact area. Elementary events of the depolarization-evoked response could be separated either at low voltages (near -50 mV) or at -20 mV in partially inactivated cells. These were of lower amplitude, narrower and of much longer duration than sparks, similar to 'lone embers' observed in the permeabilized segments. Their average amplitude was 0.19 and spatial half-width 1.3 microm. Other parameters depended on voltage. At -50 mV average duration was 111 ms and latency 185 ms. At -20 mV duration was 203 ms and latency 24 ms. Ca(2+) release current, calculated on an average of events, was nearly steady at 0.5-0.6 pA. Accordingly, simulations of the fluorescence event elicited by a subresolution source of 0.5 pA open for 100 ms had morphology similar to the experimental average. Because 0.5 pA is approximately the current measured for single RyR channels in physiological conditions, the elementary fluorescence events in rat muscle probably reflect opening of a single RyR channel. A reconstruction of cell-averaged release flux at -20 mV based on the observed distribution of latencies and calculated elementary release had qualitatively correct but slower kinetics than the release flux in prior whole-cell measurements. The qualitative agreement indicates that global Ca(2+) release flux results from summation of these discrete events. The quantitative discrepancies suggest that the partial inactivation strategy may lead to events of greater duration than those occurring physiologically in fully polarized cells.

Optical Measurement of Asian Dust over Daejeon City in 2016 by Depolarization Lidar in AD-Network

NASA Astrophysics Data System (ADS)

Bong, Park Chan; Shimizu, Atsushi; Sugimoto, Nobuo

2018-04-01

Long-term sustained heavy Asian Dust below 3 km was measured in the period of April 22-26 by 2-wavelengths depolarization lidar system. As the comparison results of Daejeon station with other stations in the AD-Net, similarly formed Asian dust had been measured at Nigata, Toyama, Matsue, and Sendai. The route of the dust was examined by HYSPLIT. More than 80 μg / m3 mass concentrations derived from the lidar measurements were compared with Air-Korea PM10 data.

Correction of Depolarizing Resonances in ELSA

NASA Astrophysics Data System (ADS)

Steier, C.; Husmann, D.

1997-05-01

The 3.5 GeV electron stretcherring ELSA (ELectron Stretcher Accelerator) at Bonn University is operational since 1987, both as a continuous beam facility for external fixed target experiments and as a partially dedicated synchrotron light source. For the external experiments an upgrade to polarized electrons is under way. One source of polarized electrons (GaAs crystal, photoeffect using circular polarized laser light) is operational. The studies of minimizing the losses in polarization degree due to crossing of depolarizing resonances that necessarily exist in circular accelerators (storagerings) just started recently. Calculations concerning different correction schemes for the depolarizing resonances in ELSA are presented, and first results of measurements are shown (done by means of a Møller polarimeter in one of the external beamlines).

Propofol and Sevoflurane Differentially Modulate Cortical Depolarization following Electric Stimulation of the Ventrobasal Thalamus.

PubMed

Kratzer, Stephan; Mattusch, Corinna; Garcia, Paul S; Schmid, Sebastian; Kochs, Eberhard; Rammes, Gerhard; Schneider, Gerhard; Kreuzer, Matthias; Haseneder, Rainer

2017-01-01

The neuronal mechanisms how anesthetics lead to loss of consciousness are unclear. Thalamocortical interactions are crucially involved in conscious perception; hence the thalamocortical network might be a promising target for anesthetic modulation of neuronal information pertaining to arousal and waking behavior. General anesthetics affect the neurophysiology of the thalamus and the cortex but the exact mechanisms of how anesthetics interfere with processing thalamocortical information remain to be elucidated. Here we investigated the effect of the anesthetic agents sevoflurane and propofol on thalamocortical network activity in vitro . We used voltage-sensitive dye imaging techniques to analyze the cortical depolarization in response to stimulation of the thalamic ventrobasal nucleus in brain slices from mice. Exposure to sevoflurane globally decreased cortical depolarization in a dose-dependent manner. Sevoflurane reduced the intensity and extent of cortical depolarization and delayed thalamocortical signal propagation. In contrast, propofol neither affected area nor amplitude of cortical depolarization. However, propofol exposure resulted in regional changes in spatial distribution of maximum fluorescence intensity in deep regions of the cortex. In summary, our experiments revealed substance-specific effects on the thalamocortical network. Functional changes of the neuronal network are known to be pivotally involved in the anesthetic-induced loss of consciousness. Our findings provide further evidence that the mechanisms of anesthetic-mediated loss of consciousness are drug- and pathway-specific.

Heterogeneous incidence and propagation of spreading depolarizations

PubMed Central

Kaufmann, Dan; Theriot, Jeremy J; Zyuzin, Jekaterina; Service, C Austin; Chang, Joshua C; Tang, Y Tanye; Bogdanov, Vladimir B; Multon, Sylvie; Schoenen, Jean; Ju, Y Sungtaek

2016-01-01

Spreading depolarizations are implicated in a diverse set of neurologic diseases. They are unusual forms of nervous system activity in that they propagate very slowly and approximately concentrically, apparently not respecting the anatomic, synaptic, functional, or vascular architecture of the brain. However, there is evidence that spreading depolarizations are not truly concentric, isotropic, or homogeneous, either in space or in time. Here we present evidence from KCl-induced spreading depolarizations, in mouse and rat, in vivo and in vitro, showing the great variability that these depolarizations can exhibit. This variability can help inform the mechanistic understanding of spreading depolarizations, and it has implications for their phenomenology in neurologic disease. PMID:27562866

Neutron Depolarization in Superconductors

NASA Astrophysics Data System (ADS)

Zhuchenko, N. K.

1995-04-01

The dependences of neutron depolarization on applied magnetic field are deduced along the magnetization hysteresis loop in terms of the Bean model of the critical state. The depolarization in uniaxial superconductors with the reversible magnetization, including uniaxial magnetic superconductors, is also considered. A strong depolarization is expected if the neutrons travel along the vortex lines. On calcule la dépendance en champ magnétique de la dépolarisation des neutrons le long du cycle d'hystérésis en termes du modèle critique de Bean. On considère aussi la dépolarisation dans les supraconducteurs uniaxiaux en fonction de l'aimantation réversible, y compris pour les supraconducteurs magnétiques. On attend une forte dépolarisation si les neutrons se propagent le long des vortex.

Depolarization Diffusion During Weak Suprathreshold Stimulation of Cardiac Tissue

DTIC Science & Technology

2001-10-25

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

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

PubMed

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

2012-12-03

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

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

NASA Technical Reports Server (NTRS)

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

1976-01-01

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

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

NASA Technical Reports Server (NTRS)

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

1982-01-01

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

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.

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.

Triple-wavelength lidar observations of the linear depolarization ratio of dried marine particles

NASA Astrophysics Data System (ADS)

Haarig, Moritz; Ansmann, Albert; Baars, Holger; Engelmann, Ronny; Althausen, Dietrich; Bohlmann, Stephanie; Gasteiger, Josef; Farrell, David

2018-04-01

For aerosol typing with lidar, sea salt particles are usually assumed to be spherical with a consequently low depolarization ratio. Evidence of dried marine particles at the top of the humid marine aerosol layer with a depolarization ratio up to 0.1 has been found at predominately maritime locations on Barbados and in the Southern Atlantic. The depolarization ratio for these probably cubic sea salt particles has been measured at three wavelengths (355, 532 and 1064 nm) simultaneously for the first time and compared to model simulations.

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

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

PubMed

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

2015-04-23

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

Oxygen availability and spreading depolarizations provide complementary prognostic information in neuromonitoring of aneurysmal subarachnoid hemorrhage patients.

PubMed

Winkler, Maren Kl; Dengler, Nora; Hecht, Nils; Hartings, Jed A; Kang, Eun J; Major, Sebastian; Martus, Peter; Vajkoczy, Peter; Woitzik, Johannes; Dreier, Jens P

2017-05-01

Multimodal neuromonitoring in neurocritical care increasingly includes electrocorticography to measure epileptic events and spreading depolarizations. Spreading depolarization causes spreading depression of activity (=isoelectricity) in electrically active tissue. If the depression is long-lasting, further spreading depolarizations occur in still isoelectric tissue where no activity can be suppressed. Such spreading depolarizations are termed isoelectric and are assumed to indicate energy compromise. However, experimental and clinical recordings suggest that long-lasting spreading depolarization-induced depression and isoelectric spreading depolarizations are often recorded outside of the actual ischemic zones, allowing the remote diagnosis of delayed cerebral ischemia after aneurysmal subarachnoid hemorrhage. Here, we analyzed simultaneous electrocorticography and tissue partial pressure of oxygen recording in 33 aneurysmal subarachnoid hemorrhage patients. Multiple regression showed that both peak total depression duration per recording day and mean baseline tissue partial pressure of oxygen were independent predictors of outcome. Moreover, tissue partial pressure of oxygen preceding spreading depolarization was similar and differences in tissue partial pressure of oxygen responses to spreading depolarization were only subtle between isoelectric spreading depolarizations and spreading depressions. This further supports that, similar to clustering of spreading depolarizations, long spreading depolarization-induced periods of isoelectricity are useful to detect energy compromise remotely, which is valuable because the exact location of future developing pathology is unknown at the time when the neurosurgeon implants recording devices.

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

NASA Astrophysics Data System (ADS)

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

2013-01-01

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

Proton transport polarization and depolarization of hydroxyapatite ceramics

NASA Astrophysics Data System (ADS)

Nakamura, Satoshi; Takeda, Hiroaki; Yamashita, Kimihiro

2001-05-01

Polarization of sintered hydroxyapatite (HAp) ceramics by application of an external dc field at higher temperature was analyzed by thermally stimulated depolarization current (TSDC) measurements. The mechanisms for the polarization and depolarization of HAp were discussed in relation to the instability of the protons in the hydroxide groups. The TSDC spectra consisted of broad peaks, while the ferroelectric substances such as the BaTiO3 ceramics exhibited a sharp peak. Although the maximum current density of 7.87 nA cm-2 for the HAp polarized at 400 °C under 1.0 kV cm-1 was approximately 1/12 lower than that of BaTiO3, the polarization charge of 14.9 μC cm-2 was almost twice as large as that of BaTiO3. Considering the activation energy of 0.72-0.89 eV for the depolarization, it was revealed that the polarization of HAp was ascribed to the migration of protons in the columnar OH- channels with a micrometer-order distance. It was also found that the polarization charge was large and long enough to enhance the biological reactivity of HAp ceramics for biomedical implants.

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.

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

Neutron depolarization effects in a high-Tc superconductor (abstract)

NASA Astrophysics Data System (ADS)

Nunes, A. C.; Pickart, S. J.; Crow, L.; Goyette, R.; McGuire, T. R.; Shinde, S.; Shaw, T. M.

1988-11-01

Using the polarized beam small-angle neutron scattering spectrometer at the Rhode Island Nuclear Science Center Reactor, we have observed significant depolarization of a neutron beam by passage through polycrystalline high-Tc superconductors, specifically 123 Y-Ba-Cu-O prepared and characterized at the IBM Watson Research Center. We believe that this technique will prove useful in studying aspects of these materials, such as the penetration depth of shielding currents, the presence and structure of trapped flux vortices, and grain size effects on the supercurrent distribution in polycrystalline samples. The two samples showed sharp transitions at 87 and 89 K, and have been studied at temperatures of 77 K; the second sample has also been studied at 4 K. The transition to the superconducting state was monitored by the shift in resonant frequency of a coil surrounding the sample. No measurable depolarization was observed in either sample at 77 K in both the field-cooled and zero-field-cooled states, using applied fields of 0 (nominal), 54, and 1400 Oe. This negative result may be connected with the fact that the material is still in the reversible region as indicated by susceptibility measurements, but it allows an estimate of the upper bound of possible inhomogeneous internal fields, assuming a distance scale for the superconducting regions. For the 10-μm grain size suggested by photomicrographs, this upper bound for the field turns out to be 1.2 kOe, which seems reasonable. At 4 K a significant depolarization was observed when the sample was cooled in low fields and a field of 1400 Oe was subsequently applied. This result suggests that flux lines are penetrating the sample. Further investigations are being carried out to determine the field and temperature dependence of the depolarization, and attempts will be made to model it quantitatively in terms of possible internal field distributions. We are also searching for possible diffraction effects from ordered vortex

[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. Copyright © 2013 Elsevier España, S.L. All rights reserved.

Total internal reflection fluorescence anisotropy imaging microscopy: setup, calibration, and data processing for protein polymerization measurements in living cells

NASA Astrophysics Data System (ADS)

Ströhl, Florian; Wong, Hovy H. W.; Holt, Christine E.; Kaminski, Clemens F.

2018-01-01

Fluorescence anisotropy imaging microscopy (FAIM) measures the depolarization properties of fluorophores to deduce molecular changes in their environment. For successful FAIM, several design principles have to be considered and a thorough system-specific calibration protocol is paramount. One important calibration parameter is the G factor, which describes the system-induced errors for different polarization states of light. The determination and calibration of the G factor is discussed in detail in this article. We present a novel measurement strategy, which is particularly suitable for FAIM with high numerical aperture objectives operating in TIRF illumination mode. The method makes use of evanescent fields that excite the sample with a polarization direction perpendicular to the image plane. Furthermore, we have developed an ImageJ/Fiji plugin, AniCalc, for FAIM data processing. We demonstrate the capabilities of our TIRF-FAIM system by measuring β -actin polymerization in human embryonic kidney cells and in retinal neurons.

Triple-wavelength depolarization-ratio profiling of Saharan dust over Barbados during SALTRACE in 2013 and 2014

NASA Astrophysics Data System (ADS)

Haarig, Moritz; Ansmann, Albert; Althausen, Dietrich; Klepel, André; Groß, Silke; Freudenthaler, Volker; Toledano, Carlos; Mamouri, Rodanthi-Elisavet; Farrell, David A.; Prescod, Damien A.; Marinou, Eleni; Burton, Sharon P.; Gasteiger, Josef; Engelmann, Ronny; Baars, Holger

2017-09-01

Triple-wavelength polarization lidar measurements in Saharan dust layers were performed at Barbados (13.1° N, 59.6° W), 5000-8000 km west of the Saharan dust sources, in the framework of the Saharan Aerosol Long-range Transport and Aerosol-Cloud-Interaction Experiment (SALTRACE-1, June-July 2013, SALTRACE-3, June-July 2014). Three case studies are discussed. High quality was achieved by comparing the dust linear depolarization ratio profiles measured at 355, 532, and 1064 nm with respective dual-wavelength (355, 532 nm) depolarization ratio profiles measured with a reference lidar. A unique case of long-range transported dust over more than 12 000 km is presented. Saharan dust plumes crossing Barbados were measured with an airborne triple-wavelength polarization lidar over Missouri in the midwestern United States 7 days later. Similar dust optical properties and depolarization features were observed over both sites indicating almost unchanged dust properties within this 1 week of travel from the Caribbean to the United States. The main results of the triple-wavelength polarization lidar observations in the Caribbean in the summer seasons of 2013 and 2014 are summarized. On average, the particle linear depolarization ratios for aged Saharan dust were found to be 0.252 ± 0.030 at 355 nm, 0.280 ± 0.020 at 532 nm, and 0.225 ± 0.022 at 1064 nm after approximately 1 week of transport over the tropical Atlantic. Based on published simulation studies we present an attempt to explain the spectral features of the depolarization ratio of irregularly shaped mineral dust particles, and conclude that most of the irregularly shaped coarse-mode dust particles (particles with diameters > 1 µm) have sizes around 1.5-2 µm. The SALTRACE results are also set into the context of the SAMUM-1 (Morocco, 2006) and SAMUM-2 (Cabo Verde, 2008) depolarization ratio studies. Again, only minor changes in the dust depolarization characteristics were observed on the way from the Saharan dust

Recording, analysis, and interpretation of spreading depolarizations in neurointensive care: Review and recommendations of the COSBID research group.

PubMed

Dreier, Jens P; Fabricius, Martin; Ayata, Cenk; Sakowitz, Oliver W; William Shuttleworth, C; Dohmen, Christian; Graf, Rudolf; Vajkoczy, Peter; Helbok, Raimund; Suzuki, Michiyasu; Schiefecker, Alois J; Major, Sebastian; Winkler, Maren Kl; Kang, Eun-Jeung; Milakara, Denny; Oliveira-Ferreira, Ana I; Reiffurth, Clemens; Revankar, Gajanan S; Sugimoto, Kazutaka; Dengler, Nora F; Hecht, Nils; Foreman, Brandon; Feyen, Bart; Kondziella, Daniel; Friberg, Christian K; Piilgaard, Henning; Rosenthal, Eric S; Westover, M Brandon; Maslarova, Anna; Santos, Edgar; Hertle, Daniel; Sánchez-Porras, Renán; Jewell, Sharon L; Balança, Baptiste; Platz, Johannes; Hinzman, Jason M; Lückl, Janos; Schoknecht, Karl; Schöll, Michael; Drenckhahn, Christoph; Feuerstein, Delphine; Eriksen, Nina; Horst, Viktor; Bretz, Julia S; Jahnke, Paul; Scheel, Michael; Bohner, Georg; Rostrup, Egill; Pakkenberg, Bente; Heinemann, Uwe; Claassen, Jan; Carlson, Andrew P; Kowoll, Christina M; Lublinsky, Svetlana; Chassidim, Yoash; Shelef, Ilan; Friedman, Alon; Brinker, Gerrit; Reiner, Michael; Kirov, Sergei A; Andrew, R David; Farkas, Eszter; Güresir, Erdem; Vatter, Hartmut; Chung, Lee S; Brennan, K C; Lieutaud, Thomas; Marinesco, Stephane; Maas, Andrew Ir; Sahuquillo, Juan; Dahlem, Markus A; Richter, Frank; Herreras, Oscar; Boutelle, Martyn G; Okonkwo, David O; Bullock, M Ross; Witte, Otto W; Martus, Peter; van den Maagdenberg, Arn Mjm; Ferrari, Michel D; Dijkhuizen, Rick M; Shutter, Lori A; Andaluz, Norberto; Schulte, André P; MacVicar, Brian; Watanabe, Tomas; Woitzik, Johannes; Lauritzen, Martin; Strong, Anthony J; Hartings, Jed A

2017-05-01

Spreading depolarizations (SD) are waves of abrupt, near-complete breakdown of neuronal transmembrane ion gradients, are the largest possible pathophysiologic disruption of viable cerebral gray matter, and are a crucial mechanism of lesion development. Spreading depolarizations are increasingly recorded during multimodal neuromonitoring in neurocritical care as a causal biomarker providing a diagnostic summary measure of metabolic failure and excitotoxic injury. Focal ischemia causes spreading depolarization within minutes. Further spreading depolarizations arise for hours to days due to energy supply-demand mismatch in viable tissue. Spreading depolarizations exacerbate neuronal injury through prolonged ionic breakdown and spreading depolarization-related hypoperfusion (spreading ischemia). Local duration of the depolarization indicates local tissue energy status and risk of injury. Regional electrocorticographic monitoring affords even remote detection of injury because spreading depolarizations propagate widely from ischemic or metabolically stressed zones; characteristic patterns, including temporal clusters of spreading depolarizations and persistent depression of spontaneous cortical activity, can be recognized and quantified. Here, we describe the experimental basis for interpreting these patterns and illustrate their translation to human disease. We further provide consensus recommendations for electrocorticographic methods to record, classify, and score spreading depolarizations and associated spreading depressions. These methods offer distinct advantages over other neuromonitoring modalities and allow for future refinement through less invasive and more automated approaches.

Recording, analysis, and interpretation of spreading depolarizations in neurointensive care: Review and recommendations of the COSBID research group

PubMed Central

Fabricius, Martin; Ayata, Cenk; Sakowitz, Oliver W; William Shuttleworth, C; Dohmen, Christian; Graf, Rudolf; Vajkoczy, Peter; Helbok, Raimund; Suzuki, Michiyasu; Schiefecker, Alois J; Major, Sebastian; Winkler, Maren KL; Kang, Eun-Jeung; Milakara, Denny; Oliveira-Ferreira, Ana I; Reiffurth, Clemens; Revankar, Gajanan S; Sugimoto, Kazutaka; Dengler, Nora F; Hecht, Nils; Foreman, Brandon; Feyen, Bart; Kondziella, Daniel; Friberg, Christian K; Piilgaard, Henning; Rosenthal, Eric S; Westover, M Brandon; Maslarova, Anna; Santos, Edgar; Hertle, Daniel; Sánchez-Porras, Renán; Jewell, Sharon L; Balança, Baptiste; Platz, Johannes; Hinzman, Jason M; Lückl, Janos; Schoknecht, Karl; Schöll, Michael; Drenckhahn, Christoph; Feuerstein, Delphine; Eriksen, Nina; Horst, Viktor; Bretz, Julia S; Jahnke, Paul; Scheel, Michael; Bohner, Georg; Rostrup, Egill; Pakkenberg, Bente; Heinemann, Uwe; Claassen, Jan; Carlson, Andrew P; Kowoll, Christina M; Lublinsky, Svetlana; Chassidim, Yoash; Shelef, Ilan; Friedman, Alon; Brinker, Gerrit; Reiner, Michael; Kirov, Sergei A; Andrew, R David; Farkas, Eszter; Güresir, Erdem; Vatter, Hartmut; Chung, Lee S; Brennan, KC; Lieutaud, Thomas; Marinesco, Stephane; Maas, Andrew IR; Sahuquillo, Juan; Dahlem, Markus A; Richter, Frank; Herreras, Oscar; Boutelle, Martyn G; Okonkwo, David O; Bullock, M Ross; Witte, Otto W; Martus, Peter; van den Maagdenberg, Arn MJM; Ferrari, Michel D; Dijkhuizen, Rick M; Shutter, Lori A; Andaluz, Norberto; Schulte, André P; MacVicar, Brian; Watanabe, Tomas; Woitzik, Johannes; Lauritzen, Martin; Strong, Anthony J; Hartings, Jed A

2016-01-01

Spreading depolarizations (SD) are waves of abrupt, near-complete breakdown of neuronal transmembrane ion gradients, are the largest possible pathophysiologic disruption of viable cerebral gray matter, and are a crucial mechanism of lesion development. Spreading depolarizations are increasingly recorded during multimodal neuromonitoring in neurocritical care as a causal biomarker providing a diagnostic summary measure of metabolic failure and excitotoxic injury. Focal ischemia causes spreading depolarization within minutes. Further spreading depolarizations arise for hours to days due to energy supply-demand mismatch in viable tissue. Spreading depolarizations exacerbate neuronal injury through prolonged ionic breakdown and spreading depolarization-related hypoperfusion (spreading ischemia). Local duration of the depolarization indicates local tissue energy status and risk of injury. Regional electrocorticographic monitoring affords even remote detection of injury because spreading depolarizations propagate widely from ischemic or metabolically stressed zones; characteristic patterns, including temporal clusters of spreading depolarizations and persistent depression of spontaneous cortical activity, can be recognized and quantified. Here, we describe the experimental basis for interpreting these patterns and illustrate their translation to human disease. We further provide consensus recommendations for electrocorticographic methods to record, classify, and score spreading depolarizations and associated spreading depressions. These methods offer distinct advantages over other neuromonitoring modalities and allow for future refinement through less invasive and more automated approaches. PMID:27317657

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.

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.

Interaction between depolarization effects, interface layer, and fatigue behavior in PZT thin film capacitors

NASA Astrophysics Data System (ADS)

Böttger, U.; Waser, R.

2017-07-01

The existence of non-ferroelectric regions in ferroelectric thin films evokes depolarization effects leading to a tilt of the P(E) hysteresis loop. The analysis of measured hysteresis of lead zirconate titanate (PZT) thin films is used to determine a depolarization factor which contains quantitative information about interfacial layers as well as ferroelectrically passive zones in the bulk. The derived interfacial capacitance is smaller than that estimated from conventional extrapolation techniques. In addition, the concept of depolarization is used for the investigation of fatigue behavior of PZT thin films indicating that the mechanism of seed inhibition, which is responsible for the effect, occurs in the entire film.

Fluorescence Determination of Tryptophan Side-Chain Accessibility and Dynamics in Triple-Helical Collagen-Like Peptides

PubMed Central

Simon-Lukasik, Kristine V.; Persikov, Anton V.; Brodsky, Barbara; Ramshaw, John A. M.; Laws, William R.; Alexander Ross, J. B.; Ludescher, Richard D.

2003-01-01

We report tryptophan fluorescence measurements of emission intensity, iodide quenching, and anisotropy that describe the environment and dynamics at X and Y sites in stable collagen-like peptides of sequence (Gly-X-Y)n. About 90% of tryptophans at both sites have similar solvent exposed fluorescence properties and a lifetime of 8.5–9 ns. Analysis of anisotropy decays using an associative model indicates that these long lifetime populations undergo rapid depolarizing motion with a 0.5 ns correlation time; however, the extent of fast motion at the Y site is considerably less than the essentially unrestricted motion at the X site. About 10% of tryptophans at both sites have a shorter (∼3 ns) lifetime indicating proximity to a protein quenching group; these minor populations are immobile on the peptide surface, depolarizing only by overall trimer rotation. Iodide quenching indicates that tryptophans at the X site are more accessible to solvent. Side chains at X sites are more solvent accessible and considerably more mobile than residues at Y sites and can more readily fluctuate among alternate intermolecular interactions in collagen fibrils. This fluorescence analysis of collagen-like peptides lays a foundation for studies on the structure, dynamics, and function of collagen and of triple-helical junctions in gelatin gels. PMID:12524302

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

Laser-excited fluorescence for measuring atmospheric pollution

NASA Technical Reports Server (NTRS)

Menzies, R. T.

1975-01-01

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

UV-VIS depolarization from Arizona Test Dust particles at exact backscattering angle

NASA Astrophysics Data System (ADS)

Miffre, Alain; Mehri, Tahar; Francis, Mirvatte; Rairoux, Patrick

2016-01-01

In this paper, a controlled laboratory experiment is performed to accurately evaluate the depolarization from mineral dust particles in the exact backward scattering direction (ϴ=180.0±0.2°). The experiment is carried out at two wavelengths simultaneously (λ=355 nm, λ=532 nm), on a determined size and shape distribution of Arizona Test Dust (ATD) particles, used as a proxy for mineral dust particles. After validating the set-up on spherical water droplets, two determined ATD-particle size distributions, representative of mineral dust after long-range transport, are generated to accurately retrieve the UV-VIS depolarization from ATD-particles at exact backscattering angle, which is new. The measured depolarization reaches at most 37.5% at λ=355 nm (35.5% at λ=532 nm), and depends on the particle size distribution. Moreover, these laboratory findings agree with T-matrix numerical simulations, at least for a determined particle size distribution and at a determined wavelength, showing the ability of the spheroidal model to reproduce mineral dust particles in the exact backward scattering direction. However, the spectral dependence of the measured depolarization could not be reproduced with the spheroidal model, even for not evenly distributed aspect ratios. Hence, these laboratory findings can be used to evaluate the applicability of the spheroidal model in the backward scattering direction and moreover, to invert UV-VIS polarization lidar returns, which is useful for radiative transfer and climatology, in which mineral dust particles are strongly involved.

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

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.

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.

Micro-scale temperature measurement method using fluorescence polarization

NASA Astrophysics Data System (ADS)

Tatsumi, K.; Hsu, C.-H.; Suzuki, A.; Nakabe, K.

2016-09-01

A novel method that can measure the fluid temperature in microscopic scale by measuring the fluorescence polarization is described in this paper. The measurement technique is not influenced by the quenching effects which appears in conventional LIF methods and is believed to show a higher reliability in temperature measurements. Experiment was performed using a microchannel flow and fluorescent molecule probes, and the effects of the fluid temperature, fluid viscosity, measurement time, and pH of the solution on the measured fluorescence polarization degree are discussed to understand the basic characteristics of the present method. The results showed that fluorescence polarization is considerably less sensible to these quenching factors. A good correlation with the fluid temperature, on the other hand, was obtained and agreed well with the theoretical values confirming the feasibility of the method.

First observation of the depolarization of Thomson scattering radiation by a fusion plasma

NASA Astrophysics Data System (ADS)

Giudicotti, L.; Kempenaars, M.; McCormack, O.; Flanagan, J.; Pasqualotto, R.; contributors, JET

2018-04-01

We report the first experimental observation of the depolarization of the Thomson scattering (TS) radiation, a relativistic effect expected to occur in very high {{T}e} plasmas and never observed so far in a fusion machine. A set of unused optical fibers in the collection optics of the high resolution Thomson scattering system of JET has been used to detect the depolarized TS radiation during a JET campaign with {{T}e}≤slant 8 keV . A linear polarizer with the axis perpendicular to the direction of the incident E-field was placed in front of a fiber optic pair observing a region close to the plasma core, while another fiber pair with no polariser simultaneously observed an adjacent plasma region. The measured intensity ratio was found to be consistent with the theory, taking into account sensitivity coefficients of the two measurement channels determined with post-experiment calibrations and Raman scattering. This depolarization effect is at the basis of polarimetric TS, a different and complementary method for the analysis of TS spectra that can provide significant advantages for {{T}e} measurements in very hot plasmas such as in ITER ≤ft({{T}e}≤slant 40 keV \\right) .

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

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.

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.

Measuring fluorescence polarization with a dichrometer.

PubMed

Sutherland, John C

2017-09-01

A method for obtaining fluorescence polarization data from an instrument designed to measure circular and linear dichroism is compared with a previously reported approach. The new method places a polarizer between the sample and a detector mounted perpendicular to the direction of the incident beam and results in determination of the fluorescence polarization ratio, whereas the previous method does not use a polarizer and yields the fluorescence anisotropy. A similar analysis with the detector located axially with the excitation beam demonstrates that there is no frequency modulated signal due to fluorescence polarization in the absence of a polarizer. Copyright © 2017. Published by Elsevier Inc.

Measuring fluorescence polarization with a dichrometer

DOE PAGES

Sutherland, John C.

2017-04-06

In this article, a method for obtaining fluorescence polarization data from an instrument designed to measure circular and linear dichroism is compared with a previously reported approach. The new method places a polarizer between the sample and a detector mounted perpendicular to the direction of the incident beam and results in determination of the fluorescence polarization ratio, whereas the previous method does not use a polarizer and yields the fluorescence anisotropy. A similar analysis with the detector located axially with the excitation beam demonstrates that there is no frequency modulated signal due to fluorescence polarization in the absence of amore » polarizer.« less

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

Constrained Analysis of Fluorescence Anisotropy Decay:Application to Experimental Protein Dynamics

PubMed Central

Feinstein, Efraim; Deikus, Gintaras; Rusinova, Elena; Rachofsky, Edward L.; Ross, J. B. Alexander; Laws, William R.

2003-01-01

Hydrodynamic properties as well as structural dynamics of proteins can be investigated by the well-established experimental method of fluorescence anisotropy decay. Successful use of this method depends on determination of the correct kinetic model, the extent of cross-correlation between parameters in the fitting function, and differences between the timescales of the depolarizing motions and the fluorophore's fluorescence lifetime. We have tested the utility of an independently measured steady-state anisotropy value as a constraint during data analysis to reduce parameter cross correlation and to increase the timescales over which anisotropy decay parameters can be recovered accurately for two calcium-binding proteins. Mutant rat F102W parvalbumin was used as a model system because its single tryptophan residue exhibits monoexponential fluorescence intensity and anisotropy decay kinetics. Cod parvalbumin, a protein with a single tryptophan residue that exhibits multiexponential fluorescence decay kinetics, was also examined as a more complex model. Anisotropy decays were measured for both proteins as a function of solution viscosity to vary hydrodynamic parameters. The use of the steady-state anisotropy as a constraint significantly improved the precision and accuracy of recovered parameters for both proteins, particularly for viscosities at which the protein's rotational correlation time was much longer than the fluorescence lifetime. Thus, basic hydrodynamic properties of larger biomolecules can now be determined with more precision and accuracy by fluorescence anisotropy decay. PMID:12524313

Monitoring changes in membrane polarity, membrane integrity, and intracellular ion concentrations in Streptococcus pneumoniae using fluorescent dyes.

PubMed

Clementi, Emily A; Marks, Laura R; Roche-Håkansson, Hazeline; Håkansson, Anders P

2014-02-17

Membrane depolarization and ion fluxes are events that have been studied extensively in biological systems due to their ability to profoundly impact cellular functions, including energetics and signal transductions. While both fluorescent and electrophysiological methods, including electrode usage and patch-clamping, have been well developed for measuring these events in eukaryotic cells, methodology for measuring similar events in microorganisms have proven more challenging to develop given their small size in combination with the more complex outer surface of bacteria shielding the membrane. During our studies of death-initiation in Streptococcus pneumoniae (pneumococcus), we wanted to elucidate the role of membrane events, including changes in polarity, integrity, and intracellular ion concentrations. Searching the literature, we found that very few studies exist. Other investigators had monitored radioisotope uptake or equilibrium to measure ion fluxes and membrane potential and a limited number of studies, mostly in Gram-negative organisms, had seen some success using carbocyanine or oxonol fluorescent dyes to measure membrane potential, or loading bacteria with cell-permeant acetoxymethyl (AM) ester versions of ion-sensitive fluorescent indicator dyes. We therefore established and optimized protocols for measuring membrane potential, rupture, and ion-transport in the Gram-positive organism S. pneumoniae. We developed protocols using the bis-oxonol dye DiBAC4(3) and the cell-impermeant dye propidium iodide to measure membrane depolarization and rupture, respectively, as well as methods to optimally load the pneumococci with the AM esters of the ratiometric dyes Fura-2, PBFI, and BCECF to detect changes in intracellular concentrations of Ca(2+), K(+), and H(+), respectively, using a fluorescence-detection plate reader. These protocols are the first of their kind for the pneumococcus and the majority of these dyes have not been used in any other bacterial

Responses of sun-induced chlorophyll fluorescence to biological and environmental variations measured with a versatile Fluorescence Auto-Measurement Equipment (FAME)

NASA Astrophysics Data System (ADS)

Gu, L.

2017-12-01

In this study, we examine responses of sun-induced chlorophyll fluorescence to biological and environmental variations measured with a versatile Fluorescence Auto-Measurement Equipment (FAME). FAME was developed to automatically and continuously measure chlorophyll fluorescence (F) of a leaf, plant or canopy in both laboratory and field environments, excited by either artificial light source or sunlight. FAME is controlled by a datalogger and allows simultaneous measurements of environmental variables complementary to the F signals. A built-in communication system allows FAME to be remotely monitored and data-downloaded. Radiance and irradiance calibrations can be done online. FAME has been applied in a variety of environments, allowing an investigation of biological and environmental controls on F emission.

Polarization changes at Lyot depolarizer output for different types of input beams.

PubMed

de Sande, J Carlos G; Piquero, Gemma; Teijeiro, Cristina

2012-03-01

Lyot depolarizers are optical devices made of birefringent materials used for producing unpolarized beams from totally polarized incident light. The depolarization is produced for polychromatic input beams due to the different phase introduced by the Lyot depolarizer for each wavelength. The effect of this device on other types of incident fields is investigated. In particular two cases are analyzed: (i) monochromatic and nonuniformly polarized incident beams and (ii) incident light synthesized by superposition of two monochromatic orthogonally polarized beams with different wavelengths. In the last case, it is theoretically and experimentally shown that the Lyot depolarizer increases the degree of polarization instead of depolarizes.

Millisecond infrared laser pulses depolarize and elicit action potentials on in-vitro dorsal root ganglion neurons

PubMed Central

Paris, Lambert; Marc, Isabelle; Charlot, Benoit; Dumas, Michel; Valmier, Jean; Bardin, Fabrice

2017-01-01

This work focuses on the optical stimulation of dorsal root ganglion (DRG) neurons through infrared laser light stimulation. We show that a few millisecond laser pulse at 1875 nm induces a membrane depolarization, which was observed by the patch-clamp technique. This stimulation led to action potentials firing on a minority of neurons beyond an energy threshold. A depolarization without action potential was observed for the majority of DRG neurons, even beyond the action potential energy threshold. The use of ruthenium red, a thermal channel blocker, stops the action potential generation, but has no effects on membrane depolarization. Local temperature measurements reveal that the depolarization amplitude is sensitive to the amplitude of the temperature rise as well as to the time rate of change of temperature, but in a way which may not fully follow a photothermal capacitive mechanism, suggesting that more complex mechanisms are involved. PMID:29082085

Formalin produces depolarizations in human airway smooth muscle in vitro.

PubMed

Richards, Ira S; DeHate, Robin B

2006-03-01

Respiratory irritants may result in airway smooth muscle (ASM) depolarization and bronchoconstriction. We examined the effect of formalin on membrane potentials in human ASM in two types of in vitro preparations: strip preparations, which contain functional sensory and motor nerve endings and cultured cells, which lack these nerve endings due to the tissue dissociation process. Depolarizations occurred in atropine-treated strip preparations in response to formalin exposures, but not in similarly-treated cultured cells, suggesting a role for non-cholinergic mediators in formalin-induced depolarization. It is suggested that formalin may act as an irritant to produce bronchoconstriction that is mediated by the release of endogenous substance P (SP) from peripheral sensory nerve endings. This is supported by our observation that exogenous SP produced depolarizations of a magnitude similar to those produced by formalin in both strip preparations and cultured cells. In addition, capsaicin, which releases endogenous SP from nerve endings, produced depolarizations of a magnitude similar to formalin in strip preparations, but was without effect in cultured cells.

Twomey Effect in Subtropical Stratocumulus Clouds from UV Depolarization LIDAR

NASA Astrophysics Data System (ADS)

de Graaf, Martin; Brown, Jessica; Donovan, David

2018-04-01

Marine stratocumulus clouds are important climate regulators, reflecting sunlight over a dark ocean background. A UV-depolarization lidar on Ascension, a small remote island in the south Atlantic, measured cloud droplet sizes and number concentration using an inversion method based on Monte Carlo (MC) modelling of multiple scattering in idealised semiadiabatic clouds. The droplet size and number concentration weremodulated due to smoke from the African continent, measured by the same instrument.

Effects of interleukin-1ß on cortical spreading depolarization and cerebral vasculature

PubMed Central

Eitner, Annett; Leuchtweis, Johannes; Bauer, Reinhard; Lehmenkühler, Alfred; Schaible, Hans-Georg

2016-01-01

During brain damage and ischemia, the cytokine interleukin-1ß is rapidly upregulated due to activation of inflammasomes. We studied whether interleukin-1ß influences cortical spreading depolarization, and whether lipopolysaccharide, often used for microglial stimulation, influences cortical spreading depolarizations. In anaesthetized rats, cortical spreading depolarizations were elicited by microinjection of KCl. Interleukin-1ß, the IL-1 receptor 1 antagonist, the GABAA receptor blocker bicuculline, and lipopolysaccharide were administered either alone or combined (interleukin-1ß + IL-1 receptor 1 antagonist; interleukin-1ß + bicuculline; lipopolysaccharide + IL-1 receptor 1 antagonist) into a local cortical treatment area. Using microelectrodes, cortical spreading depolarizations were recorded in a non-treatment and in the treatment area. Plasma extravasation in cortical grey matter was assessed with Evans blue. Local application of interleukin-1ß reduced cortical spreading depolarization amplitudes in the treatment area, but not at a high dose. This reduction was prevented by IL-1 receptor 1 antagonist and by bicuculline. However, interleukin-1ß induced pronounced plasma extravasation independently on cortical spreading depolarizations. Application of lipopolysaccharide reduced cortical spreading depolarization amplitudes but prolonged their duration; EEG activity was still present. These effects were also blocked by IL-1 receptor 1 antagonist. Interleukin-1ß evokes changes of neuronal activity and of vascular functions. Thus, although the reduction of cortical spreading depolarization amplitudes at lower doses of interleukin-1ß may reduce deleterious effects of cortical spreading depolarizations, the sum of interleukin-1ß effects on excitability and on the vasculature rather promote brain damaging mechanisms. PMID:27037093

Susceptibility of Primary Sensory Cortex to Spreading Depolarizations.

PubMed

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

2016-04-27

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. 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. Finally, we show a

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

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.

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.

Methods and apparatus for using gas and liquid phase cathodic depolarizers

NASA Technical Reports Server (NTRS)

Murphy, Oliver J. (Inventor); Hitchens, G. Duncan (Inventor)

1998-01-01

The invention provides methods for using gas and liquid phase cathodic depolarizers in an electrochemical cell having a cation exchange membrane in intimate contact with the anode and cathode. The electrochemical conversion of cathodic depolarizers at the cathode lowers the cell potential necessary to achieve a desired electrochemical conversion, such as ozone evolution, at the anode. When gaseous cathodic depolarizers, such as oxygen, are used, a gas diffusion cathode having the cation exchange membrane bonded thereto is preferred. When liquid phase cathodic depolarizers are used, the cathode may be a flow-by electrode, flow-through electrode, packed-bed electrode or a fluidized-bed electrode in intimate contact with the cation exchange membrane.

Connecting active to passive fluorescence with photosynthesis: a method for evaluating remote sensing measurements of Chl fluorescence.

PubMed

Magney, Troy S; Frankenberg, Christian; Fisher, Joshua B; Sun, Ying; North, Gretchen B; Davis, Thomas S; Kornfeld, Ari; Siebke, Katharina

2017-09-01

Recent advances in the retrieval of Chl fluorescence from space using passive methods (solar-induced Chl fluorescence, SIF) promise improved mapping of plant photosynthesis globally. However, unresolved issues related to the spatial, spectral, and temporal dynamics of vegetation fluorescence complicate our ability to interpret SIF measurements. We developed an instrument to measure leaf-level gas exchange simultaneously with pulse-amplitude modulation (PAM) and spectrally resolved fluorescence over the same field of view - allowing us to investigate the relationships between active and passive fluorescence with photosynthesis. Strongly correlated, slope-dependent relationships were observed between measured spectra across all wavelengths (F λ , 670-850 nm) and PAM fluorescence parameters under a range of actinic light intensities (steady-state fluorescence yields, F t ) and saturation pulses (maximal fluorescence yields, F m ). Our results suggest that this method can accurately reproduce the full Chl emission spectra - capturing the spectral dynamics associated with changes in the yields of fluorescence, photochemical (ΦPSII), and nonphotochemical quenching (NPQ). We discuss how this method may establish a link between photosynthetic capacity and the mechanistic drivers of wavelength-specific fluorescence emission during changes in environmental conditions (light, temperature, humidity). Our emphasis is on future research directions linking spectral fluorescence to photosynthesis, ΦPSII, and NPQ. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

Atrioventricular depolarization differences identify coronary artery anomalies in Kawasaki disease.

PubMed

Cortez, Daniel; Sharma, Nandita; Jone, Pei-Ni

2017-03-01

Kawasaki disease (KD) is the leading cause of acquired heart disease in children. Signal average electrocardiogram changes in patients during the acute phase of KD with coronary artery anomalies (CAA) include depolarization changes. We set out to determine if 12-lead-derived atrioventricular depolarization differences can identify CAA in patients with KD. A blinded, retrospective case-control study of patients with KD was performed. Deep Q waves, corrected QT-intervals (QTc), spatial QRS-T angles, T-wave vector magnitudes (RMS-T), and a novel parameter for assessment of atrioventricular depolarization difference (the spatial PR angle) and a two dimensional PR angle were assessed. Comparisons between groups were performed to test for significant differences. One hundred one patients with KD were evaluated, with 68 having CAA (67.3%, mean age 3.6 ± 3.0 years, 82.6% male), and 32 without CAA (31.7%, mean age 2.7 ± 3.2 years, 70.4% male). The spatial PR angle significantly discriminated KD patients with CAA from those without, 59.7° ± 31.1° versus 41.6° ± 11.5° (p < .001). A spatial PR angle cutoff value of 56.9° gave positive/negative predictive values and odds ratios of 93.8%, 43.5%, and 11.5% (95% confidence interval (CI) 2.6-52.2). The two dimensional PR angle either below 7° or above 92° gave positive/negative predictive values and odds ratios of 100.0%, 38.8%, and 21.1% (95% CI 1.2-362.8). No other parameters significantly differentiated the groups. Atrioventricular depolarization differences, measured by the spatial or two dimensional PR angle differentiate KD patients with CAA versus those without. © 2016 Wiley Periodicals, Inc.

Determination of the orientation of fluorescent labels relative to myosin S1 in solution from time-resolved fluorescence anisotropy experiments

NASA Astrophysics Data System (ADS)

van der Heide, Uulke A.; Gerritsen, Hans C.; Trayer, Ian P.; Levine, Yehudi K.

1992-04-01

The time-resolved fluorescence anisotropy of myosin S1 covalently labeled with Eosin-5- maleimide and 1,5-I-AEDANS was measured in solution. Each probe was specifically attached at one SH-group on the S1. The two most reactive SH sites on the heavy chain of the myosin S1 were used. The fluorescence anisotropy was measured at different excitation wavelengths. In this way, several absorption moments were utilized, each having a distinct orientation in the frame of the dye. The orientations of the transition moments in the dyes were determined in a separate experiment using an angle resolved fluorescence depolarization experiment on dyes embedded in stretched matrices of PVA polymers. The anisotropy decay curves exhibit fast (<3 ns) and slow (> 100 ns) components. The slow decay components reflect the motion of the large protein molecules. The fast anisotropy decay are attributed to a fast, but restricted, motion of the bound dye relative to the protein as experiments on free dyes in solution reveal subnanosecond anisotropy decays. The anisotropy decays have been analyzed in terms of a model which describes the restricted motion of the dye molecule relative to the protein and the overall rotation of the dye-protein complex in solution. An important element in the model is the incorporation of the orientational distribution of the dye relative to the protein. The observed anisotropy decays were analyzed using a global target approach in which the experimental data obtained at different excitation wavelengths are fitted simultaneously to the theoretical model. It is important to note that the orientational distribution of the dye relative to the protein, as well as the rotational correlation times of the motions for a dye attached to a given binding site, are independent of the excitation wavelength used. This leads to a reduction in the number of independent parameters optimized by the nonlinear least squares procedure. The orientational distribution of the dye

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.

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.

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.

Depolarization Alters Phenotype, Maintains Plasticity of Predifferentiated Mesenchymal Stem Cells

PubMed Central

Sundelacruz, Sarah; Levin, Michael

2013-01-01

Although adult stem cell transplantation has been implemented as a therapy for tissue repair, it is limited by the availability of functional adult stem cells. A potential approach to generate stem and progenitor cells may be to modulate the differentiated status of somatic cells. Therefore, there is a need for a better understanding of how the differentiated phenotype of mature cells is regulated. We hypothesize that bioelectric signaling plays an important role in the maintenance of the differentiated state, as it is a functional regulator of the differentiation process in various cells and tissues. In this study, we asked whether the mature phenotype of osteoblasts and adipocytes derived from human mesenchymal stem cells (hMSCs) could be altered by modulation of their membrane potential. hMSC-derived osteoblasts and adipocytes were depolarized by treatment with ouabain, a Na+/K+ ATPase inhibitor, or by treatment with high concentrations of extracellular K+. To characterize the effect of voltage modulation on the differentiated state, the depolarized cells were evaluated for (1) the loss of differentiation markers; (2) the up-regulation of stemness markers and stem properties; and (3) differences in gene expression profiles in response to voltage modulation. hMSC-derived osteoblasts and adipocytes exhibited significant down-regulation of bone and fat tissue markers in response to depolarization, despite the presence of differentiation-inducing soluble factors, suggesting that bioelectric signaling overrides biochemical signaling in the maintenance of cell state. Suppression of the osteoblast or adipocyte phenotype was not accompanied by up-regulation of genes associated with the stem state. Thus, depolarization does not activate the stem cell genetic signature and, therefore, does not induce a full reprogramming event. However, after transdifferentiating the depolarized cells to evaluate for multi-lineage potential, depolarized osteoblasts demonstrated improved

The physostigmine depolarization potentiating effect of salicylate in frog skeletal muscle.

PubMed

Varga, E; Kovács, L; Szücs, G; Illés, B

1975-01-01

1) The frog's sartorius muscle was depolarized depending on the degree of concentration 2--4 times more intensely by physostigmine salicylate than by physostigmine sulphate. 2) In normal Ringer's solution, 1 mM physostigmine salicylate decreased the sensitivity of the membrane to potassium depolarization by about 90%. Under similar experimental conditions, physostigmine sulphate and Na salicylate, respectively, decrease the sensitivity of the membrane to potassium depolarization by about 30%. 3) The difference manifested in the depolarizing effect of salicylate and other physostigmine salts (chloride, sulphate, phosphate, formiate, acetate, monochloracetate, benzoate and para-oxy-benzoate) is expressed already at 1 mM concentration (about 10-fold), if the muscle had been equilibrated in chloride-free glucuronate or sulphate milieu. 4) The depolarization develops slowly. It takes 30--60 minutes for the new steady state to develop even in the superficial sartorius fibres. If depolarization has reached its maximum on an average 100 mV, the membrane potential remains unchanged for hours. 5) Depolarization ensues at an unchanged degree in the presence of Na-free (choline) Ringer as well as in the presence of 2X10(-8) g/ml tetrodotoxin; therefore, it is not a Na-dependent process. 6) Under the influence of 1 mM physostigmine salicylate the membrane's resistance to the inward potassium current increased about twofold, while the increase was only 15% to the outward potassium current. It is assumed that the salicylate anion is characteristically capable of potentiating the decreasing effect of physostigmine on potassium permeability, though the role of the metabolic effect of salicylate cannot be excluded.

Fluorescence lifetime as a new parameter in analytical cytology measurements

NASA Astrophysics Data System (ADS)

Steinkamp, John A.; Deka, Chiranjit; Lehnert, Bruce E.; Crissman, Harry A.

1996-05-01

A phase-sensitive flow cytometer has been developed to quantify fluorescence decay lifetimes on fluorochrome-labeled cells/particles. This instrument combines flow cytometry (FCM) and frequency-domain fluorescence spectroscopy measurement principles to provide unique capabilities for making phase-resolved lifetime measurements, while preserving conventional FCM capabilities. Cells are analyzed as they intersect a high-frequency, intensity-modulated (sine wave) laser excitation beam. Fluorescence signals are processed by conventional and phase-sensitive signal detection electronics and displayed as frequency distribution histograms. In this study we describe results of fluorescence intensity and lifetime measurements on fluorescently labeled particles, cells, and chromosomes. Examples of measurements on intrinsic cellular autofluorescence, cells labeled with immunofluorescence markers for cell- surface antigens, mitochondria stains, and on cellular DNA and protein binding fluorochromes will be presented to illustrate unique differences in measured lifetimes and changes caused by fluorescence quenching. This innovative technology will be used to probe fluorochrome/molecular interactions in the microenvironment of cells/chromosomes as a new parameter and thus expand the researchers' understanding of biochemical processes and structural features at the cellular and molecular level.

Neuronal Depolarization Drives Increased Dopamine Synaptic Vesicle Loading via VGLUT

PubMed Central

Aguilar, Jenny I.; Dunn, Matthew; Mingote, Susana; Karam, Caline S.; Farino, Zachary J.; Sonders, Mark S.; Choi, Se Joon; Grygoruk, Anna; Zhang, Yuchao; Cela, Carolina; Choi, Ben Jiwon; Flores, Jorge; Freyberg, Robin J.; McCabe, Brian D.; Mosharov, Eugene V.; Krantz, David E.; Javitch, Jonathan A.; Sulzer, David; Sames, Dalibor; Rayport, Stephen; Freyberg, Zachary

2017-01-01

SUMMARY The ability of presynaptic dopamine terminals to tune neurotransmitter release to meet the demands of neuronal activity is critical to neurotransmission. Although vesicle content has been assumed to be static, in vitro data increasingly suggest that cell activity modulates vesicle content. Here, we use a coordinated genetic, pharmacological, and imaging approach in Drosophila to study the presynaptic machinery responsible for these vesicular processes in vivo. We show that cell depolarization increases synaptic vesicle dopamine content prior to release via vesicular hyperacidification. This depolarization-induced hyperacidification is mediated by the vesicular glutamate transporter (VGLUT). Remarkably, both depolarization-induced dopamine vesicle hyperacidification and its dependence on VGLUT2 are seen in ventral midbrain dopamine neurons in the mouse. Together, these data suggest that in response to depolarization, dopamine vesicles utilize a cascade of vesicular transporters to dynamically increase the vesicular pH gradient, thereby increasing dopamine vesicle content. PMID:28823729

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.

Depolarizing Effects of Daikenchuto on Interstitial Cells of Cajal from Mouse Small Intestine

PubMed Central

Kim, Hyungwoo; Kim, Hyun Jung; Yang, Dongki; Jung, Myeong Ho; Kim, Byung Joo

2017-01-01

Background: Daikenchuto (DKT; TJ-100, TU-100), a traditional herbal medicineis used in modern medicine to treat gastrointestinal (GI) functional disorders. Interstitial cells of Cajal (ICCs) are the pacemaker cells of the GI tract and play important roles in the regulation of GI motility. Objective: The objective of this study was to investigate the effects of DKT on the pacemaker potentials (PPs) of cultured ICCs from murine small intestine. Materials and Methods: Enzymatic digestions were used to dissociate ICCs from mouse small intestine tissues. All experiments on ICCs were performed after 12 h of culture. The whole-cell patch-clamp configuration was used to record ICC PPs (current clamp mode). All experiments were performed at 30-32°C. Results: In current-clamp modeDKT depolarized and concentration-dependently decreased the amplitudes of PPs. Y25130 (a 5-HT3 receptor antagonist) or SB269970 (a 5-HT7 receptor antagonist) did not block DKT-induced PP depolarization, but RS39604 (a 5-HT4 receptor antagonist) did. Methoctramine (a muscarinic M2 receptor antagonist) failed to block DKT-induced PP depolarization, but pretreating 4-diphenylacetoxy-N-methylpiperidine methiodide (a muscarinic M3 receptor antagonist) facilitated blockade of DKT-induced PP depolarization. Pretreatment with an external Ca2+-free solution or thapsigargin abolished PPsand under these conditions, DKT did not induce PP depolarization. Furthermore Ginseng radix and Zingiberis rhizomes depolarized PPs, whereas Zanthoxyli fructus fruit (the third component of DKT) hyperpolarized PPs. Conclusion: These results suggest that DKT depolarizes ICC PPs in an internal or external Ca2+-dependent manner by stimulating 5-HT4 and M3 receptors. Furthermore, the authors suspect that the component in DKT largely responsible for depolarization is probably also a component of Ginseng radix and Zingiberis rhizomes. SUMMARY Daikenchuto (DKT) depolarized and concentration-dependently decreased the amplitudes of

Depolarizing Effects of Daikenchuto on Interstitial Cells of Cajal from Mouse Small Intestine.

PubMed

Kim, Hyungwoo; Kim, Hyun Jung; Yang, Dongki; Jung, Myeong Ho; Kim, Byung Joo

2017-01-01

Daikenchuto (DKT; TJ-100, TU-100), a traditional herbal medicineis used in modern medicine to treat gastrointestinal (GI) functional disorders. Interstitial cells of Cajal (ICCs) are the pacemaker cells of the GI tract and play important roles in the regulation of GI motility. The objective of this study was to investigate the effects of DKT on the pacemaker potentials (PPs) of cultured ICCs from murine small intestine. Enzymatic digestions were used to dissociate ICCs from mouse small intestine tissues. All experiments on ICCs were performed after 12 h of culture. The whole-cell patch-clamp configuration was used to record ICC PPs (current clamp mode). All experiments were performed at 30-32°C. In current-clamp modeDKT depolarized and concentration-dependently decreased the amplitudes of PPs. Y25130 (a 5-HT 3 receptor antagonist) or SB269970 (a 5-HT 7 receptor antagonist) did not block DKT-induced PP depolarization, but RS39604 (a 5-HT 4 receptor antagonist) did. Methoctramine (a muscarinic M 2 receptor antagonist) failed to block DKT-induced PP depolarization, but pretreating 4-diphenylacetoxy-N-methylpiperidine methiodide (a muscarinic M 3 receptor antagonist) facilitated blockade of DKT-induced PP depolarization. Pretreatment with an external Ca 2+ -free solution or thapsigargin abolished PPsand under these conditions, DKT did not induce PP depolarization. Furthermore Ginseng radix and Zingiberis rhizomes depolarized PPs, whereas Zanthoxyli fructus fruit (the third component of DKT) hyperpolarized PPs. These results suggest that DKT depolarizes ICC PPs in an internal or external Ca 2+ -dependent manner by stimulating 5-HT 4 and M 3 receptors. Furthermore, the authors suspect that the component in DKT largely responsible for depolarization is probably also a component of Ginseng radix and Zingiberis rhizomes. Daikenchuto (DKT) depolarized and concentration-dependently decreased the amplitudes of pacemaker potentials (PPs)Y25130 (a 5-HT 3 receptor antagonist) or

A scattering model for rain depolarization

NASA Technical Reports Server (NTRS)

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

1973-01-01

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

Depolarizing collisions with hydrogen: Neutral and singly ionized alkaline earths

DOE Office of Scientific and Technical Information (OSTI.GOV)

Manso Sainz, Rafael; Ramos, Andrés Asensio; Bueno, Javier Trujillo

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 ≤ 10,000more » 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.« less

Neuronal Depolarization Drives Increased Dopamine Synaptic Vesicle Loading via VGLUT.

PubMed

Aguilar, Jenny I; Dunn, Matthew; Mingote, Susana; Karam, Caline S; Farino, Zachary J; Sonders, Mark S; Choi, Se Joon; Grygoruk, Anna; Zhang, Yuchao; Cela, Carolina; Choi, Ben Jiwon; Flores, Jorge; Freyberg, Robin J; McCabe, Brian D; Mosharov, Eugene V; Krantz, David E; Javitch, Jonathan A; Sulzer, David; Sames, Dalibor; Rayport, Stephen; Freyberg, Zachary

2017-08-30

The ability of presynaptic dopamine terminals to tune neurotransmitter release to meet the demands of neuronal activity is critical to neurotransmission. Although vesicle content has been assumed to be static, in vitro data increasingly suggest that cell activity modulates vesicle content. Here, we use a coordinated genetic, pharmacological, and imaging approach in Drosophila to study the presynaptic machinery responsible for these vesicular processes in vivo. We show that cell depolarization increases synaptic vesicle dopamine content prior to release via vesicular hyperacidification. This depolarization-induced hyperacidification is mediated by the vesicular glutamate transporter (VGLUT). Remarkably, both depolarization-induced dopamine vesicle hyperacidification and its dependence on VGLUT2 are seen in ventral midbrain dopamine neurons in the mouse. Together, these data suggest that in response to depolarization, dopamine vesicles utilize a cascade of vesicular transporters to dynamically increase the vesicular pH gradient, thereby increasing dopamine vesicle content. Copyright © 2017 Elsevier Inc. All rights reserved.

Observation of Exciton-Exciton Interaction Mediated Valley Depolarization in Monolayer MoSe2.

PubMed

Mahmood, Fahad; Alpichshev, Zhanybek; Lee, Yi-Hsien; Kong, Jing; Gedik, Nuh

2018-01-10

The valley pseudospin in monolayer transition metal dichalcogenides (TMDs) has been proposed as a new way to manipulate information in various optoelectronic devices. This relies on a large valley polarization that remains stable over long time scales (hundreds of nanoseconds). However, time-resolved measurements report valley lifetimes of only a few picoseconds. This has been attributed to mechanisms such as phonon-mediated intervalley scattering and a precession of the valley pseudospin through electron-hole exchange. Here we use transient spin grating to directly measure the valley depolarization lifetime in monolayer MoSe 2 . We find a fast valley decay rate that scales linearly with the excitation density at different temperatures. This establishes the presence of strong exciton-exciton Coulomb exchange interactions enhancing the valley depolarization. Our work highlights the microscopic processes inhibiting the efficient use of the exciton valley pseudospin in monolayer TMDs.

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.

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.

Pharmacological modulation of spreading depolarizations.

PubMed

Sánchez-Porras, Renán; Zheng, Zelong; Sakowitz, Oliver W

2015-01-01

Spreading depolarization (SD) is a wave of almost complete depolarization of the neuronal and glial cells. Nowadays there is sufficient evidence demonstrating its pathophysiological effect in migraine with aura, transient global amnesia, stroke, subarachnoid hemorrhage, intracerebral hemorrhage, and traumatic brain injury. In these cases, occurrence of SD has been associated with functional neuronal damage, neuronal necrosis, neurological degeneration, and poor clinical outcome. Animal models show that SD can be modulated by drugs that interfere with its initiation and propagation. There are many pharmacological targets that may help to suppress SD occurrence, such as Na⁺, K⁺, Cl⁻, and Ca²⁺ channels; Na⁺/K⁺ -ATPase; gap junctions; and ligand-based receptors, for example, adrenergic, serotonin, sigma-1, calcitonin gene-related peptide, GABAA, and glutamate receptors. In this regard, N-methyl-d-aspartate (NMDA) receptor blockers, in particular, ketamine, have shown promising results. Therefore, theoretically pharmacologic modulation of SD could help diminish its pathological effects.

Measuring partial fluorescence yield using filtered detectors.

PubMed

Boyko, T D; Green, R J; Moewes, A; Regier, T Z

2014-07-01

Typically, X-ray absorption near-edge structure measurements aim to probe the linear attenuation coefficient. These measurements are often carried out using partial fluorescence yield techniques that rely on detectors having photon energy discrimination improving the sensitivity and the signal-to-background ratio of the measured spectra. However, measuring the partial fluorescence yield in the soft X-ray regime with reasonable efficiency requires solid-state detectors, which have limitations due to the inherent dead-time while measuring. Alternatively, many of the available detectors that are not energy dispersive do not suffer from photon count rate limitations. A filter placed in front of one of these detectors will make the energy-dependent efficiency non-linear, thereby changing the responsivity of the detector. It is shown that using an array of filtered X-ray detectors is a viable method for measuring soft X-ray partial fluorescence yield spectra without dead-time. The feasibility of this technique is further demonstrated using α-Fe2O3 as an example and it is shown that this detector technology could vastly improve the photon collection efficiency at synchrotrons and that these detectors will allow experiments to be completed with a much lower photon flux reducing X-ray-induced damage.

A depolarization and attenuation experiment using the COMSTAR and CTS satellites

NASA Technical Reports Server (NTRS)

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

1978-01-01

An experiment for measuring precipitation attenuation and depolarization on the CTS 11.7 and the COMSTAR 19.04 and 28.56 GHz downlinks is described. Attenuation scaling, effective path length, and the relationship between isolation and attenuation are discussed. Attenuation and effective path data are presented for the months of July, August, and September, 1977.

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.

Losses and depolarization of ultracold neutrons on neutron guide and storage materials

NASA Astrophysics Data System (ADS)

Bondar, V.; Chesnevskaya, S.; Daum, M.; Franke, B.; Geltenbort, P.; Göltl, L.; Gutsmiedl, E.; Karch, J.; Kasprzak, M.; Kessler, G.; Kirch, K.; Koch, H.-C.; Kraft, A.; Lauer, T.; Lauss, B.; Pierre, E.; Pignol, G.; Reggiani, D.; Schmidt-Wellenburg, P.; Sobolev, Yu.; Zechlau, T.; Zsigmond, G.

2017-09-01

At Institut Laue-Langevin (ILL) and Paul Scherrer Institute (PSI), we have measured the losses and depolarization probabilities of ultracold neutrons on various materials: (i) nickel-molybdenum alloys with weight percentages of 82/18, 85/15, 88/12, 91/9, and 94/6 and natural nickel Ni100, (ii) nickel-vanadium NiV93/7, (iii) copper, and (iv) deuterated polystyrene (dPS). For the different samples, storage-time constants up to ˜460 s were obtained at room temperature. The corresponding loss parameters for ultracold neutrons, η , varied between 1.0 ×10-4 and 2.2 ×10-4 . All η values are in agreement with theory except for dPS, where anomalous losses at room temperature were established with four standard deviations. The depolarization probabilities per wall collision β measured with unprecedented sensitivity varied between 0.7 ×10-6 and 9.0 ×10-6 . Our depolarization result for copper differs from other experiments by 4.4 and 15.8 standard deviations. The β values of the paramagnetic NiMo alloys over molybdenum content show an increase of β with increasing Mo content. This is in disagreement with expectations from literature. Finally, ferromagnetic behavior of NiMo alloys at room temperature was found for molybdenum contents of 6.5 at.% or less and paramagnetic behavior for more than 8.7 at.%. This may contribute to solving an ambiguity in literature.

Depolarization of Pulsar Radio Emission.

PubMed

Lyutikov

1999-11-01

We show that intensity-dependent depolarization of single pulses may be due to the nonlinear decay of the "upper" ordinary (O) mode into an unpolarized extraordinary mode and a backward-propagating wave. The decay occurs in the innermost parts of the pulsar magnetosphere for obliquely propagating O waves.

Linear Depolarization of Lidar Returns by Aged Smoke Particles

NASA Technical Reports Server (NTRS)

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

2016-01-01

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

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

Chlorovirus-mediated membrane depolarization of Chlorella alters secondary active transport of solutes.

PubMed

Agarkova, Irina; Dunigan, David; Gurnon, James; Greiner, Timo; Barres, Julia; Thiel, Gerhard; Van Etten, James L

2008-12-01

Paramecium bursaria chlorella virus 1 (PBCV-1) is the prototype of a family of large, double-stranded DNA, plaque-forming viruses that infect certain eukaryotic chlorella-like green algae from the genus Chlorovirus. PBCV-1 infection results in rapid host membrane depolarization and potassium ion release. One interesting feature of certain chloroviruses is that they code for functional potassium ion-selective channel proteins (Kcv) that are considered responsible for the host membrane depolarization and, as a consequence, the efflux of potassium ions. This report examines the relationship between cellular depolarization and solute uptake. Annotation of the virus host Chlorella strain NC64A genome revealed 482 putative transporter-encoding genes; 224 are secondary active transporters. Solute uptake experiments using seven radioactive compounds revealed that virus infection alters the transport of all the solutes. However, the degree of inhibition varied depending on the solute. Experiments with nystatin, a drug known to depolarize cell membranes, produced changes in solute uptake that are similar but not identical to those that occurred during virus infection. Therefore, these studies indicate that chlorovirus infection causes a rapid and sustained depolarization of the host plasma membrane and that this depolarization leads to the inhibition of secondary active transporters that changes solute uptake.

Chlorovirus-Mediated Membrane Depolarization of Chlorella Alters Secondary Active Transport of Solutes▿

PubMed Central

Agarkova, Irina; Dunigan, David; Gurnon, James; Greiner, Timo; Barres, Julia; Thiel, Gerhard; Van Etten, James L.

2008-01-01

Paramecium bursaria chlorella virus 1 (PBCV-1) is the prototype of a family of large, double-stranded DNA, plaque-forming viruses that infect certain eukaryotic chlorella-like green algae from the genus Chlorovirus. PBCV-1 infection results in rapid host membrane depolarization and potassium ion release. One interesting feature of certain chloroviruses is that they code for functional potassium ion-selective channel proteins (Kcv) that are considered responsible for the host membrane depolarization and, as a consequence, the efflux of potassium ions. This report examines the relationship between cellular depolarization and solute uptake. Annotation of the virus host Chlorella strain NC64A genome revealed 482 putative transporter-encoding genes; 224 are secondary active transporters. Solute uptake experiments using seven radioactive compounds revealed that virus infection alters the transport of all the solutes. However, the degree of inhibition varied depending on the solute. Experiments with nystatin, a drug known to depolarize cell membranes, produced changes in solute uptake that are similar but not identical to those that occurred during virus infection. Therefore, these studies indicate that chlorovirus infection causes a rapid and sustained depolarization of the host plasma membrane and that this depolarization leads to the inhibition of secondary active transporters that changes solute uptake. PMID:18842725

Time-resolved polarization imaging by pump-probe (stimulated emission) fluorescence microscopy.

PubMed Central

Buehler, C; Dong, C Y; So, P T; French, T; Gratton, E

2000-01-01

We report the application of pump-probe fluorescence microscopy in time-resolved polarization imaging. We derived the equations governing the pump-probe stimulated emission process and characterized the pump and probe laser power levels for signal saturation. Our emphasis is to use this novel methodology to image polarization properties of fluorophores across entire cells. As a feasibility study, we imaged a 15-microm orange latex sphere and found that there is depolarization that is possibly due to energy transfer among fluorescent molecules inside the sphere. We also imaged a mouse fibroblast labeled with CellTracker Orange CMTMR (5-(and-6)-(((4-chloromethyl)benzoyl)amino)tetramethyl-rhodamine). We observed that Orange CMTMR complexed with gluthathione rotates fast, indicating the relatively low fluid-phase viscosity of the cytoplasmic microenvironment as seen by Orange CMTMR. The measured rotational correlation time ranged from approximately 30 to approximately 150 ps. This work demonstrates the effectiveness of stimulated emission measurements in acquiring high-resolution, time-resolved polarization information across the entire cell. PMID:10866979

X-ray fluorescence measurements of dissolved gas and cavitation

DOE PAGES

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

2016-09-28

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

Quantification of Material Fluorescence and Light Scattering Cross Sections Using Ratiometric Bandwidth-Varied Polarized Resonance Synchronous Spectroscopy.

PubMed

Xu, Joanna Xiuzhu; Hu, Juan; Zhang, Dongmao

2018-05-25

Presented herein is the ratiometric bandwidth-varied polarized resonance synchronous spectroscopy (BVPRS2) method for quantification of material optical activity spectra. These include the sample light absorption and scattering cross-section spectrum, the scattering depolarization spectrum, and the fluorescence emission cross-section and depolarization spectrum in the wavelength region where the sample both absorbs and emits. This ratiometric BVPRS2 spectroscopic method is a self-contained technique capable of quantitatively decoupling material fluorescence and light scattering signal contribution to its ratiometric BVPRS2 spectra through the linear curve-fitting of the ratiometric BVPRS2 signal as a function of the wavelength bandwidth used in the PRS2 measurements. Example applications of this new spectroscopic method are demonstrated with materials that can be approximated as pure scatterers, simultaneous photon absorbers/emitters, simultaneous photon absorbers/scatterers, and finally simultaneous photon absorbers/scatterers/emitters. Because the only instruments needed for this ratiometric BVPRS2 technique are the conventional UV-vis spectrophotometer and spectrofluorometer, this work should open doors for routine decomposition of material UV-vis extinction spectrum into its absorption and scattering component spectra. The methodology and insights provided in this work should be of broad significance to all chemical research that involves photon/matter interactions.

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

2008-02-04

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.

Depolarization Lidar Determination Of Cloud-Base Microphysical Properties

NASA Astrophysics Data System (ADS)

Donovan, D. P.; Klein Baltink, H.; Henzing, J. S.; de Roode, S.; Siebesma, A. P.

2016-06-01

The links between multiple-scattering induced depolarization and cloud microphysical properties (e.g. cloud particle number density, effective radius, water content) have long been recognised. Previous efforts to use depolarization information in a quantitative manner to retrieve cloud microphysical cloud properties have also been undertaken but with limited scope and, arguably, success. In this work we present a retrieval procedure applicable to liquid stratus clouds with (quasi-)linear LWC profiles and (quasi-)constant number density profiles in the cloud-base region. This set of assumptions allows us to employ a fast and robust inversion procedure based on a lookup-table approach applied to extensive lidar Monte-Carlo multiple-scattering calculations. An example validation case is presented where the results of the inversion procedure are compared with simultaneous cloud radar observations. In non-drizzling conditions it was found, in general, that the lidar- only inversion results can be used to predict the radar reflectivity within the radar calibration uncertainty (2-3 dBZ). Results of a comparison between ground-based aerosol number concentration and lidar-derived cloud base number considerations are also presented. The observed relationship between the two quantities is seen to be consistent with the results of previous studies based on aircraft-based in situ measurements.

Intestine pH measurements using fluorescence imaging: an in-vivo preliminary study

NASA Astrophysics Data System (ADS)

Marechal, Xavier-Marie; Mordon, Serge R.; Devoisselle, Jean-Marie; Begu, Sylvie; Mathieu, D.; Buys, Bruno; Dhelin, Guy; Lesage, Jean C.; Neviere, Remi; Chopin, Claude

1999-02-01

Measurement of gastrointestinal intramucosal pH has been recognized as an important factor in the detection of hypoxia-induced dysfunctions. However, current pH measurement techniques are limited in terms of time and spatial resolution. A major advance in accurate pH measurement was the development of the ratiometric fluorescent indicator dye, 2',7'-bis(carboxyethyl)-4,5- carboxyfluorescein (BCECF). This study aimed to demonstrate the feasibility of fluorescence imaging technique to measure in vivo the pH of intestine. The intestine was inserted in an optical chamber placed under a microscope. Animals were injected i.v. with the pH-sensitive fluorescent dye BCECF. Fluorescence was visualized by illuminating the intestine alternately at 490 and 470 nm. The emitted fluorescence was directed to an intensified camera. The ratio of emitted fluorescence at excitation wavelengths of 490 and 470 nm was measured, corrected and converted to pH by constructing a calibration curve. The pH controls were performed with a pH microelectrode correlated with venous blood gas sampling. We concluded that accurate pH measurements of rat intestine can be obtained by fluorescence imaging using BCECF. This technology could be easily adapted for endoscopic pH measurement.

Bi0.5Na0.5TiO3:ZnO lead-free piezoelectric composites with deferred thermal depolarization

NASA Astrophysics Data System (ADS)

Zhang, Ji; Pan, Zhao; Nie, Peng-Xiao; Cui, Yu-Shuang; Yang, Bin; Chen, Jun; Zhang, Shan-Tao

2015-06-01

Bi0.5Na0.5TiO3 (BNT) is among the most promising lead-free piezoelectric candidates. However, depolarization of BNT is a longstanding obstacle for practical applications. Here, we report that piezoelectric composites of Bi0.5Na0.5TiO3:xZnO (BNT:xZnO, where x is the mole ratio of ZnO to BNT) have deferred thermal depolarization. With increasing x from 0 to 0.4, the observed depolarization temperature (Td) tends to be deferred near x = 0.3, as confirmed by temperature dependent dielectric, ferroelectric, and piezoelectric measurements. As the result, the piezoelectric properties of the composites can be well maintained even after the poled composites are annealed at 125 °C. It is proposed that the charges stemming from ZnO can be orderly distributed to form a local field, which can keep the poling state of BNT, thus suppress the depolarization, even after the external poling filed is removed. These results may pave the way for applications of BNT-based piezoceramics and significantly improve our understanding of the depolarization mechanism by optimizing the performance of lead-free piezoelectrics.

Depolarization in liquid-crystal televisions

NASA Astrophysics Data System (ADS)

Pezzaniti, Larry J.; McClain, Stephen C.; Chipman, Russell A.; Lu, Shih-Yau

1993-12-01

TVT-6000 liquid crystal television (LCTV) polarization properties have been mapped as a function of biased voltage to the pixel and angle of incidence by a Mueller-matrix imaging polarimeter at 632.8 nm. Operating without polarizers the LCTV shows between 2% to 9% depolarization depending on angle of incidence, the incident polarization state, and the pixel bias voltage.

Excitation Anisotropy in Laser-Induced-Fluorescence Spectroscopy —High-Intensity, Broad-Line Excitation

NASA Astrophysics Data System (ADS)

Hirabayashi, Atsumu; Nambu, Yoshihiro; Fujimoto, Takashi

1986-10-01

The problem of excitation anisotropy in laser-induced-fluorescence spectroscopy (LIFS) was investigated for the intense excitation case under the broad-line condition. The depolarization coefficient for the fluorescence light was derived in the intense-excitation limit (linearly-polarized or unpolarized light excitation) and the results are presented in tables. In the region of intermediate intensity, between the weak and intense-excitation limits, the master equation was solved for a specific example of atomic transitions and its result is compared with experimental results.

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…

Comparison of calibration and standardization approaches for fluorescence guided imaging systems to benchtop fluorescence measurements in cellular systems

NASA Astrophysics Data System (ADS)

Litorja, Maritoni; DeRose, Paul

2018-02-01

Fluorescence measurements are a staple in biomedicine, from research and discovery to more recently, for fluorescenceguided imaging systems for diagnostics and surgery. Measurement validation for clinical imagers is a challenge as it is applied to many different optical systems and probe through matrices with different optical properties in a demanding field environment. In this paper we will present approaches to fluorescence calibration for a field system, in comparison to those used in laboratory instruments for cell measurements or benchtop fluorometers. We will present the common challenges and differences, and lessons from the standardization effort of laboratory fluorescence measurements. We will discuss the conceptually different pathways to measurement traceability, between counting moles of substance and measuring light.

The leading role of mitochondrial depolarization in the mechanism of glutamate-induced disruptions in Ca2+ homeostasis.

PubMed

Khodorov, B I; Storozhevykh, T P; Surin, A M; Yuryavichyus, A I; Sorokina, E G; Borodin, A V; Vinskaya, N P; Khaspekov, L G; Pinelis, V G

2002-01-01

Data obtained in studies of the nature of the correlation which we have previously observed [10,17] between mitochondrial depolarization and the level of disruption of Ca2+ homeostasis in cultivated brain neuronsare summarized. Experiments were performed on cultured cerebellar granule cells loaded with Fura-2-AM or rhodamine 123 to measure changes in cytoplasmic Ca2+ and mitochondrial potential during pathogenic treatments of the cells. Prolonged exposure to 100 microM glutamate induced a reversible increase in [Ca2+]i, which was accompanied by only a small degree of mitochondrial depolarization. A sharp increase in this mitochondrial depolarization, induced by addition of 3 mM NaCN or 300 microM dinitrophenol (DNP) to the glutamate-containing solution, resulted in further increase in [Ca2+]i, due to blockade of electrophoretic mitochondrial Ca2+ uptake. Prolonged exposure to CN- or DNP in the post-glutamate period maintained [Ca2+]i at a high level until the metabolic inhibitors were removed. In most cells, this plateau was characterized by low sensitivity to removal of external Ca2+, demonstrating that the mechanisms of Ca2+ release from neurons were disrupted. Addition of oligomycin, a blocker of mitochondrial ATP synthase/ATPase, to the solution containing glutamate and CN- or DNP eliminated the post-glutamate plateau. Parallel experiments with direct measurements of intracellular ATP levels ([ATP]) showed that profound mitochondrial depolarization induced by CN- or DNP sharply enhanced the drop in ATP due to glutamate, while oligomycin significantly weakened this effect of the metabolic inhibitors. Analysis of these data led to the conclusion that blockade of mitochondrial Ca2+ uptake and inhibition of ATP synthesis resulted from mitochondrial depolarization and plays a key role in the mechanism disrupting [Ca2+]i homeostasis after toxic exposure to glutamate.

Nd/sup 3 +/ fluorescence quantum-efficiency measurements with photoacoustics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rosencwaig, A.; Hildum, E.A.

1981-04-01

We have investigated the use of photoacoustic techniques for obtaining absolute values of fluorescence quantum efficiencies in lightly doped Nd/sup 3 +/ laser materials. We have found that surface absorptions play an important role in gas-microphone measurements, and that thermal profiles are important in piezoelectric measurements. We have obtained fluorescence quantum efficiencies for Nd/sup 3 +/ in yttrium aluminum garnet, and in silicate and borate glasses that are in good agreement with lifetime measurements and Judd-Ofelt calculations.

Rapid changes in synaptic vesicle cytochemistry after depolarization of cultured cholinergic sympathetic neurons

PubMed Central

1985-01-01

Sympathetic neurons taken from rat superior cervical ganglia and grown in culture acquire cholinergic function under certain conditions. These cholinergic sympathetic neurons, however, retain a number of adrenergic properties, including the enzymes involved in the synthesis of norepinephrine (NE) and the storage of measurable amounts of NE. These neurons also retain a high affinity uptake system for NE; despite this, the majority of the synaptic vesicles remain clear even after incubation in catecholamines. The present study shows, however, that if these neurons are depolarized before incubation in catecholamine, the synaptic vesicles acquire dense cores indicative of amine storage. These manipulations are successful when cholinergic function is induced with either a medium that contains human placental serum and embryo extract or with heart-conditioned medium, and when the catecholamine is either NE or 5-hydroxydopamine. In some experiments, neurons are grown at low densities and shown to have cholinergic function by electrophysiological criteria. After incubation in NE, only 6% of the synaptic vesicles have dense cores. In contrast, similar neurons depolarized (80 mM K+) before incubation in catecholamine contain 82% dense-cored vesicles. These results are confirmed in network cultures where the percentage of dense-cored vesicles is increased 2.5 to 6.5 times by depolarizing the neurons before incubation with catecholamine. In both single neurons and in network cultures, the vesicle reloading is inhibited by reducing vesicle release during depolarization with an increased Mg++/Ca++ ratio or by blocking NE uptake either at the plasma membrane (desipramine) or at the vesicle membrane (reserpine). In addition, choline appears to play a competitive role because its presence during incubation in NE or after reloading results in decreased numbers of dense-cored vesicles. We conclude that the depolarization step preceding catecholamine incubation acts to empty the

Marine fluorescence from high spectrally resolved satellite measurements

NASA Astrophysics Data System (ADS)

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

2014-05-01

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

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

NASA Astrophysics Data System (ADS)

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

2012-02-01

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

Dual Double-Wedge Pseudo-Depolarizer with Anamorphic PSF

NASA Technical Reports Server (NTRS)

Hill, Peter; Thompson, Patrick

2012-01-01

A polarized scene, which may occur at oblique illumination angles, creates a radiometric signal that varies as a function of viewing angle. One common optical component that is used to minimize such an effect is a polarization scrambler or depolarizer. As part of the CLARREO mission, the SOLARIS instrument project at Goddard Space Flight Center has developed a new class of polarization scramblers using a dual double-wedge pseudo-depolarizer that produces an anamorphic point spread function (PSF). The SOLARIS instrument uses two Wollaston type scramblers in series, each with a distinct wedge angle, to image a pseudo-depolarized scene that is free of eigenstates. Since each wedge is distinct, the scrambler is able to produce an anamorphic PSF that maintains high spatial resolution in one dimension by sacrificing the spatial resolution in the other dimension. This scrambler geometry is ideal for 1-D imagers, such as pushbroom slit spectrometers, which require high spectral resolution, high spatial resolution, and low sensitivity to polarized light. Moreover, the geometry is applicable to a wide range of scientific instruments that require both high SNR (signal-to-noise ratio) and low sensitivity to polarized scenes

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. © 2014 The International Union of Biochemistry and Molecular Biology.

System and method for measuring fluorescence of a sample

DOEpatents

Riot, Vincent J

2015-03-24

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

Rising intracellular zinc by membrane depolarization and glucose in insulin-secreting clonal HIT-T15 beta cells.

PubMed

Slepchenko, Kira G; Li, Yang V

2012-01-01

Zinc (Zn(2+)) appears to be intimately involved in insulin metabolism since insulin secretion is correlated with zinc secretion in response to glucose stimulation, but little is known about the regulation of zinc homeostasis in pancreatic beta-cells. This study set out to identify the intracellular zinc transient by imaging free cytosolic zinc in HIT-T15 beta-cells with fluorescent zinc indicators. We observed that membrane depolarization by KCl (30-60 mM) was able to induce a rapid increase in cytosolic concentration of zinc. Multiple zinc transients of similar magnitude were elicited during repeated stimulations. The amplitude of zinc responses was not affected by the removal of extracellular calcium or zinc. However, the half-time of the rising slope was significantly slower after removing extracellular zinc with zinc chelator CaEDTA, suggesting that extracellular zinc affect the initial rising phase of zinc response. Glucose (10 mM) induced substantial and progressive increases in intracellular zinc concentration in a similar way as KCl, with variation in the onset and the duration of zinc mobilization. It is known that the depolarization of beta-cell membrane is coupled with the secretion of insulin. Rising intracellular zinc concentration may act as a critical signaling factor in insulin metabolism of pancreatic beta-cells.

Rising Intracellular Zinc by Membrane Depolarization and Glucose in Insulin-Secreting Clonal HIT-T15 Beta Cells

PubMed Central

Slepchenko, Kira G.; Li, Yang V.

2012-01-01

Zinc (Zn2+) appears to be intimately involved in insulin metabolism since insulin secretion is correlated with zinc secretion in response to glucose stimulation, but little is known about the regulation of zinc homeostasis in pancreatic beta-cells. This study set out to identify the intracellular zinc transient by imaging free cytosolic zinc in HIT-T15 beta-cells with fluorescent zinc indicators. We observed that membrane depolarization by KCl (30–60 mM) was able to induce a rapid increase in cytosolic concentration of zinc. Multiple zinc transients of similar magnitude were elicited during repeated stimulations. The amplitude of zinc responses was not affected by the removal of extracellular calcium or zinc. However, the half-time of the rising slope was significantly slower after removing extracellular zinc with zinc chelator CaEDTA, suggesting that extracellular zinc affect the initial rising phase of zinc response. Glucose (10 mM) induced substantial and progressive increases in intracellular zinc concentration in a similar way as KCl, with variation in the onset and the duration of zinc mobilization. It is known that the depolarization of beta-cell membrane is coupled with the secretion of insulin. Rising intracellular zinc concentration may act as a critical signaling factor in insulin metabolism of pancreatic beta-cells. PMID:22536213

Time-resolved fluorescence polarization spectroscopy of visible and near infrared dyes in picosecond dynamics

NASA Astrophysics Data System (ADS)

Pu, Yang; Alfano, Robert R.

2015-03-01

Near-infrared (NIR) dyes absorb and emit light within the range from 700 to 900 nm have several benefits in biological studies for one- and/or two-photon excitation for deeper penetration of tissues. These molecules undergo vibrational and rotational motion in the relaxation of the excited electronic states, Due to the less than ideal anisotropy behavior of NIR dyes stemming from the fluorophores elongated structures and short fluorescence lifetime in picosecond range, no significant efforts have been made to recognize the theory of these dyes in time-resolved polarization dynamics. In this study, the depolarization of the fluorescence due to emission from rotational deactivation in solution will be measured with the excitation of a linearly polarized femtosecond laser pulse and a streak camera. The theory, experiment and application of the ultrafast fluorescence polarization dynamics and anisotropy are illustrated with examples of two of the most important medical based dyes. One is NIR dye, namely Indocyanine Green (ICG) and is compared with Fluorescein which is in visible range with much longer lifetime. A set of first-order linear differential equations was developed to model fluorescence polarization dynamics of NIR dye in picosecond range. Using this model, the important parameters of ultrafast polarization spectroscopy were identified: risetime, initial time, fluorescence lifetime, and rotation times.

The NPe6 fluorescence measurements by using a fluorescence sensing system for skin photosensitivity risk assessment after photodynamic therapy

NASA Astrophysics Data System (ADS)

Ohtani, Keishi; Usuda, Jitsu; Ogawa, Emiyu; Maehara, Sachio; Imai, Kentaro; Inoue, Tatsuya; Hagiwara, Masaru; Kakihana, Masatoshi; Kajiwara, Naohiro; Ohira, Tatsuo; Arai, Tsunenori; Ikeda, Norihiko

2018-02-01

Background: Skin photosensitivity is a major side effect of photodynamic therapy (PDT). It is induced by the photosensitizer remaining in the skin. It is usually rapidly metabolized by the liver, but the pharmacokinetic profile varies widely among individuals. This makes it difficult to predict the incidence of skin photosensitivity. Therefore, we conducted a prospective study to investigate whether the NPe6 fluorescence intensity in skin after NPe6-PDT could be measured safely in human patients using a fluorescence sensing system for judging the risk of skin photosensitivity. Methods: The NPe6 fluorescence measurements using a constructed fluorescence sensing system at the inside of the arm were acquired prior to and 5 and 10 minutes after NPe6 administration as well as at the time of PDT (4-5 hours after administration), at discharge (2 or 3 days after PDT), and at 1 or 2 weeks after PDT. Participants were interviewed as to whether they had any complications at 2 weeks after PDT. Results: Nine male patients and one female patient entered this study. All of the measurements of NPe6 fluorescence in the skin could be obtained without any complications. The spectral peak was detected at the time of discharge (2-3 days after administration) in most cases and it decreased at 1 or 2 weeks after PDT. Conclusions: The fluorescence of NPe6 in the skin could be detected feasibly using the fluorescence sensing system in human patients. Measuring fluorescence intensity in the skin might be useful to predict the incidence of skin photosensitivity after PDT.

Lidar measurements of boundary layer depolarization and CCSEM-EDX compositional analysis of airborne particles on collocated passive samplers throughout the forest canopy during the 2016 airborne pollen season at UMBS, Pellston, MI

NASA Astrophysics Data System (ADS)

Wozniak, M. C.; Steiner, A.; Ault, A. P.; Kort, E. A.; Lersch, T.; Casuccio, G.

2017-12-01

Observations of airborne pollen are typically made with volumetric samplers that obtain a time-averaged pollen concentration at a single point. While spatial variations in surface pollen concentrations may be known with these samplers given multiple sampling sites, real-time boundary layer transport of pollen grains cannot be determined except by particle dispersion or tracer transport models. Recently, light detection and ranging (lidar) techniques, such as depolarization, have been used to measure pollen transport and optical properties throughout the boundary layer over time. Here, we use a ground-based micro-pulse lidar (MPL) to observe boundary layer vertical profiles before, during and after the peak anemophilous (wind-driven) pollen season. The lidar depolarization ratio is measured in tandem with the normalized R-squared backscatter (NRB) intensity to determine the contribution of aspherical particles to the scatterers present throughout the boundary layer. Measurements are taken from April 15 - July 12, 2016 at the University of Michigan Biological Station (UMBS) PROPHET outdoor research lab and tower within a largely forested region. UMBS is dominated by Acer rubrum, Betula papyrifera, Pinus resinosa, Quercus rubra and Pinus strobus, all of which began flowering on 4/19, 5/3, 5/25, 5/25 and 6/14, respectively. Temperature, relative humidity and wind speed measured on site determine daytime conditions conducive to pollen dispersion from flowers. Lidar depolarization ratios between 0.08-0.14 and higher are observed in the daytime boundary layer on days shortly after the flowering dates of the aforementioned species, elevated above the background level of 0.06 or less. Lidar observations are supplemented with aerosol compositional analysis determined by computer-controlled scanning electron microscopy and energy-dispersive X-ray spectroscopy (CCSEM-EDX) on passive sampler data from below, within and above the forest canopy at PROPHET tower. Particles are

Substance P Depolarizes Lamprey Spinal Cord Neurons by Inhibiting Background Potassium Channels.

PubMed

Thörn Pérez, Carolina; Hill, Russell H; Grillner, Sten

2015-01-01

Substance P is endogenously released in the adult lamprey spinal cord and accelerates the burst frequency of fictive locomotion. This is achieved by multiple effects on interneurons and motoneurons, including an attenuation of calcium currents, potentiation of NMDA currents and reduction of the reciprocal inhibition. While substance P also depolarizes spinal cord neurons, the underlying mechanism has not been resolved. Here we show that effects of substance P on background K+ channels are the main source for this depolarization. Hyperpolarizing steps induced inward currents during whole-cell voltage clamp that were reduced by substance P. These background K+ channels are pH sensitive and are selectively blocked by anandamide and AVE1231. These blockers counteracted the effect of substance P on these channels and the resting membrane potential depolarization in spinal cord neurons. Thus, we have shown now that substance P inhibits background K+ channels that in turn induce depolarization, which is likely to contribute to the frequency increase observed with substance P during fictive locomotion.

Elastodynamic metasurface: Depolarization of mechanical waves and time effects

DOE Office of Scientific and Technical Information (OSTI.GOV)

Boutin, Claude, E-mail: claude.boutin@entpe.fr; Schwan, Logan; Dietz, Matthew S.

2015-02-14

We report the concept of microstructured surfaces with inner resonance in the field of elastodynamics, so-called elastodynamic metasurfaces. Such metasurfaces allow for wavefield manipulation of mechanical waves by tuning the boundary conditions at specific frequencies. In particular, they can be used to depolarize elastic waves without introducing heterogeneities in the medium itself; the physical means to do so in homogeneous elastic media used to remain, surprisingly, an open question while depolarization is commonplace in electromagnetism. The principle relies on the anisotropic behaviour of a subwavelength array of resonators: Their subwavelength configuration confines the Bragg interferences scattered by resonators into amore » boundary layer. The effective behaviour of the resonating array is expressed with homogenization as an unconventional impedance, the frequency-dependence, and anisotropy of which lead to depolarization and time effects. The concept of the elastodynamic metasurface is tested experimentally and results bear testament to its efficacy and robustness. Elastodynamic metasurfaces are easily realized and analytically predictable, opening new possibilities in tomography techniques, ultrasonics, geophysics, vibration control, materials and structure design.« less

Integrating fluorescence and interactance measurements to improve apple maturity assessment

NASA Astrophysics Data System (ADS)

Noh, Hyun Kwon; Lu, Renfu

2006-10-01

Fluorescence and reflectance (or interactance) are promising techniques for measuring fruit quality and condition. Our previous research showed that a hyperspectral imaging technique integrating fluorescence and reflectance could improve predictions of selected quality parameters compared to single sensing techniques. The objective of this research was to use a low cost spectrometer for rapid acquisition of fluorescence and interactance spectra from apples and develop an algorithm integrating the two types of data for predicting skin and flesh color, fruit firmness, starch index, soluble solids content, and titratable acid. Experiments were performed to measure UV light induced transient fluorescence and interactance spectra from 'Golden Delicious' apples that were harvested over a period of four weeks during the 2005 harvest season. Standard destructive tests were performed to measure maturity parameters from the apples. Principal component (PC) analysis was applied to the interactance and fluorescence data. A back-propagation feedforward neural network with the inputs of PC data was used to predict individual maturity parameters. Interactance mode was consistently better than fluorescence mode in predicting the maturity parameters. Integrating interactance and fluorescence improved predictions of all parameters except flesh chroma; values of the correlation coefficient for firmness, soluble solids content, starch index, and skin and flesh hue were 0.77, 0.77, 0.89, 0.99, and 0.96 respectively, with the corresponding standard errors of 6.93 N, 0.90%, 0.97 g/L, 0.013 rad, and 0.013 rad. These results represented 4.1% to 23.5% improvements in terms of standard error, in comparison with the better results from the two single sensing methods. Integrating interactance and fluorescence can better assess apple maturity and quality.

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.

Dopamine modulates an intrinsic mGluR5-mediated depolarization underlying prefrontal persistent activity

PubMed Central

Sidiropoulou, Kyriaki; Lu, Fang-Min; Fowler, Melissa A.; Xiao, Rui; Phillips, Christopher; Ozkan, Emin D.; Zhu, Michael X.; White, Francis J.; Cooper, Donald C.

2009-01-01

Intrinsic properties of neurons that enable them to maintain depolarized, persistently activated states in the absence of sustained input are poorly understood. In short-term memory tasks, individual prefrontal cortical (PFC) neurons are capable of maintaining persistent action potential output during delay periods between informative cues and behavioral responses. Dopamine and drugs of abuse alter PFC function and working memory possibly by modulating intrinsic neuronal properties. Here we use patch-clamp recording of layer 5 PFC pyramidal neurons to identify an action potential burst-evoked intrinsic mGluR5-mediated postsynaptic depolarization that initiates an activated state. Depolarization occurs in the absence of recurrent synaptic activity and is reduced by a postsynaptic dopamine D1/5 receptor pathway. The depolarization is substantially diminished following behavioral sensitization to cocaine; moreover the D1/5 receptor modulation is lost. We propose the burst-evoked intrinsic depolarization to be a novel form of short-term cellular memory that is modulated by dopamine and cocaine experience. PMID:19169252

Ellipsometric study of peptide layers - island-like character, depolarization and quasi-absorption

NASA Astrophysics Data System (ADS)

Pápa, Z.; Ramakrishnan, S.; Martin, M.; Cloitre, T.; Zimányi, L.; Tóth, Z.; Gergely, C.; Budai, J.

2017-11-01

In this work, the ellipsometric measurements of small molecular size polypeptides deposited onto silicon are analyzed. Results of ellipsometric evaluation procedures based on transparent layer, absorbing layer and discontinuous layer approaches are compared. Although these models result in similar fitting quality and can predict the amount of the deposited material, the gained optical properties can be rather different due to the different assumptions of the models. To choose the physically correct results, independent measurements as atomic force microscopy or transmission measurement of peptide solutions are necessary. It is shown that the measured ellipsometric depolarization can provide also useful information about the sample properties.

Analysis of laser-induced-fluorescence carbon monoxide measurements in turbulent nonpremixed flames

NASA Astrophysics Data System (ADS)

Mokhov, A. V.; Levinsky, H. B.; van der Meij, C. E.; Jacobs, R. A. A. M.

1995-10-01

The influence of fluctuating concentrations and temperature on the laser-induced-fluorescence (LIF) measurement of CO in turbulent flames is described, under conditions in which the fluorescence and the temperature are measured independently. The analysis shows that correlations between CO concentration and temperature can bias the averaged mole fraction extracted from LIF measurements. The magnitude of the bias can exceed the order of the average CO mole fraction. Further, LIF measurements of CO concentrations in a turbulent, nonpremixed, natural gas flame are described. The averaged CO mole fractions are derived from the fluorescence measurements by the use of flame temperatures independently measured by coherent anti-Stokes Raman spectroscopy. Analysis of the fluctuations in measured temperature and fluorescence indicates that temperature and CO concentrations in flame regions with intensive mixing are indeed correlated. In the flame regions where burnout of CO has ceased, the LIF measurements of the CO mole fraction correspond to the probe measurements in exhaust.

Fluorescent measurements in whole blood and plasma using red-emitting dyes

NASA Astrophysics Data System (ADS)

Abugo, Omoefe O.; Herman, Petr; Lakowicz, Joseph R.

2000-04-01

We have determined the fluorescence characteristics of albumin blue 670 and Rhodamine 800 in plasma and blood in order to test the feasibility of making direct fluorescence sensing measurements in blood. These dyes were used because of their absorption in the red/NIR where absorption by hemoglobin is minimized. Front face illumination and detection was used to minimize absorption and scattering during measurement. Fluorescence emission was observed for these dyes in plasma and blood. Attenuation of the fluorescence emission was observed in blood because of hemoglobin absorption. Using frequency domain fluorometry, we recovered the expected lifetime parameters for both dyes in blood and plasma. We were able to quantify HSA concentrations using changes in the mean lifetime of AB670, a dye previously shown to bind preferentially to HSA. Rh800 concentrations in plasma and blood were also determined using modulation sensing. Anisotropy measurements revealed high Anisotropy for these dyes in plasma and blood. It also showed an increase in the anisotropy of AB670 with increase in HSA concentration in the presence of red blood cells. These results indicate that qualitative and quantitative fluorescence measurements can be made directly in blood without the need to process the blood.

Substance P Depolarizes Lamprey Spinal Cord Neurons by Inhibiting Background Potassium Channels

PubMed Central

Thörn Pérez, Carolina; Hill, Russell H.; Grillner, Sten

2015-01-01

Substance P is endogenously released in the adult lamprey spinal cord and accelerates the burst frequency of fictive locomotion. This is achieved by multiple effects on interneurons and motoneurons, including an attenuation of calcium currents, potentiation of NMDA currents and reduction of the reciprocal inhibition. While substance P also depolarizes spinal cord neurons, the underlying mechanism has not been resolved. Here we show that effects of substance P on background K+ channels are the main source for this depolarization. Hyperpolarizing steps induced inward currents during whole-cell voltage clamp that were reduced by substance P. These background K+ channels are pH sensitive and are selectively blocked by anandamide and AVE1231. These blockers counteracted the effect of substance P on these channels and the resting membrane potential depolarization in spinal cord neurons. Thus, we have shown now that substance P inhibits background K+ channels that in turn induce depolarization, which is likely to contribute to the frequency increase observed with substance P during fictive locomotion. PMID:26197458

[Ionic mechanisms of depolarization responses induced by glutamate application to nerve cells of Helix pomatia].

PubMed

Gerasimov, V D

1982-01-01

The reversal potentials for transmembrane ionic currents induced by glutamate were measured in different D-neurons of the snail Helix pomatia. The first group of neurons had a mean reversal potential--10.6 +/- 1.2 mV and the second one--40.0 +/- 0.6 mV. Under normal conditions glutamate evoked spike discharges in the first group of neurons but not in the second one. At higher concentrations of glutamate the amplitude of D-responses in the latter group increased only to a certain level, not reaching the critical level for cell firing. Decrease in external Cl concentration led to a shift of their reversal potential in depolarizing direction. Ionic mechanisms of depolarizing responses induced by glutamate in these groups of neurons are discussed.

3D fluorescence anisotropy imaging using selective plane illumination microscopy.

PubMed

Hedde, Per Niklas; Ranjit, Suman; Gratton, Enrico

2015-08-24

Fluorescence anisotropy imaging is a popular method to visualize changes in organization and conformation of biomolecules within cells and tissues. In such an experiment, depolarization effects resulting from differences in orientation, proximity and rotational mobility of fluorescently labeled molecules are probed with high spatial resolution. Fluorescence anisotropy is typically imaged using laser scanning and epifluorescence-based approaches. Unfortunately, those techniques are limited in either axial resolution, image acquisition speed, or by photobleaching. In the last decade, however, selective plane illumination microscopy has emerged as the preferred choice for three-dimensional time lapse imaging combining axial sectioning capability with fast, camera-based image acquisition, and minimal light exposure. We demonstrate how selective plane illumination microscopy can be utilized for three-dimensional fluorescence anisotropy imaging of live cells. We further examined the formation of focal adhesions by three-dimensional time lapse anisotropy imaging of CHO-K1 cells expressing an EGFP-paxillin fusion protein.

RGK protein-mediated impairment of slow depolarization- dependent Ca2+ entry into developing myotubes

PubMed Central

Romberg, Christin F; Beqollari, Donald; Meza, Ulises; Bannister, Roger A

2014-01-01

Three physiological functions have been described for the skeletal muscle 1,4-dihydropyridine receptor (CaV1.1): (1) voltage-sensor for excitation-contraction (EC) coupling, (2) L-type Ca2+ channel, and (3) voltage-sensor for slow depolarization-dependent Ca2+ entry. Members of the RGK (Rad, Rem, Rem2, Gem/Kir) family of monomeric GTP-binding proteins are potent inhibitors of the former two functions of CaV1.1. However, it is not known whether the latter function that has been attributed to CaV1.1 is subject to modulation by RGK proteins. Thus, the purpose of this study was to determine whether Rad, Gem and/or Rem inhibit the slowly developing, persistent Ca2+ entry that is dependent on the voltage-sensing capability of CaV1.1. As a means to investigate this question, Venus fluorescent protein-fused RGK proteins (V-Rad, V-Rem and V-Gem) were overexpressed in “normal” mouse myotubes. We observed that such overexpression of V-Rad, V-Rem or V-Gem in myotubes caused marked changes in morphology of the cells. As shown previously for YFP-Rem, both L-type current and EC coupling were also impaired greatly in myotubes expressing either V-Rad or V-Gem. The reductions in L-type current and EC coupling were paralleled by reductions in depolarization-induced Ca2+ entry. Our observations provide the first evidence of modulation of this enigmatic Ca2+ entry pathway peculiar to skeletal muscle. PMID:24476902

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.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Hongtao; Salthouse, Christopher D., E-mail: salthouse@ecs.umass.edu; Center for Personalized Health Monitoring, University of Massachusetts, Amherst, Massachusetts 01003

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 themore » 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.« less

The continuum of spreading depolarizations in acute cortical lesion development: Examining Leão’s legacy

PubMed Central

Shuttleworth, C William; Kirov, Sergei A; Ayata, Cenk; Hinzman, Jason M; Foreman, Brandon; Andrew, R David; Boutelle, Martyn G; Brennan, KC; Carlson, Andrew P; Dahlem, Markus A; Drenckhahn, Christoph; Dohmen, Christian; Fabricius, Martin; Farkas, Eszter; Feuerstein, Delphine; Graf, Rudolf; Helbok, Raimund; Lauritzen, Martin; Major, Sebastian; Oliveira-Ferreira, Ana I; Richter, Frank; Rosenthal, Eric S; Sakowitz, Oliver W; Sánchez-Porras, Renán; Santos, Edgar; Schöll, Michael; Strong, Anthony J; Urbach, Anja; Westover, M Brandon; Winkler, Maren KL; Witte, Otto W; Woitzik, Johannes; Dreier, Jens P

2016-01-01

A modern understanding of how cerebral cortical lesions develop after acute brain injury is based on Aristides Leão’s historic discoveries of spreading depression and asphyxial/anoxic depolarization. Treated as separate entities for decades, we now appreciate that these events define a continuum of spreading mass depolarizations, a concept that is central to understanding their pathologic effects. Within minutes of acute severe ischemia, the onset of persistent depolarization triggers the breakdown of ion homeostasis and development of cytotoxic edema. These persistent changes are diagnosed as diffusion restriction in magnetic resonance imaging and define the ischemic core. In delayed lesion growth, transient spreading depolarizations arise spontaneously in the ischemic penumbra and induce further persistent depolarization and excitotoxic damage, progressively expanding the ischemic core. The causal role of these waves in lesion development has been proven by real-time monitoring of electrophysiology, blood flow, and cytotoxic edema. The spreading depolarization continuum further applies to other models of acute cortical lesions, suggesting that it is a universal principle of cortical lesion development. These pathophysiologic concepts establish a working hypothesis for translation to human disease, where complex patterns of depolarizations are observed in acute brain injury and appear to mediate and signal ongoing secondary damage. PMID:27328690

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.

Dynamic measurement of fluorescent proteins spectral distribution on virus infected cells

NASA Astrophysics Data System (ADS)

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

2006-09-01

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

Excitation anisotropy in laser-induced-fluorescence spectroscopy: Broad-line excitation case

NASA Astrophysics Data System (ADS)

Hirabayashi, A.; Nambu, Y.; Fujimoto, T.

1986-01-01

Treatment of excitation anisotropy for Laser-Induced-Fluorescence Spectroscopy (LIFS) is extended to the intense excitation case. The depolarization coefficient is derived for intense excitation limit (linearly-polarized or unpolarized light excitation), and the result is presented in tables. For the region of intermediate intensity between the weak and intense excitation limits, the master equation is solved for specific example of transitions and its result is compared with experiment.

Tachykininergic slow depolarization of motoneurones evoked by descending fibres in the neonatal rat spinal cord.

PubMed Central

Kurihara, T; Yoshioka, K; Otsuka, M

1995-01-01

1. In the isolated spinal cord of the neonatal rat, repetitive electrical stimulation of the upper cervical region elicited a prolonged depolarization of lumbar motoneurones (L3-5) lasting 1-2 min, which was recorded extracellularly from ventral roots, or intracellularly. 2. This depolarizing response was markedly depressed by the excitatory amino acid receptor antagonists D-(-)-2-amino-5-phosphonovaleric acid (D-APV, 30 microM) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM). The remaining response was further depressed by a 5-hydroxytryptamine (5-HT) receptor antagonist, ketanserin (3 microM). 3. In the presence of these antagonists, a small part of the depolarizing response of slow time course remained, and this response was partially blocked by the tachykinin NK1 receptor antagonists GR71251 (0.3-5 microM) and RP67580 (0.3-1 microM). In contrast, RP68651 (0.3-1 microM), the inactive enantiomer of RP67580, had no effect on the depolarizing response. 4. The slow depolarizing response in the presence of D-APV, CNQX and ketanserin was markedly potentiated by a peptidase inhibitor, thiorphan (1 microM). 5. This descending fibre-evoked slow depolarization became smaller after prolonged treatment (5-7 h) with 5,7-dihydroxytryptamine (10 microM), a neurotoxin for 5-HT neurones. Under such conditions, the effects of thiorphan and GR71251 on the slow depolarization were virtually absent. 6. Under the action of D-APV, CNQX and ketanserin, applications of tachykinins, substance P and neurokinin A produced depolarizing responses of lumbar motoneurones, and the responses were depressed by GR71251 and potentiated by thiorphan. 7. These results suggest that tachykinins contained in serotonergic fibres serve as neurotransmitters mediating the descending fibre-evoked slow excitatory postsynaptic potentials in motoneurones. PMID:7562617

Tachykininergic slow depolarization of motoneurones evoked by descending fibres in the neonatal rat spinal cord.

PubMed

Kurihara, T; Yoshioka, K; Otsuka, M

1995-06-15

1. In the isolated spinal cord of the neonatal rat, repetitive electrical stimulation of the upper cervical region elicited a prolonged depolarization of lumbar motoneurones (L3-5) lasting 1-2 min, which was recorded extracellularly from ventral roots, or intracellularly. 2. This depolarizing response was markedly depressed by the excitatory amino acid receptor antagonists D-(-)-2-amino-5-phosphonovaleric acid (D-APV, 30 microM) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM). The remaining response was further depressed by a 5-hydroxytryptamine (5-HT) receptor antagonist, ketanserin (3 microM). 3. In the presence of these antagonists, a small part of the depolarizing response of slow time course remained, and this response was partially blocked by the tachykinin NK1 receptor antagonists GR71251 (0.3-5 microM) and RP67580 (0.3-1 microM). In contrast, RP68651 (0.3-1 microM), the inactive enantiomer of RP67580, had no effect on the depolarizing response. 4. The slow depolarizing response in the presence of D-APV, CNQX and ketanserin was markedly potentiated by a peptidase inhibitor, thiorphan (1 microM). 5. This descending fibre-evoked slow depolarization became smaller after prolonged treatment (5-7 h) with 5,7-dihydroxytryptamine (10 microM), a neurotoxin for 5-HT neurones. Under such conditions, the effects of thiorphan and GR71251 on the slow depolarization were virtually absent. 6. Under the action of D-APV, CNQX and ketanserin, applications of tachykinins, substance P and neurokinin A produced depolarizing responses of lumbar motoneurones, and the responses were depressed by GR71251 and potentiated by thiorphan. 7. These results suggest that tachykinins contained in serotonergic fibres serve as neurotransmitters mediating the descending fibre-evoked slow excitatory postsynaptic potentials in motoneurones.

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

PubMed

Feng, Ying; Huang, Shi-hua

2007-12-01

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

Two regulatory RNA elements affect TisB-dependent depolarization and persister formation.

PubMed

Berghoff, Bork A; Hoekzema, Mirthe; Aulbach, Lena; Wagner, E Gerhart H

2017-03-01

Bacterial survival strategies involve phenotypic diversity which is generated by regulatory factors and noisy expression of effector proteins. The question of how bacteria exploit regulatory RNAs to make decisions between phenotypes is central to a general understanding of these universal regulators. We investigated the TisB/IstR-1 toxin-antitoxin system of Escherichia coli to appreciate the role of the RNA antitoxin IstR-1 in TisB-dependent depolarization of the inner membrane and persister formation. Persisters are phenotypic variants that have become transiently drug-tolerant by arresting growth. The RNA antitoxin IstR-1 sets a threshold for TisB-dependent depolarization under DNA-damaging conditions, resulting in two sub-populations: polarized and depolarized cells. Furthermore, our data indicate that an inhibitory 5' UTR structure in the tisB mRNA serves as a regulatory RNA element that delays TisB translation to avoid inappropriate depolarization when DNA damage is low. Investigation of the persister sub-population further revealed that both regulatory RNA elements affect persister levels as well as persistence time. This work provides an intriguing example of how bacteria exploit regulatory RNAs to control phenotypic heterogeneity. © 2016 John Wiley & Sons Ltd.

Epi-illumination optical design for fluorescence polarization measurements in flow systems.

PubMed Central

Eisert, W G; Beisker, W

1980-01-01

An epi-illumination design for fluorescence polarization measurements is introduced in flow cytometry with the optical axis orthogonally aligned to the cell stream. Various optical components and designs are discussed with respect to their influence on polarization measurements. Using the epi-configuration, paired measurements with the direction of polarization of the exciting light changed orthogonally are proposed for the compensation of system anisotropies and electronic mismatch. Large aperture corrections are employed for the excitation as well as for the emission pathway. Additional parameters such as fluorescence at 90 degrees, multiangle light scattering, and high precision cell-sizing by internally calibrated time of the flight measurements, as described previously, remain available with the design proposed here. Fluorescent latex microspheres, stained intracellular DNA, and algae have been used to test performance. PMID:7023562

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.

Inhibitory phosphorylation of GSK-3 by CaMKII couples depolarization to neuronal survival.

PubMed

Song, Bin; Lai, Bingquan; Zheng, Zhihao; Zhang, Yuying; Luo, Jingyan; Wang, Chong; Chen, Yuan; Woodgett, James R; Li, Mingtao

2010-12-24

Glycogen synthase kinase-3 (GSK-3) plays a critical role in neuronal apoptosis. The two mammalian isoforms of the kinase, GSK-3α and GSK-3β, are inhibited by phosphorylation at Ser-21 and Ser-9, respectively. Depolarization, which is vital for neuronal survival, causes both an increase in Ser-21/9 phosphorylation and an inhibition of GSK-3α/β. However, the role of GSK-3 phosphorylation in depolarization-dependent neuron survival and the signaling pathway contributing to GSK-3 phosphorylation during depolarization remain largely unknown. Using several approaches, we showed that both isoforms of GSK-3 are important for mediating neuronal apoptosis. Nonphosphorylatable GSK-3α/β mutants (S21A/S9A) promoted apoptosis, whereas a peptide encompassing Ser-9 of GSK-3β protected neurons in a phosphorylation-dependent manner; these results indicate a critical role for Ser-21/9 phosphorylation on depolarization-dependent neuron survival. We found that Ser-21/9 phosphorylation of GSK-3 was mediated by Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) but not by Akt/PKB, PKA, or p90(RSK). CaMKII associated with and phosphorylated GSK-3α/β. Furthermore, the pro-survival effect of CaMKII was mediated by GSK-3 phosphorylation and inactivation. These findings identify a novel Ca(2+)/calmodulin/CaMKII/GSK-3 pathway that couples depolarization to neuronal survival.

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.

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.

Stability and morphology of Ag nanoplatelets probed by depolarized dynamic light scattering

NASA Astrophysics Data System (ADS)

Zimbone, M.; Contino, A.; Maccarrone, G.; Musumeci, P.; Lo Faro, M. J.; Calcagno, L.

2018-06-01

The stability of silver nanoplatelet (NP) suspensions prepared with different concentrations of trisodium citrate (TSC) was studied by depolarized dynamic light scattering (DDLS) and UV–vis spectrometry. The morphology of the nanoparticles, as well as the color and stability of the sols, are tuned by the concentration of the capping agent. The nanoparticles prepared with high TSC concentration (>10‑4 M) are blue triangular NPs showing a slight truncation of the tips with aging. When low TSC concentrations are used, the color of the sols changes from blue to yellow with aging time and a strong modification of the morphology occurs: the nanoparticle shape changes from triangular to spherical. Remarkably, they show a high degree of anisotropy. The aging process was followed by the UV–vis spectra and by measuring the rotational diffusion coefficient by DDLS, providing information on the nanoparticle size and shape evolution. The high intensity of depolarized signal and the high value of rotational diffusion coefficient suggest that the aging process increases the thickness and the roughness of the nanoparticles

Origin and voltage dependence of asparagine-induced depolarization in intestinal cells of Xenopus embryo.

PubMed Central

Bergman, C; Bergman, J

1985-01-01

The kinetics and voltage dependence of asparagine (Asn)-induced depolarization in endoderm cells from Xenopus laevis embryos were analysed using current-clamp techniques. The depolarization is assumed to reflect the activation of an amino acid membrane carrier; it is accompanied by a slight increase in membrane resistance and cannot be explained by only the electrogenic character of the Asn carrier. It is proposed that the Asn depolarization arises, at least in part, from the decrease of the permeability ratio PK/PNa indirectly associated with the Na-coupled amino acid uptake. At room temperature (20-23 degrees C) the Asn response develops according to a single exponential function whose time constant is correlated with the final level of depolarization. Both amplitude and rise time of the depolarization are sensitive to variations of membrane potential and changes in Asn or Na external concentrations. Lowering the temperature decreases the amplitude of the Asn depolarization and increases its rise time with a Q10 factor of two; the kinetics remain of the Michaelis-Menten type, with a marked decrease in delta Emax and no change in Km. When the holding potential is altered by depolarizing and hyperpolarizing currents, the Asn response varies according to a bell-shaped characteristic presenting an optimum near the normal resting level. Membrane depolarizations induced by Na/K-pump inhibitors or high external K concentrations reduce the size of the Asn response; repolarizing the cell by current injection does not reverse the inhibitory effect of external K ions. Hyperpolarizing the membrane with a K-free Ringer solution increases the amplitude of the Asn response. In all these cases a decrease in delta Emax accounts for the apparent voltage sensitivity of the carrier mechanism. When induced by alterations of [K]o, an additional change in Km is observed, suggesting a K/Na-competitive inhibition of the Asn carrier. The results are discussed in terms of the amino acid

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.

Homogenization via the strong-permittivity-fluctuation theory with nonzero depolarization volume

NASA Astrophysics Data System (ADS)

Mackay, Tom G.

2004-08-01

The depolarization dyadic provides the scattering response of a single inclusion particle embedded within a homogenous background medium. These dyadics play a central role in formalisms used to estimate the effective constitutive parameters of homogenized composite mediums (HCMs). Conventionally, the inclusion particle is taken to be vanishingly small; this allows the pointwise singularity of the dyadic Green function associated with the background medium to be employed as the depolarization dyadic. A more accurate approach is pursued in this communication by taking into account the nonzero spatial extent of inclusion particles. Depolarization dyadics corresponding to inclusion particles of nonzero volume are incorporated within the strong-permittivity-fluctuation theory (SPFT). The linear dimensions of inclusion particles are assumed to be small relative to the electromagnetic wavelength(s) and the SPFT correlation length. The influence of the size of inclusion particles upon SPFT estimates of the HCM constitutive parameters is investigated for anisotropic dielectric HCMs.In particular, the interplay between correlation length and inclusion size is explored.

Hypothermia for Patients Requiring Evacuation of Subdural Hematoma: Effect on Spreading Depolarizations

DTIC Science & Technology

2017-10-01

AWARD NUMBER: W81XWH-16-C-0161 TITLE: Hypothermia for Patients Requiring Evacuation of Subdural Hematoma: Effect on Spreading Depolarizations...4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER W81XWH-16-C-0161 Hypothermia for Patients Requiring Evacuation of Subdural Hematoma: Effect on Spreading...in a sub-study of the HOPES trial to assess the effects of hypothermia on the pathologic mechanism of spreading depolarizations (SD). HOPES is a

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.

Permissive effect of dexamethasone on the increase of proenkephalin mRNA induced by depolarization of chromaffin cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Naranjo, J.R.; Mocchetti, I.; Schwartz, J.P.

1986-03-01

In cultured bovine chromaffin cells, changes in the dynamic state of enkephalin stores elicited experimentally were studied by measuring cellular proenkephalin mRNA, as well as enkephalin precursors and authentic enkephalin content of cells and culture media. In parallel, tyrosine hydroxylase mRNA and catecholamine cell content were also determined. Low concentrations (0.5-100 pM) of dexamethasone increased the cell contents of proenkephalin mRNA and enkephalin-containing peptides. High concentrations of the hormone(1 ..mu..M) were required to increase the cell contents of tyrosine hydroxylase mRNA and catecholamines. Depolarization of the cells with 10 ..mu..M veratridine resulted in a depletion of enkephalin and catecholamine storesmore » after 24 hr. The enkephalin, but not the catecholamine, content was restored by 48 hr. An increase in proenkephalin mRNA content might account for the recovery; this increase was curtailed by tetrodotoxin and enhanced by 10 pM dexamethasone. Tyrosine hydroxylase mRNA content was not significantly modified by depolarization, even in the presence of 1 ..mu..M dexamethasone. Aldosterone, progesterone, testosterone, or estradiol (1 ..mu..M) failed to change proenkephalin mRNA. Hence, dexamethasone appears to exert a specific permissive action on the stimulation of the proenkephalin gene elicited by depolarization. Though the catecholamines and enkephalins are localized in the same chromaffin granules and are coreleased by depolarization, the genes coding for the processes that are rate limiting in the production of these neuromodulators can be differentially regulated.« less

Endogenous cannabinoids mediate retrograde signals from depolarized postsynaptic neurons to presynaptic terminals.

PubMed

Ohno-Shosaku, T; Maejima, T; Kano, M

2001-03-01

Endogenous cannabinoids are considered to function as diffusible and short-lived modulators that may transmit signals retrogradely from postsynaptic to presynaptic neurons. To evaluate this possibility, we have made a paired whole-cell recording from cultured hippocampal neurons with inhibitory synaptic connections. In about 60% of pairs, a cannabinoid agonist greatly reduced the release of the inhibitory neurotransmitter GABA from presynaptic terminals. In most of such pairs but not in those insensitive to the agonist, depolarization of postsynaptic neurons and the resultant elevation of intracellular Ca2+ concentration caused transient suppression of inhibitory synaptic currents, which is mainly due to reduction of GABA release. This depolarization-induced suppression was completely blocked by selective cannabinoid antagonists. Our results reveal that endogenous cannabinoids mediate retrograde signals from depolarized postsynaptic neurons to presynaptic terminals to cause the reduction of transmitter release.

Mobility measurement by analysis of fluorescence photobleaching recovery kinetics.

PubMed Central

Axelrod, D; Koppel, D E; Schlessinger, J; Elson, E; Webb, W W

1976-01-01

Fluorescence photobleaching recovery (FPR) denotes a method for measuring two-dimensional lateral mobility of fluorescent particles, for example, the motion of fluorescently labeled molecules in approximately 10 mum2 regions of a single cell surface. A small spot on the fluorescent surface is photobleached by a brief exposure to an intense focused laser beam, and the subsequent recovery of the fluorescence is monitored by the same, but attenuated, laser beam. Recovery occurs by replenishment of intact fluorophore in the bleached spot by lateral transport from the surrounding surface. We present the theoretical basis and some practical guidelines for simple, rigorous analysis of FPR experiments. Information obtainable from FPR experiments includes: (a) identification of transport process type, i.e. the admixture of random diffusion and uniform directed flow; (b) determination of the absolute mobility coefficient, i.e. the diffusion constant and/or flow velocity; and (c) the fraction of total fluorophore which is mobile. To illustrate the experimental method and to verify the theory for diffusion, we describe some model experiments on aqueous solutions of rhodamine 6G. PMID:786399

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.

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.

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

PubMed Central

Digman, Michelle A.; Gratton, Enrico; Storti, Barbara; Beltram, Fabio

2013-01-01

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. PMID:22517076

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

NASA Astrophysics Data System (ADS)

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

2014-01-01

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

Possible effects of depolarizing GABAA conductance on the neuronal input-output relationship: a modeling study.

PubMed

Morita, Kenji; Tsumoto, Kunichika; Aihara, Kazuyuki

2005-06-01

Recent in vitro experiments revealed that the GABAA reversal potential is about 10 mV higher than the resting potential in mature mammalian neocortical pyramidal cells; thus GABAergic inputs could have facilitatory, rather than inhibitory, effects on action potential generation under certain conditions. However, how the relationship between excitatory input conductances and the output firing rate is modulated by such depolarizing GABAergic inputs under in vivo circumstances has not yet been understood. We examine herewith the input-output relationship in a simple conductance-based model of cortical neurons with the depolarized GABAA reversal potential, and show that a tonic depolarizing GABAergic conductance up to a certain amount does not change the relationship between a tonic glutamatergic driving conductance and the output firing rate, whereas a higher GABAergic conductance prevents spike generation. When the tonic glutamatergic and GABAergic conductances are replaced by in vivo-like highly fluctuating inputs, on the other hand, the effect of depolarizing GABAergic inputs on the input-output relationship critically depends on the degree of coincidence between glutamatergic input events and GABAergic ones. Although a wide range of depolarizing GABAergic inputs hardly changes the firing rate of a neuron driven by noncoincident glutamatergic inputs, a certain range of these inputs considerably decreases the firing rate if a large number of driving glutamatergic inputs are coincident with them. These results raise the possibility that the depolarized GABAA reversal potential is not a paradoxical mystery, but is instead a sophisticated device for discriminative firing rate modulation.

Precision analysis for standard deviation measurements of immobile single fluorescent molecule images.

PubMed

DeSantis, Michael C; DeCenzo, Shawn H; Li, Je-Luen; Wang, Y M

2010-03-29

Standard deviation measurements of intensity profiles of stationary single fluorescent molecules are useful for studying axial localization, molecular orientation, and a fluorescence imaging system's spatial resolution. Here we report on the analysis of the precision of standard deviation measurements of intensity profiles of single fluorescent molecules imaged using an EMCCD camera.We have developed an analytical expression for the standard deviation measurement error of a single image which is a function of the total number of detected photons, the background photon noise, and the camera pixel size. The theoretical results agree well with the experimental, simulation, and numerical integration results. Using this expression, we show that single-molecule standard deviation measurements offer nanometer precision for a large range of experimental parameters.

Ab Initio Calculations of Ultrashort Carrier Dynamics in Two-Dimensional Materials: Valley Depolarization in Single-Layer WSe2

NASA Astrophysics Data System (ADS)

Molina-Sánchez, Alejandro; Sangalli, Davide; Wirtz, Ludger; Marini, Andrea

2017-08-01

In single-layer WSe$_2$, a paradigmatic semiconducting transition metal dichalcogenide, a circularly polarized laser field can selectively excite electronic transitions in one of the inequivalent $K^{\\pm}$ valleys. Such selective valley population corresponds to a pseudospin polarization. This can be used as a degree of freedom in a valleytronic device provided that the time scale for its depolarization is sufficiently large. Yet, the mechanism behind the valley depolarization still remains heavily debated. Recent time-dependent Kerr experiments have provided an accurate way to visualize the valley dynamics by measuring the rotation of a linearly polarized probe pulse applied after a circularly polarized pump pulse. We present here a clear, accurate and parameter-free description of the valley dynamics. By using an atomistic, ab initio, approach we fully disclose the elemental mechanisms that dictate the depolarization effects. Our results are in excellent agreement with recent time-dependent Kerr experiments. We explain the Kerr dynamics and its temperature dependence in terms of electron-phonon mediated processes that induce spin-flip inter-valley transitions.

CaMKII-dependent endoplasmic reticulum fission by whisker stimulation and during cortical spreading depolarization.

PubMed

Kucharz, Krzysztof; Lauritzen, Martin

2018-04-01

Cortical spreading depolarization waves, the cause underlying migraine aura, are also the markers and mechanism of pathology in the acutely injured human brain. Propagation of spreading depolarization wave uniquely depends on the interaction between presynaptic and postsynaptic glutamate N-methyl-d-aspartate receptors (NMDARs). In the normally perfused brain, even a single wave causes a massive depolarization of neurons and glia, which results in transient loss of neuronal function and depression of the ongoing electrocorticographic activity. Endoplasmic reticulum is the cellular organelle of particular importance for modulation of neurotransmission. Neuronal endoplasmic reticulum structure is assumed to be persistently continuous in neurons, but is rapidly lost within 1 to 2 min of global cerebral ischaemia, i.e. the organelle disintegrates by fission. This phenomenon appears to be timed with the cardiac arrest-induced cortical spreading depolarizations, rather than ensuing cell death. To what extent NMDAR-dependent processes may trigger neuronal endoplasmic reticulum fission and whether fission is reversible in the normally perfused brain is unknown. We used two-photon microscopy to examine neuronal endoplasmic reticulum structural dynamics during whisker stimulation and cortical spreading depolarizations in vivo. Somatosensory stimulation triggered loss of endoplasmic reticulum continuity, a likely outcome of constriction and fission, in dendritic spines within less than 10 s of stimulation, which was spontaneously reversible and recovery to normal took 5 min. The endoplasmic reticulum fission was inhibited by blockade of NMDAR and Ca2+/calmodulin-dependent protein kinase II (CaMKII) activated downstream of the NMDARs, whereas inhibition of guanosine triphosphate hydrolases hindered regain of endoplasmic reticulum continuity, i.e. fusion. In contrast to somatosensory stimulation, endoplasmic reticulum fission during spreading depolarization was widespread and

Measuring Photosynthetic Response to Drought Stress using Active and Passive Fluorescence

NASA Astrophysics Data System (ADS)

Helm, L.; Lerdau, M.; Wang, W.; Yang, X.

2017-12-01

Photosynthesis, the endothermic reactions involving the absorption of light and fixation and reduction of carbon dioxide by plants, plays important roles in carbon and water cycles, food security, and even weather and climate patterns. Solar radiation provides the energy for photosynthesis, but often plants absorb more solar energy than they can use to reduce carbon dioxide. This excess energy, which is briefly stored as high-energy electrons in the chloroplast, must be removed or damage to the leaf's photosynthetic machinery will occur. One important energy dissipation pathway is for the high energy electrons to return to their lower valance state and, in doing so, release radiation (fluorescence). This fluorescence (known as solar induced fluorescence (SIF) has been found to strongly correlate with gross photosynthesis. Recent advances in the remote sensing of SIF allow for large-scale real-time estimation of photosynthesis. In a warming climate with more frequent stress, remote sensing is necessary for measuring the spatial and temporal variability of photosynthesis. However, the mechanisms that link SIF and photosynthesis are unclear, particularly how the relationship may or may not change under stress. We present data from leaf-level measurements of gas exchange, pulse amplitude modulation (PAM) fluorescence, and SIF in two major tree species in North America. Water-stressed and well-watered plants were compared to determine how SIF and carbon dioxide exchange are modulated by drought diurnally and seasonally. Secondly, photosynthesis and fluorescence under high and low oxygen concentrations were compared to determine how photorespiration alters the relationship between SIF and gross photosynthesis. We find a strong correlation between SIF and steady-state fluorescence measured with conventional PAM fluorometry. Our results also indicate that drought-stress modulates the SIF-photosynthesis relationship, and this may be driven by drought-induced changes in

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

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

NASA Technical Reports Server (NTRS)

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

1991-01-01

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.

Hyperpolarizing and age-dependent depolarizing responses of cultured locus coeruleus neurons to noradrenaline.

PubMed

Finlayson, P G; Marshall, K C

1984-08-01

The electrical activity and responses to noradrenaline (NA) of locus coeruleus (LC) neurons have been studied in organotypic cultures using intracellular recording. Most LC neurons were predominantly quiescent, though occasional bursts of activity were observed; a few cells were tonically active at rates of 0.5-5/s. In most cells tested, iontophoretic application of NA evoked responses which were initially hyperpolarizing, sometimes followed by a depolarizing phase and frequently followed by a period of increased excitatory synaptic activity. The enhanced synaptic activity appeared to be an indirect effect since it was blocked by bath application of tetrodotoxin (TTX). In the presence of TTX, responses to NA of all but one cell were simple hyperpolarizations or biphasic (hyperpolarization/depolarization) responses. The presence of the depolarizing component appeared to be age-dependent, since it was frequently observed in cultures grown in vitro for less than 26 days, while neurons in older cultures exhibited only hyperpolarizing responses. If such age-dependent depolarizing responses are present in vivo, they would represent a unique example of a transmitter response which is present only during a transient developmental phase.

Strong dependence of rain-induced lidar depolarization on the illumination angle: experimental evidence and geometrical-optics interpretation.

PubMed

Roy, G; Bissonnette, L R

2001-09-20

Backscatter and depolarization lidar measurements from clouds and precipitation are reported as functions of the elevation angle of the pointing lidar direction. We recorded the data by scanning the lidar beam (Nd:YAG) at a constant angular speed of ~3.5 degrees /s while operating at a repetition rate of 10 Hz. We show that in rain there is an evident and at times spectacular dependence on the elevation angle. That dependence appears to be sensitive to raindrop size. We have developed a three-dimensional polarization-dependent ray-tracing algorithm to calculate the backscatter and the depolarization ratio by large nonspherical droplets. We have applied it to raindrop shapes derived from existing static and dynamic (oscillating) models. We show that many of the observed complex backscatter and depolarization features can be interpreted to a good extent by geometrical optics. These results suggest that there is a definite need for more extensive calculations of the scattering phase matrix elements for large deformed raindrops as functions of the direction of illumination. Obvious applications are retrieval of information on the liquid-solid phase of precipitation and on the size and the vibration state of raindrops.

Remote temperature measurements in femto-liter volumes using dual-focus-Fluorescence Correlation Spectroscopy.

PubMed

Müller, Claus B; Weiss, Kerstin; Loman, Anastasia; Enderlein, Jörg; Richtering, Walter

2009-05-07

Remote temperature measurements in microfluidic devices with micrometer spatial resolution are important for many applications in biology, biochemistry and chemistry. The most popular methods use the temperature-dependent fluorescence lifetime of Rhodamine B, or the temperature-dependent size of thermosensitive materials such as microgel particles. Here, we use the recently developed method of dual-focus fluorescence correlation spectroscopy (2fFCS) for measuring the absolute diffusion coefficient of small fluorescent molecules at nanomolar concentrations and show how these data can be used for remote temperature measurements on a micrometer scale. We perform comparative temperature measurements using all three methods and show that the accuracy of 2fFCS is comparable or even better than that achievable with Rhodamine B fluorescence lifetime measurements. The temperature dependent microgel swelling leads to an enhanced accuracy within a narrow temperature range around the volume phase transition temperature, but requires the availability of specific microgels, whereas 2fFCS is applicable under very general conditions.

Mitochondrial aquaporin-8 knockdown in human hepatoma HepG2 cells causes ROS-induced mitochondrial depolarization and loss of viability

DOE Office of Scientific and Technical Information (OSTI.GOV)

Marchissio, Maria Julia; Francés, Daniel Eleazar Antonio; Carnovale, Cristina Ester

Human aquaporin-8 (AQP8) channels facilitate the diffusional transport of H{sub 2}O{sub 2} across membranes. Since AQP8 is expressed in hepatic inner mitochondrial membranes, we studied whether mitochondrial AQP8 (mtAQP8) knockdown in human hepatoma HepG2 cells impairs mitochondrial H{sub 2}O{sub 2} release, which may lead to organelle dysfunction and cell death. We confirmed AQP8 expression in HepG2 inner mitochondrial membranes and found that 72 h after cell transfection with siRNAs targeting two different regions of the human AQP8 molecule, mtAQP8 protein specifically decreased by around 60% (p < 0.05). Studies in isolated mtAQP8-knockdown mitochondria showed that H{sub 2}O{sub 2} release, assessedmore » by Amplex Red, was reduced by about 45% (p < 0.05), an effect not observed in digitonin-permeabilized mitochondria. mtAQP8-knockdown cells showed an increase in mitochondrial ROS, assessed by dichlorodihydrofluorescein diacetate (+ 120%, p < 0.05) and loss of mitochondrial membrane potential (− 80%, p < 0.05), assessed by tetramethylrhodamine-coupled quantitative fluorescence microscopy. The mitochondria-targeted antioxidant MitoTempol prevented ROS accumulation and dissipation of mitochondrial membrane potential. Cyclosporin A, a mitochondrial permeability transition pore blocker, also abolished the mtAQP8 knockdown-induced mitochondrial depolarization. Besides, the loss of viability in mtAQP8 knockdown cells verified by MTT assay, LDH leakage, and trypan blue exclusion test could be prevented by cyclosporin A. Our data on human hepatoma HepG2 cells suggest that mtAQP8 facilitates mitochondrial H{sub 2}O{sub 2} release and that its defective expression causes ROS-induced mitochondrial depolarization via the mitochondrial permeability transition mechanism, and cell death. -- Highlights: ► Aquaporin-8 is expressed in mitochondria of human hepatoma HepG2 cells. ► Aquaporin-8 knockdown impairs mitochondrial H{sub 2}O{sub 2} release and increases ROS. �

Cell-free measurements of brightness of fluorescently labeled antibodies

PubMed Central

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

2017-01-01

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

Short- and long-term functional plasticity of white matter induced by oligodendrocyte depolarization in the hippocampus.

PubMed

Yamazaki, Yoshihiko; Fujiwara, Hiroki; Kaneko, Kenya; Hozumi, Yasukazu; Xu, Ming; Ikenaka, Kazuhiro; Fujii, Satoshi; Tanaka, Kenji F

2014-08-01

Plastic changes in white matter have received considerable attention in relation to normal cognitive function and learning. Oligodendrocytes and myelin, which constitute the white matter in the central nervous system, can respond to neuronal activity with prolonged depolarization of membrane potential and/or an increase in the intracellular Ca(2+) concentration. Depolarization of oligodendrocytes increases the conduction velocity of an action potential along axons myelinated by the depolarized oligodendrocytes, indicating that white matter shows functional plasticity, as well as structural plasticity. However, the properties and mechanism of oligodendrocyte depolarization-induced functional plastic changes in white matter are largely unknown. Here, we investigated the functional plasticity of white matter in the hippocampus using mice with oligodendrocytes expressing channelrhodopsin-2. Using extracellular recordings of compound action potentials at the alveus of the hippocampus, we demonstrated that light-evoked depolarization of oligodendrocytes induced early- and late-onset facilitation of axonal conduction that was dependent on the magnitude of oligodendrocyte depolarization; the former lasted for approximately 10 min, whereas the latter continued for up to 3 h. Using whole-cell recordings from CA1 pyramidal cells and recordings of antidromic action potentials, we found that the early-onset short-lasting component included the synchronization of action potentials. Moreover, pharmacological analysis demonstrated that the activation of Ba(2+) -sensitive K(+) channels was involved in early- and late-onset facilitation, whereas 4-aminopyridine-sensitive K(+) channels were only involved in the early-onset component. These results demonstrate that oligodendrocyte depolarization induces short- and long-term functional plastic changes in the white matter of the hippocampus and plays active roles in brain functions. © 2014 Wiley Periodicals, Inc.

2D fluorescence spectra measurement of six kinds of bioagents simulants by short range Lidar

NASA Astrophysics Data System (ADS)

Sanpedro, Man

2018-02-01

Pantoea agglomerans (Pan), Staphylococcus aureus (Sta), Bacillus globigii (BG) and Escherichia coli (EH), these four kinds of bioagents simulants of were cultured and then their growth curves were measured, the generation time was 0.99h, 0.835h, 1.07h and 1.909h, respectively. A small short range fluorescence lidar working at wavelengths of 266nm and 355nm was designed and used to measure the two-dimensional fluorescence spectra of bioagents simulants in the amino acid segment and NADH segment, respectively. In a controllable fluorescence measurement chamber, the two-dimensional fluorescence spectra of vegetative liquid bacterial aerosols as well as BSA and OVA, two protein toxinic simulants were measured with a resolution of 4nm. The two-dimensional fluorescence spectral shape of Pan, Sta, EH and BG, BSA and OVA were consistent with the standard fluorescent component tryptophan in the amino acid band with FWHM of 60nm, but the central wavelength of the fluorescence spectra of these simulants blue/purple shifted obviously as affected by the external biochemical environment, concentration and ratio of different bacterial internal fluorophores, so the energy level between the excited state and the ground state of the fluorescence molecule increased. Differently, weak NADH fluorescence spectra with 100nm FWHM inside the four vegetative bacteria aerosols were detected, but Rayleigh scattering, Raman scattering contribution of water, nitrogen in the fluorescence spectra could not be effectively extracted. The second - order derivative fluorescence spectra of four simulants showed that the high - order processing and recognition of the fluorescence spectra was feasible.

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

NASA Astrophysics Data System (ADS)

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

2013-12-01

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

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. Copyright © 2013 International Society for Advancement of Cytometry.

Rain depolarization and attenuation measurements at 11.7, 19.04, and 28.56 GHz - A description of the experiment and some preliminary results

NASA Technical Reports Server (NTRS)

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

1977-01-01

The considered investigation made use of the Communications Technology Satellite (CTS) downlink and the beacons carried by the Comstar satellites. The general behavior of rain attenuation and depolarization is illustrated with the aid of data from a storm which took place on July 15, 1976. The effect of the rain on the copolarized signal is indicated in a graph. Another graph shows the behavior of the cross-polarized signal component. Phase effects are also considered together with statistical curves for attenuation. The considered data from CTS indicate that, at least during summer convective storms, attenuation at 11.7 GHz is much more severe than anticipated. Attenuation may be a more serious impediment to dual polarized satellite links at this frequency than is depolarization.

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.

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

Fluorescence errors in integrating sphere measurements of remote phosphor type LED light sources

NASA Astrophysics Data System (ADS)

Keppens, A.; Zong, Y.; Podobedov, V. B.; Nadal, M. E.; Hanselaer, P.; Ohno, Y.

2011-05-01

The relative spectral radiant flux error caused by phosphor fluorescence during integrating sphere measurements is investigated both theoretically and experimentally. Integrating sphere and goniophotometer measurements are compared and used for model validation, while a case study provides additional clarification. Criteria for reducing fluorescence errors to a degree of negligibility as well as a fluorescence error correction method based on simple matrix algebra are presented. Only remote phosphor type LED light sources are studied because of their large phosphor surfaces and high application potential in general lighting.

Site-specific multipoint fluorescence measurement system with end-capped optical fibers.

PubMed

Song, Woosub; Moon, Sucbei; Lee, Byoung-Cheol; Park, Chul-Seung; Kim, Dug Young; Kwon, Hyuk Sang

2011-07-10

We present the development and implementation of a spatially and spectrally resolved multipoint fluorescence correlation spectroscopy (FCS) system utilizing multiple end-capped optical fibers and an inexpensive laser source. Specially prepared end-capped optical fibers placed in an image plane were used to both collect fluorescence signals from the sample and to deliver signals to the detectors. The placement of independently selected optical fibers on the image plane was done by monitoring the end-capped fiber tips at the focus using a CCD, and fluorescence from specific positions of a sample were collected by an end-capped fiber, which could accurately represent light intensities or spectral data without incurring any disturbance. A fast multipoint spectroscopy system with a time resolution of ∼1.5 ms was then implemented using a prism and an electron multiplying charge coupled device with a pixel binning for the region of interest. The accuracy of our proposed system was subsequently confirmed by experimental results, based on an FCS analysis of microspheres in distilled water. We expect that the proposed multipoint site-specific fluorescence measurement system can be used as an inexpensive fluorescence measurement tool to study many intracellular and molecular dynamics in cell biology. © 2011 Optical Society of America

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.

Laser-induced fluorescence measurement of the oil film thickness in an internal combustion engine

NASA Astrophysics Data System (ADS)

Ostroski, Greg M.; Ghandhi, Jaal B.

1997-11-01

The use of a fluorescent dopant molecule to enhance the natural fluorescence of motor oils, and allow quantitative determination of temperature and film thickens in internal combustion engines has been investigated. Measurement of the fluorescence as a function of temperature were made with neat Mobil 1, and solutions of the dopant BTBP in mineral oil and Mobil 1. The fluorescence yield of neat Mobil 1 was found to vary by 30 percent over the temperature range explored, but the spectral characteristics, as measured with bandpass filters, were unaffected by temperature. The BTBP fluorescence was found to increase significantly with temperature, and it was found the narrower regions in the spectrum increased proportionally more than the fluorescence collected over the entire spectrum, allowing a determination of temperature to be made which can then be used to correct for the change in fluorescence yield. Solutions in Mobil 1 showed a smaller increase than that observed in mineral oil.

Quantum measurement and orientation tracking of fluorescent nanodiamonds inside living cells

NASA Astrophysics Data System (ADS)

McGuinness, L. P.; Yan, Y.; Stacey, A.; Simpson, D. A.; Hall, L. T.; MacLaurin, D.; Prawer, S.; Mulvaney, P.; Wrachtrup, J.; Caruso, F.; Scholten, R. E.; Hollenberg, L. C. L.

2011-06-01

Fluorescent particles are routinely used to probe biological processes. The quantum properties of single spins within fluorescent particles have been explored in the field of nanoscale magnetometry, but not yet in biological environments. Here, we demonstrate optically detected magnetic resonance of individual fluorescent nanodiamond nitrogen-vacancy centres inside living human HeLa cells, and measure their location, orientation, spin levels and spin coherence times with nanoscale precision. Quantum coherence was measured through Rabi and spin-echo sequences over long (>10 h) periods, and orientation was tracked with effective 1° angular precision over acquisition times of 89 ms. The quantum spin levels served as fingerprints, allowing individual centres with identical fluorescence to be identified and tracked simultaneously. Furthermore, monitoring decoherence rates in response to changes in the local environment may provide new information about intracellular processes. The experiments reported here demonstrate the viability of controlled single spin probes for nanomagnetometry in biological systems, opening up a host of new possibilities for quantum-based imaging in the life sciences.

Characterization of ventricular depolarization and repolarization changes in a porcine model of myocardial infarction.

PubMed

Romero, Daniel; Ringborn, Michael; Demidova, Marina; Koul, Sasha; Laguna, Pablo; Platonov, Pyotr G; Pueyo, Esther

2012-12-01

In this study, several electrocardiogram (ECG)-derived indices corresponding to both ventricular depolarization and repolarization were evaluated during acute myocardial ischemia in an experimental model of myocardial infarction produced by 40 min coronary balloon inflation in 13 pigs. Significant changes were rapidly observed from minute 4 after the start of coronary occlusion, achieving their maximum values between 11 and 22 min for depolarization and between 9 and 12 min for repolarization indices, respectively. Subsequently, these maximum changes started to decrease during the latter part of the occlusion. Depolarization changes associated with the second half of the QRS complex showed a significant but inverse correlation with the myocardium at risk (MaR) estimated by scintigraphic images. The correlation between MaR and changes of the downward slope of the QRS complex, [Formula: see text], evaluated at the two more relevant peaks observed during the occlusion, was r = -0.75, p < 0.01 and r = -0.79, p < 0.01 for the positive and negative deflections observed in [Formula: see text], temporal evolution, respectively. Repolarization changes, analyzed by evaluation of ST segment elevation at the main observed positive peak, also showed negative, however non-significant correlation with MaR: r = -0.34, p = 0.28. Our results suggest that changes evaluated in the latter part of the depolarization, such as those described by [Formula: see text], which are influenced by R-wave amplitude, QRS width and ST level variations simultaneously, correlate better with the amount of ischemia than other indices evaluated in the earlier part of depolarization or during the ST segment.

Geometry of generalized depolarizing channels

NASA Astrophysics Data System (ADS)

Burrell, Christian K.

2009-10-01

A generalized depolarizing channel acts on an N -dimensional quantum system to compress the “Bloch ball” in N2-1 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=2d (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.

Caffeine does not affect susceptibility to cortical spreading depolarization in mice.

PubMed

Yalcin, Nilufer; Chen, Shih-Pin; Yu, Esther S; Liu, Tzu-Ting; Yen, Jiin-Cherng; Atalay, Yahya B; Qin, Tao; Celik, Furkan; van den Maagdenberg, Arn Mjm; Moskowitz, Michael A; Ayata, Cenk; Eikermann-Haerter, Katharina

2018-01-01

Several factors that modulate migraine, a common primary headache disorder, also affect susceptibility to cortical spreading depolarization (CSD). CSD is a wave of neuronal and glial depolarization and thought to underlie the migraine aura and possibly headache. Here, we tested whether caffeine, known to alleviate or trigger headache after acute exposure or chronic use/withdrawal, respectively, modulates CSD. We injected C57BL/6J mice with caffeine (30, 60, or 120 mg/kg; i.p.) once (acute) or twice per day for one or two weeks (chronic). Susceptibility to CSD was evaluated by measuring the electrical CSD threshold and by assessing KCl-induced CSD. Simultaneous laser Doppler flowmetry was used to assess CSD-induced cortical blood flow changes. Recordings were performed 15 min after caffeine/vehicle administration, or 24 h after the last dose of chronic caffeine in the withdrawal group. The latter paradigm was also tested in mice carrying the familial hemiplegic migraine type 1 R192Q missense mutation, considered a valid migraine model. Neither acute/chronic administration nor withdrawal of caffeine affected CSD susceptibility or related cortical blood flow changes, either in WT or R192Q mice. Hence, adverse or beneficial effects of caffeine on headache seem unrelated to CSD pathophysiology, consistent with the non-migrainous clinical presentation of caffeine-related headache.

Kv1.1 channelopathy abolishes presynaptic spike width modulation by subthreshold somatic depolarization

PubMed Central

Vivekananda, Umesh; Novak, Pavel; Bello, Oscar D.; Korchev, Yuri E.; Krishnakumar, Shyam S.; Volynski, Kirill E.; Kullmann, Dimitri M.

2017-01-01

Although action potentials propagate along axons in an all-or-none manner, subthreshold membrane potential fluctuations at the soma affect neurotransmitter release from synaptic boutons. An important mechanism underlying analog–digital modulation is depolarization-mediated inactivation of presynaptic Kv1-family potassium channels, leading to action potential broadening and increased calcium influx. Previous studies have relied heavily on recordings from blebs formed after axon transection, which may exaggerate the passive propagation of somatic depolarization. We recorded instead from small boutons supplied by intact axons identified with scanning ion conductance microscopy in primary hippocampal cultures and asked how distinct potassium channels interact in determining the basal spike width and its modulation by subthreshold somatic depolarization. Pharmacological or genetic deletion of Kv1.1 broadened presynaptic spikes without preventing further prolongation by brief depolarizing somatic prepulses. A heterozygous mouse model of episodic ataxia type 1 harboring a dominant Kv1.1 mutation had a similar broadening effect on basal spike shape as deletion of Kv1.1; however, spike modulation by somatic prepulses was abolished. These results argue that the Kv1.1 subunit is not necessary for subthreshold modulation of spike width. However, a disease-associated mutant subunit prevents the interplay of analog and digital transmission, possibly by disrupting the normal stoichiometry of presynaptic potassium channels. PMID:28193892

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

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.

The anomalous depolarization anisotropy in the central backscattering area for turbid medium with Mie scatterers

NASA Astrophysics Data System (ADS)

Wang, Xuezhen; Lai, Jiancheng; Song, Yang; Li, Zhenhua

2018-05-01

It is generally recognized that circularly polarized light is preferentially maintained over linearly polarized light in turbid medium with Mie scatterers. However, in this work, the anomalous depolarization anisotropy is reported in the backscattering area near the point of illumination. Both experimental and Monte Carlo simulations show preferential retention of linear polarization states compared to circular polarization states in a specific backscattering area. Further analysis indicates that the anomalous depolarization behavior in the specific area is induced by lateral scattering events, which own low circular polarization memory. In addition, it is also found that the size of the anomalous depolarization area is related to the transport mean free path of the turbid medium.

Capacitance measurements of regulated exocytosis in mouse taste cells.

PubMed

Vandenbeuch, Aurelie; Zorec, Robert; Kinnamon, Sue C

2010-11-03

Exocytosis, consisting of the merger of vesicle and plasma membrane, is a common mechanism used by different types of nucleated cells to release their vesicular contents. Taste cells possess vesicles containing various neurotransmitters to communicate with adjacent taste cells and afferent nerve fibers. However, whether these vesicles engage in exocytosis on a stimulus is not known. Since vesicle membrane merger with the plasma membrane is reflected in plasma membrane area fluctuations, we measured membrane capacitance (C(m)), a parameter linearly related to membrane surface area. To investigate whether taste cells undergo regulated exocytosis, we used the compensated tight-seal whole-cell recording technique to monitor depolarization-induced changes in C(m) in the different types of taste cells. To identify taste cell types, mice expressing green fluorescent protein from the TRPM5 promoter or from the GAD67 promoter were used to discriminate type II and type III taste cells, respectively. Moreover, the cell types were also identified by monitoring their voltage-current properties. The results demonstrate that only type III taste cells show significant depolarization-induced increases in C(m), which were correlated to the voltage-activated calcium currents. The results suggest that type III, but neither type II nor type I cells exhibit depolarization-induced regulated exocytosis to release transmitter and activate gustatory afferent nerve fibers.

Instantaneous temperature field measurements using planar laser-induced fluorescence.

PubMed

Seitzman, J M; Kychakoff, G; Hanson, R K

1985-09-01

A single-pulse, laser-induced-fluorescence diagnostic for the measurement of two-dimensional temperature fields in combustion flows is described. The method uses sheet illumination from a tunable laser to excite planar laserinduced fluorescence in a stable tracer molecule, seeded at constant mole fraction into the flow field. The temporal resolution of this technique is determined by the laser pulse length. Experimental results are presented for a rodstabilized, premixed methane-air flame, using the Q(1) (22) line of the nitric oxide A(2) Sigma(+) (v = 0) ? X(2)II((1/2))(v = 0) transition (lambda approximately 225.6 nm).

MiniFluo fluorescence sensor, advances in FDOM Ocean Measurements

NASA Astrophysics Data System (ADS)

Cyr, Frédéric; Tedetti, Marc; Goutx, Madeleine

2017-04-01

As part of the European project "Next generation Low-Cost Multifunctional Web Enabled Ocean Sensor Systems Empowering Marine, Maritime and Fisheries Management (NeXOS)", we developed the MiniFluo, a glider-compatible optical sensor for measurements of fluorescent dissolved organic matter (FDOM). In situ applications of the MiniFluo are presented here. The configuration used targets both natural (Tryptophan) and an anthropogenic (Phenanthrene) DOM fluorophores. Observations from three glider campaigns in the NW Mediterranean (Fall 2015 and Spring and Summer 2016) are presented. It is shown that the use of the Minifluo highlights new features of DOM dynamics in the region. For example, the Tryptophan (an amino-acid traditionally used as a tracer for waste waters) is found here closely related to open sea Chl-a fluorescence. Differences between Chl-a and Tryptophan fluorescence also give subtle information on seasonal changes in ecosystem structure and DOM release that could not be observed with traditional glider measurements. The study also highlights the presence of phenanthrene (an anthropogenic polycyclic aromatic hydrocarbon (PAH) in the surface and sub-surface waters of the Mediterranean. Implications of these finding will be put in the context of both the Mediterranean Sea DOM dynamics and also the ocean carbon cycle, from which the Dissolved Organic Carbon pool remains qualitatively unknown.

Characterization of two distinct depolarization-activated K+ currents in isolated adult rat ventricular myocytes

PubMed Central

1991-01-01

Depolarization-activated outward K+ currents in isolated adult rat ventricular myocytes were characterized using the whole-cell variation of the patch-clamp recording technique. During brief depolarizations to potentials positive to -40 mV, Ca(2+)-independent outward K+ currents in these cells rise to a transient peak, followed by a slower decay to an apparent plateau. The analyses completed here reveal that the observed outward current waveforms result from the activation of two kinetically distinct voltage-dependent K+ currents: one that activates and inactivates rapidly, and one that activates and inactivates slowly, on membrane depolarization. These currents are referred to here as Ito (transient outward) and IK (delayed rectifier), respectively, because their properties are similar (although not identical) to these K+ current types in other cells. Although the voltage dependences of Ito and IK activation are similar, Ito activates approximately 10-fold and inactivates approximately 30-fold more rapidly than IK at all test potentials. In the composite current waveforms measured during brief depolarizations, therefore, the peak current predominantly reflects Ito, whereas IK is the primary determinant of the plateau. There are also marked differences in the voltage dependences of steady-state inactivation of these two K+ currents: IK undergoes steady-state inactivation at all potentials positive to -120 mV, and is 50% inactivated at -69 mV; Ito, in contrast, is insensitive to steady-state inactivation at membrane potentials negative to -50 mV. In addition, Ito recovers from steady-state inactivation faster than IK: at -90 mV, for example, approximately 70% recovery from the inactivation produced at -20 mV is observed within 20 ms for Ito; IK recovers approximately 25-fold more slowly. The pharmacological properties of Ito and IK are also distinct: 4-aminopyridine preferentially attenuates Ito, and tetraethylammonium suppresses predominantly IK. The voltage- and

Geometry of generalized depolarizing channels

DOE Office of Scientific and Technical Information (OSTI.GOV)

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 formsmore » a simplex.« less

Recovery of intrinsic fluorescence from single-point interstitial measurements for quantification of doxorubicin concentration.

PubMed

Baran, Timothy M; Foster, Thomas H

2013-10-01

We developed a method for the recovery of intrinsic fluorescence from single-point measurements in highly scattering and absorbing samples without a priori knowledge of the sample optical properties. The goal of the study was to demonstrate accurate recovery of fluorophore concentration in samples with widely varying background optical properties, while simultaneously recovering the optical properties. Tissue-simulating phantoms containing doxorubicin, MnTPPS, and Intralipid-20% were created, and fluorescence measurements were performed using a single isotropic probe. The resulting spectra were analyzed using a forward-adjoint fluorescence model in order to recover the fluorophore concentration and background optical properties. We demonstrated recovery of doxorubicin concentration with a mean error of 11.8%. The concentration of the background absorber was recovered with an average error of 23.2% and the scattering spectrum was recovered with a mean error of 19.8%. This method will allow for the determination of local concentrations of fluorescent drugs, such as doxorubicin, from minimally invasive fluorescence measurements. This is particularly interesting in the context of transarterial chemoembolization (TACE) treatment of liver cancer. © 2013 Wiley Periodicals, Inc.

Feasibility Demonstration of Exciplex Fluorescence Measurements in Evaporating Laminar Sprays of Diesel Fuel

DTIC Science & Technology

2011-05-15

code) 1 FEASIBILITY DEMONSTRATION OF EXCIPLEX FLUORESCENCE MEASUREMENTS IN EVAPORATING LAMINAR SPRAYS OF DIESEL FUEL Final Technical Report Grant...fluorescence is found to increase with temperature up to 538 K and then declines. Fluorescence from the liquid phase, i.e. the exciplex (Naphthalene+TMPD...to have as well characterized a description of the spray environment and assess conclusively the potential of the exciplex approach for more

Sequential pictorial presentation of neural interaction in the retina. 2. The depolarizing and hyperpolarizing bipolar cells at rod terminals.

PubMed

Sjöstrand, F S

2002-01-01

Each rod is connected to one depolarizing and one hyperpolarizing bipolar cell. The synaptic connections of cone processes to each bipolar cell and presynaptically to the two rod-bipolar cell synapses establishes conditions for lateral interaction at this level. Thus, the cones raise the threshold for bipolar cell depolarization which is the basis for spatial brightness contrast enhancement and consequently for high visual acuity (Sjöstrand, 2001a). The cones facilitate ganglion cell depolarization by the bipolar cells and cone input prevents horizontal cell blocking of depolarization of the depolarizing bipolar cell, extending rod vision to low illumination. The combination of reduced cone input and transient hyperpolarization of the hyperpolarizing bipolar cell at onset of a light stimulus facilitates ganglion cell depolarization extensively at onset of the stimulus while no corresponding enhancement applies to the ganglion cell response at cessation of the stimulus, possibly establishing conditions for discrimination between on- vs. off-signals in the visual centre. Reduced cone input and hyperpolarization of the hyperpolarizing bipolar cell at onset of a light stimulus accounts for Granit's (1941) 'preexcitatory inhibition'. Presynaptic inhibition maintains transmitter concentration low in the synaptic gap at rod-bipolar cell and bipolar cell-ganglion cell synapses, securing proportional and amplified postsynaptic responses at these synapses. Perfect timing of variations in facilitatory and inhibitory input to the ganglion cell confines the duration of ganglion cell depolarization at onset and at cessation of a light stimulus to that of a single synaptic transmission.

Oxygen-dependent delayed fluorescence measured in skin after topical application of 5-aminolevulinic acid.

PubMed

Harms, Floor A; de Boon, Wadim M I; Balestra, Gianmarco M; Bodmer, Sander I A; Johannes, Tanja; Stolker, Robert J; Mik, Egbert G

2011-10-01

Mitochondrial oxygen tension can be measured in vivo by means of oxygen-dependent quenching of delayed fluorescence of protoporphyrin IX (PpIX). Here we demonstrate that delayed fluorescence is readily observed from skin in rat and man after topical application of the PpIX precursor 5-aminolevulinic acid (ALA). Delayed fluorescence lifetimes respond to changes in inspired oxygen fraction and blood supply. The signals contain lifetime distributions and the fitting of rectangular distributions to the data appears more adequate than mono-exponential fitting. The use of topically applied ALA for delayed fluorescence lifetime measurements might pave the way for clinical use of this technique. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Aerosol Classification from High Spectral Resolution Lidar Measurements

NASA Astrophysics Data System (ADS)

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

2015-12-01

The NASA Langley airborne High Spectral Resolution Lidars, HSRL-1 and HSRL-2, have acquired large datasets of vertically resolved aerosol extinction, backscatter, and depolarization during >30 airborne field missions since 2006. The lidar measurements of aerosol intensive parameters like lidar ratio and color ratio embed information about intrinsic aerosol properties, and are combined to qualitatively classify HSRL aerosol measurements into aerosol types. Knowledge of aerosol type is important for assessing aerosol radiative forcing, and can provide useful information for source attribution studies. However, atmospheric aerosol is frequently not a single pure type, but instead is a mixture, which affects the optical and radiative properties of the aerosol. We show that aerosol intensive parameters measured by lidar can be understood using mixing rules for cases of external mixing. Beyond coarse classification and mixing between classes, variations in the lidar aerosol intensive parameters provide additional insight into aerosol processes and composition. This is illustrated by depolarization measurements at three wavelengths, 355 nm, 532 nm, and 1064 nm, made by HSRL-2. Particle depolarization ratio is an indicator of non-spherical particles. Three cases each have a significantly different spectral dependence of the depolarization ratio, related to the size of the depolarizing particles. For two dust cases, large non-spherical particles account for the depolarization of the lidar light. The spectral dependence reflects the size distribution of these particles and reveals differences in the transport histories of the two plumes. For a smoke case, the depolarization is inferred to be due to the presence of small coated soot aggregates. Interestingly, the depolarization at 355 nm is similar for this smoke case compared to the dust cases, having potential implications for the upcoming EarthCARE satellite, which will measure particle depolarization ratio only at 355 nm.

Measurement and interpretation of fluorescence polarisations in phospholipid dispersions.

PubMed

Bashford, C L; Morgan, C G; Radda, G K

1976-03-05

An instrument that measures the temperature dependence of fluorescence polarisation and intensity directly and continuously is described. The behaviour of four fluorescent probes bound to a number of well characterised model systems was then examined. The motional properties of the probes were determined from the polarisation and intensity data and were found to be sensitive to the crystalline-liquid crystalline phase transitions in phospholipid vesicles of dimyristoly and dipalmitoly phosphatidylcholine. Binary mixture of dilauroyl and dipalmitoyl phosphatidylcholine show lateral phase separation and in this system the probes parition preferentially into the more 'fluid' phase. In systems that have been reported to contain 'short range order' or 'liquid clustering', such as dioleoyl phosphatidylcholine and liquid paraffin, the motion of the probes was found to have anomalous Arrhenius behaviour consistent with the idea that homogeneous phases were not being sampled. The significance of these findings for the interpretation of the behaviour of fluorescent probes bound to natural membranes is discussed.

A review of depolarization modeling for earth-space radio paths at frequencies above 10 GHz

NASA Technical Reports Server (NTRS)

Bostian, C. W.; Stutzman, W. L.; Gaines, J. M.

1982-01-01

A review is presented of models for the depolarization, caused by scattering from raindrops and ice crystals, that limits the performance of dual-polarized satellite communication systems at frequencies above 10 GHz. The physical mechanisms of depolarization as well as theoretical formulations and empirical data are examined. Three theoretical models, the transmission, attenuation-derived, and scaling models, are described and their relative merits are considered.

Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons.

PubMed

Villarreal, Seth; Lee, Sung Hoon; Wu, Ling-Gang

2017-09-04

During endocytosis, fused synaptic vesicles are retrieved at nerve terminals, allowing for vesicle recycling and thus the maintenance of synaptic transmission during repetitive nerve firing. Impaired endocytosis in pathological conditions leads to decreases in synaptic strength and brain functions. Here, we describe methods used to measure synaptic vesicle endocytosis at the mammalian hippocampal synapse in neuronal culture. We monitored synaptic vesicle protein endocytosis by fusing a synaptic vesicular membrane protein, including synaptophysin and VAMP2/synaptobrevin, at the vesicular lumenal side, with pHluorin, a pH-sensitive green fluorescent protein that increases its fluorescence intensity as the pH increases. During exocytosis, vesicular lumen pH increases, whereas during endocytosis vesicular lumen pH is re-acidified. Thus, an increase of pHluorin fluorescence intensity indicates fusion, whereas a decrease indicates endocytosis of the labelled synaptic vesicle protein. In addition to using the pHluorin imaging method to record endocytosis, we monitored vesicular membrane endocytosis by electron microscopy (EM) measurements of Horseradish peroxidase (HRP) uptake by vesicles. Finally, we monitored the formation of nerve terminal membrane pits at various times after high potassium-induced depolarization. The time course of HRP uptake and membrane pit formation indicates the time course of endocytosis.

Time-resolved UV-excited microarray reader for fluorescence energy transfer (FRET) measurements

NASA Astrophysics Data System (ADS)

Orellana, Adelina; Hokkanen, Ari P.; Pastinen, Tomi; Takkinen, Kristina; Soderlund, Hans

2001-05-01

Analytical systems based on immunochemistry are largely used in medical diagnostics and in biotechnology. There is a significant pressure to develop the present assay formats to become easier to use, faster, and less reagent consuming. Further developments towards high density array--like multianalyte measurement systems would be valuable. To this aim we have studied the applicability of fluorescence resonance energy transfer and time-resolved fluorescence resonance energy transfer in immunoassays on microspots and in microwells. We have used engineered recombinant antibodies detecting the pentameric protein CRP as a model analyte system, and tested different assay formats. We describe also the construction of a time-resolved scanning epifluorometer with which we could measure the FRET interaction between the slow fluorescence decay from europium chelates and its energy transfer to the rapidly decaying fluorophore Cy5.

Intracellular ca2+ stores could participate to abscisic acid-induced depolarization and stomatal closure in Arabidopsis thaliana

PubMed Central

Meimoun, Patrice; Vidal, Guillaume; Bohrer, Anne-Sophie; Lehner, Arnaud; Tran, Daniel; Briand, Joël; Bouteau, François

2009-01-01

In Arabidopsis thaliana cell suspension,abscisic acid (aBa) induces changes in cytosolic calcium concentration ([Ca2+]cyt) which are the trigger for aBa-induced plasma membrane anion current activation, H+-aTPase inhibition, and subsequent plasma membrane depolarization. In the present study, we took advantage of this model to analyze the implication of intracellular Ca2+ stores in aBa signal transduction through electrophysiological current measurements, cytosolic Ca2+ activity measurements with the apoaequorin Ca2+ reporter protein and external pH measurement. Intracellular Ca2+ stores involvement was determined by using specific inhibitors of CICR channels: the cADP-ribose/ryanodine receptor (Br-cADPR and dantrolene) and of the inositol trisphosphate receptor (U73122). In addition experiments were performed on epidermal strips of A. thaliana leaves to monitor stomatal closure in response to ABA in presence of the same pharmacology. Our data provide evidence that ryanodine receptor and inositol trisphosphate receptor could be involved in ABA-induced (1) Ca2+ release in the cytosol, (2) anion channel activation and H+-ATPase inhibition leading to plasma membrane depolarization and (3) stomatal closure. Intracellular Ca2+ release could thus contribute to the control of early events in the ABA signal transduction pathway in A. thaliana. PMID:19847112

Calcium-induced calcium release in rod photoreceptor terminals boosts synaptic transmission during maintained depolarization

PubMed Central

Cadetti, Lucia; Bryson, Eric J.; Ciccone, Cory A.; Rabl, Katalin; Thoreson, Wallace B.

2008-01-01

We examined the contribution of calcium-induced calcium release (CICR) to synaptic transmission from rod photoreceptor terminals. Whole-cell recording and confocal calcium imaging experiments were conducted on rods with intact synaptic terminals in a retinal slice preparation from salamander. Low concentrations of ryanodine stimulated calcium increases in rod terminals, consistent with the presence of ryanodine receptors. Application of strong depolarizing steps (−70 to −10 mV) exceeding 200 ms or longer in duration evoked a wave of calcium that spread across the synaptic terminals of voltage-clamped rods. This secondary calcium increase was blocked by high concentrations of ryanodine, indicating it was due to CICR. Ryanodine (50 μM) had no significant effect on rod calcium current (Ica) although it slightly diminished rod light-evoked voltage responses. Bath application of 50 μM ryanodine strongly inhibited light-evoked currents in horizontal cells. Whether applied extracellularly or delivered into the rod cell through the patch pipette, ryanodine (50 μM) also inhibited excitatory post-synaptic currents (EPSCs) evoked in horizontal cells by depolarizing steps applied to rods. Ryanodine caused a preferential reduction in the later portions of EPSCs evoked by depolarizing steps of 200 ms or longer. These results indicate that CICR enhances calcium increases in rod terminals evoked by sustained depolarization, which in turn acts to boost synaptic exocytosis from rods. PMID:16819987

Nanoscale measurements of proton tracks using fluorescent nuclear track detectors

PubMed Central

Sawakuchi, Gabriel O.; Ferreira, Felisberto A.; McFadden, Conor H.; Hallacy, Timothy M.; Granville, Dal A.; Sahoo, Narayan; Akselrod, Mark S.

2016-01-01

Purpose: The authors describe a method in which fluorescence nuclear track detectors (FNTDs), novel track detectors with nanoscale spatial resolution, are used to determine the linear energy transfer (LET) of individual proton tracks from proton therapy beams by allowing visualization and 3D reconstruction of such tracks. Methods: FNTDs were exposed to proton therapy beams with nominal energies ranging from 100 to 250 MeV. Proton track images were then recorded by confocal microscopy of the FNTDs. Proton tracks in the FNTD images were fit by using a Gaussian function to extract fluorescence amplitudes. Histograms of fluorescence amplitudes were then compared with LET spectra. Results: The authors successfully used FNTDs to register individual proton tracks from high-energy proton therapy beams, allowing reconstruction of 3D images of proton tracks along with delta rays. The track amplitudes from FNTDs could be used to parameterize LET spectra, allowing the LET of individual proton tracks from therapeutic proton beams to be determined. Conclusions: FNTDs can be used to directly visualize proton tracks and their delta rays at the nanoscale level. Because the track intensities in the FNTDs correlate with LET, they could be used further to measure LET of individual proton tracks. This method may be useful for measuring nanoscale radiation quantities and for measuring the LET of individual proton tracks in radiation biology experiments. PMID:27147359

Nanoscale measurements of proton tracks using fluorescent nuclear track detectors

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sawakuchi, Gabriel O., E-mail: gsawakuchi@mdanderson.org; Sahoo, Narayan; Ferreira, Felisberto A.

Purpose: The authors describe a method in which fluorescence nuclear track detectors (FNTDs), novel track detectors with nanoscale spatial resolution, are used to determine the linear energy transfer (LET) of individual proton tracks from proton therapy beams by allowing visualization and 3D reconstruction of such tracks. Methods: FNTDs were exposed to proton therapy beams with nominal energies ranging from 100 to 250 MeV. Proton track images were then recorded by confocal microscopy of the FNTDs. Proton tracks in the FNTD images were fit by using a Gaussian function to extract fluorescence amplitudes. Histograms of fluorescence amplitudes were then compared withmore » LET spectra. Results: The authors successfully used FNTDs to register individual proton tracks from high-energy proton therapy beams, allowing reconstruction of 3D images of proton tracks along with delta rays. The track amplitudes from FNTDs could be used to parameterize LET spectra, allowing the LET of individual proton tracks from therapeutic proton beams to be determined. Conclusions: FNTDs can be used to directly visualize proton tracks and their delta rays at the nanoscale level. Because the track intensities in the FNTDs correlate with LET, they could be used further to measure LET of individual proton tracks. This method may be useful for measuring nanoscale radiation quantities and for measuring the LET of individual proton tracks in radiation biology experiments.« less

Landscape of the PARKIN-dependent ubiquitylome in response to mitochondrial depolarization.

PubMed

Sarraf, Shireen A; Raman, Malavika; Guarani-Pereira, Virginia; Sowa, Mathew E; Huttlin, Edward L; Gygi, Steven P; Harper, J Wade

2013-04-18

The PARKIN ubiquitin ligase (also known as PARK2) and its regulatory kinase PINK1 (also known as PARK6), often mutated in familial early-onset Parkinson's disease, have central roles in mitochondrial homeostasis and mitophagy. Whereas PARKIN is recruited to the mitochondrial outer membrane (MOM) upon depolarization via PINK1 action and can ubiquitylate porin, mitofusin and Miro proteins on the MOM, the full repertoire of PARKIN substrates--the PARKIN-dependent ubiquitylome--remains poorly defined. Here we use quantitative diGly capture proteomics (diGly) to elucidate the ubiquitylation site specificity and topology of PARKIN-dependent target modification in response to mitochondrial depolarization. Hundreds of dynamically regulated ubiquitylation sites in dozens of proteins were identified, with strong enrichment for MOM proteins, indicating that PARKIN dramatically alters the ubiquitylation status of the mitochondrial proteome. Using complementary interaction proteomics, we found depolarization-dependent PARKIN association with numerous MOM targets, autophagy receptors, and the proteasome. Mutation of the PARKIN active site residue C431, which has been found mutated in Parkinson's disease patients, largely disrupts these associations. Structural and topological analysis revealed extensive conservation of PARKIN-dependent ubiquitylation sites on cytoplasmic domains in vertebrate and Drosophila melanogaster MOM proteins. These studies provide a resource for understanding how the PINK1-PARKIN pathway re-sculpts the proteome to support mitochondrial homeostasis.

Landscape of the PARKIN-dependent ubiquitylome in response to mitochondrial depolarization

PubMed Central

Sarraf, Shireen A.; Raman, Malavika; Guarani-Pereira, Virginia; Sowa, Mathew E.; Huttlin, Edward L.; Gygi, Steven P.; Harper, J. Wade

2013-01-01

The PARKIN (PARK2) ubiquitin ligase and its regulatory kinase PINK1 (PARK6), often mutated in familial early onset Parkinson’s Disease (PD), play central roles in mitochondrial homeostasis and mitophagy.1–3 While PARKIN is recruited to the mitochondrial outer membrane (MOM) upon depolarization via PINK1 action and can ubiquitylate Porin, Mitofusin, and Miro proteins on the MOM,1,4–11 the full repertoire of PARKIN substrates – the PARKIN-dependent ubiquitylome - remains poorly defined. Here we employ quantitative diGLY capture proteomics12,13 to elucidate the ubiquitylation site-specificity and topology of PARKIN-dependent target modification in response to mitochondrial depolarization. Hundreds of dynamically regulated ubiquitylation sites in dozens of proteins were identified, with strong enrichment for MOM proteins, indicating that PARKIN dramatically alters the ubiquitylation status of the mitochondrial proteome. Using complementary interaction proteomics, we found depolarization-dependent PARKIN association with numerous MOM targets, autophagy receptors, and the proteasome. Mutation of PARKIN’s active site residue C431, which has been found mutated in PD patients, largely disrupts these associations. Structural and topological analysis revealed extensive conservation of PARKIN-dependent ubiquitylation sites on cytoplasmic domains in vertebrate and D. melanogaster MOM proteins. These studies provide a resource for understanding how the PINK1-PARKIN pathway re-sculpts the proteome to support mitochondrial homeostasis. PMID:23503661

Simultaneous two-dimensional laser-induced-fluorescence measurements of argon ions.

PubMed

Hansen, A K; Galante, Matthew; McCarren, Dustin; Sears, Stephanie; Scime, E E

2010-10-01

Recent laser upgrades on the Hot Helicon Experiment at West Virginia University have enabled multiplexed simultaneous measurements of the ion velocity distribution function at a single location, expanding our capabilities in laser-induced fluorescence diagnostics. The laser output is split into two beams, each modulated with an optical chopper and injected perpendicular and parallel to the magnetic field. Light from the crossing point of the beams is transported to a narrow-band photomultiplier tube filtered at the fluorescence wavelength and monitored by two lock-in amplifiers, each referenced to one of the two chopper frequencies.

Recovery of intrinsic fluorescence from single-point interstitial measurements for quantification of doxorubicin concentration

PubMed Central

Baran, Timothy M.; Foster, Thomas H.

2014-01-01

Background and Objective We developed a method for the recovery of intrinsic fluorescence from single-point measurements in highly scattering and absorbing samples without a priori knowledge of the sample optical properties. The goal of the study was to demonstrate accurate recovery of fluorophore concentration in samples with widely varying background optical properties, while simultaneously recovering the optical properties. Materials and Methods Tissue-simulating phantoms containing doxorubicin, MnTPPS, and Intralipid-20% were created, and fluorescence measurements were performed using a single isotropic probe. The resulting spectra were analyzed using a forward-adjoint fluorescence model in order to recover the fluorophore concentration and background optical properties. Results We demonstrated recovery of doxorubicin concentration with a mean error of 11.8%. The concentration of the background absorber was recovered with an average error of 23.2% and the scattering spectrum was recovered with a mean error of 19.8%. Conclusion This method will allow for the determination of local concentrations of fluorescent drugs, such as doxorubicin, from minimally invasive fluorescence measurements. This is particularly interesting in the context of transarterial chemoembolization (TACE) treatment of liver cancer. PMID:24037853

Effects of melting layer on Ku-band signal depolarization

NASA Astrophysics Data System (ADS)

Sarkar, Thumree; Das, Saurabh; Maitra, Animesh

2014-09-01

Propagation effects on Ku-band over an earth-space path is carried out at Kolkata, India, a tropical location, by receiving a Ku-band signal with horizontal plane polarization transmitted from the geostationary satellite NSS-6 (at 95°E). The amplitude of co-polar attenuation has been monitored along with the measurements of rain rate, rain drop size distribution and height profile of rain rate. The cross-polar enhancement of the signal is also monitored by receiving the same signal in orthogonal direction with another identical receiver. The experimental observations are used to study the effect of melting layer on both co-polar attenuation and cross-polar enhancement for the rain events observed during 2012-2013. Melting layer is indicated by the bright band signature in vertical profile of rain rate. The ground based drop size measurements indicate that the stratiform rain has more number of small drops whereas convective rain composed of large rain drops. The results indicate that the depolarization due to melting layer is more dominant compared to that due to the drop deformation mechanism at low rain rates.

The induction of apoptosis by methotrexate in activated lymphocytes as indicated by fluorescence hyperpolarization: a possible model for predicting methotrexate therapy for rheumatoid arthritis patients.

PubMed

Herman, Shoshy; Zurgil, Naomi; Langevitz, Pnina; Ehrenfeld, Michael; Deutsch, Mordechai

2003-04-01

The objectives of this study were to test the in vitro response of healthy non-activated, activated, and rheumatoid arthritis (RA) lymphocytes to methotrexate (MTX), and design an in vitro model for predicting the efficiency of MTX treatment for RA patients. Considering the RA profile of clonal-expanded CD4(+) T cells, phytohemagglutinin-activated mononuclear cells taken from healthy donors were incubated with different concentrations of MTX. The MTX-immunosuppressive effect was tested by fluorescence intensity measurements, including PI assay and annexin V assay. For simple detection, we used the Individual Cell Scanner (IC-S), which enables the measurement of early events in individual cells. Healthy mononuclear cells (MNC), and MNC derived from RA patients, were tested by the IC-S while utilizing fluorescence polarization (FP) measurements of fluorescein diacetate (FDA) as an established marker of activation or suppression. In healthy activated MNC, we found that MTX, through its early incubation period, interferes with the activation signal obtained by PHA and exerts an apoptotic signal, which is noted by increases in the FP. Comparing our model to six long-standing RA patients and five newly-diagnosed patients revealed significant differences in the FP measurements, including fluorescence depolarization as an early established measurement of lymphocyte activation, and hyperpolarization as a measurement of an early immunosuppressive effect. We conclude that MTX, an effective therapy for RA patients, could easily be tested by fluorescence polarization measurements of FDA before (or during) clinical use in order to predict its efficiency on a specific RA patient. Moreover, the FP measurements can be used for the diagnosis, and making timing and dosage decisions.

Single Molecule Fluorescence Measurements of Complex Systems

NASA Astrophysics Data System (ADS)

Sadegh, Sanaz

Single molecule methods are powerful tools for investigating the properties of complex systems that are generally concealed by ensemble measurements. Here we use single molecule fluorescent measurements to study two different complex systems: 1/ƒ noise in quantum dots and diffusion of the membrane proteins in live cells. The power spectrum of quantum dot (QD) fluorescence exhibits 1/ƒ beta noise, related to the intermittency of these nanosystems. As in other systems exhibiting 1/ƒ noise, this power spectrum is not integrable at low frequencies, which appears to imply infinite total power. We report measurements of individual QDs that address this long-standing paradox. We find that the level of 1/ƒbeta noise for QDs decays with the observation time. We show that the traditional description of the power spectrum with a single exponent is incomplete and three additional critical exponents characterize the dependence on experimental time. A broad range of membrane proteins display anomalous diffusion on the cell surface. Different methods provide evidence for obstructed subdiffusion and diffusion on a fractal space, but the underlying structure inducing anomalous diffusion has never been visualized due to experimental challenges. We addressed this problem by imaging the cortical actin at high resolution while simultaneously tracking individual membrane proteins in live mammalian cells. Our data show that actin introduces barriers leading to compartmentalization of the plasma membrane and that membrane proteins are transiently confined within actin fences. Furthermore, superresolution imaging shows that the cortical actin is organized into a self-similar fractal.

Depolarized haze of nano-porous AAO film via porosity and aspect control

NASA Astrophysics Data System (ADS)

Tseng, Chun-Wei; Lin, Yung-Hsiang; Cheng, Chih-Hsien; Lin, Gong-Ru

2018-01-01

Multiple scattering induced haze and depolarization effects of nano-porous AAO films controlled by detuning the porosity and aspect ratio of the nano holes are investigated. The nano-porous AAO film with its porosity increasing from 12.6% to 19.3% enhances the scattering of the incident laser beam with its maximal scattering angle enlarged from 5° to 8° under TM-mode incidence and from 6° to 10° under TE-mode incidence. Because of multiple scattering within the porous holes of the AAO, the depolarization on the reflected beam by transferring its electric field from horizontal to the vertical such that the polarization ratio is degraded with a randomized haze. The porosity of AAO surface broadens from 12.6% to 19.3% when increasing the bias voltage from 40 to 60 V during the second-step of the electro-chemical anodization process, which essentially adjusts the polarization ratio under TM-mode and TE-mode incidences raise from 0.31 to 0.35 and from 0.32 to 0.48, respectively. The depolarized haze of the nano-porous AAO film is correlated with its porosity and aspect ratio controlled by the pore size and etched depth of the AAO. Under TM-mode incidence, the simulated polarization ratio increases from 0.35 to 0.38, which correlates well with experimental results. In contrast, the experiment result slightly deviates from the theoretical prediction as the TE-mode field interacts more surface area than the TM-mode field does. Such a nano-porous AAO exhibits tunable depolarized haze via the control porosity and aspect ratio, which is particularly suitable to serve as the catalytic buffer for synthesizing the hydrophobic and hazed solar energy converters.

Activation of cathepsin L contributes to the irreversible depolarization induced by oxygen and glucose deprivation in rat hippocampal CA1 neurons.

PubMed

Kikuta, Shogo; Murai, Yoshinaka; Tanaka, Eiichiro

2017-01-01

Oxygen and glucose deprivation (OGD) elicits a rapid and irreversible depolarization with a latency of ∼5min in intracellular recordings of hippocampal CA1 neurons in rat slice preparations. In the present study, we examined the role of cathepsin L in the OGD-induced depolarization. OGD-induced depolarizations were irreversible as no recovery of membrane potential was observed. The membrane potential reached 0mV when oxygen and glucose were reintroduced immediately after the onset of the OGD-induced rapid depolarization. The OGD-induced depolarizations became reversible when the slice preparations were pre-incubated with cathepsin L inhibitors (types I and IV at 0.3-2nM and 0.3-30nM, respectively). Moreover, pre-incubation with these cathepsin inhibitors prevented the morphological changes, including swelling of the cell soma and fragmentation of dendrites, observed in control neurons after OGD. These findings suggest that the activation of cathepsin L contributes to the irreversible depolarization produced by OGD. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Shifts in the fluorescence lifetime of EGFP during bacterial phagocytosis measured by phase-sensitive flow cytometry

NASA Astrophysics Data System (ADS)

Li, Wenyan; Houston, Kevin D.; Houston, Jessica P.

2017-01-01

Phase-sensitive flow cytometry (PSFC) is a technique in which fluorescence excited state decay times are measured as fluorescently labeled cells rapidly transit a finely focused, frequency-modulated laser beam. With PSFC the fluorescence lifetime is taken as a cytometric parameter to differentiate intracellular events that are challenging to distinguish with standard flow cytometry. For example PSFC can report changes in protein conformation, expression, interactions, and movement, as well as differences in intracellular microenvironments. This contribution focuses on the latter case by taking PSFC measurements of macrophage cells when inoculated with enhanced green fluorescent protein (EGFP)-expressing E. coli. During progressive internalization of EGFP-E. coli, fluorescence lifetimes were acquired and compared to control groups. It was hypothesized that fluorescence lifetimes would correlate well with phagocytosis because phagosomes become acidified and the average fluorescence lifetime of EGFP is known to be affected by pH. We confirmed that average EGFP lifetimes consistently decreased (3 to 2 ns) with inoculation time. The broad significance of this work is the demonstration of how high-throughput fluorescence lifetime measurements correlate well to changes that are not easily tracked by intensity-only cytometry, which is affected by heterogeneous protein expression, cell-to-cell differences in phagosome formation, and number of bacterium engulfed.

Measurement of two-photon-absorption spectra through nonlinear fluorescence produced by a line-shaped excitation beam.

PubMed

Hasani, E; Parravicini, J; Tartara, L; Tomaselli, A; Tomassini, D

2018-05-01

We propose an innovative experimental approach to estimate the two-photon absorption (TPA) spectrum of a fluorescent material. Our method develops the standard indirect fluorescence-based method for the TPA measurement by employing a line-shaped excitation beam, generating a line-shaped fluorescence emission. Such a configuration, which requires a relatively high amount of optical power, permits to have a greatly increased fluorescence signal, thus avoiding the photon counterdetection devices usually used in these measurements, and allowing to employ detectors such as charge-coupled device (CCD) cameras. The method is finally tested on a fluorescent isothiocyanate sample, whose TPA spectrum, which is measured with the proposed technique, is compared with the TPA spectra reported in the literature, confirming the validity of our experimental approach. © 2018 The Authors Journal of Microscopy © 2018 Royal Microscopical Society.

Complete stress-induced depolarization of relaxor ferroelectric crystals without transition through a non-polar phase

NASA Astrophysics Data System (ADS)

Shkuratov, Sergey I.; Baird, Jason; Antipov, Vladimir G.; Hackenberger, Wesley; Luo, Jun; Zhang, Shujun; Lynch, Christopher S.; Chase, Jay B.; Jo, Hwan R.; Roberts, Christopher C.

2018-03-01

The development of relaxor ferroelectric single crystal technology is driven by the ability to tailor ferroelectric properties through domain engineering not achievable in polycrystalline materials. In this study, three types of domain-engineered rhombohedral Pb(In1/2Nb1/2)O3-Pb(Mg1/3Nb2/3)O3-PbTiO3 crystals were subjected to transverse high strain rate loading. The experimental results indicate that the domain configuration has a significant effect on the stress-induced depolarization and the associated charge released. A complete depolarization of the single-domain crystals with 3m symmetry is observed, while multidomain crystals with 4mm and mm2 symmetries retain a fraction of their initial remanent polarization. The complete depolarization of single-domain crystals is unique without transition to a non-polar phase, with a stress-induced charge density of 0.48 C/m2. This is up to three times higher than that of the multidomain crystals and PbZrxTi1-xO3 ferroelectric ceramics that are critical for ultrahigh-power transducer applications. The main offering of this work is to propose a detailed mechanism for complete stress-induced depolarization in ferroelectric crystals which does not involve an intermediate transformation to a non-polar phase.

Rapid kill of malaria parasites by artemisinin and semi-synthetic endoperoxides involves ROS-dependent depolarization of the membrane potential

PubMed Central

Antoine, Thomas; Fisher, Nicholas; Amewu, Richard; O'Neill, Paul M.; Ward, Stephen A.; Biagini, Giancarlo A.

2014-01-01

Objectives Artemisinin and artemisinin semi-synthetic derivatives (collectively known as endoperoxides) are first-line antimalarials for the treatment of uncomplicated and severe malaria. Endoperoxides display very fast killing rates and are generally recalcitrant to parasite resistance development. These key pharmacodynamic features are a result of a complex mechanism of action, the details of which lack consensus. Here, we report on the primary physiological events leading to parasite death. Methods Parasite mitochondrial (ΔΨm) and plasma membrane (ΔΨp) electrochemical potentials were measured using real-time single-cell imaging following exposure to pharmacologically relevant concentrations of endoperoxides (artemisinin, dihydroartemisinin, artesunate and the synthetic tetraoxane RKA182). In addition, mitochondrial electron transport chain components NADH:quinone oxidoreductase (alternative complex I), bc1 (complex III) and cytochrome oxidase (complex IV) were investigated to determine their functional sensitivity to the various endoperoxides. Results Parasite exposure to endoperoxides resulted in rapid depolarization of parasite ΔΨm and ΔΨp. The rate of depolarization was decreased in the presence of a reactive oxygen species (ROS) scavenger and Fe3+ chelators. Depolarization of ΔΨm by endoperoxides is not believed to be through the inhibition of mitochondrial electron transport chain components, owing to the lack of significant inhibition when assayed directly. Conclusions The depolarization of ΔΨm and ΔΨp is shown to be mediated via the generation of ROS that are initiated by iron bioactivation of endoperoxides and/or catalysed by iron-dependent oxidative stress. These data are discussed in the context of current hypotheses concerning the mode of action of endoperoxides. PMID:24335485

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

In Vivo Mitochondrial Oxygen Tension Measured by a Delayed Fluorescence Lifetime Technique

PubMed Central

Mik, Egbert G.; Johannes, Tanja; Zuurbier, Coert J.; Heinen, Andre; Houben-Weerts, Judith H. P. M.; Balestra, Gianmarco M.; Stap, Jan; Beek, Johan F.; Ince, Can

2008-01-01

Mitochondrial oxygen tension (mitoPO2) is a key parameter for cellular function, which is considered to be affected under various pathophysiological circumstances. Although many techniques for assessing in vivo oxygenation are available, no technique for measuring mitoPO2 in vivo exists. Here we report in vivo measurement of mitoPO2 and the recovery of mitoPO2 histograms in rat liver by a novel optical technique under normal and pathological circumstances. The technique is based on oxygen-dependent quenching of the delayed fluorescence lifetime of protoporphyrin IX. Application of 5-aminolevulinic acid enhanced mitochondrial protoporphyrin IX levels and induced oxygen-dependent delayed fluorescence in various tissues, without affecting mitochondrial respiration. Using fluorescence microscopy, we demonstrate in isolated hepatocytes that the signal is of mitochondrial origin. The delayed fluorescence lifetime was calibrated in isolated hepatocytes and isolated perfused livers. Ultimately, the technique was applied to measure mitoPO2 in rat liver in vivo. The results demonstrate mitoPO2 values of ∼30–40 mmHg. mitoPO2 was highly sensitive to small changes in inspired oxygen concentration around atmospheric oxygen level. Ischemia-reperfusion interventions showed altered mitoPO2 distribution, which flattened overall compared to baseline conditions. The reported technology is scalable from microscopic to macroscopic applications, and its reliance on an endogenous compound greatly enhances its potential field of applications. PMID:18641065

Measurement of Hydroxyl Radicals in Plasma Pencil by Laser Induced Fluorescence

DTIC Science & Technology

2013-07-01

31st ICPIG, July 14-19, 2013, Granada , Spain Topic number 6 Measurement of hydroxyl radicals in plasma pencil by laser induced fluorescence J...International Conference on Phenomena in Ionized Gases (31st) (ICPIG) Held in Granada , Spain on 14-19 July 2013, The original document contains color images. 14...Prescribed by ANSI Std Z39-18 31st ICPIG, July 14-19, 2013, Granada , Spain Topic number 6 camera. The fluorescence signal was significantly stronger

DNA conformation on surfaces measured by fluorescence self-interference.

PubMed

Moiseev, Lev; Unlü, M Selim; Swan, Anna K; Goldberg, Bennett B; Cantor, Charles R

2006-02-21

The conformation of DNA molecules tethered to the surface of a microarray may significantly affect the efficiency of hybridization. Although a number of methods have been applied to determine the structure of the DNA layer, they are not very sensitive to variations in the shape of DNA molecules. Here we describe the application of an interferometric technique called spectral self-interference fluorescence microscopy to the precise measurement of the average location of a fluorescent label in a DNA layer relative to the surface and thus determine specific information on the conformation of the surface-bound DNA molecules. Using spectral self-interference fluorescence microscopy, we have estimated the shape of coiled single-stranded DNA, the average tilt of double-stranded DNA of different lengths, and the amount of hybridization. The data provide important proofs of concept for the capabilities of novel optical surface analytical methods of the molecular disposition of DNA on surfaces. The determination of DNA conformations on surfaces and hybridization behavior provide information required to move DNA interfacial applications forward and thus impact emerging clinical and biotechnological fields.

Temperature measurements of inverse micelles coated in gold nanoparticles using fluorescence

NASA Astrophysics Data System (ADS)

Daley, Chad; Forrest, James A.; Speller, Ryan; William, Toews; McVeigh, Patrick; Emrick, Todd

2009-03-01

When nanoparticles are subject to laser radiation they have the ability to efficiently absorb energy from the beam and transform this energy into heat. Photothermal therapy uses this phenomenon to irreparably damage tissue surrounding nanoparticle conjugates. Despite the promise of this technique, there is no concensus on the damage mechanism or even the local heating. Here we present an experiment designed to measure local temperatures achieved in such processes. Ligand covered Gold nanoparticles are used to stabalize inverse micelles containing fluorescence dye in the water component. The fluorescence intensity being temperature dependent provides us with a means of measuring the temperature of the micelles as a function of time immediately following a laser pulse.

Measuring Phagosome pH by Ratiometric Fluorescence Microscopy

PubMed Central

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

A depolarization and attenuation experiment using the CTS satellite. [meteorological radar

NASA Technical Reports Server (NTRS)

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

1977-01-01

Rain attenuation and depolarization data collected on the communications technology satellite 11.7 GHz downlink, and changes made in equipment following rain leak damage to the parametric amplifier are discussed. A 15 GHz radar system is described.

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.

LCLS in—photon out: fluorescence measurement of neon using soft x-rays

DOE PAGES

Obaid, Razib; Buth, Christian; Dakovski, Georgi L.; ...

2018-01-09

Here, we measured the fluorescence photon yield of neon upon soft x-ray ionization (~1200 eV) from the x-ray free-electron laser at Linac Coherent Light Source, and demonstrated the usage of a grazing incidence spectrometer with a variable line spacing grating to perform x-ray fluorescence spectroscopy on a gas phase system. Our measurements also allowed us to estimate the focal size of the beam from the theoretical description developed, in terms of the rate equation approximation accounting for photoionization shake off of neutral neon and double auger decay of single core holes.

LCLS in—photon out: fluorescence measurement of neon using soft x-rays

NASA Astrophysics Data System (ADS)

Obaid, Razib; Buth, Christian; Dakovski, Georgi L.; Beerwerth, Randolf; Holmes, Michael; Aldrich, Jeff; Lin, Ming-Fu; Minitti, Michael; Osipov, Timur; Schlotter, William; Cederbaum, Lorenz S.; Fritzsche, Stephan; Berrah, Nora

2018-02-01

We measured the fluorescence photon yield of neon upon soft x-ray ionization (∼1200 eV) from the x-ray free-electron laser at Linac Coherent Light Source, and demonstrated the usage of a grazing incidence spectrometer with a variable line spacing grating to perform x-ray fluorescence spectroscopy on a gas phase system. Our measurements also allowed us to estimate the focal size of the beam from the theoretical description developed, in terms of the rate equation approximation accounting for photoionization shake off of neutral neon and double auger decay of single core holes.

Incidence, hemodynamic, and electrical characteristics of spreading depolarization in a swine model are affected by local but not by intravenous application of magnesium.

PubMed

Santos, Edgar; León, Fiorella; Silos, Humberto; Sanchez-Porras, Renan; Shuttleworth, C William; Unterberg, Andreas; Sakowitz, Oliver W

2016-12-01

The aim was to characterize the effects of magnesium sulfate, using i.v. bolus and local administration, using intrinsic signal imaging, and on electrocorticographic activity during the induction and propagation of spreading depolarizations in the gyrencephalic porcine brain. Local application of magnesium sulfate led to a complete inhibition of spreading depolarizations. One hour after washing out the topical magnesium sulfate, re-incidence of the spreading depolarizations was observed in 50% of the hemispheres. Those spreading depolarizations showed attenuation in hemodynamic characteristics and speed in intrinsic optical signal imaging. The electrical amplitude decreased through electrocorticographic activity. Intravenous magnesium therapy showed no significant effects on spreading depolarization incidence and characteristics. © The Author(s) 2016.

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.

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

Effect of the depolarization field on coherent optical properties in semiconductor quantum dots

NASA Astrophysics Data System (ADS)

Mitsumori, Yasuyoshi; Watanabe, Shunta; Asakura, Kenta; Seki, Keisuke; Edamatsu, Keiichi; Akahane, Kouichi; Yamamoto, Naokatsu

2018-06-01

We study the photon echo spectrum of self-assembled semiconductor quantum dots using femtosecond light pulses. The spectrum shape changes from a single-peaked to a double-peaked structure as the time delay between the two excitation pulses is increased. The spectrum change is reproduced by numerical calculations, which include the depolarization field induced by the biexciton-exciton transition as well as the conventional local-field effect for the exciton-ground-state transition in a quantum dot. Our findings suggest that various optical transitions in tightly localized systems generate a depolarization field, which renormalizes the resonant frequency with a change in the polarization itself, leading to unique optical properties.

Quantitative calculations of fluorescence polarization and absorption anisotropy kinetics of double- and triple-chromophore complexes with energy transfer.

PubMed Central

Demidov, A A

1994-01-01

A new method is presented for calculation of the fluorescence depolarization and kinetics of absorption anisotropy for molecular complexes with a limited number of chromophores. The method considers absorption and emission of light by both chromophores, and also energy transfer between them, with regard to their mutual orientations. The chromophores in each individual complex are rigidly positioned. The complexes are randomly distributed and oriented in space, and there is no energy transfer between them. The new "practical" formula for absorption anisotropy and fluorescence depolarization kinetics, P(t) = [3B(t) - 1 + 2A(t)]/[3 + B(t) + 4A(t)], is derived both for double- and triple-chromophore complexes with delta-pulse excitation. The parameter B(t) is given by (a) B(t) = cos2(theta) for double-chromophore complexes, and (b) B(t) = q12(t)cos2(theta 12) + q13(t)-cos2(theta 13) + q23(t)cos2(theta 23) for triple-chromophore complexes, where q12(t) + q13(t) + q23(t) = 1. Here theta ij are the angles between the chromophore transition dipole moments in the individual molecular complex. The parameters qij(t) and A(t) are dependent on chromophore spectroscopic features and on the rates of energy transfer. PMID:7696461

Factors that reverse the persistent depolarization produced by deprivation of oxygen and glucose in rat hippocampal CA1 neurons in vitro.

PubMed

Yamamoto, S; Tanaka, E; Shoji, Y; Kudo, Y; Inokuchi, H; Higashi, H

1997-08-01

In CA1 pyramidal neurons in rat hippocampal tissue slices, superfusion with ischemia-simulating medium produced a rapid depolarization after 6 min of exposure. The membrane potential eventually reached 0 after 5 min (a persistent depolarization), even when oxygen and glucose were reintroduced. The role of various ions in the reversal of this persistent depolarization after reintroduction of oxygen and glucose was investigated. The peak of the persistent depolarization was decreased in solutions containing reduced Na+ or Ca2+ and in solutions containing Co2+ or Ni2+. In contrast, the depolarization was not affected by reduction of external K+ or Cl- or by addition of tetrodotoxin (TTX), flunarizine, or nifedipine. These results suggest that sustained Na+ and Ca2+ influxes produce the persistent depolarization. The membrane potential recovered after reintroduction of oxygen and glucose in low Ca2+, low Cl-, or K+-rich medium and in TTX- or tetraethylammonium-containing medium, but not in low Na+ or low K+ medium and in flunarizine- or nifedipine-containing medium. Either reduction in extracellular Ca2+ or addition of Co2+ was the most effective in promoting recovery from the persistent depolarization, suggesting that Ca2+ influx has a key role in causing the membrane dysfunction. The peak of the persistent depolarization was reduced by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), DL-2-amino-5-phosphonopentanoic acid (AP5), DL-amino-3-phosphonopropionic acid (AP3), or DL-amino-4-phosphonobutyric acid, suggesting that activation of non-N-methyl-D-aspartate (non-NMDA), NMDA, and metabotropic glutamate (Glu) receptors is involved in the generation and maintenance of the persistent depolarization. Among these Glu receptor antagonists, only CNQX or AP5 was able to reduce dose dependently the level of depolarization, suggesting that Ca2+ influx via both alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid/kainate type II receptors and NMDA receptors contributes to the

Fluorescence microscopy for measuring fibril angles in pine tracheids

Treesearch

Ralph O. Marts

1955-01-01

Observation and measurement of fibril angles in increment cores or similar small samples from living pine trees was facilitated by the use of fluorescence microscopy. Although some autofluorescence was present, brighter images could be obtained by staining the specimens with a 0.1% aqueous solution of a fluorochrome (Calcozine flavine TG extra concentrated, Calcozine...

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

NASA Astrophysics Data System (ADS)

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

2013-11-01

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

Plant-Stress Measurements Using Laser-Induced Fluorescence Excitation: Poland Experiment

DOE Office of Scientific and Technical Information (OSTI.GOV)

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 primarymore » 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.« less

Improvement of graphite crystal analyzer for light elements on X-ray fluorescence holography measurement

NASA Astrophysics Data System (ADS)

Happo, Naohisa; Hada, Takuma; Kubota, Atsushi; Ebisu, Yoshihiro; Hosokawa, Shinya; Kimura, Koji; Tajiri, Hiroo; Matsushita, Tomohiro; Hayashi, Kouichi

2018-05-01

Using a graphite crystal analyzer, focused monochromatic fluorescent X-rays can be obtained on an X-ray fluorescence holography (XFH) measurement. To measure the holograms of elements lighter than Ti, we improved a cylindrical-type crystal analyzer and constructed a small C-shaped analyzer. Using the constructed C-shaped analyzer, a Ca Kα hologram of a fluorite single crystal was obtained, from which we reconstructed a clear atomic image. The XFH measurements for the K, Ca, and Sc elements become possible using the presently constructed analyzer.

5-Aminolevulinic acid-induced protoporphyrin IX fluorescence in meningioma: qualitative and quantitative measurements in vivo.

PubMed

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

2014-03-01

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

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

PubMed Central

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

2014-01-01

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

Relationship between depolarization-induced force responses and Ca2+ content in skeletal muscle fibres of rat and toad.

PubMed

Owen, V J; Lamb, G D; Stephenson, D G; Fryer, M W

1997-02-01

1. The relationship between the total Ca2+ content of a muscle fibre and the magnitude of the force response to depolarization was examined in mechanically skinned fibres from the iliofibularis muscle of the toad and the extensor digitorum longus muscle of the rat. The response to depolarization in each skinned fibre was assessed either at the endogenous level of Ca2+ content or after depleting the fibre of Ca2+ to some degree. Ca2+ content was determined by a fibre lysing technique. 2. In both muscle types, the total Ca2+ content could be reduced from the endogenous level of approximately 1.3 mmol l-1 (expressed relative to intact fibre volume) to approximately 0.25 mmol l-1 by either depolarization or caffeine application in the presence of Ca2+ chelators, showing that the great majority of the Ca2+ was stored in the sarcoplasmic reticulum (SR). Chelation of Ca2+ in the transverse tubular (T-) system, either by exposure of fibres to EGTA before skinning or by permeabilizing the T-system with saponin after skinning, reduced the lower limit of Ca2+ content to < or = 0.12 mmol l-1, indicating that 10-20% of the total fibre Ca2+ resided in the T-system. 3. In toad fibres, both the peak and the area (i.e. time integral) of the force response to depolarization were reduced by any reduction in SR Ca2+ content, with both decreasing to zero in an approximately linear manner as the SR Ca2+ content was reduced to < 15% of the endogenous level. In rat fibres, the peak size of the force response was less affected by small decreases in SR content, but both the peak and area of the response decreased to zero with greater depletion. In partially depleted toad fibres, inhibition of SR Ca2+ uptake potentiated the force response to depolarization almost 2-fold. 4. The results show that in this skinned fibre preparation: (a) T-system depolarization and caffeine application can each virtually fully deplete the SR of Ca2+, irrespective of any putative inhibitory effect of SR depletion

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. 2013 Published by Elsevier Inc.

Measurement of Zeta-Potential at Microchannel Wall by a Nanoscale Laser Induced Fluorescence Imaging

NASA Astrophysics Data System (ADS)

Kazoe, Yutaka; Sato, Yohei

A nanoscale laser induced fluorescence imaging was proposed by using fluorescent dye and the evanescent wave with total internal reflection of a laser beam. The present study focused on the two-dimensional measurement of zeta-potential at the microchannel wall, which is an electrostatic potential at the wall surface and a dominant parameter of electroosmotic flow. The evanescent wave, which decays exponentially from the wall, was used as an excitation light of the fluorescent dye. The fluorescent intensity detected by a CCD camera is closely related to the zeta-potential. Two kinds of fluorescent dye solution at different ionic concentrations were injected into a T-shaped microchannel, and formed a mixing flow field in the junction area. The two-dimensional distribution of zeta-potential at the microchannel wall in the pressure-driven flow field was measured. The obtained zeta-potential distribution has a transverse gradient toward the mixing flow field and was changed by the difference in the averaged velocity of pressure-driven flow. To understand the ion motion in the mixing flow field, the three-dimensional flow structure was analyzed by the velocity measurement using micron-resolution particle image velocimetry and the numerical simulation. It is concluded that the two-dimensional distribution of zeta-potential at the microchannel wall was dependent on the ion motion in the flow field, which was governed by the convection and molecular diffusion.

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.

Detection of saharan mineral dust aerosol transport over brazilian northeast through a depolarization lidar

NASA Astrophysics Data System (ADS)

Guedes, Anderson G.; Landulfo, Eduardo; Montilla-Rosero, Elena; Lopes, Fábio J. S.; Hoelzemann, Judith J.; Fernandez, José Henrique; Silva, Marcos P. A.; Santos, Renata S. S.; Guerrero-Rascado, Juan L.; Alados-Arboledas, Lucas

2018-04-01

In this study we present results of linear volume depolarization ratio profiles obtained by a depolarization lidar in operation in Natal, Brazil. The DUSTER system has 4 channels, namely: 1064, 532 s/p and 355 nm. This system is calibrated with a half-wave plate using the Δ90° methodology. The data obtained from this system is correlated with AERONET sunphotometer data, and, when available, CALIPSO satellite data. In addition a trajectory model (HYSPLIT) is used to calculate backward trajectories to assess the origin of the dust polluted air parcels. The objective is to create a transport database of Saharan dust.

Spreading Depression, Spreading Depolarizations, and the Cerebral Vasculature

PubMed Central

Ayata, Cenk; Lauritzen, Martin

2015-01-01

Spreading depression (SD) is a transient wave of near-complete neuronal and glial depolarization associated with massive transmembrane ionic and water shifts. It is evolutionarily conserved in the central nervous systems of a wide variety of species from locust to human. The depolarization spreads slowly at a rate of only millimeters per minute by way of grey matter contiguity, irrespective of functional or vascular divisions, and lasts up to a minute in otherwise normal tissue. As such, SD is a radically different breed of electrophysiological activity compared with everyday neural activity, such as action potentials and synaptic transmission. Seventy years after its discovery by Leão, the mechanisms of SD and its profound metabolic and hemodynamic effects are still debated. What we did learn of consequence, however, is that SD plays a central role in the pathophysiology of a number of diseases including migraine, ischemic stroke, intracranial hemorrhage, and traumatic brain injury. An intriguing overlap among them is that they are all neurovascular disorders. Therefore, the interplay between neurons and vascular elements is critical for our understanding of the impact of this homeostatic breakdown in patients. The challenges of translating experimental data into human pathophysiology notwithstanding, this review provides a detailed account of bidirectional interactions between brain parenchyma and the cerebral vasculature during SD and puts this in the context of neurovascular diseases. PMID:26133935

Multiple scattering effects on the Linear Depolarization Ratio (LDR) measured during CaPE by a Ka-band air-borne radar

NASA Technical Reports Server (NTRS)

Iguchi, Toshio; Meneghini, Robert

1993-01-01

Air-borne radar measurements of thunderstorms were made as part of the CaPE (Convection and Precipitation/Electrification) experiment in Florida in July 1991. The radar has two channels, X-band (10 GHz) and Ka-band (34.5 GHz), and is capable of measuring cross-polarized returns as well as co-polarized returns. In stratiform rain, the cross-polarized components can be observed only at the bright band region and from the surface reflection. The linear depolarization ratios (LDR's) measured at X-band and Ka-band at the bright band are nearly equal. In convective rain, however, the LDR in Ka-band often exceeds the X-band LDR by several dB, and sometimes by more than 10 dB, reaching LDR values of up to -5 dB over heavy convective rain. For randomly oriented hydrometeors, such high LDR values cannot be explained by single scattering from non-spherical scattering particles alone. Because the LDR by single backscatter depends weakly on the wavelength, the difference between the Ka-band and X-band LDR's suggests that multiple scattering effects prevail in the Ka-band LDR. In order to test this inference, the magnitude of the cross-polarized component created by double scattering was calculated using the parameters of the airborne radar, which for both frequencies has beamwidths of 5.1 degrees and pulse widths of 0.5 microsecond. Uniform rain beyond the range of 3 km is assumed.

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.

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.

Depolarizing Actions of Hydrogen Sulfide on Hypothalamic Paraventricular Nucleus Neurons

PubMed Central

Khademullah, C. Sahara; Ferguson, Alastair V.

2013-01-01

Hydrogen sulfide (H2S) is a novel neurotransmitter that has been shown to influence cardiovascular functions as well and corticotrophin hormone (CRH) secretion. Since the paraventricular nucleus of the hypothalamus (PVN) is a central relay center for autonomic and endocrine functions, we sought to investigate the effects of H2S on the neuronal population of the PVN. Whole cell current clamp recordings were acquired from the PVN neurons and sodium hydrosulfide hydrate (NaHS) was bath applied at various concentrations (0.1, 1, 10, and 50 mM). NaHS (1, 10, and 50 mM) elicited a concentration-response relationship from the majority of recorded neurons, with almost exclusively depolarizing effects following administration. Cells responded and recovered from NaHS administration quickly and the effects were repeatable. Input differences from baseline and during the NaHS-induced depolarization uncovered a biphasic response, implicating both a potassium and non-selective cation conductance. The results from the neuronal population of the PVN shed light on the possible physiological role that H2S has in autonomic and endocrine function. PMID:23691233

Modeling of transdermal fluorescence measurements from first-in-human clinical trials for renal function determination using fluorescent tracer agent MB-102

NASA Astrophysics Data System (ADS)

Shultz, Kimberly M.; Debreczeny, Martin P.; Dorshow, Richard B.; Keating, Jennifer E.; Bechtel, Kate L.

2017-02-01

The fluorescent tracer agent 3,6-diamino-2,5-bisN-[(1R)-1-carboxy-2-hydroxyethyl]carbamoylpyrazine, designated MB-102, is cleared from the body solely by the kidneys. A prototype noninvasive fluorescence detection device has been developed for monitoring transdermal fluorescence after bolus intravenous injection of MB-102 in order to measure kidney function. A mathematical model of the detected fluorescence signal was created for evaluation of observed variations in agent kinetics across body locations and for analysis of candidate instrument geometries. The model comprises pharmacokinetics of agent distribution within body compartments, local diffusion of the agent within the skin, Monte Carlo photon transport through tissue, and ray tracing of the instrument optics. Data from eight human subjects with normal renal function and a range of skin colors shows good agreement with simulated data. Body site dependence of equilibration kinetics was explored using the model to find the local vasculature-to-interstitial diffusion time constant, blood volume fraction, and interstitial volume fraction. Finally, candidate instrument geometries were evaluated using the model. While an increase in source-detector separation was found to increase sensitivity to tissue optical properties, it reduced the relative intensity of the background signal with minimal effect on the measured equilibration kinetics.

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

Impossible Dreams, Impossible Choices, and Thoughts about Depolarizing the Debate

ERIC Educational Resources Information Center

Morrow, Susan L.; Beckstead, A. Lee; Hayes, Jeffrey A.; Haldeman, Douglas C.

2004-01-01

The titles of the reactions to this major contribution alone set the stage for further exploration of the issues regarding the hopes and dreams of same-sex attracted (SSA) clients in religious conflict and their therapists, issues of choice, and whether or not it is possible - or even appropriate - to depolarize the current debate (Gonsiorek,…

Time-resolved and temperature tuneable measurements of fluorescent intensity using a smartphone fluorimeter.

PubMed

Hossain, Md Arafat; Canning, John; Yu, Zhikang; Ast, Sandra; Rutledge, Peter J; Wong, Joseph K-H; Jamalipour, Abbas; Crossley, Maxwell J

2017-05-30

A smartphone fluorimeter capable of time-based fluorescence intensity measurements at various temperatures is reported. Excitation is provided by an integrated UV LED (λ ex = 370 nm) and detection obtained using the in-built CMOS camera. A Peltier is integrated to allow measurements of the intensity over T = 10 to 40 °C. All components are controlled using a smartphone battery powered Arduino microcontroller and a customised Android application that allows sequential fluorescence imaging and quantification every δt = 4 seconds. The temperature dependence of fluorescence intensity for four emitters (rhodamine B, rhodamine 6G, 5,10,15,20-tetraphenylporphyrin and 6-(1,4,8,11-tetraazacyclotetradecane)2-ethyl-naphthalimide) are characterised. The normalised fluorescence intensity over time of the latter chemosensor dye complex in the presence of Zn 2+ is observed to accelerate with an increasing rate constant, k = 1.94 min -1 at T = 15 °C and k = 3.64 min -1 at T = 30 °C, approaching a factor of ∼2 with only a change in temperature of ΔT = 15 °C. Thermally tuning these twist and bend associated rates to optimise sensor approaches and device applications is proposed.

Investigation of the Feasibility of Using Laser Induced Fluorescence for Concentration Measurements by Diatomic Sulfur.

DTIC Science & Technology

1982-12-01

GEP/PH/82D-1O INVESTIGATION OF THE FEASIBILITY OF USING LASER INDUCED FLUORESCENCE FOR CONCENTRATION MEASUREMENTS OF DIATOMIC SULFUR THESIS AFIT/GEP...FEASIBILITY OF USING LASER INDUCED FLUORESCENCE FOR CONCENTRATION MEASUREMENTS OF DIATOMIC SULFUR THESIS Presented to the Faculty of the School of...December 1982 SPecial Approved for public release; distribution unlimited Preface This thesis presented a rare opportunity and a formidable challenge

Selective depolarization of the muscle membrane in frog nerve-muscle preparations by a chromatographically purified extract of the dinoflagellate Ostreopsis lenticularis

PubMed Central

Meunier, Frédéric A; Mercado, José A; Molgó, Jordi; Tosteson, Thomas R; Escalona de Motta, Gladys

1997-01-01

The actions of a chromatographically identified extract of the marine dinoflagellate Ostreopsis lenticularis, named ostreotoxin-3 (OTX-3), were studied on frog isolated neuromuscular preparations. OTX-3 (1–10 μg ml−1) applied to cutaneous pectoris nerve-muscle preparations depolarized skeletal muscle fibres and caused spontaneous contractions. The depolarization was neither reversed by prolonged washing nor by (+)-tubocurarine. OTX-3 decreased the amplitude of miniature end plate potentials (m.e.p.ps) but did not affect their frequency. Extracellular recording of compound action potentials revealed that OTX-3 affected neither excitability nor conduction along intramuscular nerve branches. End-plate potentials (e.p.ps) elicited by nerve stimulation were reduced in amplitude by OTX-3 and even showed reversed polarity in junctions deeply depolarized by the toxin. Membrane depolarization induced by OTX-3 was decreased about 70% in muscles pretreated for 30 min with 10 μM tetrodotoxin. In contrast, muscles pretreated with 5 μM μ-conotoxin GIIIA were completely insensitive to OTX-3-induced depolarization. OTX-3 did not affect e.p.p. amplitude and the quantal content of e.p.ps in junctions in which muscle depolarization was abolished by μ-conotoxin GIIIA. OTX-3 is a novel type of sodium-channel activating toxin that discriminates between nerve and skeletal muscle membranes. PMID:9249261

Relationship between depolarization-induced force responses and Ca2+ content in skeletal muscle fibres of rat and toad.

PubMed Central

Owen, V J; Lamb, G D; Stephenson, D G; Fryer, M W

1997-01-01

1. The relationship between the total Ca2+ content of a muscle fibre and the magnitude of the force response to depolarization was examined in mechanically skinned fibres from the iliofibularis muscle of the toad and the extensor digitorum longus muscle of the rat. The response to depolarization in each skinned fibre was assessed either at the endogenous level of Ca2+ content or after depleting the fibre of Ca2+ to some degree. Ca2+ content was determined by a fibre lysing technique. 2. In both muscle types, the total Ca2+ content could be reduced from the endogenous level of approximately 1.3 mmol l-1 (expressed relative to intact fibre volume) to approximately 0.25 mmol l-1 by either depolarization or caffeine application in the presence of Ca2+ chelators, showing that the great majority of the Ca2+ was stored in the sarcoplasmic reticulum (SR). Chelation of Ca2+ in the transverse tubular (T-) system, either by exposure of fibres to EGTA before skinning or by permeabilizing the T-system with saponin after skinning, reduced the lower limit of Ca2+ content to < or = 0.12 mmol l-1, indicating that 10-20% of the total fibre Ca2+ resided in the T-system. 3. In toad fibres, both the peak and the area (i.e. time integral) of the force response to depolarization were reduced by any reduction in SR Ca2+ content, with both decreasing to zero in an approximately linear manner as the SR Ca2+ content was reduced to < 15% of the endogenous level. In rat fibres, the peak size of the force response was less affected by small decreases in SR content, but both the peak and area of the response decreased to zero with greater depletion. In partially depleted toad fibres, inhibition of SR Ca2+ uptake potentiated the force response to depolarization almost 2-fold. 4. The results show that in this skinned fibre preparation: (a) T-system depolarization and caffeine application can each virtually fully deplete the SR of Ca2+, irrespective of any putative inhibitory effect of SR depletion

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.

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.

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.

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

NASA Astrophysics Data System (ADS)

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

2004-03-01

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

Absolute intensity measurements of impurity emissions in a shock tunnel and their consequences for laser-induced fluorescence experiments

NASA Technical Reports Server (NTRS)

Palma, P. C.; Houwing, A. F. P.; Sandeman, R. J.

1993-01-01

Absolute intensity measurements of impurity emissions in a shock tunnel nozzle flow are presented. The impurity emission intensities were measured with a photomultiplier and optical multichannel analyzer and calibrated against an intensity standard. The various metallic contaminants were identified and their intensities measured in the spectral regions 290 to 330 nm and 375 to 385 nm. A comparison with calculated fluorescence intensities for predissociated laser-induced fluorescence signals is made. It is found that the emission background is negligible for most fluorescence experiments.

Anion Channel Inhibitor NPPB-Inhibited Fluoride Accumulation in Tea Plant (Camellia sinensis) Is Related to the Regulation of Ca2+, CaM and Depolarization of Plasma Membrane Potential

PubMed Central

Zhang, Xian-Chen; Gao, Hong-Jian; Yang, Tian-Yuan; Wu, Hong-Hong; Wang, Yu-Mei; Zhang, Zheng-Zhu; Wan, Xiao-Chun

2016-01-01

Tea plant is known to be a hyper-accumulator of fluoride (F). Over-intake of F has been shown to have adverse effects on human health, e.g., dental fluorosis. Thus, understanding the mechanisms fluoride accumulation and developing potential approaches to decrease F uptake in tea plants might be beneficial for human health. In the present study, we found that pretreatment with the anion channel inhibitor NPPB reduced F accumulation in tea plants. Simultaneously, we observed that NPPB triggered Ca2+ efflux from mature zone of tea root and significantly increased relative CaM in tea roots. Besides, pretreatment with the Ca2+ chelator (EGTA) and CaM antagonists (CPZ and TFP) suppressed NPPB-elevated cytosolic Ca2+ fluorescence intensity and CaM concentration in tea roots, respectively. Interestingly, NPPB-inhibited F accumulation was found to be significantly alleviated in tea plants pretreated with either Ca2+ chelator (EGTA) or CaM antagonists (CPZ and TFP). In addition, NPPB significantly depolarized membrane potential transiently and we argue that the net Ca2+ and H+ efflux across the plasma membrane contributed to the restoration of membrane potential. Overall, our results suggest that regulation of Ca2+-CaM and plasma membrane potential depolarization are involved in NPPB-inhibited F accumulation in tea plants. PMID:26742036

Differential 3D Mueller-matrix mapping of optically anisotropic depolarizing biological layers

NASA Astrophysics Data System (ADS)

Ushenko, O. G.; Grytsyuk, M.; Ushenko, V. O.; Bodnar, G. B.; Vanchulyak, O.; Meglinskiy, I.

2018-01-01

The paper consists of two parts. The first part is devoted to the short theoretical basics of the method of differential Mueller-matrix description of properties of partially depolarizing layers. It was provided the experimentally measured maps of differential matrix of the 2nd order of polycrystalline structure of the histological section of rectum wall tissue. It was defined the values of statistical moments of the1st-4th orders, which characterize the distribution of matrix elements. In the second part of the paper it was provided the data of statistic analysis of birefringence and dichroism of the histological sections of connecting component of vagina wall tissue (normal and with prolapse). It were defined the objective criteria of differential diagnostics of pathologies of vagina wall.

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.

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

PubMed

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

2017-01-01

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

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.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

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

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.

Unmixing of fluorescence spectra to resolve quantitative time-series measurements of gene expression in plate readers.

PubMed

Lichten, Catherine A; White, Rachel; Clark, Ivan B N; Swain, Peter S

2014-02-03

To connect gene expression with cellular physiology, we need to follow levels of proteins over time. Experiments typically use variants of Green Fluorescent Protein (GFP), and time-series measurements require specialist expertise if single cells are to be followed. Fluorescence plate readers, however, a standard in many laboratories, can in principle provide similar data, albeit at a mean, population level. Nevertheless, extracting the average fluorescence per cell is challenging because autofluorescence can be substantial. Here we propose a general method for correcting plate reader measurements of fluorescent proteins that uses spectral unmixing and determines both the fluorescence per cell and the errors on that fluorescence. Combined with strain collections, such as the GFP fusion collection for budding yeast, our methodology allows quantitative measurements of protein levels of up to hundreds of genes and therefore provides complementary data to high throughput studies of transcription. We illustrate the method by following the induction of the GAL genes in Saccharomyces cerevisiae for over 20 hours in different sugars and argue that the order of appearance of the Leloir enzymes may be to reduce build-up of the toxic intermediate galactose-1-phosphate. Further, we quantify protein levels of over 40 genes, again over 20 hours, after cells experience a change in carbon source (from glycerol to glucose). Our methodology is sensitive, scalable, and should be applicable to other organisms. By allowing quantitative measurements on a per cell basis over tens of hours and over hundreds of genes, it should increase our understanding of the dynamic changes that drive cellular behaviour.

Unmixing of fluorescence spectra to resolve quantitative time-series measurements of gene expression in plate readers

PubMed Central

2014-01-01

Background To connect gene expression with cellular physiology, we need to follow levels of proteins over time. Experiments typically use variants of Green Fluorescent Protein (GFP), and time-series measurements require specialist expertise if single cells are to be followed. Fluorescence plate readers, however, a standard in many laboratories, can in principle provide similar data, albeit at a mean, population level. Nevertheless, extracting the average fluorescence per cell is challenging because autofluorescence can be substantial. Results Here we propose a general method for correcting plate reader measurements of fluorescent proteins that uses spectral unmixing and determines both the fluorescence per cell and the errors on that fluorescence. Combined with strain collections, such as the GFP fusion collection for budding yeast, our methodology allows quantitative measurements of protein levels of up to hundreds of genes and therefore provides complementary data to high throughput studies of transcription. We illustrate the method by following the induction of the GAL genes in Saccharomyces cerevisiae for over 20 hours in different sugars and argue that the order of appearance of the Leloir enzymes may be to reduce build-up of the toxic intermediate galactose-1-phosphate. Further, we quantify protein levels of over 40 genes, again over 20 hours, after cells experience a change in carbon source (from glycerol to glucose). Conclusions Our methodology is sensitive, scalable, and should be applicable to other organisms. By allowing quantitative measurements on a per cell basis over tens of hours and over hundreds of genes, it should increase our understanding of the dynamic changes that drive cellular behaviour. PMID:24495318

Noninvasive Measurement of Bacterial Intracellular pH on a Single-Cell Level with Green Fluorescent Protein and Fluorescence Ratio Imaging Microscopy

PubMed Central

Olsen, Katja N.; Budde, Birgitte B.; Siegumfeldt, Henrik; Rechinger, K. Björn; Jakobsen, Mogens; Ingmer, Hanne

2002-01-01

We show that a pH-sensitive derivative of the green fluorescent protein, designated ratiometric GFP, can be used to measure intracellular pH (pHi) in both gram-positive and gram-negative bacterial cells. In cells expressing ratiometric GFP, the excitation ratio (fluorescence intensity at 410 and 430 nm) is correlated to the pHi, allowing fast and noninvasive determination of pHi that is ideally suited for direct analysis of individual bacterial cells present in complex environments. PMID:12147523

The release of acetylcholine from post-ganglionic cell bodies in response to depolarization.

PubMed Central

Johnson, D A; Pilar, G

1980-01-01

1. Acetylcholine (Ach) release from parasympathetic ganglia cell somata was investigated in denervated avian ciliary ganglia. Three days after the input to the ganglion (the oculomotor nerve) was sectioned, all presynaptic nerve terminals had degenerated. 2. Denervated ganglia were shown to contain endogenous ACh and to be capable of synthesizing [3H]ACh from [3H]choline added to the incubation medium. 3. In response to depolarization induced by incubation in 50 mM-[K+]o, denervated ganglia released [3H]ACh into bath effluents in amounts approximately 15% of the non-denervated contralateral control. This release was shown to be Ca2+ dependent in both intact and denervated ganglia. 4. Antidromic electrical stimulation of ciliary nerves also elicited [3H]ACh release. Nicotine (1 microgram/microliter.) depolarized denervated ciliary ganglion cells and evoked release of the transmitter and this release was antagonized by curare. 5. It is concluded that the ganglionic cell bodies sysnthesized ACh and released the transmitter in response to K+ depolarization, antidromic stimulation and cholinergic agonists, despite the lack of morphological specializations usually associated with stimulus-induced release of neurotransmitter. The evidence suggests the existence of a mechanism of transmitter release which is Ca2+ dependent, probably from a cytoplasmic pool and therefore distinct from the usual vesicular release at the nerve terminal. Images Plate 1 Plate 2 PMID:6247485

Membrane potentials in pinched-off presynaptic nerve ternimals monitored with a fluorescent probe: evidence that synaptosomes have potassium diffusion potentials.

PubMed Central

Blaustein, M P; Goldring, J M

1975-01-01

1. Some physiological properties of tissue fractions from rat brain homogenates have been examined. Of the three fractions studied (presynaptic nerve terminals, mitochondria and fragmented membranes), only the nerve terminals (synaptosomes) have the ability to accumulate 42K from physiological salt solutions. 2. The ability to accumulate and retain K is lost if synaptosomes are exposed to very hypotonic solutions. The K uptake and total K content is reduced by ouabain and by inhibitors of glycolysis and oxidative phosphorylation. 3. These results suggest that synaptosomes in physiological saline accumulate K against a concentration gradient, and may have K diffusion potentials across their surface membranes. The voltage-sensitive fluorescent probe, 3,3'-dipentyl 2,2'-oxacarbocyanine (CC5), was used to test this possibility. 4. In the squid axon, the fluorescent emission of CC5 is directly proportional to membrane potential; depolarization causes an increase in fluorescence. 5. The fluorescence of synaptosomes ('synaptosome fluorescence') treated with CC5 is increased when [K]o is increased or [K]o is reduced; replacement of external Na by Li or choline has little effect on the synaptosome fluorescence. In quantitative terms, synaptosome fluorescence is proportional to log ([K]o plus 0-05[Na]o). Rb is about as effective as K in enhancing synaptosome fluorescence; Cs is about 1/4 as effective. The effect of increased [K]o is reversible. 6. The fluorescence data provide corroborative evidence that there is normally a large K gradient ([K]o smaller than [I]i) across the synaptosome surface membrane. The data suggest the [K]i may be in excess of 100 mM. 7. Replacement of Cl- by methylsulphate did not significantly affect the relationship between synaptosome fluorescence and [K]o, nor did removal of external Ca. 8. The fluorescence of CC5-treated mitochondria, membrane fragmnets, or lysed synaptosomes is unaffected by changes in the K concentration of the medium. 9

Crayfish neuromuscular facilitation activated by constant presynaptic action potentials and depolarizing pulses.

PubMed

Zucker, R S

1974-08-01

1. Experiments were conducted to test the hypothesis that facilitation of transmitter release in response to repetitive stimulation of the exciter motor axon to the crayfish claw opener muscle is due to an increase in the amplitude or duration of the action potential in presynaptic terminals. No consistent changes were found in the nerve terminal potential (n.t.p.) recorded extracellularly at synaptic sites on the surface of muscle fibres.2. Apparent changes in n.t.p. are attributed to three causes.(i) Some recordings are shown to be contaminated by non-specific muscle responses which grow during facilitation.(ii) Some averaged n.t.p.s exhibit opposite changes in amplitude and duration which suggest a change in the synchrony of presynaptic nerve impulses at different frequencies.(iii) Some changes in n.t.p. are blocked by gamma-methyl glutamate, an antagonist of the post-synaptic receptor, which suggests that these changes are caused by small muscle movements.3. The only change in n.t.p. believed to represent an actual change in the intracellular signal is a reduction in n.t.p. amplitude to the second of two stimuli separated by a brief interval.4. Tetra-ethyl ammonium ions increase synaptic transmission about 20% and prolong the n.t.p. about 15%. This result suggests that an increase in n.t.p. large enough to increase transmission by the several hundred per cent occurring during facilitation would be detected.5. The nerve terminals are electrically excitable, and most synaptic sites have a diphasic or triphasic n.t.p., which suggests that the motor neurone terminals are actively invaded by nerve impulses.6. When nerve impulses are blocked in tetrodotoxin, depolarization of nerve terminals increases the frequency of miniature excitatory junctional potentials (e.j.p.s), and a phasic e.j.p. can be evoked by large, brief depolarizing pulses. Responses to repetitive or paired depolarizations of constant amplitude and duration exhibit a facilitation similar to that of e

B cell activation. III. B cell plasma membrane depolarization and hyper- Ia antigen expression induced by receptor immunoglobulin cross-linking are coupled

PubMed Central

1983-01-01

We report investigation of the relationship between ligand-induced B cell plasma membrane depolarization and increased expression of membrane-associated, I-A subregion encoded (mI-A) antigens. Results demonstrate that equal frequencies of B cells are stimulated to undergo membrane depolarization and to increase mI-A expression in response to mitogen, anti-Ig, and thymus-independent (TI) or thymus-dependent (TD) antigens. Further, a cause-and-effect relationship between these two events is suggested by results that demonstrate that inhibition of anti- Fab--induced depolarization by valinomycin also inhibits the subsequent increase in mI-A antigen expression and "passive" (non-ligand-mediated) depolarization of murine B cells by K+ results in hyper-mI-A antigen expression. Based upon these results we hypothesize that antigen- mediated receptor cross-linking results in signal transduction via membrane depolarization, which is resultant in increased mI-A antigen synthesis and cell surface expression. This increase in mI-A antigen density may render the B cell more receptive to subsequent interaction with I-region-restricted helper T cells. PMID:6415207

An integrated platform for surface forces measurements and fluorescence correlation spectroscopy

NASA Astrophysics Data System (ADS)

Mukhopadhyay, Ashis; Zhao, Jiang; Bae, Sung Chul; Granick, Steve

2003-06-01

We describe an apparatus to measure the diffusion of dilute fluorophores in molecularly thin liquid films within a surface forces apparatus (SFA). The design is a significant modification of the traditional SFA in that it allows one to combine nanorheology with the single-molecule sensitive technique of fluorescence correlation spectroscopy. The primary enabling idea was to place a miniaturized SFA onto the stage of an optical microscope equipped with a long working distance objective and illuminated by a femtosecond laser. A secondary enabling idea was that the silver coating on the backside of mica, normally used in the traditional SFA design for interferometric measurements of the film thickness, was replaced by multilayer dielectric coatings that allowed simultaneous interferometry and fluorescence measurements in different regions of the optical spectrum. To illustrate the utility of this instrument, we contrast the translational diffusion of rhodamine dye molecules (in the solvent, 1,2-propane diol), in the unconfined bulk state and confined between mica sheets to the thickness 2.5 nm. The diffusion coefficient is found to decrease by 2 orders of magnitude under confinement.

Micro-Droplet Detection Method for Measuring the Concentration of Alkaline Phosphatase-Labeled Nanoparticles in Fluorescence Microscopy

PubMed Central

Li, Rufeng; Wang, Yibei; Xu, Hong; Fei, Baowei; Qin, Binjie

2017-01-01

This paper developed and evaluated a quantitative image analysis method to measure the concentration of the nanoparticles on which alkaline phosphatase (AP) was immobilized. These AP-labeled nanoparticles are widely used as signal markers for tagging biomolecules at nanometer and sub-nanometer scales. The AP-labeled nanoparticle concentration measurement can then be directly used to quantitatively analyze the biomolecular concentration. Micro-droplets are mono-dispersed micro-reactors that can be used to encapsulate and detect AP-labeled nanoparticles. Micro-droplets include both empty micro-droplets and fluorescent micro-droplets, while fluorescent micro-droplets are generated from the fluorescence reaction between the APs adhering to a single nanoparticle and corresponding fluorogenic substrates within droplets. By detecting micro-droplets and calculating the proportion of fluorescent micro-droplets to the overall micro-droplets, we can calculate the AP-labeled nanoparticle concentration. The proposed micro-droplet detection method includes the following steps: (1) Gaussian filtering to remove the noise of overall fluorescent targets, (2) a contrast-limited, adaptive histogram equalization processing to enhance the contrast of weakly luminescent micro-droplets, (3) an red maximizing inter-class variance thresholding method (OTSU) to segment the enhanced image for getting the binary map of the overall micro-droplets, (4) a circular Hough transform (CHT) method to detect overall micro-droplets and (5) an intensity-mean-based thresholding segmentation method to extract the fluorescent micro-droplets. The experimental results of fluorescent micro-droplet images show that the average accuracy of our micro-droplet detection method is 0.9586; the average true positive rate is 0.9502; and the average false positive rate is 0.0073. The detection method can be successfully applied to measure AP-labeled nanoparticle concentration in fluorescence microscopy. PMID:29160812

Micro-Droplet Detection Method for Measuring the Concentration of Alkaline Phosphatase-Labeled Nanoparticles in Fluorescence Microscopy.

PubMed

Li, Rufeng; Wang, Yibei; Xu, Hong; Fei, Baowei; Qin, Binjie

2017-11-21

This paper developed and evaluated a quantitative image analysis method to measure the concentration of the nanoparticles on which alkaline phosphatase (AP) was immobilized. These AP-labeled nanoparticles are widely used as signal markers for tagging biomolecules at nanometer and sub-nanometer scales. The AP-labeled nanoparticle concentration measurement can then be directly used to quantitatively analyze the biomolecular concentration. Micro-droplets are mono-dispersed micro-reactors that can be used to encapsulate and detect AP-labeled nanoparticles. Micro-droplets include both empty micro-droplets and fluorescent micro-droplets, while fluorescent micro-droplets are generated from the fluorescence reaction between the APs adhering to a single nanoparticle and corresponding fluorogenic substrates within droplets. By detecting micro-droplets and calculating the proportion of fluorescent micro-droplets to the overall micro-droplets, we can calculate the AP-labeled nanoparticle concentration. The proposed micro-droplet detection method includes the following steps: (1) Gaussian filtering to remove the noise of overall fluorescent targets, (2) a contrast-limited, adaptive histogram equalization processing to enhance the contrast of weakly luminescent micro-droplets, (3) an red maximizing inter-class variance thresholding method (OTSU) to segment the enhanced image for getting the binary map of the overall micro-droplets, (4) a circular Hough transform (CHT) method to detect overall micro-droplets and (5) an intensity-mean-based thresholding segmentation method to extract the fluorescent micro-droplets. The experimental results of fluorescent micro-droplet images show that the average accuracy of our micro-droplet detection method is 0.9586; the average true positive rate is 0.9502; and the average false positive rate is 0.0073. The detection method can be successfully applied to measure AP-labeled nanoparticle concentration in fluorescence microscopy.

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

NASA Astrophysics Data System (ADS)

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

2016-09-01

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

Induction of necrosis and apoptosis to KB cancer cells by sanguinarine is associated with reactive oxygen species production and mitochondrial membrane depolarization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chang, M.-C.; Chan, C.-P.; Wang, Y.-J.

2007-01-15

Sanguinarine is a benzopheanthridine alkaloid present in the root of Sanguinaria canadensis L. and Chellidonium majus L. In this study, sanguinarine (2 and 3 {mu}M) exhibited cytotoxicity to KB cancer cells by decreasing MTT reduction to 83% and 52% of control after 24-h of exposure. Sanguinarine also inhibited the colony forming capacity (> 52-58%) and growth of KB cancer cells at concentrations higher than 0.5-1 {mu}M. Short-term exposure to sanguinarine (> 0.5 {mu}M) effectively suppressed the adhesion of KB cells to collagen and fibronectin (FN). Sanguinarine (2 and 3 {mu}M) induced evident apoptosis as indicated by an increase in sub-G0/G1more » populations, which was detected after 6-h of exposure. Only a slight increase in cells arresting in S-phase and G2/M was noted. Induction of KB cell apoptosis and necrosis by sanguinarine (2 and 3 {mu}M) was further confirmed by Annexin V-PI dual staining flow cytometry and the presence of DNA fragmentation. The cytotoxicity by sanguinarine was accompanied by an increase in production of reactive oxygen species (ROS) and depolarization of mitochondrial membrane potential as indicated by single cell flow cytometric analysis of DCF and rhodamine fluorescence. NAC (1 and 3 mM) and catalase (2000 U/ml) prevented the sanguinarine-induced ROS production and cytotoxicity, whereas dimethylthiourea (DMT) showed no marked preventive effect. These results suggest that sanguinarine has anticarcinogenic properties with induction of ROS production and mitochondrial membrane depolarization, which mediate cancer cell death.« less

The Use Of Fluorescence Quenching To Measure Oxygen Concentration

NASA Astrophysics Data System (ADS)

Cox, M. E.; Dunn, B.

1986-01-01

The method of fluorescence quenching is used to measure the concentration of molecular oxygen. The method is rapid, reversible, and does not consume oxygen. The method may provide the basis for a unique biomedical sensor. The key to developing such a device lies in the choice of a fluorophor/polymer composite matrix having the desired optical and transport properties. Experimental results will be presented for certain parameters essential for assessing device development. The properties of interest include the kinetics of fluorescence quenching, the biomolecular rate constants, the temperature dependence of oxygen solubility and diffusivity in the composite matrix, and the oxygen diffusion coefficient. Poly(dimethyl siloxane) [PDMS] was chosen as the polymer host because it is biocompatible, hydrophobic, has a high diffusivity for the simple gases, and is easily bonded to fused silica. 9,10-diphenyl anthracene [9,10-D] was selected since it is readily soluble in a number of organic solvents, has an excitation spectrum in the near UV, an emission spectrum in the visible, a long fluorescence lifetime, and a high quantum yield. When incorporated into PDMS, the optical spectra of 9,10-D does not alter appreciably. The response time of the device is determined by the solution/diffusion kinetics of oxygen in PDMS. The solubility of oxygen in PDMS decreases with increasing temperature and an enthalpy of solution of off = -3.0 kcal/mole. (1) The diffusion of oxyzen in PDMS is found to obey an Arrhenius relation over the temperature range of 5'C to 450C with D = Do exp (-ED/RT) (2) where Do = 0.115 cm2/s (3) and ED = 4.77 kcal/mole. (4) Results of these studies indicate that an appropriate device, based on a fluorophor/polymer composite, for the measurement of oxygen concentration should be sensitive over those ranges which are important for physiological monitoring.

Measurement of fluorophore concentrations and fluorescence quantum yield in tissue-simulating phantoms using three diffusion models of steady-state spatially resolved fluorescence.

PubMed

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

2003-12-21

Steady-state diffusion theory models of fluorescence in tissue have been investigated for recovering fluorophore concentrations and fluorescence quantum yield. Spatially resolved fluorescence, excitation and emission reflectance Carlo simulations, and measured using a multi-fibre probe on tissue-simulating phantoms containing either aluminium phthalocyanine tetrasulfonate (AlPcS4), Photofrin meso-tetra-(4-sulfonatophenyl)-porphine dihydrochloride The accuracy of the fluorophore concentration and fluorescence quantum yield recovered by three different models of spatially resolved fluorescence were compared. The models were based on: (a) weighted difference of the excitation and emission reflectance, (b) fluorescence due to a point excitation source or (c) fluorescence due to a pencil beam excitation source. When literature values for the fluorescence quantum yield were used for each of the fluorophores, the fluorophore absorption coefficient (and hence concentration) at the excitation wavelength (mu(a,x,f)) was recovered with a root-mean-square accuracy of 11.4% using the point source model of fluorescence and 8.0% using the more complicated pencil beam excitation model. The accuracy was calculated over a broad range of optical properties and fluorophore concentrations. The weighted difference of reflectance model performed poorly, with a root-mean-square error in concentration of about 50%. Monte Carlo simulations suggest that there are some situations where the weighted difference of reflectance is as accurate as the other two models, although this was not confirmed experimentally. Estimates of the fluorescence quantum yield in multiple scattering media were also made by determining mu(a,x,f) independently from the fitted absorption spectrum and applying the various diffusion theory models. The fluorescence quantum yields for AlPcS4 and TPPS4 were calculated to be 0.59 +/- 0.03 and 0.121 +/- 0.001 respectively using the point source model, and 0.63 +/- 0.03 and 0

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

Lenticular cytoprotection, part 2: link between glycogen synthase kinase-3β, epithelial to mesenchymal transition, and mitochondrial depolarization.

PubMed

Neelam, Sudha; Brooks, Morgan M; Cammarata, Patrick R

2014-01-01

The inhibition of GSK-3β blocks mitochondrial membrane permeability transition (mMPT) for HLE-B3 cells in atmospheric oxygen. GSK-3β, as part of a multifactorial complex, also regulates nuclear levels of β-catenin, a known coordinator of cell survival and adhesion. The purpose of these studies was to demonstrate a novel, but likely disadvantageous, link between β-catenin's influence on the expression of the pro-survival protein, vascular endothelial growth factor (VEGF), resulting in enhanced lens epithelial cell mitochondrial protection against depolarization and nuclear β-catenin as an inducer of epithelial to mesenchymal transition (EMT). Virally transformed human lens epithelial cells (HLE-B3) were treated with SB216763, a specific inhibitor of GSK-3β catalytic activity and XAV939, a specific β-catenin inhibitor that bars the translocation of β-catenin from cytoplasm to the nucleus. Western blot analysis was employed to detect the levels of cytoplasmic and nuclear β-catenin and phospho-β-catenin, pBcl-2 and the EMT proteins, α-smooth muscle actin (α-SMA), and fibronectin. ELISA was used to measure the levels of VEGF in cell culture supernatants. JC-1 analysis was performed to analyze the influence of either SB216763 or XAV939 on mitochondrial depolarization. Cultured lens epithelial cells maintained in hypoxia (1% oxygen) and subsequently reintroduced into atmospheric oxygen and treated with the GSK-3β inhibitor SB216763 illustrated a marked inhibition of phosphorylation of glycogen synthase (downstream substrate of GSK-3β) and significant increase in nuclear translocation of β-catenin. The augmented nuclear β-catenin levels positively correlated with increased expression of α-SMA and fibronectin, both marker proteins indicative of EMT. The enhanced nuclear β-catenin activity also elicited increased VEGF and pBcl-2 expression, resulting in increased resistance to mitochondrial depolarization. Treatment of the cells with the �

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…

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

Fluorescent optical position sensor

DOEpatents

Weiss, Jonathan D.

2005-11-15

A fluorescent optical position sensor and method of operation. A small excitation source side-pumps a localized region of fluorescence at an unknown position along a fluorescent waveguide. As the fluorescent light travels down the waveguide, the intensity of fluorescent light decreases due to absorption. By measuring with one (or two) photodetectors the attenuated intensity of fluorescent light emitted from one (or both) ends of the waveguide, the position of the excitation source relative to the waveguide can be determined by comparing the measured light intensity to a calibrated response curve or mathematical model. Alternatively, excitation light can be pumped into an end of the waveguide, which generates an exponentially-decaying continuous source of fluorescent light along the length of the waveguide. The position of a photodetector oriented to view the side of the waveguide can be uniquely determined by measuring the intensity of the fluorescent light emitted radially at that location.

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...

Ca(2+)-activated anion channels and membrane depolarizations induced by blue light and cold in Arabidopsis seedlings

NASA Technical Reports Server (NTRS)

Lewis, B. D.; Karlin-Neumann, G.; Davis, R. W.; Spalding, E. P.; Evans, M. L. (Principal Investigator)

1997-01-01

The activation of an anion channel in the plasma membrane of Arabidopsis thaliana hypocotyls by blue light (BL) is believed to be a signal-transducing event leading to growth inhibition. Here we report that the open probability of this particular anion channel depends on cytoplasmic Ca2+ ([Ca2+]cyt) within the concentration range of 1 to 10 microM, raising the possibility that BL activates the anion channel by increasing [Ca2+]cyt. Arabidopsis seedlings cytoplasmically expressing aequorin were generated to test this possibility. Aequorin luminescence did not increase during or after BL, providing evidence that Ca2+ does not play a second-messenger role in the activation of anion channels. However, cold shock simultaneously triggered a large increase in [Ca2+]cyt and a 110-mV transient depolarization of the plasma membrane. A blocker of the anion channel, 5-nitro-2-(3-phenylpropylamino)-benzoic acid, blocked 61% of the cold-induced depolarization without affecting the increase in [Ca2+]cyt. These data led us to propose that cold shock opens Ca2+ channels at the plasma membrane, allowing an inward, depolarizing Ca2+ current. The resulting large increase in [Ca2+]cyt activates the anion channel, which further depolarizes the membrane. Although an increase in [Ca2+]cyt may activate anion channels in response to cold, it appears that BL does so via a Ca(2+)-independent pathway.

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.

Isoflurane depolarizes bronchopulmonary C neurons by inhibiting transient A-type and delayed rectifier potassium channels.

PubMed

Zhang, Zhenxiong; Zhuang, Jianguo; Zhang, Cancan; Xu, Fadi

2013-04-01

Inhalation of isoflurane (ISO), a widely used volatile anesthetic, can produce clinical tachypnea. In dogs, this response is reportedly mediated by bronchopulmonary C-fibers (PCFs), but the relevant mechanisms remain unclear. Activation of transient A-type potassium current (IA) channels and delayed rectifier potassium current (IK) channels hyperpolarizes neurons, and inhibition of both channels by ISO increases neural firing. Due to the presence of these channels in the cell bodies of rat PCFs, we determined whether ISO could stimulate PCFs to produce tachypnea in anesthetized rats, and, if so, whether this response resulted from ISO-induced depolarization of the pulmonary C neurons via the inhibition of IA and IK. We recorded ventilatory responses to 5% ISO exposure in anesthetized rats before and after blocking PCF conduction and the responses of pulmonary C neurons (extracellularly recorded) to ISO exposure. ISO-induced (1mM) changes in pulmonary C neuron membrane potential and IA/IK were tested using the perforated patch clamp technique. We found that: (1) ISO inhalation evoked a brief tachypnea (∼7s) and that this response disappeared after blocking PCF conduction; (2) the ISO significantly elevated (by 138%) the firing rate of most pulmonary C neurons (17 out of 21) in the nodose ganglion; and (3) ISO perfusion depolarized the pulmonary C neurons in the vitro and inhibited both IA and IK, and this evoked-depolarization was largely diminished after blocking both IA and IK. Our results suggest that ISO is able to stimulate PCFs to elicit tachypnea in rats, at least partly, via inhibiting IA and IK, thereby depolarizing the pulmonary C neurons. Copyright © 2013. Published by Elsevier B.V.

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.

Multiple scattering in chiral media: border effects, reduced depolarization, and sensitivity limit

NASA Astrophysics Data System (ADS)

Delplancke, Francoise; Badoz, Jacques P.; Boccara, A. Claude

1997-10-01

Suspensions of polystyrene latex beads in chiral solutions were investigated. The rotatory power, induced by solubilized sucrose, in near-forward scattering was measured via a method using polarization modulation by photo-elastic modulator. The sensitivity of the measurement was enhanced and optimized in order to measure sucrose concentrations as low as 5 mg/ml in a cell 5 mm thick only. Different concentrations and diameters of latex particles were used in combination with different sucrose concentrations going from 1 mg/ml up to saturation. The experiments showed that the apparent rotatory power is enhanced by multiple scattering, that depolarization effects are less important with highly concentrated sucrose solutions and that attention has to be paid to cell border effects in order to avoid important artifacts, in case of highly scattering suspensions. Qualitative and theoretical explanations of those observations are presented. One possible application of this method is to measure the sugar content in human blood, in vivo, non-invasively, through the skin. The concentration to be evaluated is at the sensitivity limit. So any artifact has to be removed carefully, e.g. skin cell birefringence or chirality.

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

PubMed

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

2017-02-01

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

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.

Measurement of drug-target engagement in live cells by two-photon fluorescence anisotropy imaging.

PubMed

Vinegoni, Claudio; Fumene Feruglio, Paolo; Brand, Christian; Lee, Sungon; Nibbs, Antoinette E; Stapleton, Shawn; Shah, Sunil; Gryczynski, Ignacy; Reiner, Thomas; Mazitschek, Ralph; Weissleder, Ralph

2017-07-01

The ability to directly image and quantify drug-target engagement and drug distribution with subcellular resolution in live cells and whole organisms is a prerequisite to establishing accurate models of the kinetics and dynamics of drug action. Such methods would thus have far-reaching applications in drug development and molecular pharmacology. We recently presented one such technique based on fluorescence anisotropy, a spectroscopic method based on polarization light analysis and capable of measuring the binding interaction between molecules. Our technique allows the direct characterization of target engagement of fluorescently labeled drugs, using fluorophores with a fluorescence lifetime larger than the rotational correlation of the bound complex. Here we describe an optimized protocol for simultaneous dual-channel two-photon fluorescence anisotropy microscopy acquisition to perform drug-target measurements. We also provide the necessary software to implement stream processing to visualize images and to calculate quantitative parameters. The assembly and characterization part of the protocol can be implemented in 1 d. Sample preparation, characterization and imaging of drug binding can be completed in 2 d. Although currently adapted to an Olympus FV1000MPE microscope, the protocol can be extended to other commercial or custom-built microscopes.

Activity-associated miRNA are packaged in Map1b-enriched exosomes released from depolarized neurons.

PubMed

Goldie, Belinda J; Dun, Matthew D; Lin, Minjie; Smith, Nathan D; Verrills, Nicole M; Dayas, Christopher V; Cairns, Murray J

2014-08-01

Rapid input-restricted change in gene expression is an important aspect of synaptic plasticity requiring complex mechanisms of post-transcriptional mRNA trafficking and regulation. Small non-coding miRNA are uniquely poised to support these functions by providing a nucleic-acid-based specificity component for universal-sequence-dependent RNA binding complexes. We investigated the subcellular distribution of these molecules in resting and potassium chloride depolarized human neuroblasts, and found both selective enrichment and depletion in neurites. Depolarization was associated with a neurite-restricted decrease in miRNA expression; a subset of these molecules was recovered from the depolarization medium in nuclease resistant extracellular exosomes. These vesicles were enriched with primate specific miRNA and the synaptic-plasticity-associated protein MAP1b. These findings further support a role for miRNA as neural plasticity regulators, as they are compartmentalized in neurons and undergo activity-associated redistribution or release into the extracellular matrix. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

CaMKII and CaMKIV mediate distinct prosurvival signaling pathways in response to depolarization in neurons

PubMed Central

Bok, Jinwoong; Wang, Qiong; Huang, Jie; Green, Steven H.

2007-01-01

By fusing the CaMKII inhibitory peptide AIP to GFP, we constructed a specific and effective CaMKII inhibitor, GFP-AIP. Expression of GFP-AIP and/or dominant-inhibitory CaMKIV in cultured neonatal rat spiral ganglion neurons (SGNs) shows that CaMKII and CaMKIV act additively and in parallel, to mediate the prosurvival effect of depolarization. Depolarization or expression of constitutively-active CaMKII functionally inactivates Bad, indicating that this is one means by which CaMKII promotes neuronal survival. CaMKIV, but not CaMKII, requires CREB to promote SGN survival, consistent with the exclusively nuclear localization of CaMKIV and indicating that the principal prosurvival function of CaMKIV is activation of CREB. Consistent with this, a constitutively-active CREB construct that provides a high level of CREB activity promotes SGN survival, although low levels of CREB activity did not do so. Also, in apoptotic SGNs, activation of CREB by depolarization is disabled, presumably as part of a cellular commitment to apoptosis. PMID:17651987

Retrieval of the non-depolarizing components of depolarizing Mueller matrices by using symmetry conditions and least squares minimization

NASA Astrophysics Data System (ADS)

Kuntman, Ertan; Canillas, Adolf; Arteaga, Oriol

2017-11-01

Experimental Mueller matrices contain certain amount of uncertainty in their elements and these uncertainties can create difficulties for decomposition methods based on analytic solutions. In an earlier paper [1], we proposed a decomposition method for depolarizing Mueller matrices by using certain symmetry conditions. However, because of the experimental error, that method creates over-determined systems with non-unique solutions. Here we propose to use least squares minimization approach in order to improve the accuracy of our results. In this method, we are taking into account the number of independent parameters of the corresponding symmetry and the rank constraints on the component matrices to decide on our fitting model. This approach is illustrated with experimental Mueller matrices that include material media with different Mueller symmetries.

Contribution of corner reflections from oriented ice crystals to backscattering and depolarization characteristics for off-zenith lidar profiling

NASA Astrophysics Data System (ADS)

Borovoi, Anatoli G.; Konoshonkin, Alexander V.; Kustova, Natalia V.; Veselovskii, Igor A.

2018-06-01

Backscattering Mueller matrix and the depolarization and color ratios for quasi-horizontally oriented hexagonal ice plates have been calculated within the framework of the physical optics approximation. In the case of a tilted lidar, the dependence of the color and depolarization ratios on polarization of the incident light has been analyzed. It is shown that the corner reflection effect inherent to the pristine hexagonal ice crystals results in sharp peaks of both the backscattering cross section and depolarization ratio at the lidar tilts of about 30° off zenith. The experimental results obtained recently by Veselovskii et al. [13] at the lidar tilt of 43° have been interpreted as a partial manifestation of the corner reflection effect. The retrieval of the vertical profile of the ice crystal fraction consisting of quasi-horizontally oriented hexagonal plates has been demonstrated.

Backscatter laser depolarization studies of simulated stratospheric aerosols - Crystallized sulfuric acid droplets

NASA Technical Reports Server (NTRS)

Sassen, Kenneth; Zhao, Hongjie; Yu, Bing-Kun

1989-01-01

The optical depolarizing properties of simulated stratospheric aerosols were studied in laboratory laser (0.633 micrometer) backscattering experiments for application to polarization lidar observations. Clouds composed of sulfuric acid solution droplets, some treated with ammonia gas, were observed during evaporation. The results indicate that the formation of minute ammonium sulfate particles from the evaporation of acid droplets produces linear depolarization ratios of beta equivalent to 0.02, but beta equivalent to 0.10 to 0.15 are generated from aged acid cloud aerosols and acid droplet crystalization effects following the introduction of ammonia gas into the chamber. It is concluded that partially crystallized sulfuric acid droplets are a likely candidate for explaining the lidar beta equivalent to 0.10 values that have been observed in the lower stratosphere in the absence of the relatively strong backscattering from homogeneous sulfuric acid droplet (beta equivalent to 0) or ice crystal (beta equivalent to 0.5) clouds.

Backscatter laser depolarization studies of simulated stratospheric aerosols: Crystallized sulfuric acid droplets

NASA Technical Reports Server (NTRS)

Sassen, Kenneth; Zhao, Hongjie; Yu, Bing-Kun

1988-01-01

The optical depolarizing properties of simulated stratospheric aerosols were studied in laboratory laser (0.633 micrometer) backscattering experiments for application to polarization lidar observations. Clouds composed of sulfuric acid solution droplets, some treated with ammonia gas, were observed during evaporation. The results indicate that the formation of minute ammonium sulfate particles from the evaporation of acid droplets produces linear depolarization ratios of beta equivalent to 0.02, but beta equivalent to 0.10 to 0.15 are generated from aged acid cloud aerosols and acid droplet crystallization effects following the introduction of ammonia gas into the chamber. It is concluded that partially crystallized sulfuric acid droplets are a likely candidate for explaining the lidar beta equivalent to 0.10 values that have been observed in the lower stratosphere in the absence of the relatively strong backscattering from homogeneous sulfuric acid droplet (beta equivalent to 0) or ice crystal (beta equivalent to 0.5) clouds.

Glucocorticoid interactions with ethanol effects on depolarization-induced calcium influx in brain synaptosomes.

PubMed

Sze, P Y

1996-04-01

Depolarization-induced Ca2+ influx in brain synaptosomes is known to be inhibited by ethanol and stimulated by glucocorticoids. The present study was undertaken to characterize the interactions of corticosterone (CORT) with ethanol effects on 45Ca2+ uptake in synaptosomes depolarized by high K+ (70 mM). CORT was shown to antagonize the inhibitory effects of ethanol on the fast-phase component of the K(+)-induced 45Ca2+ uptake (the initial 3 s following depolarization). Glucocorticoid antagonism of ethanol inhibition of the 45Ca2+ uptake exhibited a high degree of steroid specificity; steroids with glucocorticoid activity including cortisol, dexamethasone and triamcinolone were effective, whereas gonadal steroids and excitatory neuroactive steroid metabolites were ineffective. From the shift of concentration-response relationships when CORT and ethanol were present in combination, the interaction between steroid stimulation and ethanol inhibition of 45Ca2+ uptake occurred in an additive manner over the range of their effective concentrations. Parallel to 45Ca2+ uptake, the binding sites for [3H]PN 200-110 were reduced by ethanol and increased by CORT. These opposite effects on [3H]dihydropyridine labeled sites were found also to antagonize each other, and the antagonism again occurred in an additive relationship. These results demonstrate that glucocorticoids antagonized ethanol inhibition of voltage-dependent Ca2+ channel activity in brain synaptosomes, and support the notion that these steroids may be among the endogenous factors that modulate neuronal sensitivity to ethanol.

Conditional-sampling spectrograph detection system for fluorescence measurements of individual airborne biological particles

NASA Astrophysics Data System (ADS)

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

1996-03-01

We report the design and operation of a prototype conditional-sampling spectrograph detection system that can record the fluorescence spectra of individual, micrometer-sized aerosols as they traverse an intense 488-nm intracavity laser beam. The instrument's image-intensified CCD detector is gated by elastic scattering or by undispersed fluorescence from particles that enter the spectrograph's field of view. It records spectra only from particles with preselected scattering-fluorescence levels (a fiber-optic-photomultiplier subsystem provides the gating signal). This conditional-sampling procedure reduces data-handling rates and increases the signal-to-noise ratio by restricting the system's exposures to brief periods when aerosols traverse the beam. We demonstrate these advantages by reliably capturing spectra from individual fluorescent microspheres dispersed in an airstream. The conditional-sampling procedure also permits some discrimination among different types of particles, so that spectra may be recorded from the few interesting particles present in a cloud of background aerosol. We demonstrate such discrimination by measuring spectra from selected fluorescent microspheres in a mixture of two types of microspheres, and from bacterial spores in a mixture of spores and nonfluorescent kaolin particles.

Mapping the local organization of cell membranes using excitation-polarization-resolved confocal fluorescence microscopy.

PubMed

Kress, Alla; Wang, Xiao; Ranchon, Hubert; Savatier, Julien; Rigneault, Hervé; Ferrand, Patrick; Brasselet, Sophie

2013-07-02

Fluorescence anisotropy and linear dichroism imaging have been widely used for imaging biomolecular orientational distributions in protein aggregates, fibrillar structures of cells, and cell membranes. However, these techniques do not give access to complete orientational order information in a whole image, because their use is limited to parts of the sample where the average orientation of molecules is known a priori. Fluorescence anisotropy is also highly sensitive to depolarization mechanisms such as those induced by fluorescence energy transfer. A fully excitation-polarization-resolved fluorescence microscopy imaging that relies on the use of a tunable incident polarization and a nonpolarized detection is able to circumvent these limitations. We have developed such a technique in confocal epifluorescence microscopy, giving access to new regions of study in the complex and heterogeneous molecular organization of cell membranes. Using this technique, we demonstrate morphological changes at the subdiffraction scale in labeled COS-7 cell membranes whose cytoskeleton is perturbed. Molecular orientational order is also seen to be affected by cholesterol depletion, reflecting the strong interplay between lipid-packing regions and their nearby cytoskeleton. This noninvasive optical technique can reveal local organization in cell membranes when used as a complement to existing methods such as generalized polarization. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Fluorescence lifetime endoscopy using TCSPC for the measurement of FRET in live cells

PubMed Central

Fruhwirth, Gilbert O.; Ameer-Beg, Simon; Cook, Richard; Watson, Timothy; Ng, Tony; Festy, Frederic

2010-01-01

Development of remote imaging for diagnostic purposes has progressed dramatically since endoscopy began in the 1960’s. The recent advent of a clinically licensed intensity-based fluorescence micro-endoscopic instrument has offered the prospect of real-time cellular resolution imaging. However, interrogating protein-protein interactions deep inside living tissue requires precise fluorescence lifetime measurements to derive the Förster resonance energy transfer between two tagged fluorescent markers. We developed a new instrument combining remote fiber endoscopic cellular-resolution imaging with TCSPC-FLIM technology to interrogate and discriminate mixed fluorochrome labeled beads and expressible GFP/TagRFP tags within live cells. Endoscopic-FLIM (e-FLIM) data was validated by comparison with data acquired via conventional FLIM and e-FLIM was found to be accurate for both bright bead and dim live cell samples. The fiber based micro-endoscope allowed remote imaging of 4 µm and 10 µm beads within a thick Matrigel matrix with confident fluorophore discrimination using lifetime information. More importantly, this new technique enabled us to reliably measure protein-protein interactions in live cells embedded in a 3D matrix, as demonstrated by the dimerization of the fluorescent protein-tagged membrane receptor CXCR4. This cell-based application successfully demonstrated the suitability and great potential of this new technique for in vivo pre-clinical biomedical and possibly human clinical applications. PMID:20588974

Measurements of KrF laser-induced O2 fluorescence in high-temperature atmospheric air

NASA Technical Reports Server (NTRS)

Grinstead, Jay H.; Laufer, Gabriel; Mcdaniel, James C., Jr.

1993-01-01

Conditions for obtaining laser-induced O2 fluorescence using a tunable KrF laser has been determined theoretically and experimentally. With this laser source, O2 rotational temperature measurement is possible even in the absence of vibrational equilibrium. Temperature measurement using a two-line excitation scheme has been demonstrated in a high-temperature atmospheric-air furnace. A measurement uncertainty of 10.7 percent for the temperature range 1325-1725 K was realized. At atmospheric pressure, O2 LIF measurements are possible for air temperatures above 1250 K. Interference from OH fluorescence in reacting flows can be avoided by the proper selection of O2 transitions. Depletion of the ground state population by the incident laser is negligible for intensities below 7.5 x 10 to the 6th W/sq cm/per cm.

Real Time Measures of Prestin Charge and Fluorescence during Plasma Membrane Trafficking Reveal Sub-Tetrameric Activity

PubMed Central

Bian, Shumin; Navaratnam, Dhasakumar; Santos-Sacchi, Joseph

2013-01-01

Prestin (SLC26a5) is the outer hair cell integral membrane motor protein that drives cochlear amplification, and has been described as an obligate tetramer. We studied in real time the delivery of YFP-prestin to the plasma membrane of cells from a tetracycline-inducible cell line. Following the release of temperature block to reinstate trans Golgi network delivery of the integral membrane protein, we measured nonlinear capacitance (NLC) and membrane fluorescence during voltage clamp. Prestin was delivered exponentially to the plasma membrane with a time constant of less than 10 minutes, with both electrical and fluorescence methods showing high temporal correlation. However, based on disparity between estimates of prestin density derived from either fluorescence or NLC, we conclude that sub-tetrameric forms of prestin contribute to our electrical and fluorescence measures. Thus, in agreement with previous observations we find that functional prestin is not an obligate tetramer. PMID:23762468

Particle-fluorescence spectrometer for real-time single-particle measurements of atmospheric organic carbon and biological aerosol.

PubMed

Pan, Yong-Le; Pinnick, Ronald G; Hill, Steven C; Chang, Richard K

2009-01-15

A particle-fluorescence spectrometer (PFS) for real-time measurements of single-particle UV-laser-induced fluorescence (UV-LIF) excited with a pulsed (263-nm) laser is reported. The dispersed UV-LIF spectra are measured by a 32-anode PMT detector with spectral coverage from 280-600 nm. The PFS represents a significant improvement over our previous apparatus [Pinnick et al., Atmos. Environ. 2004, 38, 1657] and can (1) measure fluorescence spectra of bacterial particles having light-scattering sizes as small as 1 microm (previously limited to about 3 microm) and so can measure particles with size in the range of 1-10 microm, (2) measure each particle's elastic scattering which can be used to estimate particle size (not available previously), (3) measure single-particle fluorescence spectra with a laser and detector that can record spectra as fast as 90,000/s, although the highest rates we have found experimentally in atmospheric measurements is only several hundred per second (previously limited by detectors to only 25/s), and (4) provide a time stamp for a data block of spectra with time resolution from 10 ms to 10 min. In addition, the PFS has been modified to be more robust, transportable, and smaller. The use of an aerodynamic-focusing sheath inlet nozzle assembly has improved the sample rate. The PFS has been employed to measure UV-LIF spectra from individual atmospheric particles during October-December 2006 and January-May 2008 in New Haven, CT, and during January-May 2007 in Las Cruces, NM.

Depolarized inactivation overcomes impaired activation to produce DRG neuron hyperexcitability in a Nav1.7 mutation in a patient with distal limb pain.

PubMed

Huang, Jianying; Yang, Yang; Dib-Hajj, Sulayman D; van Es, Michael; Zhao, Peng; Salomon, Jody; Drenth, Joost P H; Waxman, Stephen G

2014-09-10

Sodium channel Nav1.7, encoded by SCN9A, is expressed in DRG neurons and regulates their excitability. Genetic and functional studies have established a critical contribution of Nav1.7 to human pain disorders. We have now characterized a novel Nav1.7 mutation (R1279P) from a female human subject with distal limb pain, in which depolarized fast inactivation overrides impaired activation to produce hyperexcitability and spontaneous firing in DRG neurons. Whole-cell voltage-clamp recordings in human embryonic kidney (HEK) 293 cells demonstrated that R1279P significantly depolarizes steady-state fast-, slow-, and closed-state inactivation. It accelerates deactivation, decelerates inactivation, and facilitates repriming. The mutation increases ramp currents in response to slow depolarizations. Our voltage-clamp analysis showed that R1279P depolarizes channel activation, a change that was supported by our multistate structural modeling. Because this mutation confers both gain-of-function and loss-of-function attributes on the Nav1.7 channel, we tested the impact of R1279P expression on DRG neuron excitability. Current-clamp studies reveal that R1279P depolarizes resting membrane potential, decreases current threshold, and increases firing frequency of evoked action potentials within small DRG neurons. The populations of spontaneously firing and repetitively firing neurons were increased by expressing R1279P. These observations indicate that the dominant proexcitatory gating changes associated with this mutation, including depolarized steady-state fast-, slow-, and closed-state inactivation, faster repriming, and larger ramp currents, override the depolarizing shift of activation, to produce hyperexcitability and spontaneous firing of nociceptive neurons that underlie pain. Copyright © 2014 the authors 0270-6474/14/3412328-13$15.00/0.

Measurement of the Fluorescence Lifetime of Chlorophyll a In Vivo

PubMed Central

Singhal, G. S.; Rabinowitch, E.

1969-01-01

New measurements have been made of fluorescence lifetime (τ) of chlorophyll a in the algae Chlorella pyrenoidosa, Porphyridium cruentum, Anacystis nidulans, and in spinach chloroplast. τ-values of 0.6 and 0.7 nsec were obtained with green plants. Anacystis and Porphyridium gave a τ of 0.5 nsec. The previously described two stage decay of fluorescence in vivo in these organisms could not be confirmed. This observation could have been caused by a second wave of light emission from the exciting hydrogen lamp (not detected in earlier work). The lifetimes found in this study (calculated, as before, by the method of convolution integrals) were close to those found by other observers for “low” excitation intensities; the value first reported from this laboratory (1.0-1.7 nsec) may have corresponded to “high” excitation intensity. PMID:5778187

Use of an automated fluorescent microsphere method to measure regional blood flow in the fetal lamb.

PubMed

Tan, W; Riggs, K W; Thies, R L; Rurak, D W

1997-08-01

We have developed a method for measuring regional blood flow by means of fluorescent microspheres in all organs and tissues of the fetal lamb, including brain, heart, lung, liver, gut, spleen, kidney, adrenal, brown fat, skin, muscle, bone, and placenta. Five different fluorescent-labeled microspheres were used: blue (B), yellow-green (Y), orange (O), red (R), and crimson (C). An automated, 96-well microplate fluorescent reader (bottom reading) was chosen for the assay because of the rapidity and high throughput that it offers. Tissue samples were digested by 4 M ethanolic KOH. The sedimentation method and dye extraction with Cellosolve acetate, as previously reported by others, were used for the sample processing. The bones were crushed and allowed to directly soak in Cellosolve acetate to extract the dye. The relationship between microsphere number and fluorescent intensity was linear over a broad range of microsphere numbers (80-20,000/mL). The coefficients of variation of within-run and between-run precision were 3.39 +/- 1.10% and 4.54 +/- 1.10%, respectively. Recovery of microspheres from tissues and blood averaged 94.3 +/- 2.5% and was not dependent on microsphere number. The spillover of the fluorescent signals into adjacent colors was 4.0 +/- 0.1% for O to Y, 8.1 +/- 0.4% for O to R, and 9.1 +/- 0.5% for R to C, and these values were constant over a wide range in concentrations of the microsphere pairs. No evidence was obtained for quenching of the emission of one fluorophore via photon absorption by another fluorophore. The measurements of regional blood flow obtained with fluorescent microspheres in three chronically instrumented fetal lambs at approximately 140 days gestation were similar to the flow estimates obtained using radioactive microspheres in four other fetal lambs at the same gestational age. The fluorescent method is thus a viable alternative to the radioactive technique for the measurement of regional blood flow to all fetal organs and

Thermal Stress-Induced Depolarization Loss in Conventional and Panda-Shaped Photonic Crystal Fiber Lasers

NASA Astrophysics Data System (ADS)

Mousavi, Seyedeh Laleh; Sabaeian, Mohammad

2016-10-01

We report on the modeling of the depolarization loss in the conventional and panda-shaped photonic crystal fiber lasers (PCFLs) due to the self-heating of the fiber, which we call it thermal stress-induced depolarization loss (TSIDL). We first calculated the temperature distribution over the fiber cross sections and then calculated the thermal stresses/strains as a function of heat load per meter. Thermal stress-induced birefringence (TSIB), which is defined as | n x - n y |, in the core and cladding regions was calculated. Finally, TSIDL was calculated for the conventional and panda-shaped PCFLs as a function of fiber length and, respectively, saturated values of 22 and 25 % were obtained which were independent of heat load per meter. For panda-shaped PCFLs, prior to being saturated, an oscillating and damping behavior against the fiber length was seen where in some lengths reached 35 %. The results are close to an experimental value of 30 % reported for a pulsed PCFL (Limpert et al., Opt Express 12:1313-1319, 2004) where the authors reported a degree of polarization of 70 % (i.e., a depolarization of 30 %). The most important result of this work is a saturation behavior of TSIDL at long-enough lengths of the fiber laser which is independent of heat load per meter. To our knowledge, this the first report of TSIBL for PCFLs.

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.

Depolarization of the Electrogenic Transmembrane Electropotential of Zea mays L. by Bipolaris (Helminthosporium) maydis Race T Toxin, Azide, Cyanide, Dodecyl Succinic Acid, or Cold Temperature 1

PubMed Central

Mertz, Stuart M.; Arntzen, Charles J.

1978-01-01

The transmembrane electrical potential of root cells of Zea mays L. cv. W64A in a modified 1× Higinbotham solution was partially depolarized by semipurified toxin obtained from Bipolaris (Helminthosporium) maydis race T. At a given toxin concentration depolarization of Texas cytoplasm cells was much greater than for normal cytoplasm cells. This observation correlated directly to the differential host susceptibility to the fungus. The time course and magnitude of depolarization were dependent on toxin concentration; at high concentration the electropotential difference change was rapid. Cortex cells depolarized more slowly than epidermal cells indicating that the toxin slowly permeated intercellular regions. Toxin concentrations which affected electropotential difference were of the same magnitude as those required to inhibit root growth, ion uptake, and mitochondrial processes. Azide, cyanide, and cold temperature (5 C) gave the same partial depolarization as did the toxin. Dodecyl succinic acid caused complete depolarization. These and other data indicate that one of the primary actions of the toxin is to inhibit electrogenic ion pumps in the plasmalemma. PMID:16660605

Reducing background contributions in fluorescence fluctuation time-traces for single-molecule measurements in solution.

PubMed

Földes-Papp, Zeno; Liao, Shih-Chu Jeff; You, Tiefeng; Barbieri, Beniamino

2009-08-01

We first report on the development of new microscope means that reduce background contributions in fluorescence fluctuation methods: i) excitation shutter, ii) electronic switches, and iii) early and late time-gating. The elements allow for measuring molecules at low analyte concentrations. We first found conditions of early and late time-gating with time-correlated single-photon counting that made the fluorescence signal as bright as possible compared with the fluctuations in the background count rate in a diffraction-limited optical set-up. We measured about a 140-fold increase in the amplitude of autocorrelated fluorescence fluctuations at the lowest analyte concentration of about 15 pM, which gave a signal-to-background advantage of more than two-orders of magnitude. The results of this original article pave the way for single-molecule detection in solution and in live cells without immobilization or hydrodynamic/electrokinetic focusing at longer observation times than are currently available.

Chloride Cotransporters as a Molecular Mechanism underlying Spreading Depolarization-Induced Dendritic Beading.

PubMed

Steffensen, Annette B; Sword, Jeremy; Croom, Deborah; Kirov, Sergei A; MacAulay, Nanna

2015-09-02

Spreading depolarizations (SDs) are waves of sustained neuronal and glial depolarization that propagate massive disruptions of ion gradients through the brain. SD is associated with migraine aura and recently recognized as a novel mechanism of injury in stroke and brain trauma patients. SD leads to neuronal swelling as assessed in real time with two-photon laser scanning microscopy (2PLSM). Pyramidal neurons do not express aquaporins and thus display low inherent water permeability, yet SD rapidly induces focal swelling (beading) along the dendritic shaft by unidentified molecular mechanisms. To address this issue, we induced SD in murine hippocampal slices by focal KCl microinjection and visualized the ensuing beading of dendrites expressing EGFP by 2PLSM. We confirmed that dendritic beading failed to arise during large (100 mOsm) hyposmotic challenges, underscoring that neuronal swelling does not occur as a simple osmotic event. SD-induced dendritic beading was not prevented by pharmacological interference with the cytoskeleton, supporting the notion that dendritic beading may result entirely from excessive water influx. Dendritic beading was strictly dependent on the presence of Cl(-), and, accordingly, combined blockade of Cl(-)-coupled transporters led to a significant reduction in dendritic beading without interfering with SD. Furthermore, our in vivo data showed a strong inhibition of dendritic beading during pharmacological blockage of these cotransporters. We propose that SD-induced dendritic beading takes place as a consequence of the altered driving forces and thus activity for these cotransporters, which by transport of water during their translocation mechanism may generate dendritic beading independently of osmotic forces. Spreading depolarization occurs during pathological conditions such as stroke, brain injury, and migraine and is characterized as a wave of massive ion translocation between intracellular and extracellular space in association with

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

Depolarization of the conductance-voltage relationship in the NaV1.5 mutant, E1784K, is due to altered fast inactivation.

PubMed

Peters, Colin H; Yu, Alec; Zhu, Wandi; Silva, Jonathan R; Ruben, Peter C

2017-01-01

E1784K is the most common mixed long QT syndrome/Brugada syndrome mutant in the cardiac voltage-gated sodium channel NaV1.5. E1784K shifts the midpoint of the channel conductance-voltage relationship to more depolarized membrane potentials and accelerates the rate of channel fast inactivation. The depolarizing shift in the midpoint of the conductance curve in E1784K is exacerbated by low extracellular pH. We tested whether the E1784K mutant shifts the channel conductance curve to more depolarized membrane potentials by affecting the channel voltage-sensors. We measured ionic currents and gating currents at pH 7.4 and pH 6.0 in Xenopus laevis oocytes. Contrary to our expectation, the movement of gating charges is shifted to more hyperpolarized membrane potentials by E1784K. Voltage-clamp fluorimetry experiments show that this gating charge shift is due to the movement of the DIVS4 voltage-sensor being shifted to more hyperpolarized membrane potentials. Using a model and experiments on fast inactivation-deficient channels, we show that changes to the rate and voltage-dependence of fast inactivation are sufficient to shift the conductance curve in E1784K. Our results localize the effects of E1784K to DIVS4, and provide novel insight into the role of the DIV-VSD in regulating the voltage-dependencies of activation and fast inactivation.

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.

"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…

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.

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.

FLEX: an imaging spectrometer for measurement of vegetation fluorescence

NASA Astrophysics Data System (ADS)

Smorenburg, Kees; Visser, Huib; Court, Andrew; Stoll, Marc Ph.

2017-11-01

Detection of vegetation fluorescence gives information about plant functioning, stress and vitality. During the past decades several ground based laser fluorosensors have been developed to investigate these processes and to demonstrate the value of this technique. FLEX (= FLuorescense EXplorer) is a space mission to measure the fluorescence of vegetation on earth over large areas from space. Such a mission would greatly improve the understanding and enhance the capability to quantify e.g. the role of terrestrial vegetation in global carbon sequestration. Because the fluorescence signal, which is excited by solar irradiation is low with respect to the reflected sunlight the signal from a satellite is proposed to be measured in the solar Fraunhofer lines, where the reflection signal is much reduced. The heart of FLEX is a high resolution imaging spectrometer with 2 channels: channel 1 around the Fraunhofer lines at ‡ = 397 nm, ‡= 423 nm and/or ‡ = 434 nm and channel 2 around the Fraunhofer line at ‡ = 656 nm. The required spectral resolution will depend on the linewidth (0.02-0.3 nm). A first definition of the field of view is 8.4 degrees, leading from an 800 km satellite altitude to a swath of about 120 km. For detection a 1024x1024 pixel frame transfer CCD detector is proposed, with a pixel dimension of 13 x 13 ‡ mm2. The maximum footprint is about 500x500m2. The optical configuration contains a scan mirror for solar calibration, for pointing the FOV in swath direction and for freezing the observed ground scene up to a few seconds to increase the signal to noise performance. At this moment the concept of FLEX is elaborated in a feasibility study. Both the scientific and instrument requirements are updated and the concept is studied in detail. Besides a development plan for FLEX is made. In this paper the idea and the headlines of FLEX are described.

Using depolarization to quantify ice nucleating particle concentrations: a new method

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zenker, Jake; Collier, Kristen N.; Xu, Guanglang

We have developed a new method to determine ice nucleating particle (INP) concentrations observed by the Texas A&M University continuous flow diffusion chamber (CFDC) under a wide range of operating conditions. In this study, we evaluate differences in particle optical properties detected by the Cloud and Aerosol Spectrometer with POLarization (CASPOL) to differentiate between ice crystals, droplets, and aerosols. The depolarization signal from the CASPOL instrument is used to determine the occurrence of water droplet breakthrough (WDBT) conditions in the CFDC. The standard procedure for determining INP concentration is to count all particles that have grown beyond a nominal sizemore » cutoff as ice crystals. During WDBT this procedure overestimates INP concentration, because large droplets are miscounted as ice crystals. Here we design a new analysis method based on depolarization ratio that can extend the range of operating conditions of the CFDC. The method agrees reasonably well with the traditional method under non-WDBT conditions with a mean percent error of ±32.1 %. Additionally, a comparison with the Colorado State University CFDC shows that the new analysis method can be used reliably during WDBT conditions.« less

Using depolarization to quantify ice nucleating particle concentrations: a new method

DOE PAGES

Zenker, Jake; Collier, Kristen N.; Xu, Guanglang; ...

2017-12-01

We have developed a new method to determine ice nucleating particle (INP) concentrations observed by the Texas A&M University continuous flow diffusion chamber (CFDC) under a wide range of operating conditions. In this study, we evaluate differences in particle optical properties detected by the Cloud and Aerosol Spectrometer with POLarization (CASPOL) to differentiate between ice crystals, droplets, and aerosols. The depolarization signal from the CASPOL instrument is used to determine the occurrence of water droplet breakthrough (WDBT) conditions in the CFDC. The standard procedure for determining INP concentration is to count all particles that have grown beyond a nominal sizemore » cutoff as ice crystals. During WDBT this procedure overestimates INP concentration, because large droplets are miscounted as ice crystals. Here we design a new analysis method based on depolarization ratio that can extend the range of operating conditions of the CFDC. The method agrees reasonably well with the traditional method under non-WDBT conditions with a mean percent error of ±32.1 %. Additionally, a comparison with the Colorado State University CFDC shows that the new analysis method can be used reliably during WDBT conditions.« less

Using depolarization to quantify ice nucleating particle concentrations: a new method

NASA Astrophysics Data System (ADS)

Zenker, Jake; Collier, Kristen N.; Xu, Guanglang; Yang, Ping; Levin, Ezra J. T.; Suski, Kaitlyn J.; DeMott, Paul J.; Brooks, Sarah D.

2017-12-01

We have developed a new method to determine ice nucleating particle (INP) concentrations observed by the Texas A&M University continuous flow diffusion chamber (CFDC) under a wide range of operating conditions. In this study, we evaluate differences in particle optical properties detected by the Cloud and Aerosol Spectrometer with POLarization (CASPOL) to differentiate between ice crystals, droplets, and aerosols. The depolarization signal from the CASPOL instrument is used to determine the occurrence of water droplet breakthrough (WDBT) conditions in the CFDC. The standard procedure for determining INP concentration is to count all particles that have grown beyond a nominal size cutoff as ice crystals. During WDBT this procedure overestimates INP concentration, because large droplets are miscounted as ice crystals. Here we design a new analysis method based on depolarization ratio that can extend the range of operating conditions of the CFDC. The method agrees reasonably well with the traditional method under non-WDBT conditions with a mean percent error of ±32.1 %. Additionally, a comparison with the Colorado State University CFDC shows that the new analysis method can be used reliably during WDBT conditions.

Quantitative Nucleotide Level Analysis of Regulation of Translation in Response to Depolarization of Cultured Neural Cells

PubMed Central

Dalal, Jasbir S.; Yang, Chengran; Sapkota, Darshan; Lake, Allison M.; O'Brien, David R.; Dougherty, Joseph D.

2017-01-01

Studies on regulation of gene expression have contributed substantially to understanding mechanisms for the long-term activity-dependent alterations in neural connectivity that are thought to mediate learning and memory. Most of these studies, however, have focused on the regulation of mRNA transcription. Here, we utilized high-throughput sequencing coupled with ribosome footprinting to globally characterize the regulation of translation in primary mixed neuronal-glial cultures in response to sustained depolarization. We identified substantial and complex regulation of translation, with many transcripts demonstrating changes in ribosomal occupancy independent of transcriptional changes. We also examined sequence-based mechanisms that might regulate changes in translation in response to depolarization. We found that these are partially mediated by features in the mRNA sequence—notably upstream open reading frames and secondary structure in the 5′ untranslated region—both of which predict downregulation in response to depolarization. Translationally regulated transcripts are also more likely to be targets of FMRP and include genes implicated in autism in humans. Our findings support the idea that control of mRNA translation plays an important role in response to neural activity across the genome. PMID:28190998

Temperature measurements of micro-droplets using pulsed 2-color laser-induced fluorescence with MDR-enhanced energy transfer

NASA Astrophysics Data System (ADS)

Palmer, Johannes; Reddemann, Manuel A.; Kirsch, Valeri; Kneer, Reinhold

2016-12-01

In this work, a new measurement system is presented for studying temperature of micro-droplets by pulsed 2-color laser-induced fluorescence. Pulsed fluorescence excitation allows motion blur suppression and thus simultaneous measurements of droplet size, velocity and temperature. However, high excitation intensities of pulsed lasers lead to morphology-dependent resonances inside micro-droplets, which are accompanied by disruptive stimulated emission. Investigations showed that stimulated emission can be avoided by enhanced energy transfer via an additional dye. The suitability and accuracy of the new pulsed method are verified on the basis of a spectroscopic analysis and comparison to continuously excited 2-color laser-induced fluorescence.

Depolarization changes during acute myocardial ischemia by evaluation of QRS slopes: standard lead and vectorial approach.

PubMed

Romero, Daniel; Ringborn, Michael; Laguna, Pablo; Pahlm, Olle; Pueyo, Esther

2011-01-01

Diagnosis and risk stratification of patients with acute coronary syndromes can be improved by adding information from the depolarization phase (QRS complex) to the conventionally used ST-T segment changes. In this study, ischemia-induced changes in the main three slopes of the QRS complex, upward ( ℑ(US)) and downward ( ℑ(DS) ) slopes of the R wave as well as the upward ( ℑ(TS)) slope of the terminal S wave, were evaluated as to represent a robust measure of pathological changes within the depolarization phase. From ECG recordings both in a resting state (control recordings) and during percutaneous coronary intervention (PCI)-induced transmural ischemia, we developed a method for quantification of ℑ(US), ℑ(DS), and ℑ(TS) that incorporates dynamic ECG normalization so as to improve the sensitivity in the detection of ischemia-induced changes. The same method was also applied on leads obtained by projection of QRS loops onto their dominant directions. We show that ℑ(US), ℑ(DS), and ℑ(TS) present high stability in the resting state, thus providing a stable reference for ischemia characterization. Maximum relative factors of change ( ℜ(ℑ)) during PCI were found in leads derived from the QRS loop, reaching 10.5 and 13.7 times their normal variations in the control for ℑ(US) and ℑ(DS), respectively. For standard leads, the relative factors of change were 6.01 and 9.31. The ℑ(TS) index presented a similar behavior to that of ℑ(DS). The timing for the occurrence of significant changes in ℑ(US) and ℑ(DS) varied with lead, ranging from 30 s to 2 min after initiation of coronary occlusion. In the present ischemia model, relative ℑ(DS) changes were smaller than ST changes in most leads, however with only modest correlation between the two indices, suggesting they present different information about the ischemic process. We conclude that QRS slopes offer a robust tool for evaluating depolarization changes during myocardial ischemia.

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.

Temperature measurement in a compressible flow field using laser-induced iodine fluorescence

NASA Technical Reports Server (NTRS)

Fletcher, D. G.; Mcdaniel, J. C.

1987-01-01

The thermometric capability of a two-line fluorescence technique using iodine seed molecules in air is investigated analytically and verified experimentally in a known steady compressible flow field. Temperatures ranging from 165 to 295 K were measured in the flowfield using two iodine transitions accessed with a 30-GHz dye-laser scan near 543 nm. The effect of pressure broadening on temperature measurement is evaluated.

Rapid measurement of meat spoilage using fluorescence spectroscopy

NASA Astrophysics Data System (ADS)

Wu, Binlin; Dahlberg, Kevin; Gao, Xin; Smith, Jason; Bailin, Jacob

2017-02-01

Food spoilage is mainly caused by microorganisms, such as bacteria. In this study, we measure the autofluorescence in meat samples longitudinally over a week in an attempt to develop a method to rapidly detect meat spoilage using fluorescence spectroscopy. Meat food is a biological tissue, which contains intrinsic fluorophores, such as tryptophan, collagen, nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) etc. As meat spoils, it undergoes various morphological and chemical changes. The concentrations of the native fluorophores present in a sample may change. In particular, the changes in NADH and FAD are associated with microbial metabolism, which is the most important process of the bacteria in food spoilage. Such changes may be revealed by fluorescence spectroscopy and used to indicate the status of meat spoilage. Therefore, such native fluorophores may be unique, reliable and nonsubjective indicators for detection of spoiled meat. The results of the study show that the relative concentrations of all above fluorophores change as the meat samples kept in room temperature ( 19° C) spoil. The changes become more rapidly after about two days. For the meat samples kept in a freezer ( -12° C), the changes are much less or even unnoticeable over a-week-long storage.

Quantitative evaluation of the neuroprotective effects of thiopental sodium, propofol, and halothane on brain ischemia in the gerbil: effects of the anesthetics on ischemic depolarization and extracellular glutamate concentration.

PubMed

Kobayashi, Motomu; Takeda, Yoshimasa; Taninishi, Hideki; Takata, Ken; Aoe, Hisami; Morita, Kiyoshi

2007-07-01

Although propofol and thiopental are commonly used as neuroprotective agents, it has not been determined which is more neuroprotective. This study was designed to quantitatively evaluate the neuroprotective effects of thiopental, propofol, and halothane on brain ischemia by determining P50, ischemic time necessary for causing 50% neuronal damage. Gerbils were anesthetized with thiopental, propofol, or halothane and underwent 2-vessel occlusion (0, 3, 5 or 10 min). Direct current potentials were measured in bilateral CA1 regions, in which histologic evaluation was performed 5 days later. In some animals, extracellular glutamate concentrations (microdialysis) were measured during 7.5 minutes of ischemia. P50 in the thiopental, propofol, and halothane groups were estimated to be 8.4, 6.5 (P<0.05, vs. thiopental), and 5.1 (P<0.05) minutes, respectively. Durations of ischemic depolarization were equally reduced in the thiopental and propofol groups compared with that in the halothane group. Severity of neuronal damage with identical duration of ischemic depolarization was attenuated by thiopental compared with the effect of propofol. Maximum glutamate concentrations in the thiopental and propofol group were significantly reduced compared with that in the halothane groups but were comparable. By using P50, we found that the neuroprotective effect of thiopental was greater than that of propofol. Although duration of ischemic depolarization was equally reduced in thiopental and propofol groups, thiopental has a greater suppressive effect on neuronal injury during identical duration of ischemic depolarization than propofol does. Glutamate concentration during brain ischemia tended to be attenuated more by thiopental than by propofol, but it was not statistically significant.

Comparison of 68Ga- and fluorescence-labeled microspheres for measurement of relative pulmonary perfusion in anesthetized pigs.

PubMed

Braune, Anja; Scharffenberg, Martin; Naumann, Anne; Bluth, Thomas; de Abreu, Marcelo Gama; Kotzerke, Jörg

2018-06-01

We compared 68 Gallium ( 68 Ga)- and fluorescence-labeled microspheres for measurement of pulmonary perfusion distribution in anesthetized pigs without lung injury. In two mechanically ventilated pigs, the distribution of pulmonary perfusion was marked in vivo with 68 Ga- and fluorescence-labeled microspheres in supine and prone position. After each injection, the distribution of 68 Ga-labeled microspheres was measured in vivo with positron emission tomography/ computed tomography (PET/CT) in the position in which microspheres were injected and vice versa. The distribution of fluorescence-labeled microspheres was measured ex vivo . Perfusion distributions were compared between methods and postures within four lung regions and along the ventro-dorsal gradient. After each injection of 68 Ga-labeled microspheres, changes in ventro-dorsal perfusion gradients induced by repositioning were compared for volume- and mass-normalized PET/CT measurements. Regional and gradient analyses of in vivo and ex vivo measurements, respectively, consistently revealed higher pulmonary perfusion in dorsal than ventral regions in supine positioned animals. Both methods showed more pronounced perfusion gradients in supine compared to prone position. Changes in animal position were associated with alterations in the ventro-dorsal perfusion gradient when volume-, but not mass-normalization was conducted for PET/CT data. Ex vivo fluorescence- and in vivo 68 Ga-labeled microspheres measurements revealed similar perfusion distributions. Mass-normalized perfusion measurements by 68 Ga-labeled microspheres and PET/CT were not affected by positioning artifacts. Schattauer GmbH.

Measurement of contaminant removal from skin using a portable fluorescence scanning system.

PubMed

Hession, Helena; Byrne, Miriam; Cleary, S; Andersson, K G; Roed, J

2006-01-01

The residence time of particulate contamination on the human body is a factor that has an important impact on the accuracy of exposure assessment in the aftermath of an accidental release of radionuclides to the atmosphere. Measurements of particle clearance from human skin were made using an illumination system to excite fluorescence in labelled silica particles and a CCD camera and image processing system to detect this fluorescence. The illumination system consists of high-intensity light emitting diodes (LEDS) of suitable wavelengths arranged on a portable stand. The physically small size of the LEDs allows them to be positioned close to the fluorescing surface, thus maximising the fluorescent signal that can be obtained. The limit of detection was found to be 50 microg of tracer particle per cm2. Experiments were carried out to determine the clearance rates of 10 microm and 3 microm particles from the skin. Results show that, in the absence of any mechanical rubbing of the skin, the clearance of particles from the skin followed an approximately exponential decay with a half-time of 1.5-7.8 h. Skin hairiness and degree of human movement were found, in addition to particle size, to have an important influence on particle fall-off rate.

Laser-Induced Fluorescence Measurements of Translational Temperature and Relative Cycle Number by use of Optically Pumped Trace-Sodium Vapor

NASA Technical Reports Server (NTRS)

Dobson, Chris C.

1998-01-01

Sodium fluorescence induced by a narrow bandwidth tunable laser has been used to measure temperature, pressure, axial velocity and species concentrations in wind tunnels, rocket engine exhausts and the upper atmosphere. Optical pumping of the ground states of the sodium, however, can radically alter the shape of the laser induced fluorescence excitation spectrum, complicating such measurements. Here a straightforward extension of rate equations originally proposed to account for the features of the pumped spectrum is to make temperature measurements from spectra taken in pumped vapor. Also determined from the spectrum is the relative fluorescence cycle number, which has application to measurement of diffusion rate and transverse flow velocity. The accuracy of both the temperature and cycle-number measurements is comparable with that of temperature measurements made in the absence of pumping.

Laser-Induced Fluorescence Measurements of Translational Temperature and Relative Cycle Number by use of Optically Pumped Trace-Sodium Vapor

NASA Technical Reports Server (NTRS)

Dobson, Chris C.

1999-01-01

Sodium fluorescence induced by a narrow-bandwidth tunable laser has been used to measure temperature, pressure, axial velocity, and species concentrations in wind tunnels, rocket engine exhausts, and the upper atmosphere. Optical pumping of the ground states of the sodium, however, can radically alter the shape of the laser-induced fluorescence excitation spectrum, complicating such measurements. Here a straightforward extension of rate equations originally proposed to account for the features of the pumped spectrum is used to make temperature measurements from spectra taken in pumped vapor. Also determined from the spectrum is the relative fluorescence cycle number, which has application to measurement of diffusion rate and transverse flow velocity, The accuracy of both the temperature and the cycle-number measurements is comparable with that of temperature measurements made in the absence of pumping.

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

PubMed

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

2011-01-01

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

An Innovative Concept for Spacebased Lidar Measurement of Ocean Carbon Biomass

NASA Technical Reports Server (NTRS)

Hu, Yongxiang; Behrenfeld, Michael; Hostetler, Chris; Pelon, Jacques; Trepte, Charles; Hair, John; Slade, Wayne; Cetinic, Ivona; Vaughan, Mark; Lu, Xiaomei;

Measuring thermodynamic details of DNA hybridization using fluorescence.

PubMed

You, Yong; Tataurov, Andrey V; Owczarzy, Richard

2011-07-01

Modern real-time PCR systems make it easy to monitor fluorescence while temperature is varied for hundreds of samples in parallel, permitting high-throughput studies. We employed such system to investigate melting transitions of ordered nucleic acid structures into disordered random coils. Fluorescent dye and quencher were attached to oligonucleotides in such a way that changes of fluorescence intensity with temperature indicated progression of denaturation. When fluorescence melting data were compared with traditional ultraviolet optical experiments, commonly used dye/quencher combinations, like fluorescein and tetramethylrhodamine, showed substantial discrepancies. We have therefore screened 22 commercially available fluorophores and quenchers for their ability to reliably report annealing and melting transitions. Dependence of fluorescence on temperature and pH was also investigated. The optimal performance was observed using Texas Red or ROX dyes with Iowa Black RQ or Black Hole quenchers. These labels did not alter two-state nature of duplex melting process and provided accurate melting temperatures, free energies, enthalpies, and entropies. We also suggest a new strategy for determination of DNA duplex thermodynamics where concentration of a dye-labeled strand is kept constant and its complementary strand modified with a quencher is added at increasing excess. These methodological improvements will help build predictive models of nucleic acid hybridization. Copyright © 2011 Wiley Periodicals, Inc., a Wiley company.

Basic quantitative polymerase chain reaction using real-time fluorescence measurements.

PubMed

Ares, Manuel

2014-10-01

This protocol uses quantitative polymerase chain reaction (qPCR) to measure the number of DNA molecules containing a specific contiguous sequence in a sample of interest (e.g., genomic DNA or cDNA generated by reverse transcription). The sample is subjected to fluorescence-based PCR amplification and, theoretically, during each cycle, two new duplex DNA molecules are produced for each duplex DNA molecule present in the sample. The progress of the reaction during PCR is evaluated by measuring the fluorescence of dsDNA-dye complexes in real time. In the early cycles, DNA duplication is not detected because inadequate amounts of DNA are made. At a certain threshold cycle, DNA-dye complexes double each cycle for 8-10 cycles, until the DNA concentration becomes so high and the primer concentration so low that the reassociation of the product strands blocks efficient synthesis of new DNA and the reaction plateaus. There are two types of measurements: (1) the relative change of the target sequence compared to a reference sequence and (2) the determination of molecule number in the starting sample. The first requires a reference sequence, and the second requires a sample of the target sequence with known numbers of the molecules of sequence to generate a standard curve. By identifying the threshold cycle at which a sample first begins to accumulate DNA-dye complexes exponentially, an estimation of the numbers of starting molecules in the sample can be extrapolated. © 2014 Cold Spring Harbor Laboratory Press.

Fluorescence lifetime imaging ophthalmoscopy.

PubMed

Dysli, Chantal; Wolf, Sebastian; Berezin, Mikhail Y; Sauer, Lydia; Hammer, Martin; Zinkernagel, Martin S

2017-09-01

Imaging techniques based on retinal autofluorescence have found broad applications in ophthalmology because they are extremely sensitive and noninvasive. Conventional fundus autofluorescence imaging measures fluorescence intensity of endogenous retinal fluorophores. It mainly derives its signal from lipofuscin at the level of the retinal pigment epithelium. Fundus autofluorescence, however, can not only be characterized by the spatial distribution of the fluorescence intensity or emission spectrum, but also by a characteristic fluorescence lifetime function. The fluorescence lifetime is the average amount of time a fluorophore remains in the excited state following excitation. Fluorescence lifetime imaging ophthalmoscopy (FLIO) is an emerging imaging modality for in vivo measurement of lifetimes of endogenous retinal fluorophores. Recent reports in this field have contributed to our understanding of the pathophysiology of various macular and retinal diseases. Within this review, the basic concept of fluorescence lifetime imaging is provided. It includes technical background information and correlation with in vitro measurements of individual retinal metabolites. In a second part, clinical applications of fluorescence lifetime imaging and fluorescence lifetime features of selected retinal diseases such as Stargardt disease, age-related macular degeneration, choroideremia, central serous chorioretinopathy, macular holes, diabetic retinopathy, and retinal artery occlusion are discussed. Potential areas of use for fluorescence lifetime imaging ophthalmoscopy will be outlined at the end of this review. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

In situ observation of sol-gel transition of agarose aqueous solution by fluorescence measurement.

PubMed

Wang, Zheng; Yang, Kun; Li, Haining; Yuan, Chaosheng; Zhu, Xiang; Huang, Haijun; Wang, Yongqiang; Su, Lei; Fang, Yapeng

2018-06-01

Sol-gel transition behavior of agarose aqueous solution was investigated by using rheology and fluorescence measurement. On heating, the storage modulus G' decreased gradually, then deviated abruptly at the temperature of about 65°C, and finally decreased slowly again. For fluorescence measurement, the phase transition point kept almost at the temperature of 65°C, which was consistent with that in rheology measurement. Upon compression, it was indicated that the fluorescence lifetime for the probe in the agarose aqueous solution showed a dramatic change in the vicinity of the phase transition point. T vs. P phase diagram of agarose aqueous solution was constructed, which showed that the melting point was an increasing function of pressure. Based on the phase diagram, the agarose gels were prepared by cooling under atmospheric pressure and the pressure of 300MPa, respectively. From the result of the recovered samples studied by optical rheometry, it was found that agarose gel prepared under high pressure had a higher elasticity and lower viscosity index, compared with that under atmospheric pressure. It could be speculated that such kinds of properties might be attributed to the smaller pore size during gelation under high pressure. Copyright © 2018. Published by Elsevier B.V.

Optical absorption and thermally stimulated depolarization current studies of nickel chloride-doped poly(vinyl alcohol) irradiated with low-level fast neutron doses

DOE Office of Scientific and Technical Information (OSTI.GOV)

Abd El-Kader, F.H.; Ibrahim, S.S.; Attia, G.

1993-11-15

The influence of neutron irradiation on ultraviolet/visible absorption and thermally stimulated depolarization current in nickel chloride-poly(vinyl alcohol) (PVA) cast films has been investigated. The spectral measurements indicate the responsibility of the Ni[sup 2][sup +] ion in its octahedral symmetry. Dopant concentrations higher than 10 wt % NiCl[sub 2] are found to make the samples more resistant to a degradation effect caused by neutron irradiation. The thermally stimulated depolarization currents (TSDC) of pure PVA revealed the existence of the glass transition T[sub g] and space charge relaxation peaks, whereas doped-PVA samples show a new sub-T[sub g] relaxation peak. A proposed mechanismmore » is introduced to account for the neutron effects on both glass transition and space charge relaxation peaks. The peak positions, peak currents, and stored charges of the sub-T[sub g] relaxation peak are strongly affected by both the concentration of the dopant and neutron exposure doses.« less

Nuclear Resonance Fluorescence Measurements of High Explosives

DOE Office of Scientific and Technical Information (OSTI.GOV)

Caggiano, Joseph A.; Warren, Glen A.; Korbly, Steve

Pacific Northwest National Laboratory and Passport Systems have collaborated to perform Nuclear Resonance Fluorescence experiments using several high quality high-explosive simulant samples. These measurements were conducted to determine the feasibility of finding and characterizing high explosive material by NRF interrogation. Electron beams of 5.1, 5.3, 8, and 10 MeV were used to produce bremsstrahlung photon beams, which irradiated the samples. The gamma-ray spectra were collected using high-purity germanium detectors. Nitrogen-to-carbon ratios of the high-explosive simulants were extracted from the 5.1 and 5.3 MeV data and compare favorably with accepted values. Analysis of the 8 and 10 MeV data is inmore » progress; preliminary isotopic comparisons within the samples are consistent with the expected results.« less

Mechanical deformation induces depolarization of neutrophils.

PubMed

Ekpenyong, Andrew E; Toepfner, Nicole; Fiddler, Christine; Herbig, Maik; Li, Wenhong; Cojoc, Gheorghe; Summers, Charlotte; Guck, Jochen; Chilvers, Edwin R

2017-06-01

The transition of neutrophils from a resting state to a primed state is an essential requirement for their function as competent immune cells. This transition can be caused not only by chemical signals but also by mechanical perturbation. After cessation of either, these cells gradually revert to a quiescent state over 40 to 120 min. We use two biophysical tools, an optical stretcher and a novel microcirculation mimetic, to effect physiologically relevant mechanical deformations of single nonadherent human neutrophils. We establish quantitative morphological analysis and mechanical phenotyping as label-free markers of neutrophil priming. We show that continued mechanical deformation of primed cells can cause active depolarization, which occurs two orders of magnitude faster than by spontaneous depriming. This work provides a cellular-level mechanism that potentially explains recent clinical studies demonstrating the potential importance, and physiological role, of neutrophil depriming in vivo and the pathophysiological implications when this deactivation is impaired, especially in disorders such as acute lung injury.

Sequential depolarization of root cortical and stelar cells induced by an acute salt shock - implications for Na(+) and K(+) transport into xylem vessels.

PubMed

Wegner, Lars H; Stefano, Giovanni; Shabala, Lana; Rossi, Marika; Mancuso, Stefano; Shabala, Sergey

2011-05-01

Early events in NaCl-induced root ion and water transport were investigated in maize (Zea mays L) roots using a range of microelectrode and imaging techniques. Addition of 100 mm NaCl to the bath resulted in an exponential drop in root xylem pressure, rapid depolarization of trans-root potential and a transient drop in xylem K(+) activity (A(K+) ) within ∼1 min after stress onset. At this time, no detectable amounts of Na(+) were released into the xylem vessels. The observed drop in A(K+) was unexpected, given the fact that application of the physiologically relevant concentrations of Na(+) to isolated stele has caused rapid plasma membrane depolarization and a subsequent K(+) efflux from the stelar tissues. This controversy was explained by the difference in kinetics of NaCl-induced depolarization between cortical and stelar cells. As root cortical cells are first to be depolarized and lose K(+) to the environment, this is associated with some K(+) shift from the stelar symplast to the cortex, resulting in K(+) being transiently removed from the xylem. Once Na(+) is loaded into the xylem (between 1 and 5 min of root exposure to NaCl), stelar cells become more depolarized, and a gradual recovery in A(K+) occurs. © 2011 Blackwell Publishing Ltd.

Postsynaptic Depolarization Enhances GABA Drive to Dorsomedial Hypothalamic Neurons through Somatodendritic Cholecystokinin Release.

PubMed

Crosby, Karen M; Baimoukhametova, Dinara V; Bains, Jaideep S; Pittman, Quentin J

2015-09-23

Somatodendritically released peptides alter synaptic function through a variety of mechanisms, including autocrine actions that liberate retrograde transmitters. Cholecystokinin (CCK) is a neuropeptide expressed in neurons in the dorsomedial hypothalamic nucleus (DMH), a region implicated in satiety and stress. There are clear demonstrations that exogenous CCK modulates food intake and neuropeptide expression in the DMH, but there is no information on how endogenous CCK alters synaptic properties. Here, we provide the first report of somatodendritic release of CCK in the brain in male Sprague Dawley rats. CCK is released from DMH neurons in response to repeated postsynaptic depolarizations, and acts in an autocrine fashion on CCK2 receptors to enhance postsynaptic NMDA receptor function and liberate the retrograde transmitter, nitric oxide (NO). NO subsequently acts presynaptically to enhance GABA release through a soluble guanylate cyclase-mediated pathway. These data provide the first demonstration of synaptic actions of somatodendritically released CCK in the hypothalamus and reveal a new form of retrograde plasticity, depolarization-induced potentiation of inhibition. Significance statement: Somatodendritic signaling using endocannabinoids or nitric oxide to alter the efficacy of afferent transmission is well established. Despite early convincing evidence for somatodendritic release of neurohypophysial peptides in the hypothalamus, there is only limited evidence for this mode of release for other peptides. Here, we provide the first evidence for somatodendritic release of the satiety peptide cholecystokinin (CCK) in the brain. We also reveal a new form of synaptic plasticity in which postsynaptic depolarization results in enhancement of inhibition through the somatodendritic release of CCK. Copyright © 2015 the authors 0270-6474/15/3513160-11$15.00/0.

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.

Depolarization of the conductance-voltage relationship in the NaV1.5 mutant, E1784K, is due to altered fast inactivation

PubMed Central

Yu, Alec; Zhu, Wandi; Silva, Jonathan R.; Ruben, Peter C.

2017-01-01

E1784K is the most common mixed long QT syndrome/Brugada syndrome mutant in the cardiac voltage-gated sodium channel NaV1.5. E1784K shifts the midpoint of the channel conductance-voltage relationship to more depolarized membrane potentials and accelerates the rate of channel fast inactivation. The depolarizing shift in the midpoint of the conductance curve in E1784K is exacerbated by low extracellular pH. We tested whether the E1784K mutant shifts the channel conductance curve to more depolarized membrane potentials by affecting the channel voltage-sensors. We measured ionic currents and gating currents at pH 7.4 and pH 6.0 in Xenopus laevis oocytes. Contrary to our expectation, the movement of gating charges is shifted to more hyperpolarized membrane potentials by E1784K. Voltage-clamp fluorimetry experiments show that this gating charge shift is due to the movement of the DIVS4 voltage-sensor being shifted to more hyperpolarized membrane potentials. Using a model and experiments on fast inactivation-deficient channels, we show that changes to the rate and voltage-dependence of fast inactivation are sufficient to shift the conductance curve in E1784K. Our results localize the effects of E1784K to DIVS4, and provide novel insight into the role of the DIV-VSD in regulating the voltage-dependencies of activation and fast inactivation. PMID:28898267

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

A fluorescence anisotropy method for measuring protein concentration in complex cell culture media.

PubMed

Groza, Radu Constantin; Calvet, Amandine; Ryder, Alan G

2014-04-22

The rapid, quantitative analysis of the complex cell culture media used in biopharmaceutical manufacturing is of critical importance. Requirements for cell culture media composition profiling, or changes in specific analyte concentrations (e.g. amino acids in the media or product protein in the bioprocess broth) often necessitate the use of complicated analytical methods and extensive sample handling. Rapid spectroscopic methods like multi-dimensional fluorescence (MDF) spectroscopy have been successfully applied for the routine determination of compositional changes in cell culture media and bioprocess broths. Quantifying macromolecules in cell culture media is a specific challenge as there is a need to implement measurements rapidly on the prepared media. However, the use of standard fluorescence spectroscopy is complicated by the emission overlap from many media components. Here, we demonstrate how combining anisotropy measurements with standard total synchronous fluorescence spectroscopy (TSFS) provides a rapid, accurate quantitation method for cell culture media. Anisotropy provides emission resolution between large and small fluorophores while TSFS provides a robust measurement space. Model cell culture media was prepared using yeastolate (2.5 mg mL(-1)) spiked with bovine serum albumin (0 to 5 mg mL(-1)). Using this method, protein emission is clearly discriminated from background yeastolate emission, allowing for accurate bovine serum albumin (BSA) quantification over a 0.1 to 4.0 mg mL(-1) range with a limit of detection (LOD) of 13.8 μg mL(-1). Copyright © 2014. Published by Elsevier B.V.

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.

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).

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. © The Author 2014. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Cardiac Calcium ATPase Dimerization Measured by Cross-Linking and Fluorescence Energy Transfer.

PubMed

Blackwell, Daniel J; Zak, Taylor J; Robia, Seth L

2016-09-20

The cardiac sarco/endoplasmic reticulum calcium ATPase (SERCA) establishes the intracellular calcium gradient across the sarcoplasmic reticulum membrane. It has been proposed that SERCA forms homooligomers that increase the catalytic rate of calcium transport. We investigated SERCA dimerization in rabbit left ventricular myocytes using a photoactivatable cross-linker. Western blotting of cross-linked SERCA revealed higher-molecular-weight species consistent with SERCA oligomerization. Fluorescence resonance energy transfer measurements in cells transiently transfected with fluorescently labeled SERCA2a revealed that SERCA readily forms homodimers. These dimers formed in the absence or presence of the SERCA regulatory partner, phospholamban (PLB) and were unaltered by PLB phosphorylation or changes in calcium or ATP. Fluorescence lifetime data are compatible with a model in which PLB interacts with a SERCA homodimer in a stoichiometry of 1:2. Together, these results suggest that SERCA forms constitutive homodimers in live cells and that dimer formation is not modulated by SERCA conformational poise, PLB binding, or PLB phosphorylation. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Sizing protein-templated gold nanoclusters by time resolved fluorescence anisotropy decay measurements

NASA Astrophysics Data System (ADS)

Soleilhac, Antonin; Bertorelle, Franck; Antoine, Rodolphe

2018-03-01

Protein-templated gold nanoclusters (AuNCs) are very attractive due to their unique fluorescence properties. A major problem however may arise due to protein structure changes upon the nucleation of an AuNC within the protein for any future use as in vivo probes, for instance. In this work, we propose a simple and reliable fluorescence based technique measuring the hydrodynamic size of protein-templated gold nanoclusters. This technique uses the relation between the time resolved fluorescence anisotropy decay and the hydrodynamic volume, through the rotational correlation time. We determine the molecular size of protein-directed AuNCs, with protein templates of increasing sizes, e.g. insulin, lysozyme, and bovine serum albumin (BSA). The comparison of sizes obtained by other techniques (e.g. dynamic light scattering and small-angle X-ray scattering) between bare and gold clusters containing proteins allows us to address the volume changes induced either by conformational changes (for BSA) or the formation of protein dimers (for insulin and lysozyme) during cluster formation and incorporation.

Sizing protein-templated gold nanoclusters by time resolved fluorescence anisotropy decay measurements.

PubMed

Soleilhac, Antonin; Bertorelle, Franck; Antoine, Rodolphe

2018-03-15

Protein-templated gold nanoclusters (AuNCs) are very attractive due to their unique fluorescence properties. A major problem however may arise due to protein structure changes upon the nucleation of an AuNC within the protein for any future use as in vivo probes, for instance. In this work, we propose a simple and reliable fluorescence based technique measuring the hydrodynamic size of protein-templated gold nanoclusters. This technique uses the relation between the time resolved fluorescence anisotropy decay and the hydrodynamic volume, through the rotational correlation time. We determine the molecular size of protein-directed AuNCs, with protein templates of increasing sizes, e.g. insulin, lysozyme, and bovine serum albumin (BSA). The comparison of sizes obtained by other techniques (e.g. dynamic light scattering and small-angle X-ray scattering) between bare and gold clusters containing proteins allows us to address the volume changes induced either by conformational changes (for BSA) or the formation of protein dimers (for insulin and lysozyme) during cluster formation and incorporation. Copyright © 2017 Elsevier B.V. All rights reserved.

Imaging fluorescence-correlation spectroscopy for measuring fast surface diffusion at liquid/solid interfaces.

PubMed

Cooper, Justin T; Harris, Joel M

2014-08-05

The development of techniques to probe interfacial molecular transport is important for understanding and optimizing surface-based analytical methods including surface-enhanced spectroscopies, biological assays, and chemical separations. Single-molecule-fluorescence imaging and tracking has been used to measure lateral diffusion rates of fluorescent molecules at surfaces, but the technique is limited to the study of slower diffusion, where molecules must remain relatively stationary during acquisition of an image in order to build up sufficient intensity in a spot to detect and localize the molecule. Although faster time resolution can be achieved by fluorescence-correlation spectroscopy (FCS), where intensity fluctuations in a small spot are related to the motions of molecules on the surface, long-lived adsorption events arising from surface inhomogeneity can overwhelm the correlation measurement and mask the surface diffusion of the moving population. Here, we exploit a combination of these two techniques, imaging-FCS, for measurement of fast interfacial transport at a model chromatographic surface. This is accomplished by rapid imaging of the surface using an electron-multiplied-charged-coupled-device (CCD) camera, while limiting the acquisition to a small area on the camera to allow fast framing rates. The total intensity from the sampled region is autocorrelated to determine surface diffusion rates of molecules with millisecond time resolution. The technique allows electronic control over the acquisition region, which can be used to avoid strong adsorption sites and thus minimize their contribution to the measured autocorrelation decay and to vary the acquisition area to resolve surface diffusion from adsorption and desorption kinetics. As proof of concept, imaging-FCS was used to measure surface diffusion rates, interfacial populations, and adsorption-desorption rates of 1,1'-dioctadecyl-3,3,3'3'-tetramethylindocarbocyanine (DiI) on planar C18- and C1

A new anthropometric phantom for calibrating in vivo measurements of stable lead in the human leg using X-ray fluorescence

DOE Office of Scientific and Technical Information (OSTI.GOV)

Spitz, H.; Jenkins, M.; Lodwick, J.

2000-02-01

A new anthropometric phantom has been developed for calibrating in vivo measurements of stable lead deposited in bone using x-ray fluorescence. The phantom reproduces the shape of the mid shaft of the adult human leg and is fabricated using polyurethanes and calcium carbonate to produce materials that exhibit the same density, energy transmission, and calcium content as cortical bone, bone marrow, and muscle. The phantom includes a removable tibia fabricated using simulants for cortical bone and bone marrow to which a precise amount of stable lead has been added to cortical bone. The formulations used in fabricating the new anthropometricmore » phantom are much more uniform in density and composition than the conventional phantom made from Plexiglas cylinders filled with plaster-of-Paris. The energy spectrum from an x-ray fluorescence measurement of the phantom using a {sup 109}Cd source is indistinguishable from an in vivo x-ray fluorescence measurement of the human leg, demonstrating that the materials used in the phantom exhibit the same radiological properties as human tissue. Likewise, results from x-ray fluorescence measurements of the phantom exhibit the same positional dependency as the human leg and vary by approximately 36% when, for example, the phantom containing 54 ppm of stable lead in the tibia was rotated by only 15 degrees. The detection limit for a 30 min {sup 109}Cd K shell x-ray fluorescence in vivo measurement is approximately 20 ppm determined from a background measurement using the new phantom containing no added lead in the muscle, bone, or bone marrow. The new anthropometric phantom significantly improves in vivo x-ray fluorescence calibration measurements by (1) faithfully reproducing the anatomy of the human leg, (2) having components that exhibit radiological properties similar to that of human tissue, and (3) providing a realistic calibration standard that can be used for in vivo x-ray fluorescence intercomparison measurements.« less

A new and specific non-NMDA receptor antagonist, FG 9065, blocks L-AP4-evoked depolarization in rat cerebral cortex.

PubMed

Sheardown, M J

1988-04-13

L(+)-AP4 (2-amino-4-phosphonobutyrate) depolarized slices of rat cerebral cortex, when applied following a 2 min priming application of quisqualate. This response diminishes with time and is not seen after NMDA application. A new selective non-N-methyl-D-aspartate (NMDA) antagonist, 6-cyano-7-nitro-2,3-dihydroxyquinoxaline (FG 9065), inhibits the L(+)-AP4 depolarization. It is argued that the response is mediated indirectly by postsynaptic quisqualate receptors.

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.

Experimental observation of spontaneous depolarized guided acoustic-wave Brillouin scattering in side cores of a multicore fiber

NASA Astrophysics Data System (ADS)

Hayashi, Neisei; Mizuno, Yosuke; Nakamura, Kentaro; Set, Sze Yun; Yamashita, Shinji

2018-06-01

Spontaneous depolarized guided acoustic-wave Brillouin scattering (GAWBS) was experimentally observed in one of the side cores of an uncoated multicore fiber (MCF). The frequency bandwidth in the side core was up to ∼400 MHz, which is 0.5 times that in the central core. The GAWBS spectrum of the side core of the MCF included intrinsic peaks, which had different acoustic resonance frequencies from those of the central core. In addition, the spontaneous depolarized GAWBS in the central/side core was unaffected by that in the other core. These results will lead to the development of polarization/phase modulators using an MCF.

T-matrix modeling of linear depolarization by morphologically complex soot and soot-containing aerosols

NASA Astrophysics Data System (ADS)

Mishchenko, Michael I.; Liu, Li; Mackowski, Daniel W.

2013-07-01

We use state-of-the-art public-domain Fortran codes based on the T-matrix method to calculate orientation and ensemble averaged scattering matrix elements for a variety of morphologically complex black carbon (BC) and BC-containing aerosol particles, with a special emphasis on the linear depolarization ratio (LDR). We explain theoretically the quasi-Rayleigh LDR peak at side-scattering angles typical of low-density soot fractals and conclude that the measurement of this feature enables one to evaluate the compactness state of BC clusters and trace the evolution of low-density fluffy fractals into densely packed aggregates. We show that small backscattering LDRs measured with ground-based, airborne, and spaceborne lidars for fresh smoke generally agree with the values predicted theoretically for fluffy BC fractals and densely packed near-spheroidal BC aggregates. To reproduce higher lidar LDRs observed for aged smoke, one needs alternative particle models such as shape mixtures of BC spheroids or cylinders.

T-Matrix Modeling of Linear Depolarization by Morphologically Complex Soot and Soot-Containing Aerosols

NASA Technical Reports Server (NTRS)

Mishchenko, Michael I.; Liu, Li; Mackowski, Daniel W.

2013-01-01

We use state-of-the-art public-domain Fortran codes based on the T-matrix method to calculate orientation and ensemble averaged scattering matrix elements for a variety of morphologically complex black carbon (BC) and BC-containing aerosol particles, with a special emphasis on the linear depolarization ratio (LDR). We explain theoretically the quasi-Rayleigh LDR peak at side-scattering angles typical of low-density soot fractals and conclude that the measurement of this feature enables one to evaluate the compactness state of BC clusters and trace the evolution of low-density fluffy fractals into densely packed aggregates. We show that small backscattering LDRs measured with groundbased, airborne, and spaceborne lidars for fresh smoke generally agree with the values predicted theoretically for fluffy BC fractals and densely packed near-spheroidal BC aggregates. To reproduce higher lidar LDRs observed for aged smoke, one needs alternative particle models such as shape mixtures of BC spheroids or cylinders.

Low K+-induced hyperpolarizations trigger transient depolarizations and action potentials in rabbit ventricular myocytes

PubMed Central

Akuzawa-Tateyama, M; Tateyama, M; Ochi, R

1998-01-01

The effects of large reductions of [K+]o on membrane potential were studied in isolated rabbit ventricular myocytes using the whole-cell patch clamp technique.Decreasing [K+]o from the normal level of 5.4 mm to 0.1 mm increased resting membrane potential (Vrest) from −75.6 ± 0.3 to −140.3 ± 1.9 mV (means ± s.e.m; n = 127), induced irregular, transient depolarizations with mean maximal amplitudes of 19.5 ± 1.5 mV and elicited action potentials in 56.7 % of trials. The action potentials exhibited overshoots of 37.9 ± 1.5 mV (n = 72) and sustained plateaux.Addition of 0.1 mm La3+ in the presence of 0.1 mm[K+]o significantly increased Vrest but decreased the amplitude of transient depolarizations and suppressed the firing of action potentials.Replacement of external Na+ or Cl− with N-methyl-D-glucamine or aspartate, respectively, or internal dialysis with 10 mm EGTA or BAPTA had little effect on low [K+]o-induced membrane potential changes.Hyperpolarizing voltage clamp pulses to potentials between −110 and −200 mV activated irregular inward currents that increased in amplitude and frequency with increasing hyperpolarization and were depressed by 0.1 mm La3+.The generation of transient depolarizations by low [K+]o can be explained as being a consequence of decreasing the inward rectifier K+ current (IK1) and the appearance of inward currents reflecting electroporation resulting from strong electric fields across the membrane. PMID:9824717

Fluorescence of bioaerosols: mathematical model including primary fluorescing and absorbing molecules in bacteria.

PubMed

Hill, Steven C; Pan, Yong-Le; Williamson, Chatt; Santarpia, Joshua L; Hill, Hanna H

2013-09-23

This paper describes a mathematical model of fluorescent biological particles composed of bacteria, viruses, or proteins. The fluorescent and/or light absorbing molecules included in the model are amino acids (tryptophan, etc.); nucleic acids (DNA, RNA, etc.); coenzymes (nicotinamide adenine dinucleotides, flavins, and vitamins B₆ and K and variants of these); and dipicolinates. The concentrations, absorptivities, and fluorescence quantum yields are estimated from the literature, often with large uncertainties. The bioparticles in the model are spherical and homogeneous. Calculated fluorescence cross sections for particles excited at 266, 280, and 355 nm are compared with measured values from the literature for several bacteria, bacterial spores and albumins. The calculated 266- and 280-nm excited fluorescence is within a factor of 3.2 of the measurements for the vegetative cells and proteins, but overestimates the fluorescence of spores by a factor of 10 or more. This is the first reported modeling of the fluorescence of bioaerosols in which the primary fluorophores and absorbing molecules are included.

Optical fiber-based setup for in vivo measurement of the delayed fluorescence lifetime of oxygen sensors

NASA Astrophysics Data System (ADS)

Piffaretti, Filippo M.; Santhakumar, Kanappan; Forte, Eddy; van den Bergh, Hubert E.; Wagnières, Georges A.

2011-03-01

A new optical-fiber-based spectrofluorometer for in vivo or in vitro detection of delayed fluorescence is presented and characterized. This compact setup is designed so that it can be readily adapted for future clinical use. Optical excitation is done with a nitrogen laser-pumped, tunable dye laser, emitting in the UV-vis part of the spectrum. Excitation and luminescence signals are carried to and from the biological tissues under investigation, located out of the setup enclosure, by a single optical fiber. These measurements, as well as measurements performed without a fiber on in vitro samples in a thermostable quartz cell, in a controlled-atmosphere enclosure, are possible due to the efficient collection of the laser-induced luminescence light which is collected and focused on the detector with a high aperture parabolic mirror. The detection is based on a gated photomultiplier which allows for time-resolved measurements of the delayed fluorescence intensity. Thus, relevant luminescence lifetimes, typically in the sub-microsecond-to-millisecond range, can be measured with near total rejection of the sample's prompt fluorescence. The instrument spectral and temporal resolution, as well as its sensitivity, is characterized and measurement examples are presented. The primary application foreseen for this setup is the monitoring and adjustment of the light dose delivered during photodynamic therapy.

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.

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.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Belz, J.; Cao, Z.; Huentemeyer, P.

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.

Measuring the remineralization potential of different agents with quantitative light-induced fluorescence digital Biluminator.

PubMed

Kucukyilmaz, Ebru; Savas, Selcuk

2017-01-26

The aim of this study was to investigate the effectiveness of different remineralization agents by quantitative light-induced fluorescence digital BiluminatorTM (QLF-D). Artificial caries lesions were created, and the teeth were divided according to the tested materials: (i) distilled water, (ii) acidulated phosphate fluoride (APF), (iii) Curodont Repair (CR), (iv) ammonium hexafluorosilicate (SiF) and (v) ammonium hexafluorosilicate plus cetylpyridinium chloride (SiF + CPC). After treatment procedures, each of the samples was placed in artificial saliva. After demineralization and 1 and 4 weeks of remineralization procedures, fluorescence loss and lesion areas were measured with QLF-D. Data were statistically analyzed (α = 0.05). The fluorescence values of the demineralized enamel specimens treated with the various agents differed significantly compared with pretreatment values for both 1 and 4 weeks (p<0.05). At 4 weeks, the highest fluorescence gain was calculated in the CR, APF and SiF groups compared with the control (p<0.05). APF, SiF and CR groups yielded greater remineralization ability than SiF + CPC and control groups.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wu Di; Li Yuhua; Wong, Molly D.

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 signalmore » 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.« less

Comparative evaluation of performance measures for shading correction in time-lapse fluorescence microscopy.

PubMed

Liu, L; Kan, A; Leckie, C; Hodgkin, P D

2017-04-01

Time-lapse fluorescence microscopy is a valuable technology in cell biology, but it suffers from the inherent problem of intensity inhomogeneity due to uneven illumination or camera nonlinearity, known as shading artefacts. This will lead to inaccurate estimates of single-cell features such as average and total intensity. Numerous shading correction methods have been proposed to remove this effect. In order to compare the performance of different methods, many quantitative performance measures have been developed. However, there is little discussion about which performance measure should be generally applied for evaluation on real data, where the ground truth is absent. In this paper, the state-of-the-art shading correction methods and performance evaluation methods are reviewed. We implement 10 popular shading correction methods on two artificial datasets and four real ones. In order to make an objective comparison between those methods, we employ a number of quantitative performance measures. Extensive validation demonstrates that the coefficient of joint variation (CJV) is the most applicable measure in time-lapse fluorescence images. Based on this measure, we have proposed a novel shading correction method that performs better compared to well-established methods for a range of real data tested. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

The application of laser induced predissociation fluorescence to the measurement of vibrational temperatures in a shock layer flow

NASA Technical Reports Server (NTRS)

Sutton, D. J.; Houwing, A. F. P.; Palma, P. C.; Boyce, R. R.; Sandeman, R. J.; Mundt, CH.

1993-01-01

Single shot spatially and spectrally resolved laser induced predissociation fluorescence measurements in a shock layer around a cylinder in a pulsed supersonic free stream are presented. Fluorescence signals were produced using the tuned output of an argon fluoride excimer laser to excite a mixture of rovibrational transitions in molecular oxygen. The signals produced along a line inside the shock layer were focussed onto a two dimensional detector coupled to a spectrometer, thus allowing spectral and spatial resolution of the fluorescence. In this way, it was possible to detect two fluorescence signals from two different transitions simultaneously, allowing the determination of vibrational temperatures without the need for calibration. However, to minimize problems associated with low signal to noise ratios, background subtraction and spatial averaging was required. The experimental measurements are compared with theoretical inviscid shock layer calculations for nonequilibrium air. A description of the strategies employed in these calculations is also provided.

Atmospheric Fluorescence Yield

NASA Technical Reports Server (NTRS)

Adams, James H., Jr.; Christl, M. J.; Fountain, W. F.; Gregory, J. C.; Martens, K.; Sokolsky, P.; Whitaker, Ann F. (Technical Monitor)

2001-01-01

Several existing and planned experiments estimate the energies of ultra-high energy cosmic rays from air showers using the atmospheric fluorescence from these showers. Accurate knowledge of the conversion from atmospheric fluorescence to energy loss by ionizing particles in the atmosphere is key to this technique. In this paper we discuss a small balloon-borne instrument to make the first in situ measurements versus altitude of the atmospheric fluorescence yield. The instrument can also be used in the lab to investigate the dependence of the fluorescence yield in air on temperature, pressure and the concentrations of other gases that present in the atmosphere. The results can be used to explore environmental effects on and improve the accuracy of cosmic ray energy measurements for existing ground-based experiments and future space-based experiments.

Trimodal color-fluorescence-polarization endoscopy aided by a tumor selective molecular probe accurately detects flat lesions in colitis-associated cancer

NASA Astrophysics Data System (ADS)

Charanya, Tauseef; York, Timothy; Bloch, Sharon; Sudlow, Gail; Liang, Kexian; Garcia, Missael; Akers, Walter J.; Rubin, Deborah; Gruev, Viktor; Achilefu, Samuel

2014-12-01

Colitis-associated cancer (CAC) arises from premalignant flat lesions of the colon, which are difficult to detect with current endoscopic screening approaches. We have developed a complementary fluorescence and polarization reporting strategy that combines the unique biochemical and physical properties of dysplasia and cancer for real-time detection of these lesions. Using azoxymethane-dextran sodium sulfate (AOM-DSS) treated mice, which recapitulates human CAC and dysplasia, we show that an octapeptide labeled with a near-infrared (NIR) fluorescent dye selectively identified all precancerous and cancerous lesions. A new thermoresponsive sol-gel formulation allowed topical application of the molecular probe during endoscopy. This method yielded high contrast-to-noise ratios (CNR) between adenomatous tumors (20.6±1.65) and flat lesions (12.1±1.03) and surrounding uninvolved colon tissue versus CNR of inflamed tissues (1.62±0.41). Incorporation of nanowire-filtered polarization imaging into NIR fluorescence endoscopy shows a high depolarization contrast in both adenomatous tumors and flat lesions in CAC, reflecting compromised structural integrity of these tissues. Together, the real-time polarization imaging provides real-time validation of suspicious colon tissue highlighted by molecular fluorescence endoscopy.

The Role of Cell Volume in the Dynamics of Seizure, Spreading Depression, and Anoxic Depolarization

PubMed Central

Ullah, Ghanim; Wei, Yina; Dahlem, Markus A; Wechselberger, Martin; Schiff, Steven J

2015-01-01

Cell volume changes are ubiquitous in normal and pathological activity of the brain. Nevertheless, we know little of how cell volume affects neuronal dynamics. We here performed the first detailed study of the effects of cell volume on neuronal dynamics. By incorporating cell swelling together with dynamic ion concentrations and oxygen supply into Hodgkin-Huxley type spiking dynamics, we demonstrate the spontaneous transition between epileptic seizure and spreading depression states as the cell swells and contracts in response to changes in osmotic pressure. Our use of volume as an order parameter further revealed a dynamical definition for the experimentally described physiological ceiling that separates seizure from spreading depression, as well as predicted a second ceiling that demarcates spreading depression from anoxic depolarization. Our model highlights the neuroprotective role of glial K buffering against seizures and spreading depression, and provides novel insights into anoxic depolarization and the relevant cell swelling during ischemia. We argue that the dynamics of seizures, spreading depression, and anoxic depolarization lie along a continuum of the repertoire of the neuron membrane that can be understood only when the dynamic ion concentrations, oxygen homeostasis,and cell swelling in response to osmotic pressure are taken into consideration. Our results demonstrate the feasibility of a unified framework for a wide range of neuronal behaviors that may be of substantial importance in the understanding of and potentially developing universal intervention strategies for these pathological states. PMID:26273829

Depolarizing actions of GABA in immature neurons depend neither on ketone bodies nor on pyruvate.

PubMed

Tyzio, Roman; Allene, Camille; Nardou, Romain; Picardo, Michel A; Yamamoto, Sumii; Sivakumaran, Sudhir; Caiati, Maddalena D; Rheims, Sylvain; Minlebaev, Marat; Milh, Mathieu; Ferré, Pascal; Khazipov, Rustem; Romette, Jean-Louis; Lorquin, Jean; Cossart, Rosa; Khalilov, Ilgam; Nehlig, Astrid; Cherubini, Enrico; Ben-Ari, Yehezkel

2011-01-05

GABA depolarizes immature neurons because of a high [Cl(-)](i) and orchestrates giant depolarizing potential (GDP) generation. Zilberter and coworkers (Rheims et al., 2009; Holmgren et al., 2010) showed recently that the ketone body metabolite DL-3-hydroxybutyrate (DL-BHB) (4 mM), lactate (4 mM), or pyruvate (5 mM) shifted GABA actions to hyperpolarizing, suggesting that the depolarizing effects of GABA are attributable to inadequate energy supply when glucose is the sole energy source. We now report that, in rat pups (postnatal days 4-7), plasma D-BHB, lactate, and pyruvate levels are 0.9, 1.5, and 0.12 mM, respectively. Then, we show that DL-BHB (4 mM) and pyruvate (200 μM) do not affect (i) the driving force for GABA(A) receptor-mediated currents (DF(GABA)) in cell-attached single-channel recordings, (2) the resting membrane potential and reversal potential of synaptic GABA(A) receptor-mediated responses in perforated patch recordings, (3) the action potentials triggered by focal GABA applications, or (4) the GDPs determined with electrophysiological recordings and dynamic two-photon calcium imaging. Only very high nonphysiological concentrations of pyruvate (5 mM) reduced DF(GABA) and blocked GDPs. Therefore, DL-BHB does not alter GABA signals even at the high concentrations used by Zilberter and colleagues, whereas pyruvate requires exceedingly high nonphysiological concentrations to exert an effect. There is no need to alter conventional glucose enriched artificial CSF to investigate GABA signals in the developing brain.

Failure of feedback as a putative common mechanism of spreading depolarizations in migraine and stroke

NASA Astrophysics Data System (ADS)

Dahlem, Markus A.; Schneider, Felix M.; Schöll, Eckehard

2008-06-01

The stability of cortical function depends critically on proper regulation. Under conditions of migraine and stroke a breakdown of transmembrane chemical gradients can spread through cortical tissue. A concomitant component of this emergent spatio-temporal pattern is a depolarization of cells detected as slow voltage variations. The propagation velocity of ˜3mm/min indicates a contribution of diffusion. We propose a mechanism for spreading depolarizations (SD) that rests upon a nonlocal or noninstantaneous feedback in a reaction-diffusion system. Depending upon the characteristic space and time scales of the feedback, the propagation of cortical SD can be suppressed by shifting the bifurcation line, which separates the parameter regime of pulse propagation from the regime where a local disturbance dies out. The optimization of this feedback is elaborated for different control schemes and ranges of control parameters.

Noninvasive measurement of glucose concentration on human fingertip by optical coherence tomography

NASA Astrophysics Data System (ADS)

Chen, Tseng-Lin; Lo, Yu-Lung; Liao, Chia-Chi; Phan, Quoc-Hung

2018-04-01

A method is proposed for determining the glucose concentration on the human fingertip by extracting two optical parameters, namely the optical rotation angle and the depolarization index, using a Mueller optical coherence tomography technique and a genetic algorithm. The feasibility of the proposed method is demonstrated by measuring the optical rotation angle and depolarization index of aqueous glucose solutions with low and high scattering, respectively. It is shown that for both solutions, the optical rotation angle and depolarization index vary approximately linearly with the glucose concentration. As a result, the ability of the proposed method to obtain the glucose concentration by means of just two optical parameters is confirmed. The practical applicability of the proposed technique is demonstrated by measuring the optical rotation angle and depolarization index on the human fingertip of healthy volunteers under various glucose conditions.

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…

Slow Bursting Neurons of Mouse Cortical Layer 6b Are Depolarized by Hypocretin/Orexin and Major Transmitters of Arousal

PubMed Central

Wenger Combremont, Anne-Laure; Bayer, Laurence; Dupré, Anouk; Mühlethaler, Michel; Serafin, Mauro

2016-01-01

Neurons firing spontaneously in bursts in the absence of synaptic transmission have been previously recorded in different layers of cortical brain slices. It has been suggested that such neurons could contribute to the generation of alternating UP and DOWN states, a pattern of activity seen during slow-wave sleep. Here, we show that in layer 6b (L6b), known from our previous studies to contain neurons highly responsive to the wake-promoting transmitter hypocretin/orexin (hcrt/orx), there is a set of neurons, endowed with distinct intrinsic properties, which displayed a strong propensity to fire spontaneously in rhythmic bursts. In response to small depolarizing steps, they responded with a delayed firing of action potentials which, upon higher depolarizing steps, invariably inactivated and were followed by a depolarized plateau potential and a depolarizing afterpotential. These cells also displayed a strong hyperpolarization-activated rectification compatible with the presence of an Ih current. Most L6b neurons with such properties were able to fire spontaneously in bursts. Their bursting activity was of intrinsic origin as it persisted not only in presence of blockers of ionotropic glutamatergic and GABAergic receptors but also in a condition of complete synaptic blockade. However, a small number of these neurons displayed a mix of intrinsic bursting and synaptically driven recurrent UP and DOWN states. Most of the bursting L6b neurons were depolarized and excited by hcrt/orx through a direct postsynaptic mechanism that led to tonic firing and eventually inactivation. Similarly, they were directly excited by noradrenaline, histamine, dopamine, and neurotensin. Finally, the intracellular injection of these cells with dye and their subsequent Neurolucida reconstruction indicated that they were spiny non-pyramidal neurons. These results lead us to suggest that the propensity for slow rhythmic bursting of this set of L6b neurons could be directly impeded by hcrt

Right precordial-directed electrocardiographical markers identify arrhythmogenic right ventricular cardiomyopathy in the absence of conventional depolarization or repolarization abnormalities.

PubMed

Cortez, Daniel; Svensson, Anneli; Carlson, Jonas; Graw, Sharon; Sharma, Nandita; Brun, Francesca; Spezzacatene, Anita; Mestroni, Luisa; Platonov, Pyotr G

2017-10-13

Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) carries a risk of sudden death. We aimed to assess whether vectorcardiographic (VCG) parameters directed toward the right heart and a measured angle of the S-wave would help differentiate ARVD/C with otherwise normal electrocardiograms from controls. Task Force 2010 definite ARVD/C criteria were met for all patients. Those who did not fulfill Task Force depolarization or repolarization criteria (-ECG) were compared with age and gender-matched control subjects. Electrocardiogram measures of a 3-dimentional spatial QRS-T angle, a right-precordial-directed orthogonal QRS-T (RPD) angle, a root mean square of the right sided depolarizing forces (RtRMS-QRS), QRS duration (QRSd) and the corrected QT interval (QTc), and a measured angle including the upslope and downslope of the S-wave (S-wave angle) were assessed. Definite ARVD/C was present in 155 patients by 2010 Task Force criteria (41.7 ± 17.6 years, 65.2% male). -ECG ARVD/C patients (66 patients) were compared to 66 control patients (41.7 ± 17.6 years, 65.2% male). All parameters tested except the QRSd and QTc significantly differentiated -ECG ARVD/C from control patients (p < 0.004 to p < 0.001). The RPD angle and RtRMS-QRS best differentiated the groups. Combined, the 2 novel criteria gave 81.8% sensitivity, 90.9% specificity and odds ratio of 45.0 (95% confidence interval 15.8 to 128.2). ARVD/C disease process may lead to development of subtle ECG abnormalities that can be distinguishable using right-sided VCG or measured angle markers better than the spatial QRS-T angle, the QRSd or QTc, in the absence of Taskforce ECG criteria.

Effect of fluorescent particle size on the modulation efficiency of ultrasound-modulated fluorescence.

PubMed

Liu, Yuan; Yuan, Baohong; Vignola, Joseph

2012-01-01

To investigate whether the size of fluorescent particles affects the modulation efficiency of ultrasound-modulated fluorescence (UMF), we measured UMF and DC (direct current) signals of the fluorescence emission from four different sized fluorescent particles: (1) three carboxylate-modified fluorescent microspheres (FM) with diameters of 20 nm, 200 nm, and 1.0 µm and (2) streptavidin-conjugated Alexa Fluor 647 with a diameter of approximately 5 nm. The UMF and DC signals were simultaneously measured using a broadband lock-in amplifier and a narrowband amplifier, respectively. The ratio of the UMF strength to the DC signal strength is defined as the modulation efficiency. This modulation efficiency was then used to evaluate the effects of fluorophore size and concentration. Results show that the modulation efficiency was improved by approximately a factor of two when the size of the fluorescent particles is increased from 5 nm to 1 µm. In addition, the linear relationship between the UMF strength and ultrasound pressure (observed in our previous study) were maintained regardless of the fluorescent particle sizes.

Effect of fluorescent particle size on the modulation efficiency of ultrasound-modulated fluorescence

PubMed Central

Liu, Yuan; Yuan, Baohong; Vignola, Joseph

2013-01-01

To investigate whether the size of fluorescent particles affects the modulation efficiency of ultrasound-modulated fluorescence (UMF), we measured UMF and DC (direct current) signals of the fluorescence emission from four different sized fluorescent particles: (1) three carboxylate-modified fluorescent microspheres (FM) with diameters of 20 nm, 200 nm, and 1.0 µm and (2) streptavidin-conjugated Alexa Fluor 647 with a diameter of approximately 5 nm. The UMF and DC signals were simultaneously measured using a broadband lock-in amplifier and a narrowband amplifier, respectively. The ratio of the UMF strength to the DC signal strength is defined as the modulation efficiency. This modulation efficiency was then used to evaluate the effects of fluorophore size and concentration. Results show that the modulation efficiency was improved by approximately a factor of two when the size of the fluorescent particles is increased from 5 nm to 1 µm. In addition, the linear relationship between the UMF strength and ultrasound pressure (observed in our previous study) were maintained regardless of the fluorescent particle sizes. PMID:24179476

Two-Photon Excitation, Fluorescence Microscopy, and Quantitative Measurement of Two-Photon Absorption Cross Sections

NASA Astrophysics Data System (ADS)

DeArmond, Fredrick Michael

As optical microscopy techniques continue to improve, most notably the development of super-resolution optical microscopy which garnered the Nobel Prize in Chemistry in 2014, renewed emphasis has been placed on the development and use of fluorescence microscopy techniques. Of particular note is a renewed interest in multiphoton excitation due to a number of inherent properties of the technique including simplified optical filtering, increased sample penetration, and inherently confocal operation. With this renewed interest in multiphoton fluorescence microscopy, comes an increased demand for robust non-linear fluorescent markers, and characterization of the associated tool set. These factors have led to an experimental setup to allow a systematized approach for identifying and characterizing properties of fluorescent probes in the hopes that the tool set will provide researchers with additional information to guide their efforts in developing novel fluorophores suitable for use in advanced optical microscopy techniques as well as identifying trends for their synthesis. Hardware was setup around a software control system previously developed. Three experimental tool sets were set up, characterized, and applied over the course of this work. These tools include scanning multiphoton fluorescence microscope with single molecule sensitivity, an interferometric autocorrelator for precise determination of the bandwidth and pulse width of the ultrafast Titanium Sapphire excitation source, and a simplified fluorescence microscope for the measurement of two-photon absorption cross sections. Resulting values for two-photon absorption cross sections and two-photon absorption action cross sections for two standardized fluorophores, four commercially available fluorophores, and ten novel fluorophores are presented as well as absorption and emission spectra.

Measuring the Quenching of no Fluorescence Produced from the Excitation of Photo-Fragmented Nitrobenzene Using a Picosecond Laser.

NASA Astrophysics Data System (ADS)

Lue, Christopher J.; Tanjaroon, Chakree; Johnson, J. Bruce; Reeve, Scott W.; Allen, Susan D.

2013-06-01

The military is interested in using spectroscopic methods to detect nitroaromatic compounds related to explosives. Upon absorption of a UV photon, nitrobenzene can dissociate into C_6H_5O and NO. Wynn, et al. have shown that looking at NO fluorescence from the photodissociated nitrobenzene could be a possible detection method. However, the fluorescence can easily be quenched by molecular oxygen and other constituents in air. We have measured fluorescence lifetimes of the nascent NO resulting from photo-fragmented nitrobenzene using a pulsed picosecond tunable laser (pulse width ≈15 ps) by means of a two-color process. In the two-color process, photons of a particular energy dissociated the nitrobenzene while photons of a different energy probed the A^2Σ^+← X^2Π_{(1/2,3/2)} NO band system between 225-260 nm. We have performed the measurements with different background pressures of He, N_2, and air. We present the results of these measurements which indicate considerable quenching of the NO fluorescence due to oxygen. Wynn, C. M.; Palmacci, S.; Kunz, R. R.; and Rothschild, M.Opt. Express, OSA, 2010, 18, 5399-5406

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

NASA Astrophysics Data System (ADS)

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

2009-06-01

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

Near-Infrared II Fluorescence for Imaging Hindlimb Vessel Regeneration with Dynamic Tissue Perfusion Measurement

PubMed Central

Hong, Guosong; Lee, Jerry C.; Jha, Arshi; Diao, Shuo; Nakayama, Karina H.; Hou, Luqia; Doyle, Timothy C.; Robinson, Joshua T.; Antaris, Alexander L.; Dai, Hongjie; Cooke, John P.; Huang, Ngan F.

2014-01-01

Background Real-time vascular imaging that provides both anatomic and hemodynamic information could greatly facilitate the diagnosis of vascular diseases and provide accurate assessment of therapeutic effects. Here we have developed a novel fluorescence-based all-optical method, named near-infrared II (NIR-II) fluorescence imaging, to image murine hindlimb vasculature and blood flow in an experimental model of peripheral arterial disease, by exploiting fluorescence in the NIR-II region (1000–1400 nm) of photon wavelengths. Methods and Results Owing to the reduced photon scattering of NIR-II fluorescence compared to traditional NIR fluorescence imaging and thus much deeper penetration depth into the body, we demonstrated that the mouse hindlimb vasculature could be imaged with higher spatial resolution than in vivo microCT. Furthermore, imaging over 26 days revealed a significant increase in hindlimb microvascular density in response to experimentally induced ischemia within the first 8 days of the surgery (P < 0.005), which was confirmed by histological analysis of microvascular density. Moreover, the tissue perfusion in the ischemic hindlimb could be quantitatively measured by the dynamic NIR-II method, revealing the temporal kinetics of blood flow recovery that resembled microbead-based blood flowmetry and laser Doppler blood spectroscopy. Conclusions The penetration depth of millimeters, high spatial resolution and fast acquisition rate of NIR-II imaging makes it a useful imaging tool for murine models of vascular disease. PMID:24657826

Near-infrared II fluorescence for imaging hindlimb vessel regeneration with dynamic tissue perfusion measurement.

PubMed

Hong, Guosong; Lee, Jerry C; Jha, Arshi; Diao, Shuo; Nakayama, Karina H; Hou, Luqia; Doyle, Timothy C; Robinson, Joshua T; Antaris, Alexander L; Dai, Hongjie; Cooke, John P; Huang, Ngan F

2014-05-01

Real-time vascular imaging that provides both anatomic and hemodynamic information could greatly facilitate the diagnosis of vascular diseases and provide accurate assessment of therapeutic effects. Here, we have developed a novel fluorescence-based all-optical method, named near-infrared II (NIR-II) fluorescence imaging, to image murine hindlimb vasculature and blood flow in an experimental model of peripheral arterial disease, by exploiting fluorescence in the NIR-II region (1000-1400 nm) of photon wavelengths. Because of the reduced photon scattering of NIR-II fluorescence compared with traditional NIR fluorescence imaging and thus much deeper penetration depth into the body, we demonstrated that the mouse hindlimb vasculature could be imaged with higher spatial resolution than in vivo microscopic computed tomography. Furthermore, imaging during 26 days revealed a significant increase in hindlimb microvascular density in response to experimentally induced ischemia within the first 8 days of the surgery (P<0.005), which was confirmed by histological analysis of microvascular density. Moreover, the tissue perfusion in the ischemic hindlimb could be quantitatively measured by the dynamic NIR-II method, revealing the temporal kinetics of blood flow recovery that resembled microbead-based blood flowmetry and laser Doppler blood spectroscopy. The penetration depth of millimeters, high spatial resolution, and fast acquisition rate of NIR-II imaging make it a useful imaging tool for murine models of vascular disease. © 2014 American Heart Association, Inc.

3-compartment talaporfin sodium pharmacokinetic model by optimization using fluorescence measurement data from canine skin to estimate the concentration in interstitial space

NASA Astrophysics Data System (ADS)

Uno, Yuko; Ogawa, Emiyu; Aiyoshi, Eitaro; Arai, Tsunenori

2018-02-01

We constructed the 3-compartment talaporfin sodium pharmacokinetic model for canine by an optimization using the fluorescence measurement data from canine skin to estimate the concentration in the interstitial space. It is difficult to construct the 3-compartment model consisted of plasma, interstitial space, and cell because there is a lack of the dynamic information. Therefore, we proposed the methodology to construct the 3-compartment model using the measured talaporfin sodium skin fluorescence change considering originated tissue part by a histological observation. In a canine animal experiment, the talaporfin sodium concentration time history in plasma was measured by a spectrophotometer with a prepared calibration curve. The time history of talaporfin sodium Q-band fluorescence on left femoral skin of a beagle dog excited by talaporfin sodium Soret-band of 409 nm was measured in vivo by our previously constructed measurement system. The measured skin fluorescence was classified to its source, that is, specific ratio of plasma, interstitial space, and cell. We represented differential rate equations of the talaporfin sodium concentration in plasma, interstitial space, cell. The specific ratios and a converting constant to obtain absolute value of skin concentration were arranged. Minimizing the squared error of the difference between the measured fluorescence data and calculated concentration by the conjugate gradient method in MATLAB, the rate constants in the 3-compartment model were determined. The accuracy of the fitting operation was confirmed with determination coefficient of 0.98. We could construct the 3-compartment pharmacokinetic model for canine using the measured talaporfin sodium fluorescence change from canine skin.

Ocean Lidar Measurements of Beam Attenuation and a Roadmap to Accurate Phytoplankton Biomass Estimates

NASA Astrophysics Data System (ADS)

Hu, Yongxiang; Behrenfeld, Mike; Hostetler, Chris; Pelon, Jacques; Trepte, Charles; Hair, John; Slade, Wayne; Cetinic, Ivona; Vaughan, Mark; Lu, Xiaomei; Zhai, Pengwang; Weimer, Carl; Winker, David; Verhappen, Carolus C.; Butler, Carolyn; Liu, Zhaoyan; Hunt, Bill; Omar, Ali; Rodier, Sharon; Lifermann, Anne; Josset, Damien; Hou, Weilin; MacDonnell, David; Rhew, Ray

2016-06-01

Beam attenuation coefficient, c, provides an important optical index of plankton standing stocks, such as phytoplankton biomass and total particulate carbon concentration. Unfortunately, c has proven difficult to quantify through remote sensing. Here, we introduce an innovative approach for estimating c using lidar depolarization measurements and diffuse attenuation coefficients from ocean color products or lidar measurements of Brillouin scattering. The new approach is based on a theoretical formula established from Monte Carlo simulations that links the depolarization ratio of sea water to the ratio of diffuse attenuation Kd and beam attenuation C (i.e., a multiple scattering factor). On July 17, 2014, the CALIPSO satellite was tilted 30° off-nadir for one nighttime orbit in order to minimize ocean surface backscatter and demonstrate the lidar ocean subsurface measurement concept from space. Depolarization ratios of ocean subsurface backscatter are measured accurately. Beam attenuation coefficients computed from the depolarization ratio measurements compare well with empirical estimates from ocean color measurements. We further verify the beam attenuation coefficient retrievals using aircraft-based high spectral resolution lidar (HSRL) data that are collocated with in-water optical measurements.

Transient Fluorescence Spectroscopy and laser induced fluorescence lifetimes of terbium doped dipicolinic acid

NASA Astrophysics Data System (ADS)

Makoui, Anali

We have investigated the use of deep UV laser induced fluorescence for the sensitive detection and spectroscopic lifetime studies of terbium doped dipicolinic acid (DPA-Tb) and used this to study the optical characteristics of DPA which is a chemical surrounding most bacterial spores. Background absorption spectra, fluorescence spectra, and Excitation Emission Matrix (EEM) spectra were made of the DPA-Tb complex, using both fixed 266 nm wavelength and tunable (220 nm--280 nm) UV laser excitations. Of importance, the fluorescence lifetimes of the four main fluorescence peaks (488 nm, 543 nm, 581 nm, and 618 nm) of the DPA-Tb complex have been measured for the first time to our knowledge. The lifetimes of all the fluorescing lines have been measured as a function of DPA-Tb concentration, solvent pH, and solvent composition, including that for the weakest fluorescing line of DPA-Tb at 618 nm. In addition, a new spectroscopic lifetime measurement technique, which we call "Transient Fluorescence Spectroscopy", was developed. In this technique, a weak, quasi-CW, amplitude modulated UV laser (8.5 kHz) was used to measure the lifetimes of the fluorescence lines, and yields insight into energy transfer and excitation lifetimes within the system. This technique is especially useful when a high power laser is not either available or not suitable. In the latter case, this would be when a high power pulsed deep-UV laser could produce bleaching or destruction of the biological specimen. In addition, this technique simulated the excitation and fluorescence emission of the DPA-Tb using a 4-level energy model, and solved the dynamic transient rate equations to predict the temporal behavior of the DPA-Tb emitted fluorescence. Excellent agreement between the experiments and the simulation were found. This technique has the potential to provide a more accurate value for the fluorescence lifetime values. In addition, with the use of asymmetric excitation waveforms, the dynamic

Chloride channel blockers promote relaxation of TEA-induced contraction in airway smooth muscle

PubMed Central

Yim, Peter D.; Gallos, George; Perez-zoghbi, Jose F.; Trice, Jacquelyn; Zhang, Yi; Siviski, Matthew; Sonett, Joshua; Emala, Charles W.

2014-01-01

Enhanced airway smooth muscle (ASM) contraction is an important component in the pathophysiology of asthma. We have shown that ligand gated chloride channels modulate ASM contractile tone during the maintenance phase of an induced contraction, however the role of chloride flux in depolarization-induced contraction remains incompletely understood. To better understand the role of chloride flux under these conditions, muscle force (human ASM, guinea pig ASM), peripheral small airway luminal area (rat ASM) and airway smooth muscle plasma membrane electrical potentials (human cultured ASM) were measured. We found ex vivo guinea pig airway rings, human ASM strips and small peripheral airways in rat lungs slices relaxed in response to niflumic acid following depolarization-induced contraction induced by K+ channel blockade with tetraethylammonium chloride (TEA). In isolated human airway smooth muscle cells TEA induce depolarization as measured by a fluorescent indicator or whole cell patch clamp and this depolarization was reversed by niflumic acid. These findings demonstrate that ASM depolarization induced contraction is dependent on chloride channel activity. Targeting of chloride channels may be a novel approach to relax hypercontractile airway smooth muscle in bronchoconstrictive disorders. PMID:24662476

Chloride channel blockers promote relaxation of TEA-induced contraction in airway smooth muscle.

PubMed

Yim, Peter D; Gallos, George; Perez-Zoghbi, Jose F; Trice, Jacquelyn; Zhang, Yi; Siviski, Matthew; Sonett, Joshua; Emala, Charles W

2013-01-01

Enhanced airway smooth muscle (ASM) contraction is an important component in the pathophysiology of asthma. We have shown that ligand gated chloride channels modulate ASM contractile tone during the maintenance phase of an induced contraction, however the role of chloride flux in depolarization-induced contraction remains incompletely understood. To better understand the role of chloride flux under these conditions, muscle force (human ASM, guinea pig ASM), peripheral small airway luminal area (rat ASM) and airway smooth muscle plasma membrane electrical potentials (human cultured ASM) were measured. We found ex vivo guinea pig airway rings, human ASM strips and small peripheral airways in rat lungs slices relaxed in response to niflumic acid following depolarization-induced contraction induced by K(+) channel blockade with tetraethylammonium chloride (TEA). In isolated human airway smooth muscle cells TEA induce depolarization as measured by a fluorescent indicator or whole cell patch clamp and this depolarization was reversed by niflumic acid. These findings demonstrate that ASM depolarization induced contraction is dependent on chloride channel activity. Targeting of chloride channels may be a novel approach to relax hypercontractile airway smooth muscle in bronchoconstrictive disorders.

Software-based measurement of thin filament lengths: an open-source GUI for Distributed Deconvolution analysis of fluorescence images

PubMed Central

Gokhin, David S.; Fowler, Velia M.

2016-01-01

The periodically arranged thin filaments within the striated myofibrils of skeletal and cardiac muscle have precisely regulated lengths, which can change in response to developmental adaptations, pathophysiological states, and genetic perturbations. We have developed a user-friendly, open-source ImageJ plugin that provides a graphical user interface (GUI) for super-resolution measurement of thin filament lengths by applying Distributed Deconvolution (DDecon) analysis to periodic line scans collected from fluorescence images. In the workflow presented here, we demonstrate thin filament length measurement using a phalloidin-stained cryosection of mouse skeletal muscle. The DDecon plugin is also capable of measuring distances of any periodically localized fluorescent signal from the Z- or M-line, as well as distances between successive Z- or M-lines, providing a broadly applicable tool for quantitative analysis of muscle cytoarchitecture. These functionalities can also be used to analyze periodic fluorescence signals in nonmuscle cells. PMID:27644080

Fluorescence spectra of atmospheric aerosol at Adelphi, Maryland, USA: measurement and classification of single particles containing organic carbon

NASA Astrophysics Data System (ADS)

Pinnick, Ronald G.; Hill, Steven C.; Pan, Yong-Le; Chang, Richard K.

We measured laser-induced fluorescence spectra from individual supermicron-sized atmospheric particles drawn into our laboratory at Adelphi, MD, an urban site in the Washington, DC metroplex. A virtural impactor concentrator is used along with an aerodynamic-focusing-nozzle which forms, within an optical chamber, a focused aerosol jet where single aerosol particles can be interrogated on-the-fly with a pulsed 266-nm-wavelength laser. Sample rates are a few liter per minute, and are size dependent. Crossed-diode laser beams indicate when a particle is traversing the sample region and are used to trigger the UV laser to fire and the gated intensified CCD to record the fluorescence spectrum. Our breadboard fluorescence particle spectrometer measures particles in the 3-10 μm diameter size range. Typical trigger rates are a few per second. The usable spectral range is from about 295 to 605 nm. The majority of the particles have very weak fluorescence (on average 8 percent of particles have fluorescence signals above noise). The spectra were grouped using a heirarchical cluster analysis, with parameters chosen so that spectra typically cluster into 4-12 main categories. From the set of all cluster spectra we chose 8 template spectra for reanalyzing all the data. On average, 92 percent (81-94 percent) of the spectra were similar to these templates (using the same thresholds used for the cluster analysis). The major emission bands of the most commonly occurring spectra have peaks: near 460 nm (28 percent of fluorescent particles on average), a very broad hump, and may be humic acids or humic like substances; near 317 nm (on average 24 percent of fluorescent particles); near 321 and 460 nm (a double hump, 12 percent of fluorescent particles); and near 341 nm (8 percent of fluorescent particles). Some of the fluorescence in spectra peaking in the 317-341 nm range is probably from dicyclic aromatics and heterocyclics, including the amino acid tryptophan in biological

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.

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.

Measurements of IN and BIO-IN with the fast ice nucleus chamber FINCH at Mt. Zugspitze, Mt. Puy de Dôme and Jungfraujoch during fall and winter

NASA Astrophysics Data System (ADS)

Nillius, B.; Frank, F.; Bingemer, H.; Curtius, J.; Bundke, U.

2013-05-01

In this work we present IN measurements at Mt. Zugspitze, Germany, 2650 m.a.s.l., Mt. Puy de Dôme, France, 1464 m.a.s.l. and Jungfraujoch, Switzerland, 3580 m a.s.l during fall and winter 2012 with the instrument FINCH HALO (Fast Ice Nucleus Chamber for the High Altitude and LOng range research aircraft HALO). In this device the temperature and super saturation for activation of Ice Nuclei (IN) and the growth to ice crystals is obtained by mixing three gas flows of different temperatures and moisture. After the growth of IN and Cloud Condensation Nuclei (CCN) to macroscopic ice crystals and super-cooled water droplets in the development chamber, they are counted using an optical detector. The discrimination between ice and water is made by measuring the circular depolarization ratio of the backscattered laser light of each individual particle. IN are classified as biological particles by measuring their individual intrinsic-fluorescence during the winter campaigns in average 30-40 % of the IN show an intrinsic fluorescence and are supposed to be of biological origin.

Seizures, refractory status epilepticus, and depolarization block as endogenous brain activities

NASA Astrophysics Data System (ADS)

El Houssaini, Kenza; Ivanov, Anton I.; Bernard, Christophe; Jirsa, Viktor K.

2015-01-01

Epilepsy, refractory status epilepticus, and depolarization block are pathological brain activities whose mechanisms are poorly understood. Using a generic mathematical model of seizure activity, we show that these activities coexist under certain conditions spanning the range of possible brain activities. We perform a detailed bifurcation analysis and predict strategies to escape from some of the pathological states. Experimental results using rodent data provide support of the model, highlighting the concept that these pathological activities belong to the endogenous repertoire of brain activities.

Application of a pulsed laser for measurements of bathymetry and algal fluorescence.

NASA Technical Reports Server (NTRS)

Hickman, G. D.; Hogg, J. E.; Friedman, E. J.; Ghovanlou, A. H.

1973-01-01

The technique of measuring water depths with an airborne pulsed dye laser is studied, with emphasis on the degrading effect of some environmental and operational parameters on the transmitted and reflected laser signals. Extrapolation of measurements of laser stimulated fluorescence, performed as a function of both the algal cell concentration and the distance between the algae and the laser/receiver, indicate that a laser system operating from a height of 500 m should be capable of detecting chlorophyll concentrations as low as 1.0 mg/cu m.-

Protein sorption on polymer surfaces measured by fluorescence labels.

PubMed

Brynda, E; Drobník, J; Vacík, J; Kálal, J

1978-01-01

Fluorescence labeling can be used in studying protein sorption on various surfaces with a sensitivity of about 10(-8) g/cm2, commensurate with radioactive labeling. Fluorescamine proved to be the most suitable compound for studying protein sorption on hydrophilic gels, because, unlike fluoresceine isothiocyanate and dansylchloride, free fluorochrome does not interfere with measurements. Sorption properties of labeled serum albumin were tested on poly(2-hydroxyethyl methacrylate), on the copolymer of 2-hydroxyethyl methacrylate with methyl methacrylate, and on polyethylene. Labeling does not cause aggregation of the protein, but, as expected, it shifts and somewhat broadens its electrophoretic band while at the same time slightly raising its affinity toward hydrophobic surfaces.

An aircraft compatible laser induced fluorescence system - In situ and remote measurements of trace gases

NASA Technical Reports Server (NTRS)

Davis, D. D.; Philen, D.

1978-01-01

The laser-induced fluorescence technique for obtaining direct measurements of atmospheric OH and other gases is described. A narrow-band UV laser is tuned to one or more of the electronic absorption bands of a specified molecule so as to cause fluorescence from a bonding excited electronic state. The monitored wavelength is longer than the laser wavelength. Equipment, specifics for OH detection, data processing, and interference are discussed, and application of the technique to the detection of NO, SO2, and CH2O is considered.

High resolution measurements of solar induced chlorophyll fluorescence in the Fraunhofer oxigen bands

NASA Astrophysics Data System (ADS)

Mazzoni, M.; Agati, G.; Cecchi, G.; Toci, G.; Mazzinghi, P.

2017-11-01

Spectra of solar radiance reflected by leaves close to the Fraunhofer bands show the net contribution of chlorophyll fluorescence emission which adds to the reflected solar spectra. In a laboratory experiment, a low stray light, high resolution, 0.85 m double monochromator was used to filter radiation living leaves still attached to the plant in correspondence of the 687 nm and 760 nm O2 absorption bands. Reference spectra from a non fluorescent white reference were also acquired. Acquisition was performed by a Microchannel plate (MCP) intensified diode array with 512 elements. A fit of the spectral data outside the absorption lines allowed to retrieve the spectral base-line as a function of wavelength for the reference panel and the leaf. Reflectance functions were determined extending the Plascyck equation system to all the resolved lines of the oxygen absorption bands and using the base-lines for the continuum values. Fluorescence was deduced from the same equation system, using both the measured leaf and reference radiance spectra and the leaf reflectance fitting function.

Efficiency and concentration ratio measurements of fluorescent solar concentrators using a xenon measurement system.

PubMed

Heidler, K

1981-03-01

An indoor test stand for fluorescent planar concentrator-collectors (FPC) with a 1.6-kW xenon light source, irradiating a rectangular triangle (active area 400 cm(2)) with 82 mW cm(-2), and a typical homogeneity of +/-10% is described in detail. Conditions and definitions for the efficiency measurements are discussed, taking intensity and reflection effects into account. As an illustration FPC efficiency and concentration ratios are reported (best optical efficiency was 7.5% for a single-plate one-dye FPC), the influence of thickness variation is discussed, and an enhancement in solar cell efficiency is demonstrated.

Optimal fluorescence waveband determination for detecting defect cherry tomatoes using fluorescence excitation-emission matrix

USDA-ARS?s Scientific Manuscript database

A multi-spectral fluorescence imaging technique was used to detect defect cherry tomatoes. The fluorescence excitation and emission matrix was used to measure for defects, sound surface, and stem areas to determine the optimal fluorescence excitation and emission wavelengths for discrimination. Two-...