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Sample records for absorption fluorescence excitation

  1. Fluorescence, Absorption, and Excitation Spectra of Polycyclic Aromatic Hydrocarbons as a Tool for Quantitative Analysis

    ERIC Educational Resources Information Center

    Rivera-Figueroa, A. M.; Ramazan, K. A.; Finlayson-Pitts, B. J.

    2004-01-01

    A quantitative and qualitative study of the interplay between absorption, fluorescence, and excitation spectra of pollutants called polycyclic aromatic hydrocarbons (PAHs) is conducted. The study of five PAH displays the correlation of the above-mentioned properties along with the associated molecular changes.

  2. Electronically excited dipole moment of 4-aminobenzonitrile from thermochromic absorption and fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Kawski, A.; Kukliński, B.; Bojarski, P.

    2006-07-01

    The effect of temperature on absorption and fluorescence spectra of 4-aminobenzonitrile (ABN) in 1,2-dichloroethane is studied for temperature ranging from 296 K to 343 K. The analysis of absorption and fluorescence band shift on the basis of Bilot and Kawski theory [L. Bilot, A. Kawski, Z. Naturforsch. 17a (1962) 621], for the known dipole moment in the ground state μg = 5.92 D, and α/ a3 = 0.5 ( α is the polarizability and a is the Onsager interaction radius of the solute) yields for ABN: (1) the empirical Onsager interaction radius a = 3.3 Å, (2) the dipole moment in the excited S 1 state μe = 7.14 D which agrees very well with the value of μe = 7.20 D obtained by Borst et al. [D.R. Borst, T.M. Korter, D.W. Pratt, Chem. Phys. Lett. 350 (2001) 485] from Stark effect studies. Both values of μe concern free ABN molecule and differ significantly from the values of μg (8.0 D, 8.5 D and 8.3 D in cyclohexane, benzene and 1,4-dioxane, respectively) obtained by Schuddeboom et al. [W. Schuddeboom, S.A. Jonker, J.M. Warman, U. Leinhos, W. Kühnle, K.A. Zachariasse, J. Phys. Chem. 96 (1992) 10809] from the time-resolved microwave conductivity measurements which are solvent-dependent. The group moment additivity law in the case of ABN molecule is approximately applicable, both in the ground and in the excited electronic state.

  3. Excited-state intramolecular proton transfer of 2-acetylindan-1,3-dione studied by ultrafast absorption and fluorescence spectroscopy

    PubMed Central

    Verma, Pramod Kumar; Steinbacher, Andreas; Schmiedel, Alexander; Nuernberger, Patrick; Brixner, Tobias

    2015-01-01

    We employ transient absorption from the deep-UV to the visible region and fluorescence upconversion to investigate the photoinduced excited-state intramolecular proton-transfer dynamics in a biologically relevant drug molecule, 2-acetylindan-1,3-dione. The molecule is a ß-diketone which in the electronic ground state exists as exocyclic enol with an intramolecular H-bond. Upon electronic excitation at 300 nm, the first excited state of the exocyclic enol is initially populated, followed by ultrafast proton transfer (≈160 fs) to form the vibrationally hot endocyclic enol. Subsequently, solvent-induced vibrational relaxation takes place (≈10 ps) followed by decay (≈390 ps) to the corresponding ground state. PMID:26798837

  4. Fluorescence excitation and ultraviolet absorption spectra and theoretical calculations for benzocyclobutane: Vibrations and structure of its excited S{sub 1}(π,π{sup *}) electronic state

    SciTech Connect

    Shin, Hee Won; Ocola, Esther J.; Laane, Jaan; Kim, Sunghwan

    2014-01-21

    The fluorescence excitation spectra of jet-cooled benzocyclobutane have been recorded and together with its ultraviolet absorption spectra have been used to assign the vibrational frequencies for this molecule in its S{sub 1}(π,π{sup *}) electronic excited state. Theoretical calculations at the CASSCF(6,6)/aug-cc-pVTZ level of theory were carried out to compute the structure of the molecule in its excited state. The calculated structure was compared to that of the molecule in its electronic ground state as well as to the structures of related molecules in their S{sub 0} and S{sub 1}(π,π{sup *}) electronic states. In each case the decreased π bonding in the electronic excited states results in longer carbon-carbon bonds in the benzene ring. The skeletal vibrational frequencies in the electronic excited state were readily assigned and these were compared to the ground state and to the frequencies of five similar molecules. The vibrational levels in both S{sub 0} and S{sub 1}(π,π{sup *}) states were remarkably harmonic in contrast to the other bicyclic molecules. The decreases in the frequencies of the out-of-plane skeletal modes reflect the increased floppiness of these bicyclic molecules in their S{sub 1}(π,π{sup *}) excited state.

  5. Solvent Effects on the Electronic Absorption and Fluorescence Spectra of HNP: Estimation of Ground and Excited State Dipole Moments.

    PubMed

    Desai, Vani R; Hunagund, Shirajahammad M; Basanagouda, Mahantesha; Kadadevarmath, Jagadish S; Sidarai, Ashok H

    2016-07-01

    We report the effect of solvents on absorption and fluorescence spectra of biologically active 3(2H)-pyridazinone namely 5-(2-hydroxy-naphthalen-1-yl)-2-phenyl-2H-pyridazin-3-one (HNP) in different solvents at room temperature. The ground and the excited state dipole moments of HNP molecule was estimated from Lippert's, Bakshiev's and Kawski-Chamma-Viallet's equations using the solvatochromic shift method. The ground state dipole moment (μ g ) was also estimated by Guggenheim and Higasi method using the dielectric constant and refractive index of solute at different concentrations, the μ g value obtained from these two methods are comparable to the μ g value obtained by the solvatochromic shift method. The excited state dipole moment (μ e ) is greater than the ground state dipole moment (μ g ), which indicates that the excited state is more polar than the ground state. Further, we have evaluated the change in dipole moment (Δμ) from the solvatochromic shift method and on the basis of molecular-microscopic solvent polarity parameter[Formula: see text], later on the values were compared. PMID:27220623

  6. Ultrafast Time-Resolved Emission and Absorption Spectra of meso-Pyridyl Porphyrins upon Soret Band Excitation Studied by Fluorescence Up-Conversion and Transient Absorption Spectroscopy.

    PubMed

    Venkatesh, Yeduru; Venkatesan, M; Ramakrishna, B; Bangal, Prakriti Ranjan

    2016-09-01

    thermally equilibrated Qx state by vibrational cooling/relaxations of excess energy within solvent. This relaxed Qx state decays to ground as well as triplet state by 7-8 ns time scale. The femtosecond transient absorption studies of TpyPs in three different excitations at S2 (400 nm), Qy (515 nm), and Qx (590 nm) along with extensive global and target model analysis of TA data exclusively generate the true spectra of each excited species/state with their respective lifetimes along with microscopic rate constants associated with each state. The following five exponential components with lifetime values of 65-70 fs, ∼0.3-0.5 ps, ∼20 ± 2 ps, ∼7 ± 1 ns, and 1-2 μs are observed which are associated with S2, Qy, hot Qx, thermally relaxed Qx, and lowest triplet (T1) states, respectively, when excited at S2, and four (Qy, hot Qx, thermally relaxed Qx, and lowest triplet (T1) states) and three (hot Qx, thermally relaxed Qx, and lowest triplet (T1) states) states are obtained when excited at 515 nm (Qy) and 590 nm (Qx), respectively, as expected. The TA results parallel the fluorescence up-conversion studies, and both the results not only compliment each other but also unveil the ultrafast internal conversion from S2 to Qy, S2 to Qx, and Qy to Qx for all three isomers in a similar fashion with nearly equal characteristic decay times.

  7. Ultrafast Time-Resolved Emission and Absorption Spectra of meso-Pyridyl Porphyrins upon Soret Band Excitation Studied by Fluorescence Up-Conversion and Transient Absorption Spectroscopy.

    PubMed

    Venkatesh, Yeduru; Venkatesan, M; Ramakrishna, B; Bangal, Prakriti Ranjan

    2016-09-01

    thermally equilibrated Qx state by vibrational cooling/relaxations of excess energy within solvent. This relaxed Qx state decays to ground as well as triplet state by 7-8 ns time scale. The femtosecond transient absorption studies of TpyPs in three different excitations at S2 (400 nm), Qy (515 nm), and Qx (590 nm) along with extensive global and target model analysis of TA data exclusively generate the true spectra of each excited species/state with their respective lifetimes along with microscopic rate constants associated with each state. The following five exponential components with lifetime values of 65-70 fs, ∼0.3-0.5 ps, ∼20 ± 2 ps, ∼7 ± 1 ns, and 1-2 μs are observed which are associated with S2, Qy, hot Qx, thermally relaxed Qx, and lowest triplet (T1) states, respectively, when excited at S2, and four (Qy, hot Qx, thermally relaxed Qx, and lowest triplet (T1) states) and three (hot Qx, thermally relaxed Qx, and lowest triplet (T1) states) states are obtained when excited at 515 nm (Qy) and 590 nm (Qx), respectively, as expected. The TA results parallel the fluorescence up-conversion studies, and both the results not only compliment each other but also unveil the ultrafast internal conversion from S2 to Qy, S2 to Qx, and Qy to Qx for all three isomers in a similar fashion with nearly equal characteristic decay times. PMID:27494567

  8. Excitation/Detection Strategies for OH Planar Laser-Induced Fluorescence Measurements in the Presence of Interfering Fuel Signal and Absorption Effects

    NASA Technical Reports Server (NTRS)

    Heath, Christopher M.; Anderson, Robert C.; Hicks, Yolanda R.

    2011-01-01

    Planar laser-induced fluorescence (PLIF) excitation/detection methods have been applied to obtain spatial distributions of the hydroxyl [OH] reacting intermediary and hydrocarbon [HC] primary species in laminar and turbulent combustion reactions. In this report, broadband and narrowband excitation/filtering techniques are explored to identify an optimal experimental configuration yielding significant fluorescent signal with low absorption losses. The combustion environments analyzed include 1) a laminar non-premixed methane/air flame and 2) a turbulent, non-premixed Jet-A/air fueled flame within a lean flame tube combustor. Hydrocarbon-based fuel and OH were excited via the R1 (1), R1(10) and R2(7) transitions of the A(sup 2)Epsilon(+) X(sup 2)pi(1,0) band using a broadband Nd:YAG pumped optical parametric oscillator (OPO) and narrowband Nd:YAG/dye laser with ultraviolet frequency extension (UVX) package. Variables tested for influence on fluorescent signal and absorption characteristics were excitation line, laser energy, exciting linewidth, combustion reactants, and test flow conditions. Results are intended to guide the transition from a dye/UVX laser to an OPO system for performing advanced diagnostics of low-emission combustion concepts.

  9. Solvent effects on the absorption and fluorescence spectra of quinine sulphate: Estimation of ground and excited-state dipole moments

    NASA Astrophysics Data System (ADS)

    Joshi, Sunita; Pant, Debi D.

    2012-06-01

    Ground and excited state dipole moments of probe quinine sulphate (QS) was obtained using Solvatochromic shift method. Higher dipole moment is observed for excited state as compared to the ground state which is attributed to the higher polarity of excited state.

  10. Prediction of BOD, COD, and total nitrogen concentrations in a typical urban river using a fluorescence excitation-emission matrix with PARAFAC and UV absorption indices.

    PubMed

    Hur, Jin; Cho, Jinwoo

    2012-01-01

    The development of a real-time monitoring tool for the estimation of water quality is essential for efficient management of river pollution in urban areas. The Gap River in Korea is a typical urban river, which is affected by the effluent of a wastewater treatment plant (WWTP) and various anthropogenic activities. In this study, fluorescence excitation-emission matrices (EEM) with parallel factor analysis (PARAFAC) and UV absorption values at 220 nm and 254 nm were applied to evaluate the estimation capabilities for biochemical oxygen demand (BOD), chemical oxygen demand (COD), and total nitrogen (TN) concentrations of the river samples. Three components were successfully identified by the PARAFAC modeling from the fluorescence EEM data, in which each fluorophore group represents microbial humic-like (C1), terrestrial humic-like organic substances (C2), and protein-like organic substances (C3), and UV absorption indices (UV(220) and UV(254)), and the score values of the three PARAFAC components were selected as the estimation parameters for the nitrogen and the organic pollution of the river samples. Among the selected indices, UV(220), C3 and C1 exhibited the highest correlation coefficients with BOD, COD, and TN concentrations, respectively. Multiple regression analysis using UV(220) and C3 demonstrated the enhancement of the prediction capability for TN.

  11. Estimation of ground and excited state dipole moment of laser dyes C504T and C521T using solvatochromic shifts of absorption and fluorescence spectra

    NASA Astrophysics Data System (ADS)

    Basavaraja, Jana; Suresh Kumar, H. M.; Inamdar, S. R.; Wari, M. N.

    2016-02-01

    The absorption and fluorescence spectra of laser dyes: coumarin 504T (C504T) and coumarin 521T (C521T) have been recorded at room temperature in a series of non-polar and polar solvents. The spectra of these dyes showed bathochromic shift with increasing in solvent polarity indicating the involvement of π → π* transition. Kamlet-Taft and Catalan solvent parameters were used to analyze the effect of solvents on C504T and C521T molecules. The study reveals that both general solute-solvent interactions and specific interactions are operative in these two systems. The ground state dipole moment was estimated using Guggenheim's method and also by quantum mechanical calculations. The solvatochromic data were used to determine the excited state dipole moment (μe). It is observed that dipole moment value of excited state (μe) is higher than that of the ground state in both the laser dyes indicating that these dyes are more polar in nature in the excited state than in the ground state.

  12. Standoff alpha radiation detection via excited state absorption of air

    SciTech Connect

    Yao, Jimmy; Yin, Stuart Shizhuo; Brenizer, Jack; Hui, Rongqing

    2013-06-24

    A standoff alpha radiation detection technique based on the physical mechanism of excited state absorption of air molecules was explored and is presented in this paper. Instead of directly detecting the radiation via measuring the intensity of radiation induced air fluorescence, the radiation is detected via the excited state absorption of alpha radiation excited/ionized air molecules. Both theoretical analyses and experimental verifications were conducted. The experimental results confirmed that the radiation could be detected via excited state absorption of radiation excited/ionized air molecules at a 10 m standoff distance, which was consistent with the theoretical analyses.

  13. Absorption and fluorescence spectroscopy on a smartphone

    NASA Astrophysics Data System (ADS)

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

    2015-07-01

    A self-powered smartphone-based field-portable "dual" spectrometer has been developed for both absorption and fluorescence measurements. The smartphone's existing flash LED has sufficient optical irradiance to undertake absorption measurements within a 3D-printed case containing a low cost nano-imprinted polymer diffraction grating. A UV (λex ~ 370 nm) and VIS (λex ~ 450 nm) LED are wired into the circuit of the flash LED to provide an excitation source for fluorescence measurements. Using a customized app on the smartphone, measurements of absorption and fluorescence spectra are demonstrated using pH-sensitive and Zn2+-responsive probes. Detection over a 300 nm span with 0.42 nm/pixel spectral resolution is demonstrated. Despite the low cost and small size of the portable spectrometer, the results compare well with bench top instruments.

  14. Two-photon excitation in laser scanning fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Strickler, James H.; Webb, Watt W.

    1991-04-01

    Simultaneous absorption of two red photons from a strongly focused subpicosecond colliding pulse mode4ocked dye laser stimulates visible fluorescence emission from fluorophores having their normal absorption in the ultraviolet1. The quadratic increase of the two-photon excitation rate with excitation intensity restricts fluorescence emission to the focal volume thus providing the same depth resolution as does confocal microscopy. Image degradation due to out of focus backround is thus avoided. Photobleaching and most cellular photodamage are similarly confined to the focus thereby minimizing sample degredation during acquisition of the multiple sections required for 3-d image reconstruction. Fluorescence images of living cells and other thick photolabile fluorescence labled assemblies illustrate the depth discrimination of both two-photon fluorescence excitation and photobleaching. The quadratic intensity dependence of two-photon excitation allows 3-d spatially resolved photochemistry in particular the photolytic release of caged compounds such as neurotransmitters nucleotides fluorescent dyes and second messengers such as 1P3 and Ca. The two-photon release of cased ATP has been measured and release of a caged fluorescent dye has been shown. Point photobleaching and a 3-d " write once read many" optical memory have been demonstrated. Two-photon excitation of photo-initiated polymerization with a sharply focused single beam allows microfabrication of complex structures of arbitrary form. By scanning the focused beam through a liquid polymer with a UV excited initiator it is possible to harden the polymer only at the focus thereby creating

  15. Fluorescence monitor method for measuring effective absorption coefficients of molecular rovibronic transitions using tunable dye laser excitation: The case of absorber linewidth narrower than the laser linewidth applied to H/sub 2/CO

    SciTech Connect

    Fairchild, P.W.; Garland, N.L.; Howard, W.E. III; Lee, E.K.C.

    1980-10-01

    A technique for measuring ''effective'' absorption coefficients is described. It circumvents deviations from Beer's law caused when the excitation source bandwidth is larger than the absorber bandwidth. The technique employs a fluorescence cell placed after an absorption cell to selectively monitor absorption in the center region of the source line. Model calculations relating the fluorescence intensity to source and absorber line shapes indicate that this method should yield linear Beer's law plots for moderate values of k/sub 0/Nl and ..cap alpha.., where ..cap alpha.. is the ratio of the source bandwidth to the absorber bandwidth. This technique has been applied to a number of single rotational levels in the 4/sup 1//sub 0/ transition of the H/sub 2/CO A /sup 1/A/sub 2/reverse arrowX /sup 1/A/sub 1/ system using pulsed, tunable dye laser excitation. The effective absorption coefficients determined experimentally have been compared to the theoretically calculated absorption coefficients.

  16. Laser Excited Fluorescence Studies Of Black Liquor

    NASA Astrophysics Data System (ADS)

    Horvath, J. J.; Semerjian, H. G.

    1986-10-01

    Laser excited fluorescence of black liquor was investigated as a possible monitoring technique for pulping processes. A nitrogen pumped dye laser was used to examine the fluorescence spectrum of black liquor solutions. Various excitation wavelengths were used between 290 and 403 nm. Black liquor fluorescence spectra were found to vary with both excitation wavelength and black liquor concentration. Laser excited fluorescence was found to be a sensitive technique for measurement of black liquor with good detection limits and linear response over a large dynamic range.

  17. [Saltation behavior in excitation spectra of fluorescent molecules].

    PubMed

    Miao, Di; Xu, Yi-zhuang; Yang, Jun; Xu, Zhen-hua; Wu, Jin-guang

    2004-05-01

    Excitation spectra are commonly used to study relationship between molecular structure of fluorescent substances and energy transfer during the fluorescence process. It is generally taken for granted that the excitation spectrum of the sample is equivalent to its absorption spectrum, even a copy of the latter. However, exceptions have been found in many cases. Considering various factors that affect the excitation spectra of solution comprehensively, a model has been established to study the behavior of the excitation spectra. After analyzing the model mathematically, including introducing catastrophe theory, we came into the following conclusions: As far as the topological properties are concerned, the excitation spectra are the same as its absorption spectra, provided the concentration of the substance is below a threshold. However, when the concentration is beyond the threshold, the excitation spectra undergo a series of topological saltation, leading to significant a deviation from the absorption spectra. Comparative studies of both excitation and absorption spectra of naphthalene dissolved in n-hexane confirmed the above hypothesis.

  18. UV absorption, laser excited fluorescence and direct sampling in the study of the formation of polycyclic aromatic hydrocarbons in rich CH/sub 4//O/sub 2/ flames

    SciTech Connect

    Di Lorenzo, A.; D'Alessio, A.; Cinocotti, V.; Masi, S.; Menna, P.; Venitozzi, C.

    1981-01-01

    UV absorption and laser excited visible fluorescence techniques have been used to analyze systematically the early regions of atmospheric pressure, premixed flat CH/sub 4//O/sub 2/ flames. A range of CH/sub 4//O/sub 2/ ratios (R) from 0.1 to 1.27 was explored and different flow rates and inert addition were also considered for the sooting R.1.27 flame. A new sampling and collecting method allowed the authors to follow the formation of individual polycyclic aromatic hydrocarbons (PAH) immediately downstream of the reaction zone. Almost down to the reaction zone, the relative concentration of two and three ring PAH is quite high and then undergoes a rapid decline downstream, where four ring PAH are predominate; the concentrations of 5-7 ring PAH are almost negligible. The influence of the probe on the PAH concentration levels was also investigated using fluorescence emission. 12 refs.

  19. Absorption and fluorescence properties of fluorescein

    NASA Astrophysics Data System (ADS)

    Sjöback, Robert; Nygren, Jan; Kubista, Mikael

    1995-06-01

    We have characterized the protolytic equilibria of fluorescein and determined the spectroscopic properties of its protolytic forms. The protolytic constants relating the chemical activities (which at low ionic strength equal concentrations) of the cation, neutral form, anion and dianion are p K1 = 2.08, p K2 = 4.31, and p K3 = 6.43. All forms have rather high molar absorptivities being ɛ437FH 3 1 = 53 000, ɛ434FH 2 = 11 000, ɛ453FH- = 29 000 ( ɛ472FH = 29 000) and ɛ490F 2 = 76 900 M -1 cm -1 for the cation, neutral form, anion and dianion, respectively. The dianion has the most intense fluorescence with a quantum yield of 0.93 but also the anion shows considerable fluorescence with a quantum yield of 0.37. The neutral and cationic species are upon excitation converted into the anion and fluoresce with quantum yields of about 0.30 and 0.18, respectively.

  20. Laser Excited Fluorescence For Forensic Diagnostics

    NASA Astrophysics Data System (ADS)

    McKinney, Robert E.

    1986-07-01

    The application of laser excited fluorescence to the detection and identification of latent fingerprints was first accomplished ten years ago. The development of the technology has progressed rapidly with the introduction of commercial equipment by several manufacturers. Systems based on Argon-ion, Copper-vapor, and frequency-doubled Nd:YAG lasers are compared. The theoretical basis of detection by fluorescence is discussed along with the more useful techniques of dye staining. Other applications of the laser excited fluorescence in forensic investigation include gunshot residue analysis, serology, collection of trace evidence, and document examination.

  1. Temperature dependence of the excited state absorption of alexandrite

    SciTech Connect

    Shand, M.L.; Jenssen, H.P.

    1983-03-01

    The temperature dependence from 28 to 290/sup 0/C of the excited-state absorption cross section sigma /SUB 2a/ (E) in the gain wavelength region of alexandrite has been determined from the temperature dependence of the single pass gain (SPG) and of the fluorescence. sigma /SUB 2a/ (E) and the emission cross section increase with temperature at approximately the same rate.

  2. Two-photon excitation fluorescence bioassays.

    PubMed

    Hänninen, Pekka; Soukka, Jori; Soini, Juhani T

    2008-01-01

    Application of two-photon excitation of fluorescence in microscopy is one of the major discoveries of the "renaissance" of light microscopy that started in the 1980s. The technique derives its advantages from the biologically "smooth" wavelength of the excitation light and the confinement of the excitation. Difficult, and seemingly nontransparent, samples may be imaged with the technique with good resolution. Although the bioresearch has been concentrating mostly on the positive properties of the technique for imaging, the same properties may be applied successfully to nonimaging bioassays. This article focuses on the development path of two-photon excitation-based assay system. PMID:18596366

  3. Fluorescent excitation of interstellar H2

    NASA Technical Reports Server (NTRS)

    Black, John H.; Van Dishoeck, Ewine F.

    1987-01-01

    The infrared emission spectrum of H2 excited by ultraviolet absorption, followed by fluorescence, was investigated using comprehensive models of interstellar clouds for computing the spectrum and to assess the effects on the intensity to various cloud properties, such as density, size, temperature, and the intensity of the UV radiation field. It is shown that the absolute H2 IR line intensities depend primarily on the density of the cloud and the strength of the incident UV radiation, and to a much lesser exent on the temperature of the gas, the total thickness of the cloud, and the optical properties of the grains. A variety of recent observational results are discussed with reference to theoretical models. It is shown that the rich H2 emission spectrum of the reflection nebula NGC 2023 can be reproduced by a model with density of about 10,000/cu cm, temperature of about 80 K, and UV flux approximately 300 times that of the Galactic background starlight.

  4. Multiphoton cascade absorption in single molecule fluorescence saturation spectroscopy.

    PubMed

    Winckler, Pascale; Jaffiol, Rodolphe

    2013-05-01

    Saturation spectroscopy is a relevant method to investigate photophysical parameters of single fluorescent molecules. Nevertheless, the impact of a gradual increase, over a broad range, of the laser excitation on the intramolecular dynamics is not completely understood, particularly concerning their fluorescence emission (the so-called brightness). Thus, we propose a comprehensive theoretical and experimental study to interpret the unexpected evolution of the brightness with the laser power taking into account the cascade absorption of two and three photons. Furthermore, we highlight the key role played by the confocal observation volume in fluorescence saturation spectroscopy of single molecules in solution.

  5. Multiphoton fluorescence excitation: new spectral windows for biological nonlinear microscopy.

    PubMed Central

    Xu, C; Zipfel, W; Shear, J B; Williams, R M; Webb, W W

    1996-01-01

    Intrinsic, three-dimensionally resolved, microscopic imaging of dynamical structures and biochemical processes in living preparations has been realized by nonlinear laser scanning fluorescence microscopy. The search for useful two-photon and three-photon excitation spectra, motivated by the emergence of nonlinear microscopy as a powerful biophysical instrument, has now discovered a virtual artist's palette of chemical indicators, fluorescent markers, and native biological fluorophores, including NADH, flavins, and green fluorescent proteins, that are applicable to living biological preparations. More than 25 two-photon excitation spectra of ultraviolet and visible absorbing molecules reveal useful cross sections, some conveniently blue-shifted, for near-infrared absorption. Measurements of three-photon fluorophore excitation spectra now define alternative windows at relatively benign wavelengths to excite deeper ultraviolet fluorophores. The inherent optical sectioning capability of nonlinear excitation provides three-dimensional resolution for imaging and avoids out-of-focus background and photodamage. Here, the measured nonlinear excitation spectra and their photophysical characteristics that empower nonlinear laser microscopy for biological imaging are described. Images Fig. 1 Fig. 5 Fig. 7 PMID:8855254

  6. Up-converted fluorescence from photosynthetic light-harvesting complexes linearly dependent on excitation intensity.

    PubMed

    Leiger, Kristjan; Freiberg, Arvi

    2016-01-01

    Weak up-converted fluorescence related to bacteriochlorophyll a was recorded from various detergent-isolated and membrane-embedded light-harvesting pigment-protein complexes as well as from the functional membranes of photosynthetic purple bacteria under continuous-wave infrared laser excitation at 1064 nm, far outside the optically allowed singlet absorption bands of the chromophore. The fluorescence increases linearly with the excitation power, distinguishing it from the previously observed two-photon excited fluorescence upon femtosecond pulse excitation. Possible mechanisms of this excitation are discussed.

  7. Spectroscopy of α,ω-dithienyl polyenes: evidence for two-photon absorption states from fluorescence studies

    NASA Astrophysics Data System (ADS)

    Natarajan, Lalgudi V.; Kirkpatrick, Sean M.; Sutherland, Richard L.; Sowards, Laura A.; Spangler, Charles W.; Fleitz, Paul A.; Cooper, Thomas M.

    1998-10-01

    The photophysics of a series (alpha) ,(omega) -dithienyl polyenes was studied. Both one photon excited fluorescence and fluorescence originating from two photon absorption states were investigated. There is strong evidence of two photon absorption from fluorescence studies in agreement with our earlier studies involving nonlinear absorption.

  8. Excitation-resolved fluorescence tomography with simplified spherical harmonics equations

    NASA Astrophysics Data System (ADS)

    Klose, Alexander D.; Pöschinger, Thomas

    2011-03-01

    Fluorescence tomography (FT) reconstructs the three-dimensional (3D) fluorescent reporter probe distribution inside biological tissue. These probes target molecules of biological function, e.g. cell surface receptors or enzymes, and emit fluorescence light upon illumination with an external light source. The fluorescence light is detected on the tissue surface and a source reconstruction algorithm based on the simplified spherical harmonics (SPN) equations calculates the unknown 3D probe distribution inside tissue. While current FT approaches require multiple external sources at a defined wavelength range, the proposed FT method uses only a white light source with tunable wavelength selection for fluorescence stimulation and further exploits the spectral dependence of tissue absorption for the purpose of 3D tomographic reconstruction. We will show the feasibility of the proposed hyperspectral excitation-resolved fluorescence tomography method with experimental data. In addition, we will demonstrate the performance and limitations of such a method under ideal and controlled conditions by means of a digital mouse model and synthetic measurement data. Moreover, we will address issues regarding the required amount of wavelength intervals for fluorescent source reconstruction. We will explore the impact of assumed spatially uniform and nonuniform optical parameter maps on the accuracy of the fluorescence source reconstruction. Last, we propose a spectral re-scaling method for overcoming the observed limitations in reconstructing accurate source distributions in optically non-uniform tissue when assuming only uniform optical property maps for the source reconstruction process.

  9. Two-photon-excited fluorescence spectroscopy of atomic fluorine at 170 nm

    NASA Technical Reports Server (NTRS)

    Herring, G. C.; Dyer, Mark J.; Jusinski, Leonard E.; Bischel, William K.

    1988-01-01

    Two-photon-excited fluorescence spectroscopy of atomic fluorine is reported. A doubled dye laser at 286-nm is Raman shifted in H2 to 170 nm (sixth anti-Stokes order) to excite ground-state 2P(0)J fluorine atoms to the 2D(0)J level. The fluorine atoms are detected by one of two methods: observing the fluorescence decay to the 2PJ level or observing F(+) production through the absorption of an additional photon by the excited atoms. Relative two-photon absorption cross sections to and the radiative lifetimes of the 2D(0)J states are measured.

  10. Excitation-emission spectra and fluorescence quantum yields for fresh and aged biogenic secondary organic aerosols

    SciTech Connect

    Lee, Hyun Ji; Laskin, Alexander; Laskin, Julia; Nizkorodov, Sergey A.

    2013-05-10

    Certain biogenic secondary organic aerosols (SOA) become absorbent and fluorescent when exposed to reduced nitrogen compounds such as ammonia, amines and their salts. Fluorescent SOA may potentially be mistaken for biological particles by detection methods relying on fluorescence. This work quantifies the spectral distribution and effective quantum yields of fluorescence of SOA generated from two monoterpenes, limonene and a-pinene, and two different oxidants, ozone (O3) and hydroxyl radical (OH). The SOA was generated in a smog chamber, collected on substrates, and aged by exposure to ~100 ppb ammonia vapor in air saturated with water vapor. Absorption and excitation-emission matrix (EEM) spectra of aqueous extracts of aged and control SOA samples were measured, and the effective absorption coefficients and fluorescence quantum yields (~0.005 for 349 nm excitation) were determined from the data. The strongest fluorescence for the limonene-derived SOA was observed for excitation = 420+- 50 nm and emission = 475 +- 38 nm. The window of the strongest fluorescence shifted to excitation = 320 +- 25 nm and emission = 425 +- 38 nm for the a-pinene-derived SOA. Both regions overlap with the excitation-emission matrix (EEM) spectra of some of the fluorophores found in primary biological aerosols. Our study suggests that, despite the low quantum yield, the aged SOA particles should have sufficient fluorescence intensities to interfere with the fluorescence detection of common bioaerosols.

  11. Visible-wavelength two-photon excitation microscopy for fluorescent protein imaging

    NASA Astrophysics Data System (ADS)

    Yamanaka, Masahito; Saito, Kenta; Smith, Nicholas I.; Arai, Yoshiyuki; Uegaki, Kumiko; Yonemaru, Yasuo; Mochizuki, Kentaro; Kawata, Satoshi; Nagai, Takeharu; Fujita, Katsumasa

    2015-10-01

    The simultaneous observation of multiple fluorescent proteins (FPs) by optical microscopy is revealing mechanisms by which proteins and organelles control a variety of cellular functions. Here we show the use of visible-light based two-photon excitation for simultaneously imaging multiple FPs. We demonstrated that multiple fluorescent targets can be concurrently excited by the absorption of two photons from the visible wavelength range and can be applied in multicolor fluorescence imaging. The technique also allows simultaneous single-photon excitation to offer simultaneous excitation of FPs across the entire range of visible wavelengths from a single excitation source. The calculation of point spread functions shows that the visible-wavelength two-photon excitation provides the fundamental improvement of spatial resolution compared to conventional confocal microscopy.

  12. Excitation-emission spectra and fluorescence quantum yields for fresh and aged biogenic secondary organic aerosols.

    PubMed

    Lee, Hyun Ji Julie; Laskin, Alexander; Laskin, Julia; Nizkorodov, Sergey A

    2013-06-01

    Certain biogenic secondary organic aerosols (SOA) become absorbent and fluorescent when exposed to reduced nitrogen compounds such as ammonia, amines, and their salts. Fluorescent SOA may potentially be mistaken for biological particles by detection methods relying on fluorescence. This work quantifies the spectral distribution and effective quantum yields of fluorescence of water-soluble SOA generated from two monoterpenes, limonene and α-pinene, and two different oxidants, ozone (O3) and hydroxyl radical (OH). The SOA was generated in a smog chamber, collected on substrates, and aged by exposure to ∼100 ppb ammonia in air saturated with water vapor. Absorption and excitation-emission matrix (EEM) spectra of aqueous extracts of aged and control SOA samples were measured, and the effective absorption coefficients and fluorescence quantum yields (∼0.005 for 349 nm excitation) were determined from the data. The strongest fluorescence for the limonene-derived SOA was observed for λexcitation = 420 ± 50 nm and λemission = 475 ± 38 nm. The window of the strongest fluorescence shifted to λexcitation = 320 ± 25 nm and λemission = 425 ± 38 nm for the α-pinene-derived SOA. Both regions overlap with the EEM spectra of some of the fluorophores found in primary biological aerosols. Despite the low quantum yield, the aged SOA particles may have sufficient fluorescence intensities to interfere with the fluorescence detection of common bioaerosols.

  13. Theoretical investigations of absorption and fluorescence spectra of protonated pyrene.

    PubMed

    Chin, Chih-Hao; Lin, Sheng Hsien

    2016-05-25

    The equilibrium geometry and 75 vibrational normal-mode frequencies of the ground and first excited states of protonated pyrene isomers were calculated and characterized in the adiabatic representation by using the complete active space self-consistent field (CASSCF) method. Electronic absorption spectra of solid neon matrixes in the wavelength range 495-415 nm were determined by Maier et al. and they were analyzed using time-dependent density functional theory calculations (TDDFT). CASSCF calculations and absorption and emission spectra simulations by one-photon excitation equations were used to optimize the excited and ground state structures of protonated pyrene isomers. The absorption band was attributed to the S0 → S1 electronic transition in 1H-Py(+), and a band origin was used at 20580.96 cm(-1). The displaced harmonic oscillator approximation and Franck-Condon approximation were used to simulate the absorption spectrum of the (1) (1)A' ← X[combining tilde](1)A' transition of 1H-Py(+), and the main vibronic transitions were assigned for the first ππ* state. It shows that the vibronic structures were dominated by one of the eight active totally symmetric modes, with ν15 being the most crucial. This indicates that the electronic transition of the S1((1)A') state calculated in the adiabatic representation effectively includes a contribution from the adiabatic vibronic coupling through Franck-Condon factors perturbed by harmonic oscillators. The present method can adequately reproduce experimental absorption and fluorescence spectra of a gas phase. PMID:27181017

  14. Absorption and fluorescent spectral studies of imidazophenazine derivatives.

    PubMed

    Ryazanova, O A; Zozulya, V N; Voloshin, I M; Karachevtsev, V A; Makitruk, V L; Stepanian, S G

    2004-07-01

    Absorption and fluorescent spectra as well as fluorescence polarization degree of imidazo-[4,5-d]-phenazine (F1) and its two modified derivatives, 2-trifluoridemethylimidazo-[4,5-d]-phenazine (F2) and 1,2,3-triazole-[4,5-d]-phenazine (F3), were investigated in organic solvents of various polarities and hydrogen bonding abilities. Extinction coefficients of F2 and F3 are increased, their fluorescence Stokes shifts are reduced in comparison with those for unmodified imidazophenazine. For F3 a red shift of the longwave absorption band is observed by 15-20 nm. Modifications of imidazophenazine have led to a sufficient increase of fluorescence polarization degrees that enables to use F2 and F3 as promising fluorescent probes with polarization method application. The configuration, atomic charge distribution and dipole moments of the isolated dye molecules in the ground state were calculated by the DFT method. The computation has revealed that ground state dipole moments of F1, F2, and F3 differ slightly and are equal to 3.5, 3.2, and 3.7D, respectively. The changes in dipole moments upon the optical excitation for all derivatives estimated using Lippert equation were found to be Deltamu = 9 D. The energies of the electronic S1<--S0 transition in solvents of different proton donor abilities were determined, and energetic diagram illustrating the substituent effect was plotted. For nucleoside analogs of these compounds, covalently incorporated into a nucleotide chain, we have considered a possibility to use them as fluorescent reporters of hybridization of antisense oligonucleotides, as well as molecular anchors for its stabilization. PMID:15248979

  15. Ultrafast excited-state dynamics and fluorescence deactivation of near-infrared fluorescent proteins engineered from bacteriophytochromes

    PubMed Central

    Zhu, Jingyi; Shcherbakova, Daria M.; Hontani, Yusaku; Verkhusha, Vladislav V.; Kennis, John T. M.

    2015-01-01

    Near-infrared fluorescent proteins, iRFPs, are recently developed genetically encoded fluorescent probes for deep-tissue in vivo imaging. Their functions depend on the corresponding fluorescence efficiencies and electronic excited state properties. Here we report the electronic excited state deactivation dynamics of the most red-shifted iRFPs: iRFP702, iRFP713 and iRFP720. Complementary measurements by ultrafast broadband fluorescence and absorption spectroscopy show that single exponential decays of the excited state with 600 ~ 700 ps dominate in all three iRFPs, while photoinduced isomerization was completely inhibited. Significant kinetic isotope effects (KIE) were observed with a factor of ~1.8 in D2O, and are interpreted in terms of an excited-state proton transfer (ESPT) process that deactivates the excited state in competition with fluorescence and chromophore mobility. On this basis, new approaches for rational molecular engineering may be applied to iRFPs to improve their fluorescence. PMID:26246319

  16. Ultrafast excited-state dynamics and fluorescence deactivation of near-infrared fluorescent proteins engineered from bacteriophytochromes

    NASA Astrophysics Data System (ADS)

    Zhu, Jingyi; Shcherbakova, Daria M.; Hontani, Yusaku; Verkhusha, Vladislav V.; Kennis, John T. M.

    2015-08-01

    Near-infrared fluorescent proteins, iRFPs, are recently developed genetically encoded fluorescent probes for deep-tissue in vivo imaging. Their functions depend on the corresponding fluorescence efficiencies and electronic excited state properties. Here we report the electronic excited state deactivation dynamics of the most red-shifted iRFPs: iRFP702, iRFP713 and iRFP720. Complementary measurements by ultrafast broadband fluorescence and absorption spectroscopy show that single exponential decays of the excited state with 600 ~ 700 ps dominate in all three iRFPs, while photoinduced isomerization was completely inhibited. Significant kinetic isotope effects (KIE) were observed with a factor of ~1.8 in D2O, and are interpreted in terms of an excited-state proton transfer (ESPT) process that deactivates the excited state in competition with fluorescence and chromophore mobility. On this basis, new approaches for rational molecular engineering may be applied to iRFPs to improve their fluorescence.

  17. Ultrafast excited-state dynamics and fluorescence deactivation of near-infrared fluorescent proteins engineered from bacteriophytochromes.

    PubMed

    Zhu, Jingyi; Shcherbakova, Daria M; Hontani, Yusaku; Verkhusha, Vladislav V; Kennis, John T M

    2015-08-06

    Near-infrared fluorescent proteins, iRFPs, are recently developed genetically encoded fluorescent probes for deep-tissue in vivo imaging. Their functions depend on the corresponding fluorescence efficiencies and electronic excited state properties. Here we report the electronic excited state deactivation dynamics of the most red-shifted iRFPs: iRFP702, iRFP713 and iRFP720. Complementary measurements by ultrafast broadband fluorescence and absorption spectroscopy show that single exponential decays of the excited state with 600~700 ps dominate in all three iRFPs, while photoinduced isomerization was completely inhibited. Significant kinetic isotope effects (KIE) were observed with a factor of ~1.8 in D2O, and are interpreted in terms of an excited-state proton transfer (ESPT) process that deactivates the excited state in competition with fluorescence and chromophore mobility. On this basis, new approaches for rational molecular engineering may be applied to iRFPs to improve their fluorescence.

  18. Optimal fluorescence waveband determination for detecting defect cherry tomatoes using fluorescence excitation-emission matrix

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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-...

  19. Implantable CMOS imaging device with absorption filters for green fluorescence imaging

    NASA Astrophysics Data System (ADS)

    Sunaga, Yoshinori; Haruta, Makito; Takehara, Hironari; Ohta, Yasumi; Motoyama, Mayumi; Noda, Toshihiko; Sasagawa, Kiyotaka; Tokuda, Takashi; Ohta, Jun

    2014-03-01

    Green fluorescent materials such as Green Fluorescence Protein (GFP) and fluorescein are often used for observing neural activities. Thus, it is important to observe the fluorescence in a freely moving state in order to understand neural activities corresponding to behaviors. In this work, we developed an implantable CMOS imaging device for in-vivo green fluorescence imaging with efficient excitation light rejection using a combination of absorption filters. An interference filter is usually used for a fluorescence microscope in order to achieve high fluorescence imaging sensitivity. However, in the case of the implantable device, interference filters are not suitable because their transmission spectra depend on incident angle. To solve this problem we used two kinds of absorption filters that do not have angle dependence. An absorption filter consisting of yellow dye (VARYFAST YELLOW 3150) was coated on the pixel array of an image sensor. The rejection ratio of ideal excitation light (490 nm) against green fluorescence (510 nm) was 99.66%. However, the blue LED as an excitation light source has a broad emission spectrum and its intensity at 510 nm is 2.2 x 10-2 times the emission peak intensity. By coating LEDs with the emission absorption filters, the intensity of the unwanted component of the excitation light was reduced to 1.4 x 10-4. Using the combination of absorption filters, we achieved excitation light transmittance of 10-5 onto the image sensor. It is expected that high-sensitivity green fluorescence imaging of neural activities in a freely moving mouse will be possible by using this technology.

  20. Multispectral excitation based multiple fluorescent targets resolving in fluorescence molecular tomography

    NASA Astrophysics Data System (ADS)

    Zhou, Yuan; Guang, Huizhi; Pu, Huangsheng; Zhang, Jiulou; Bai, Jing; Luo, Jianwen

    2016-04-01

    Fluorescence molecular tomography (FMT) can visualize biological activities at cellular and molecular levels in vivo, and has been extensively used in drug delivery and tumor detection research of small animals. The ill-posedness of the FMT inverse problem makes it difficult to reconstruct and resolve multiple adjacent fluorescent targets that have different functional features but are labeled with the same fluorochrome. An algorithm based on independent component analysis (ICA) for multispectral excited FMT is proposed to resolve multiple fluorescent targets in this study. Fluorescent targets are excited by multispectral excitation, and the three-dimensional distribution of fluorescent yields under the excitation spectrum is reconstructed by an iterative Tikhonov regularization algorithm. Subsequently, multiple fluorescent targets are resolved from mixed fluorescence signals by employing ICA. Simulations were performed and the results demonstrate that multiple adjacent fluorescent targets can be resolved if the number of excitation wavelengths is not smaller than that of fluorescent targets with different concentrations. The algorithm obtains both independent components that provide spatial information of different fluorescent targets and spectral courses that reflect variation trends of fluorescent yields along with the excitation spectrum. By using this method, it is possible to visualize the metabolism status of drugs in different structure organs, and quantitatively depict the variation trends of fluorescent yields of each functional organ under the excitation spectrum. This method may provide a pattern for tumor detection, drug delivery and treatment monitoring in vivo.

  1. Origin of Excitation Dependent Fluorescence in Carbon Nanodots.

    PubMed

    Sharma, Arjun; Gadly, Trilochan; Gupta, Alka; Ballal, Anand; Ghosh, Sunil Kumar; Kumbhakar, Manoj

    2016-09-15

    The fascinating aspect of excitation dependent fluorescence in carbon nanodots has led to several hypotheses, starting from particle size distribution to the presence of different emissive states and even to sluggish solvent relaxation around nanodot. In this contribution we provide definitive evidence for the involvement of discrete multiple electronic states for the excitation dependent emission in carbon nanodots. The presence of different types of aggregates even at very dilute solutions used in ensemble fluorescence spectroscopy, where fluorescence intensity shows linear dependence with absorbance, is the origin of these multiple electronic states. Inhomogeneous broadening due to slow solvent relaxation leading to excitation dependent spectral shift has negligible influence in conventional solvents. PMID:27588560

  2. A Parallel Reconstruction Scheme in Fluorescence Tomography Based on Contrast of Independent Inversed Absorption Properties

    PubMed Central

    Yi, Ji; Bai, Jing

    2006-01-01

    Based on an independent forward model in fluorescent tomography, a parallel reconstructed scheme for inhomogeneous mediums with unknown absorption property is proposed in this paper. The method considers the two diffusion equations as separately describing the propagation of excited light in tissues with and without fluorescent probes inside. Then the concentration of fluorophores is obtained directly through the difference between two estimations of absorption coefficient which can be parallel inversed. In this way, the multiparameter estimation problem in fluorescent tomography is transformed into two independent single-coefficient determined schemes of diffusion optical tomography (DOT). Any algorithms proved to be efficient and effective in DOT can be directly applied here. In this study the absorption property is estimated from the independent diffusion equations by a gradient-based optimization method with finite element method (FEM) solving the forward model. Simulation results of three representative occasions show that the reconstructed method can well estimate fluorescent property and tissue absorption distribution. PMID:23165045

  3. Saturated excitation of fluorescence to quantify excitation enhancement in aperture antennas.

    PubMed

    Aouani, Heykel; Hostein, Richard; Mahboub, Oussama; Devaux, Eloïse; Rigneault, Hervé; Ebbesen, Thomas W; Wenger, Jérôme

    2012-07-30

    Fluorescence spectroscopy is widely used to probe the electromagnetic intensity amplification on optical antennas, yet measuring the excitation intensity amplification is a challenge, as the detected fluorescence signal is an intricate combination of excitation and emission. Here, we describe a novel approach to quantify the electromagnetic amplification in aperture antennas by taking advantage of the intrinsic non linear properties of the fluorescence process. Experimental measurements of the fundamental f and second harmonic 2f amplitudes of the fluorescence signal upon excitation modulation are used to quantify the electromagnetic intensity amplification with plasmonic aperture antennas.

  4. The two-photon excitation cross section of 6MAP, a fluorescent adenine analogue.

    PubMed

    Stanley, Robert J; Hou, Zhanjia; Yang, Aiping; Hawkins, Mary E

    2005-03-01

    6MAP is a fluorescent analogue of adenine that undergoes Watson-Crick base pairing and base stacking in double-stranded DNA. The one-photon absorption spectrum of 6MAP is characterized by a maximum around 330 nm with moderate quantum yield fluorescence centered at about 420 nm. To take advantage of this probe for confocal and single-molecule microscopy, it would be advantageous to be able to excite the analogue via two photons. We report the first determination of the two-photon excitation cross section and spectrum for 6MAP from 614 to 700 nm. The power dependence of the fluorescence indicates that emission results from the absorption of two photons. The one-photon and two-photon emission line shapes are identical within experimental error. A study of the concentration dependence of the fluorescence yield for one-photon excitation shows no measurable quenching up to about 5 microM. The maximum in the two-photon excitation spectrum gives a two-photon cross section, delta(TPE), of 3.4 +/- 0.1 Goeppert-Mayer (G.M.) at 659 nm, which correlates well with the one-photon absorption maximum. This compares quite favorably with cross sections of various naturally fluorescent biological molecules such as flavins and nicotiamide. In addition, we have also obtained the two-photon-induced fluorescence emission spectrum of quinine sulfate. It is approximately the same as that for one-photon excitation, suggesting that two-photon excitation of quinine sulfate may be used for calibration purposes.

  5. The two-photon excitation cross section of 6MAP, a fluorescent adenine analogue.

    PubMed

    Stanley, Robert J; Hou, Zhanjia; Yang, Aiping; Hawkins, Mary E

    2005-03-01

    6MAP is a fluorescent analogue of adenine that undergoes Watson-Crick base pairing and base stacking in double-stranded DNA. The one-photon absorption spectrum of 6MAP is characterized by a maximum around 330 nm with moderate quantum yield fluorescence centered at about 420 nm. To take advantage of this probe for confocal and single-molecule microscopy, it would be advantageous to be able to excite the analogue via two photons. We report the first determination of the two-photon excitation cross section and spectrum for 6MAP from 614 to 700 nm. The power dependence of the fluorescence indicates that emission results from the absorption of two photons. The one-photon and two-photon emission line shapes are identical within experimental error. A study of the concentration dependence of the fluorescence yield for one-photon excitation shows no measurable quenching up to about 5 microM. The maximum in the two-photon excitation spectrum gives a two-photon cross section, delta(TPE), of 3.4 +/- 0.1 Goeppert-Mayer (G.M.) at 659 nm, which correlates well with the one-photon absorption maximum. This compares quite favorably with cross sections of various naturally fluorescent biological molecules such as flavins and nicotiamide. In addition, we have also obtained the two-photon-induced fluorescence emission spectrum of quinine sulfate. It is approximately the same as that for one-photon excitation, suggesting that two-photon excitation of quinine sulfate may be used for calibration purposes. PMID:16851408

  6. Fluorescence Emission and Excitation Spectra of Photo-Fragmented Nitrobenzene.

    NASA Astrophysics Data System (ADS)

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

    2012-06-01

    Upon absorption of a UV photon, nitrobenzene readily dissociates into C_6H_5, NO_2, C_6H_5NO, O, C_6H_5O, and NO through three different channels. We have recorded high resolution emission and excitation spectra of the NO resulting from photo-fragmented nitrobenzene using a pulsed picosecond tunable laser and a nanosecond dye laser. Specifically, the lasers probed the A^2Σ^+→ X^2π(1/2,3/2) NO band system between 225-260 nm using an one or two color process. In a one color process, the same energy (wavelength) photon is used to dissociate nitrobenzene and excite NO. In a two color process, photons of a particular energy are used to dissociate the nitrobenzene while photons of a different energy are used to probe the resultant NO. We have determined the rotational and vibrational temperatures of the nascent NO. And, we have examined the effect of the relative timing of the two photons on the fluorescence spectra to extract information about the photodissociation dynamics. Lin, M.-F.; Lee, Y. T.; Ni, C.-K.; Xu, S. and Lin, M. C. J. Chem. Phys., AIP, 2007, 126.

  7. Two-photon excited fluorescence emission from hemoglobin

    NASA Astrophysics Data System (ADS)

    Sun, Qiqi; Zeng, Yan; Zhang, Wei; Zheng, Wei; Luo, Yi; Qu, Jianan Y.

    2015-03-01

    Hemoglobin, one of the most important proteins in blood, is responsible for oxygen transportation in almost all vertebrates. Recently, we discovered two-photon excited hemoglobin fluorescence and achieved label-free microvascular imaging based on the hemoglobin fluorescence. However, the mechanism of its fluorescence emission still remains unknown. In this work, we studied the two-photon excited fluorescence properties of the hemoglobin subunits, heme/hemin (iron (II)/(III) protoporphyrin IX) and globin. We first studied the properties of heme and the similar spectral and temporal characteristics of heme and hemoglobin fluorescence provide strong evidence that heme is the fluorophore in hemoglobin. Then we studied the fluorescence properties of hemin, globin and methemoglobin, and found that the hemin may have the main effect on the methemoglobin fluorescence and that globin has tryptophan fluorescence like other proteins. Finally, since heme is a centrosymmetric molecule, that the Soret band fluorescence of heme and hemoglobin was not observed in the single photon process in the previous study may be due to the parity selection rule. The discovery of heme two-photon excited fluorescence may open a new window for heme biology research, since heme as a cofactor of hemoprotein has many functions, including chemical catalysis, electron transfer and diatomic gases transportation.

  8. Theoretical study of absorption and fluorescence spectra of firefly luciferin in aqueous solutions.

    PubMed

    Hiyama, Miyabi; Akiyama, Hidefumi; Yamada, Kenta; Koga, Nobuaki

    2012-01-01

    The absorption and fluorescence spectra of firefly luciferin, which is an analog of oxyluciferin, are investigated by performing the density functional theory (DFT) calculations, especially focusing on the experimentally unassigned peaks. Time-dependent DFT calculations are performed for the excited states of firefly luciferin and its conjugate acids and bases. We find that (1) the peaks in the experimental absorption spectra correspond to the excited states of not only (6'O(-), 4COO(-)) and (6'OH, 4COO(-)), but also (6'OH, 4COOH) and (6'OH, 3H(+), 4COOH); (2) the peaks in the experimental fluorescence spectra correspond to the excited states of not only (6'O(-), 4COO(-)), but also (6'OH, 4COO(-)), (6'O(-), 4COOH), (6'OH, 4COOH) and (6'OH, 3H(+), 4COOH); (3) the unassigned peak near 400 nm in the experimental absorption spectra at pH 1 is assigned to the absorption from the equilibrium ground state to the first excited state of (6'OH, 3H(+), 4COOH); and (4) the unassigned peak at 610 nm in the experimental fluorescence spectra corresponds to the transition from the equilibrium first excited state to the ground state of (6'OH, 4COO(-)). PMID:22364397

  9. Optimal fluorescence waveband determination for detecting defective cherry tomatoes using a fluorescence excitation-emission matrix.

    PubMed

    Baek, In-Suck; Kim, Moon S; Lee, Hoosoo; Lee, Wang-Hee; Cho, Byoung-Kwan

    2014-11-14

    A multi-spectral fluorescence imaging technique was used to detect defective 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-way ANOVA revealed the optimal excitation wavelength for detecting defect areas was 410 nm. Principal component analysis (PCA) was applied to the fluorescence emission spectra of all regions at 410 nm excitation to determine the emission wavelengths for defect detection. The major emission wavelengths were 688 nm and 506 nm for the detection. Fluorescence images combined with the determined emission wavebands demonstrated the feasibility of detecting defective cherry tomatoes with >98% accuracy. Multi-spectral fluorescence imaging has potential utility in non-destructive quality sorting of cherry tomatoes.

  10. Optimal fluorescence waveband determination for detecting defective cherry tomatoes using a fluorescence excitation-emission matrix.

    PubMed

    Baek, In-Suck; Kim, Moon S; Lee, Hoosoo; Lee, Wang-Hee; Cho, Byoung-Kwan

    2014-01-01

    A multi-spectral fluorescence imaging technique was used to detect defective 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-way ANOVA revealed the optimal excitation wavelength for detecting defect areas was 410 nm. Principal component analysis (PCA) was applied to the fluorescence emission spectra of all regions at 410 nm excitation to determine the emission wavelengths for defect detection. The major emission wavelengths were 688 nm and 506 nm for the detection. Fluorescence images combined with the determined emission wavebands demonstrated the feasibility of detecting defective cherry tomatoes with >98% accuracy. Multi-spectral fluorescence imaging has potential utility in non-destructive quality sorting of cherry tomatoes. PMID:25405507

  11. Optimal Fluorescence Waveband Determination for Detecting Defective Cherry Tomatoes Using a Fluorescence Excitation-Emission Matrix

    PubMed Central

    Baek, In-Suck; Kim, Moon S.; Lee, Hoosoo; Lee, Wang-Hee; Cho, Byoung-Kwan

    2014-01-01

    A multi-spectral fluorescence imaging technique was used to detect defective 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-way ANOVA revealed the optimal excitation wavelength for detecting defect areas was 410 nm. Principal component analysis (PCA) was applied to the fluorescence emission spectra of all regions at 410 nm excitation to determine the emission wavelengths for defect detection. The major emission wavelengths were 688 nm and 506 nm for the detection. Fluorescence images combined with the determined emission wavebands demonstrated the feasibility of detecting defective cherry tomatoes with >98% accuracy. Multi-spectral fluorescence imaging has potential utility in non-destructive quality sorting of cherry tomatoes. PMID:25405507

  12. Fluorescence excitation-emission matrix spectroscopy of vitiligo skin in vivo (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Zhao, Jianhua; Richer, Vincent; Al Jasser, Mohammed; Zandi, Soodabeh; Kollias, Nikiforos; Kalia, Sunil; Zeng, Haishan; Lui, Harvey

    2016-02-01

    Fluorescence signals depend on the intensity of the exciting light, the absorption properties of the constituent molecules, and the efficiency with which the absorbed photons are converted to fluorescence emission. The optical features and appearance of vitiligo have been explained primarily on the basis of reduced epidermal pigmentation, which results in abnormal white patches on the skin. The objective of this study is to explore the fluorescence properties of vitiligo and its adjacent normal skin using fluorescence excitation-emission matrix (EEM) spectroscopy. Thirty five (35) volunteers with vitiligo were acquired using a double-grating spectrofluorometer with excitation and emission wavelengths of 260-450 nm and 300-700 nm respectively. As expected, the most pronounced difference between the spectra obtained from vitiligo lesions compared to normally pigmented skin was that the overall fluorescence was much higher in vitiligo; these differences increased at shorter wavelengths, thus matching the characteristic spectral absorption of epidermal melanin. When comparing the fluorescence spectra from vitiligo to normal skin we detected three distinct spectral bands centered at 280nm, 310nm, and 335nm. The 280nm band may possibly be related to inflammation, whereas the 335 nm band may arise from collagen or keratin cross links. The source of the 310 nm band is uncertain; it is interesting to note its proximity to the 311 nm UV lamps used for vitiligo phototherapy. These differences are accounted for not only by changes in epidermal pigment content, but also by other optically active cutaneous biomolecules.

  13. Visualization of molecular fluorescence point spread functions via remote excitation switching fluorescence microscopy

    PubMed Central

    Su, Liang; Lu, Gang; Kenens, Bart; Rocha, Susana; Fron, Eduard; Yuan, Haifeng; Chen, Chang; Van Dorpe, Pol; Roeffaers, Maarten B. J.; Mizuno, Hideaki; Hofkens, Johan; Hutchison, James A.; Uji-i, Hiroshi

    2015-01-01

    The enhancement of molecular absorption, emission and scattering processes by coupling to surface plasmon polaritons on metallic nanoparticles is a key issue in plasmonics for applications in (bio)chemical sensing, light harvesting and photocatalysis. Nevertheless, the point spread functions for single-molecule emission near metallic nanoparticles remain difficult to characterize due to fluorophore photodegradation, background emission and scattering from the plasmonic structure. Here we overcome this problem by exciting fluorophores remotely using plasmons propagating along metallic nanowires. The experiments reveal a complex array of single-molecule fluorescence point spread functions that depend not only on nanowire dimensions but also on the position and orientation of the molecular transition dipole. This work has consequences for both single-molecule regime-sensing and super-resolution imaging involving metallic nanoparticles and opens the possibilities for fast size sorting of metallic nanoparticles, and for predicting molecular orientation and binding position on metallic nanoparticles via far-field optical imaging. PMID:25687887

  14. Visualization of molecular fluorescence point spread functions via remote excitation switching fluorescence microscopy.

    PubMed

    Su, Liang; Lu, Gang; Kenens, Bart; Rocha, Susana; Fron, Eduard; Yuan, Haifeng; Chen, Chang; Van Dorpe, Pol; Roeffaers, Maarten B J; Mizuno, Hideaki; Hofkens, Johan; Hutchison, James A; Uji-I, Hiroshi

    2015-01-01

    The enhancement of molecular absorption, emission and scattering processes by coupling to surface plasmon polaritons on metallic nanoparticles is a key issue in plasmonics for applications in (bio)chemical sensing, light harvesting and photocatalysis. Nevertheless, the point spread functions for single-molecule emission near metallic nanoparticles remain difficult to characterize due to fluorophore photodegradation, background emission and scattering from the plasmonic structure. Here we overcome this problem by exciting fluorophores remotely using plasmons propagating along metallic nanowires. The experiments reveal a complex array of single-molecule fluorescence point spread functions that depend not only on nanowire dimensions but also on the position and orientation of the molecular transition dipole. This work has consequences for both single-molecule regime-sensing and super-resolution imaging involving metallic nanoparticles and opens the possibilities for fast size sorting of metallic nanoparticles, and for predicting molecular orientation and binding position on metallic nanoparticles via far-field optical imaging. PMID:25687887

  15. Evanescent optical fluorescence excitation: the role of photonic mode density

    NASA Astrophysics Data System (ADS)

    Menges, Bernhard; Sato, Akihiro; Knoll, Wolfgang

    2007-04-01

    In this paper we will discuss a few basic concepts concerning the use of evanescent optical fields for the excitation of fluorescent chromophores placed near the interface. The observation of enhanced fluorescence from chromophores excited by surface plasmon and waveguide modes will be presented and discussed. We attribute the enhancement to the near-field interaction between the chromophores and the increased photonic mode density by surface plasmon and waveguide modes. We determined limits of detection (LOD) of DNA hybridisation using the same sensor architecture by surface plasmon fluorescence spectroscopy (SPFS) and optical waveguide fluorescence spectroscopy (OWFS). Both SPFS and OWFS techniques have the same detection principle using an enhanced electromagnetic field to excite fluorophores and make it possible to monitor DNA hybridisation in real-time with high sensitivity. The relative photonic mode density of each mode was calculated under the resonance condition, and these values are reflected in the LOD values.

  16. Enhancement of resonant absorption through excitation of SPR

    NASA Astrophysics Data System (ADS)

    Giulietti, Danilo; Calcagno, L.; Curcio, Alessandro; Cutroneo, M.; Galletti, Mario; Skala, J.; Torrisi, L.; Zimbone, M.

    2016-09-01

    In this experiment the absorption of the laser radiation impinging on polymeric films with Au nanoparticles implanted in surface was studied. By varying the polarization and the incidence angle of the laser radiation on target, the role in the laser absorption of both excitation of surface plasmons and excitation of electronic plasma waves at critical density through resonant absorption was highlighted. In conditions of p-polarized laser irradiations at 1015 W /cm2 intensity, resonant absorption can be induced in films enhancing proton and ion acceleration. Plasma on-line diagnostics is based on SiC detectors. Measurements of kinetic energy of accelerated ions indicate a significant increment using p-polarized laser light with respect to no-polarized light irradiation.

  17. Development of FRET-based dual-excitation ratiometric fluorescent pH probes and their photocaged derivatives.

    PubMed

    Yuan, Lin; Lin, Weiying; Cao, Zengmei; Wang, Jiaoliang; Chen, Bin

    2012-01-23

    Dual-excitation ratiometric fluorescent probes allow the measurement of fluorescence intensities at two excitation wavelengths, which should provide a built-in correction for environmental effects. However, most of the small-molecule dual-excitation ratiometric probes that have been reported thus far have shown rather limited separation between the excitation wavelengths (20-70 nm) and/or a very small molar absorption coefficient at one of the excitation wavelengths. These shortcomings can lead to cross-excitation and thus to errors in the measurement of fluorescence intensities and ratios. Herein, we report a FRET-based molecular strategy for the construction of small-molecule dual-excitation ratiometric probes in which the donor and acceptor excitation bands exhibit large separations between the excitation wavelengths and comparable excitation intensities, which is highly desirable for determining the fluorescence intensities and signal ratios with high accuracy. Based on this strategy, we created a coumarin-rhodamine FRET platform that was then employed to develop the first class of FRET-based dual-excitation ratiometric pH probes that have two well-resolved excitation bands (excitation separations>160 nm) and comparable excitation intensities. In addition, these pH probes may be considered as in a kind of "secured ratioing mode". As a further application of these pH probes, the dual-excitation ratiometric pH probes were transformed into the first examples of photocaged dual-excitation ratiometric pH probes to improve the spatiotemporal resolution. It is expected that the modular nature of our FRET-based molecular strategy should render it applicable to other small-molecule dual-dye energy-transfer systems based on diverse fluorescent dyes for the development of a wide range of dual-excitation ratiometric probes with outstanding spectral features, including large separations between the excitation wavelengths and comparable excitation intensities.

  18. Spectral signatures of fluorescence and light absorption to identify crude oils found in the marine environment

    NASA Astrophysics Data System (ADS)

    Baszanowska, E.; Otremba, Z.

    2014-08-01

    To protect the natural marine ecosystem, it is necessary to continuously enhance knowledge of environmental contamination, including oil pollution. Therefore, to properly track the qualitative and quantitative changes in the natural components of seawater, a description of the essential spectral features describing petroleum products is necessary. This study characterises two optically-different types of crude oils (Petrobaltic and Romashkino) - substances belonging to multi-fluorophoric systems. To obtain the spectral features of crude oils, the excitation-emission spectroscopy technique was applied. The fluorescence and light absorption properties for various concentrations of oils at a stabilised temperature are described. Both excitation-emission spectra (EEMs) and absorption spectra of crude oils are discussed. Based on the EEM spectra, both excitation end emission peaks for the wavelengthindependent fluorescence maximum (Exmax/ Emmax) - characteristic points for each type of oil - were identified and compared with the literature data concerning typical marine chemical structures.

  19. Stepwise two-photon excited fluorescence from higher excited states of chlorophylls in photosynthetic antenna complexes.

    PubMed

    Leupold, Dieter; Teuchner, Klaus; Ehlert, Jürgen; Irrgang, Klaus-Dieter; Renger, Gernot; Lokstein, Heiko

    2006-09-01

    Stepwise two-photon excited fluorescence (TPEF) spectra of the photosynthetic antenna complexes PCP, CP47, CP29, and light-harvesting complex II (LHC II) were measured. TPEF emitted from higher excited states of chlorophyll (Chl) a and b was elicited via consecutive absorption of two photons in the Chl a/b Qy range induced by tunable 100-fs laser pulses. Global analyses of the TPEF line shapes with a model function for monomeric Chl a in a proteinaceous environment allow distinction between contributions from monomeric Chls a and b, strongly excitonically coupled Chls a, and Chl a/b heterodimers/-oligomers. The analyses indicate that the longest wavelength-absorbing Chl species in the Qy region of LHC II is a Chl a homodimer with additional contributions from adjacent Chl b. Likewise, in CP47 a spectral form at approximately 680 nm (that is, however, not the red-most species) is also due to strongly coupled Chls a. In contrast to LHC II, the red-most Chl subband of CP29 is due to a monomeric Chl a. The two Chls b in CP29 exhibit marked differences: a Chl b absorbing at approximately 650 nm is not excitonically coupled to other Chls. Based on this finding, the refractive index of its microenvironment can be determined to be 1.48. The second Chl b in CP29 (absorbing at approximately 640 nm) is strongly coupled to Chl a. Implications of the findings with respect to excitation energy transfer pathways and rates are discussed. Moreover, the results will be related to most recent structural analyses.

  20. Investigation of two-photon absorption properties in new A-D-A compounds emitting blue and yellow fluorescence

    NASA Astrophysics Data System (ADS)

    Jin, Fan; Cai, Zhi-Bin; Huang, Jiu-Qiang; Li, Sheng-Li; Tian, Yu-Peng

    2015-08-01

    Three new acceptor-donor-acceptor compounds (LBQ, DBQ, BYQ) were synthesized and characterized by infrared, hydrogen nuclear magnetic resonance, mass spectrometry and elemental analysis. Their photophysical properties were investigated including linear absorption, single-photon excited fluorescence, fluorescence quantum yield and two-photon absorption. These compounds in CH2Cl2 exhibit good fluorescence quantum yield which are 0.36, 0.26, and 0.25 and the largest two-photon absorption cross-section which are 48, 36, and 181 GM respectively. Under the excitation of Ti: sapphire laser with a pulse width of 140 fs, LBQ and DBQ emit blue two-photon excited fluorescence (TPEF), while BYQ emits bright yellow TPEF. BYQ has a good solubility in water and the σ can be as large as 130 GM, so it shows promising applications in many pharmaceutical and biological fields.

  1. Suppression of excited-state absorption in laser crystals

    NASA Astrophysics Data System (ADS)

    Kuznetsova, Elena; Kolesov, Roman; Kocharovskaya, Olga

    2004-10-01

    Currently, a lot of experimental effort in solid-state optics is devoted to searching for laser materials suitable for tunable lasing, primarily in UV and VUV spectral regions. Researchers mainly focus on optical crystals doped with either transition metal or rare-earth ions. The latter ones doped into wide bandgap dielectric crystals have spectrally broad vibronic emission bands associated with 4fn-15d â" 4fn interconfigurational transitions, whose energies lie mostly in UV and VUV regions of the spectrum. The transitions are electric-dipole-allowed, therefore have large absorption and emission cross-sections, and are promising for efficient tunable laser action. However, in almost all promising crystals laser action in UV and VUV is hindered or completely prohibited due to excited-state absorption (ESA), i.e. absorption from metastable laser levels to higher-energy states, which occurs at emission or/and pump wavelengths. A method of suppression of losses due to excited-state absorption (ESA) in laser crystals is proposed, based on a well-known phenomenon of electromagnetically induced transparency (EIT). Absorption from a populated excited electronic state can be reduced under the action of an additional driving coherent field, resonantly coupling the terminal state of ESA to some intermediate discrete state.

  2. Neuron absorption study and mid-IR optical excitations

    NASA Astrophysics Data System (ADS)

    Guo, Dingkai; Chen, Xing; Vadala, Shilpa; Leach, Jennie; Kostov, Yordan; Bewley, William W.; Kim, Chul-Soo; Kim, Mijin; Canedy, Chadwick L.; Merritt, Charles D.; Vurgaftman, Igor; Meyer, Jerry R.; Choa, Fow-Sen

    2012-02-01

    Neuronal optical excitation can provide non-contacting tools to explore brain circuitry and a durable stimulation interface for cardiac pacing and visual as well as auditory sensory neuronal stimulation. To obtain accurate absorption spectra, we scan the transmission of neurons in cell culture medium, and normalize it by subtracting out the absorption spectrum of the medium alone. The resulting spectra show that the main neuronal absorption peaks are in the 3000- 6000nm band, although there is a smaller peak near 1450nm. By coupling the output of a 3μm interband cascade laser (ICL) into a mid-IR fluorozirconate fiber, we can effectively deliver more than 1J/cm2 photon intensity to the excitation site for neuronal stimulation.

  3. Transient absorption spectroscopy detection of sensitized delayed fluorescence in chiral benzophenone/naphthalene systems

    NASA Astrophysics Data System (ADS)

    Bonancía, Paula; Jiménez, M. Consuelo; Miranda, Miguel A.

    2011-10-01

    Transient absorption spectroscopy has proven to be a powerful tool to investigate the formation and decay of excited singlet states upon triplet-triplet annihilation, following T-T energy transfer from a selectively excited sensitizer. Thus, upon selective excitation of benzophenone (BZP) by laser flash photolysis (LFP) at λ = 355 nm in the presence of naphthalene (NPT), a negative band centered at 340 nm has been detected, with growth and decay in the microsecond timescale. It has been assigned to the P-type NPT delayed-fluorescence. In the case of chiral BZP/NPT systems, stereodifferentiation has been observed in the kinetics of the involved photophysical processes.

  4. Photon statistics of atomic fluorescence after {pi}-pulse excitation

    SciTech Connect

    Yoshimi, Kazuyoshi; Koshino, Kazuki

    2010-09-15

    The photon statistics of atomic fluorescence after {pi}-pulse excitation is investigated in a system in which the input and output ports are connected to an atom. Since spontaneous decay during input pulse excitation occurs, the output pulse generally contains a multiphoton component with a certain probability. We quantitatively evaluate the probability of the output pulse containing multiple photons and determine the conditions for ideal single-photon generation.

  5. Vibronic Structures in Absorption and Fluorescence Spectra of Firefly Oxyluciferin in Aqueous Solutions.

    PubMed

    Hiyama, Miyabi; Noguchi, Yoshifumi; Akiyama, Hidefumi; Yamada, Kenta; Koga, Nobuaki

    2015-01-01

    To elucidate the factors determining the spectral shapes and widths of the absorption and fluorescence spectra for keto and enol oxyluciferin and their conjugate bases in aqueous solutions, the intensities of vibronic transitions between their ground and first electronic excited states were calculated for the first time via estimation of the vibrational Franck-Condon factors. The major normal modes, overtones and combination tones in absorption and fluorescence spectra are similar for all species. The theoretical full widths at half maximum of absorption spectra are 0.4-0.7 eV and those for the fluorescence spectra are 0.4-0.5 eV, except for phenolate-keto that exhibits exceptionally sharp peak widths due to the dominance of the 0-0' or 0'-0 band. These spectral shapes and widths explain many relevant features of the experimentally observed spectra. PMID:25946599

  6. Interference of fluorescence x-rays and coherent excitation of core levels

    SciTech Connect

    Ma, Y. |; Blume, M.

    1994-08-01

    The question of coherence in inelastic x-ray absorption and fluorescence processes among identical interacting atoms is studied using a simple diatomic model. Conditions for the coherence are discussed in terms of energy scales, such as the core hole life-time, instrument energy resolutions, and the splitting of the electronic levels. As in the classical Young double-slit experiment, the primary requirement is that it be impossible to determine which atom has undergone the excitation-decay process.

  7. Fluorescence spectroscopy of excitation transfer in Photosystem 1

    SciTech Connect

    Mukerji, I.

    1990-12-01

    This thesis centers on the study of excitation transfer in a photosynthetic antenna array. The spectroscopic properties of two pigment-protein complexes were investigated. These complexes, isolated from higher plants, display an unusual temperature dependent fluorescence behavior. The author have chosen to study this fluorescence behavior with respect to energy transfer to the reaction center and in an isolated intact antenna preparation. A Photosystem 1 complex, PSI-200, was isolated from spinach. We have characterized this system by both steady state and time-resolved fluorescence spectroscopy. Fluorescence polarization measurements indicate that this emission arises from pigments which absorb in the long wavelength region of the spectrum and comprise a relatively small portion of the antenna population. Comparison of spectral characteristics were made with a PSI complex isolated from the thermophilic cyanobacterium, Synechococcus, sp. To address the role of Chl b in stimulating long wavelength fluorescence and the temperature dependence of the system, we have studied the energy transfer dynamics in an antenna complex, LHC-I isolated from PSI-200. Kinetic measurements indicate that initially absorbed excitation is rapidly redistributed to longer wavelength emitting pigments within 40 ps. The temperature dependence of F685 results from increased back transfer from long wavelength emitters to F685. We suggest that changes in excitation transfer between the various emitting species and a non-radiative fluorescence quenching mechanism account for the temperature dependence of the system. 144 refs., 50 figs., 3 tabs.

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

    NASA Technical Reports Server (NTRS)

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

    1981-01-01

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

  9. Fourier transform two-dimensional fluorescence excitation spectrometer by using tandem Fabry-Pérot interferometer

    NASA Astrophysics Data System (ADS)

    Anzai, Hiroshi; Joshi, Neeraj Kumar; Fuyuki, Masanori; Wada, Akihide

    2015-01-01

    A Fourier transform two-dimensional fluorescence excitation spectrometer (FT-2DFES) was developed based on the multiplex technique using a tandem Fabry-Pérot interferometer (tandem FPI). In addition to the advantage of the multiplex technique, the main advantage of the tandem FPI is applicable to the modulation of transition with a large absorption bandwidth (larger than 100 nm) and is thus applicable to the modulation of the excitation of molecules in the condensed phase. As a demonstration of the effectiveness of FT-2DFES, we succeeded in separately observing the fluorescence excitation peaks from a mixed methanol solution of laser dyes (coumarin 480, rhodamine 6G, DCM (4-dicyanomethylene-2-methyl-6-(p-(dimethylamino)styryl)-4H-pyran), and LDS750). Furthermore, the energy transfer from rhodamine 6G to LDS750 was observed.

  10. Fourier transform two-dimensional fluorescence excitation spectrometer by using tandem Fabry-Pérot interferometer.

    PubMed

    Anzai, Hiroshi; Joshi, Neeraj Kumar; Fuyuki, Masanori; Wada, Akihide

    2015-01-01

    A Fourier transform two-dimensional fluorescence excitation spectrometer (FT-2DFES) was developed based on the multiplex technique using a tandem Fabry-Pérot interferometer (tandem FPI). In addition to the advantage of the multiplex technique, the main advantage of the tandem FPI is applicable to the modulation of transition with a large absorption bandwidth (larger than 100 nm) and is thus applicable to the modulation of the excitation of molecules in the condensed phase. As a demonstration of the effectiveness of FT-2DFES, we succeeded in separately observing the fluorescence excitation peaks from a mixed methanol solution of laser dyes (coumarin 480, rhodamine 6G, DCM (4-dicyanomethylene-2-methyl-6-(p-(dimethylamino)styryl)-4H-pyran), and LDS750). Furthermore, the energy transfer from rhodamine 6G to LDS750 was observed.

  11. Integrated ultrasonic particle positioning and low excitation light fluorescence imaging

    SciTech Connect

    Bernassau, A. L.; Al-Rawhani, M.; Beeley, J.; Cumming, D. R. S.

    2013-12-09

    A compact hybrid system has been developed to position and detect fluorescent micro-particles by combining a Single Photon Avalanche Diode (SPAD) imager with an acoustic manipulator. The detector comprises a SPAD array, light-emitting diode (LED), lenses, and optical filters. The acoustic device is formed of multiple transducers surrounding an octagonal cavity. By stimulating pairs of transducers simultaneously, an acoustic landscape is created causing fluorescent micro-particles to agglomerate into lines. The fluorescent pattern is excited by a low power LED and detected by the SPAD imager. Our technique combines particle manipulation and visualization in a compact, low power, portable setup.

  12. Excilamps: Promising sources for exciting spontaneous fluorescence of organic molecules

    NASA Astrophysics Data System (ADS)

    Krekov, G. M.; Lisenko, A. A.; Matvienko, G. G.; Sosnin, É. V.

    2009-11-01

    We present the results of real and numerical experiments devoted to investigation of the spatial gradient of the intensity of chlorophyll fluorescence in the volume of a plant leaf. It is demonstrated for the first time that potential effective sources capable of exciting spontaneous fluorescence from the photosystem of a leaf are the exciplex lamps (exilamps) that generate continuous UV radiation. Estimations using the Monte Carlo method show that the main factor responsible for a transformation of the fluorescence spectrum during the passage of radiation through the volume of the leaf is the reabsorption process.

  13. Producing a dual-fluorescent molecule by tuning the energetics of excited-state intramolecular proton transfer.

    PubMed

    Tasaki, Satomi; Momotake, Atsuya; Kanna, Yoko; Sato, Tomoo; Nishimura, Yoshinobu; Arai, Tatsuo

    2015-10-01

    We report herein the selective preparation of normal, tautomeric, and dual-fluorescent molecules with a common ESIPT core. 2'-Hydroxyacetophenone (OHAP) is known as a typical molecule that undergoes excited-state intramolecular hydrogen transfer (ESIPT) to display fluorescence emission from the excited state of the tautomer. In this study, a series of ten OHAP-cored fluorescent molecules were prepared and their excited state properties have been explored. The bathochromic shift of the π-π* absorption band with π-extensions of substituents of these molecules indicates that the excitation energy of the normal form of the OHAP unit was reduced due to the substituents, while the energy of the excited tautomer appeared to be independent of the π-extension of the substituents. When pyrene or anthracene was connected at the end (molecules 4 and 5), only normal fluorescence appeared, and the tautomer fluorescence disappeared. An anthracene derivative (molecule 10) displayed dual fluorescence, indicating that the normal and the tautomer excited states were energetically "balanced". A fluorescence lifetime analysis revealed the ESIPT reaction rate of 10 to be much slower than those of other derivatives and that the normal and tautomer forms were in equilibrium in the excited state.

  14. Synthesis and blue-violet two-photon excited fluorescence of a new organoboron compound

    NASA Astrophysics Data System (ADS)

    Cao, Duxia; Liu, Zhiqiang; Li, Guozhong; Liu, Guoqun; Zhang, Guohui

    2008-02-01

    A new A-π-D-π-A type organoboron compound, 3,6-bis(dimesitylboryl)- N-butyl-carbazole (abbreviated as BBC), with trivalent boron as electron acceptor and carbazole as electron donor and π-conjugated core, has been synthesized and its single and two-photon related photophysical properties experimentally investigated. Pumped by 720 nm laser pulses in femtosecond regime, it showed strong two-photon excited blue-violet fluorescence at 393 nm in toluene and 403 nm in THF. The measured two-photon absorption cross-section by two-photon fluorescence method in toluene and THF is 34 GM and 38 GM, respectively.

  15. Absorption and Fluorescence Spectra of Poly(p-phenylenevinylene) (PPV) Oligomers: An ab Initio Simulation

    PubMed Central

    2014-01-01

    The absorption and fluorescence spectra of poly(p-phenylenevinylene) (PPV) oligomers with up to seven repeat units were theoretically investigated using the algebraic diagrammatic construction method to second order, ADC(2), combined with the resolution-of-the-identity (RI) approach. The ground and first excited state geometries of the oligomers were fully optimized. Vertical excitation energies and oscillator strengths of the first four transitions were computed. The vibrational broadening of the absorption and fluorescence spectra was studied using a semiclassical nuclear ensemble method. After correcting for basis set and solvent effects, we achieved a balanced description of the absorption and fluorescence spectra by means of the ADC(2) approach. This fact is documented by the computed Stokes shift along the PPV series, which is in good agreement with the experimental values. The experimentally observed band width of the UV absorption and fluorescence spectra is well reproduced by the present simulations showing that the nuclear ensemble generated should be well suitable for consecutive surface hopping dynamics simulations. PMID:25415930

  16. Excited state dynamics of single metal and semiconductor nanowires studied by transient absorption microscopy

    NASA Astrophysics Data System (ADS)

    Lo, Shun S.; Shi, Hong Y.; Major, Todd A.; Petchsang, Nattasamon; Huang, Libai; Kuno, Masaru K.; Hartland, Gregory V.

    2013-03-01

    Transient absorption microscopy (TAM) is a relatively new technique that allows the study of single nanostructures with sub-picosecond time resolution. Here, we present results for CdTe and Au Nanowires (NW). For the first material, we find an interesting power dependence of the excited dynamics, suggesting that a trap-filling mechanism is responsible for the observed behaviour. Additionally, acoustic phonons were observed, which were well described using continuum elastic models.[2] Carrier diffusion along these NWs are also reported. In the case of Au NWs, the propagation of surface plasmon polaritons was investigated. The results are in agreement with previous studies performed with fluorescence based techniques.[3,4] Unlike fluorescence techniques, multiple measurements on the same nanostructures are possible with TAM allowing one-to-one comparisons under different excitation polarizations and environments. NSF Award CHE-1110560 and CHE-0946447, Univ. of Notre Dame Strategic Research Initiative. L. Huang, DOE (DE-FC02-04ER15533)

  17. [Study on the absorption and fluorescence spectra of ethylene glycol and glycerol].

    PubMed

    Xu, Hui; Zhu, Tuo; Yu, Rui-Peng

    2007-07-01

    The absorption and fluorescence spectra of ethylene glycol and glycerol solution induced by UV light were studied respectively in the present paper. The most intense absorption wavelength for both of them was located at 198 nm. Moreover, fluorescence was detected when induced by suitable UV light, and the corresponding fluorescence spectra were listed. But there is no obvious relationship found between the fluorescence intensity and the excited wavelength, and a further research should be done. From the first derivative fluorescence spectra of ethylene glycol, it was concluded that under the UV light of 210 nm, the variation speed for relative intensity proved to be the fastest. In contrast, when excited by 225 nm, the speed proved to be the slowest. In addition, based on the quantum calculation and the transition from HOMO to LUMO of electronics in one-dimensional quantum well, the authors attempted to give out the value of absorption wavelength. In consideration of the bond-length variety brought out by the chain processing, the error between the experimental and calculation values should be apprehensible, and the latter can serve as some reference value in theory.

  18. Orthogonal spectra and cross sections: Application to optimization of multi-spectral absorption and fluorescence lidar

    SciTech Connect

    Shokair, I.R.

    1997-09-01

    This report addresses the problem of selection of lidar parameters, namely wavelengths for absorption lidar and excitation fluorescence pairs for fluorescence lidar, for optimal detection of species. Orthogonal spectra and cross sections are used as mathematical representations which provide a quantitative measure of species distinguishability in mixtures. Using these quantities, a simple expression for the absolute error in calculated species concentration is derived and optimization is accomplished by variation of lidar parameters to minimize this error. It is shown that the optimum number of wavelengths for detection of a species using absorption lidar (excitation fluorescence pairs for fluorescence lidar) is the same as the number of species in the mixture. Each species present in the mixture has its own set of optimum wavelengths. There is usually some overlap in these sets. The optimization method is applied to two examples, one using absorption and the other using fluorescence lidar, for analyzing mixtures of four organic compounds. The effect of atmospheric attenuation is included in the optimization process. Although the number of optimum wavelengths might be small, it is essential to do large numbers of measurements at these wavelengths in order to maximize canceling of statistical errors.

  19. Optimizing ultrafast illumination for multiphoton-excited fluorescence imaging.

    PubMed

    Stoltzfus, Caleb R; Rebane, Aleksander

    2016-05-01

    We study the optimal conditions for high throughput two-photon excited fluorescence (2PEF) and three-photon excited fluorescence (3PEF) imaging using femtosecond lasers. We derive relations that allow maximization of the rate of imaging depending on the average power, pulse repetition rate, and noise characteristics of the laser, as well as on the size and structure of the sample. We perform our analysis using ~100 MHz, ~1 MHz and 1 kHz pulse rates and using both a tightly-focused illumination beam with diffraction-limited image resolution, as well loosely focused illumination with a relatively low image resolution, where the latter utilizes separate illumination and fluorescence detection beam paths. Our theoretical estimates agree with the experiments, which makes our approach especially useful for optimizing high throughput imaging of large samples with a field-of-view up to 10x10 cm(2). PMID:27231620

  20. Optimizing ultrafast illumination for multiphoton-excited fluorescence imaging

    PubMed Central

    Stoltzfus, Caleb R.; Rebane, Aleksander

    2016-01-01

    We study the optimal conditions for high throughput two-photon excited fluorescence (2PEF) and three-photon excited fluorescence (3PEF) imaging using femtosecond lasers. We derive relations that allow maximization of the rate of imaging depending on the average power, pulse repetition rate, and noise characteristics of the laser, as well as on the size and structure of the sample. We perform our analysis using ~100 MHz, ~1 MHz and 1 kHz pulse rates and using both a tightly-focused illumination beam with diffraction-limited image resolution, as well loosely focused illumination with a relatively low image resolution, where the latter utilizes separate illumination and fluorescence detection beam paths. Our theoretical estimates agree with the experiments, which makes our approach especially useful for optimizing high throughput imaging of large samples with a field-of-view up to 10x10 cm2. PMID:27231620

  1. Anti-Stokes Fluorescent Probe with Incoherent Excitation

    PubMed Central

    Li, Yang; Zhou, Shifeng; Dong, Guoping; Peng, Mingying; Wondraczek, Lothar; Qiu, Jianrong

    2014-01-01

    Although inorganic anti-Stokes fluorescent probes have long been developed, the operational mode of today's most advanced examples still involves the harsh requirement of coherent laser excitation, which often yields unexpected light disturbance or even photon-induced deterioration during optical imaging. Here, we demonstrate an efficient anti-Stokes fluorescent probe with incoherent excitation. We show that the probe can be operated under light-emitting diode excitation and provides tunable anti-Stokes energy shift and decay kinetics, which allow for rapid and deep tissue imaging over a very large area with negligible photodestruction. Charging of the probe can be achieved by either X-rays or ultraviolet-visible light irradiation, which enables multiplexed detection and function integration with standard X-ray medical imaging devices. PMID:24518662

  2. Fluorescence upconversion properties of a class of improved pyridinium dyes induced by two-photon absorption

    NASA Astrophysics Data System (ADS)

    Xu, Guibao; Hu, Dawei; Zhao, Xian; Shao, Zongshu; Liu, Huijun; Tian, Yupeng

    2007-06-01

    We report the fluorescence upconversion properties of a class of improved pyridinium toluene- p-sulfonates having donor- π-acceptor (D- π-A) structure under two-photon excitation at 1064 nm. The experimental results show that both the two-photon excited (TPE) fluorescence lifetime and the two-photon pumped (TPP) energy upconversion efficiency were increased with the enhancement of electron-donating capability of the donor in the molecule. It is also indicated that an overlong alkyl group tends to result in a weakened molecular conjugation, leading to a decreased two-photon absorption (TPA) cross section. By choosing the donor, we can obtain a longest fluorescence lifetime of 837 ps, a highest energy upconversion efficiency of ˜6.1%, and a maximum TPA cross-section of 8.74×10 -48 cm 4 s/photon in these dyes.

  3. Absorption Reconstruction Improves Biodistribution Assessment of Fluorescent Nanoprobes Using Hybrid Fluorescence-mediated Tomography

    PubMed Central

    Gremse, Felix; Theek, Benjamin; Kunjachan, Sijumon; Lederle, Wiltrud; Pardo, Alessa; Barth, Stefan; Lammers, Twan; Naumann, Uwe; Kiessling, Fabian

    2014-01-01

    Aim: Fluorescence-mediated tomography (FMT) holds potential for accelerating diagnostic and theranostic drug development. However, for proper quantitative fluorescence reconstruction, knowledge on optical scattering and absorption, which are highly heterogeneous in different (mouse) tissues, is required. We here describe methods to assess these parameters using co-registered micro Computed Tomography (µCT) data and nonlinear whole-animal absorption reconstruction, and evaluate their importance for assessment of the biodistribution and target site accumulation of fluorophore-labeled drug delivery systems. Methods: Besides phantoms with varying degrees of absorption, mice bearing A431 tumors were imaged 15 min and 48 h after i.v. injection of a fluorophore-labeled polymeric drug carrier (pHPMA-Dy750) using µCT-FMT. The outer shape of mice and a scattering map were derived using automated segmentation of the µCT data. Furthermore, a 3D absorption map was reconstructed from the trans-illumination data. We determined the absorption of five interactively segmented regions (heart, liver, kidney, muscle, tumor). Since blood is the main near-infrared absorber in vivo, the absorption was also estimated from the relative blood volume (rBV), determined by contrast-enhanced µCT. We compared the reconstructed absorption with the rBV-based values and analyzed the effect of using the absorption map on the fluorescence reconstruction. Results: Phantom experiments demonstrated that absorption reconstruction is possible and necessary for quantitative fluorescence reconstruction. In vivo, the reconstructed absorption showed high values in strongly blood-perfused organs such as the heart, liver and kidney. The absorption values correlated strongly with the rBV-based absorption values, confirming the accuracy of the absorption reconstruction. Usage of homogenous absorption instead of the reconstructed absorption map resulted in reduced values in the heart, liver and kidney, by

  4. Unmixing multiple adjacent fluorescent targets with multispectral excited fluorescence molecular tomography.

    PubMed

    Zhou, Yuan; Guang, Huizhi; Pu, Huangsheng; Zhang, Jiulou; Luo, Jianwen

    2016-06-20

    Fluorescence molecular tomography (FMT) can visualize biological activities at cellular and molecular levels in vivo, and has been extensively used in drug delivery and tumor detection research of small animals. The ill-posedness of the FMT inverse problem makes it difficult to reconstruct and unmix multiple adjacent fluorescent targets that have different functional features but are labeled with the same fluorochrome. A method based on independent component analysis for multispectral excited FMT was proposed in our previous study. It showed that double fluorescent targets with certain edge-to-edge distance (EED) could be unmixed by the method. In this study, the situation is promoted to unmix multiple adjacent fluorescent targets (i.e., more than two fluorescent targets and EED=0). Phantom experiments on the resolving ability of the proposed algorithm demonstrate that the algorithm performs well in unmixing multiple adjacent fluorescent targets in both lateral and axial directions. And also, we recovered the locational information of each independent fluorescent target and described the variable trends of the corresponding fluorescent targets under the excitation spectrum. This method is capable of unmixing multiple fluorescent targets with small EED but labeled with the same fluorochrome, and may be used in imaging of nonspecific probe targeting and metabolism of drugs. PMID:27409108

  5. Biocompound detection through fluorescence excitation-emission matrix analysis

    NASA Astrophysics Data System (ADS)

    Twede, David R.; Sanders, Lee C.; Wagner, Michael L.

    2004-01-01

    The excitation-emission matrix (EEM) is the luminescence spectral emission intensity of fluorescent compounds as a function of the excitation wavelength. EEMs offer the promise of an additional degree of information for enhanced compound detection and identification. Veridian has collected pure-component EEMs of amino acids (Trp, Phe, Tyr), Bacillus globigii (bg), Bacillus thuringiensis (bt,), and selected backgrounds. Also collected were EEMs of mixtures of amino acids and of bg in solution with a few backgrounds. The EEMs of pure components and mixtures were analyzed for phenomenology and for potential methods of unmixing and identifying the constituents of EEMs having mixed components of a similar nature.

  6. Biocompound detection through fluorescence excitation-emission matrix analysis

    NASA Astrophysics Data System (ADS)

    Twede, David R.; Sanders, Lee C.; Wagner, Michael L.

    2003-12-01

    The excitation-emission matrix (EEM) is the luminescence spectral emission intensity of fluorescent compounds as a function of the excitation wavelength. EEMs offer the promise of an additional degree of information for enhanced compound detection and identification. Veridian has collected pure-component EEMs of amino acids (Trp, Phe, Tyr), Bacillus globigii (bg), Bacillus thuringiensis (bt,), and selected backgrounds. Also collected were EEMs of mixtures of amino acids and of bg in solution with a few backgrounds. The EEMs of pure components and mixtures were analyzed for phenomenology and for potential methods of unmixing and identifying the constituents of EEMs having mixed components of a similar nature.

  7. [Study of the Detecting System of CH4 and SO2 Based on Spectral Absorption Method and UV Fluorescence Method].

    PubMed

    Wang, Shu-tao; Wang, Zhi-fang; Liu, Ming-hua; Wei, Meng; Chen, Dong-ying; Wang, Xing-long

    2016-01-01

    According to the spectral absorption characteristics of polluting gases and fluorescence characteristics, a time-division multiplexing detection system is designed. Through this system we can detect Methane (CH4) and sulfur dioxide (SO2) by using spectral absorption method and the SO2 can be detected by using UV fluorescence method. The system consists of four parts: a combination of a light source which could be switched, the common optical path, the air chamber and the signal processing section. The spectral absorption characteristics and fluorescence characteristics are measured first. Then the experiment of detecting CH4 and SO2 through spectral absorption method and the experiment of detecting SO2 through UV fluorescence method are conducted, respectively. Through measuring characteristics of spectral absorption and fluorescence, we get excitation wavelengths of SO2 and CH4 measured by spectral absorption method at the absorption peak are 280 nm and 1.64 μm, respectively, and the optimal excitation wavelength of SO2 measured by UV fluorescence method is 220 nm. we acquire the linear relation between the concentration of CH4 and relative intensity and the linear relation between the concentration of SO2 and output voltage after conducting the experiment of spectral absorption method, and the linearity are 98.7%, 99.2% respectively. Through the experiment of UV fluorescence method we acquire that the relation between the concentration of SO2 and the voltage is linear, and the linearity is 99.5%. Research shows that the system is able to be applied to detect the polluted gas by absorption spectrum method and UV fluorescence method. Combing these two measurement methods decreases the costing and the volume, and this system can also be used to measure the other gases. Such system has a certain value of application. PMID:27228784

  8. [Study of the Detecting System of CH4 and SO2 Based on Spectral Absorption Method and UV Fluorescence Method].

    PubMed

    Wang, Shu-tao; Wang, Zhi-fang; Liu, Ming-hua; Wei, Meng; Chen, Dong-ying; Wang, Xing-long

    2016-01-01

    According to the spectral absorption characteristics of polluting gases and fluorescence characteristics, a time-division multiplexing detection system is designed. Through this system we can detect Methane (CH4) and sulfur dioxide (SO2) by using spectral absorption method and the SO2 can be detected by using UV fluorescence method. The system consists of four parts: a combination of a light source which could be switched, the common optical path, the air chamber and the signal processing section. The spectral absorption characteristics and fluorescence characteristics are measured first. Then the experiment of detecting CH4 and SO2 through spectral absorption method and the experiment of detecting SO2 through UV fluorescence method are conducted, respectively. Through measuring characteristics of spectral absorption and fluorescence, we get excitation wavelengths of SO2 and CH4 measured by spectral absorption method at the absorption peak are 280 nm and 1.64 μm, respectively, and the optimal excitation wavelength of SO2 measured by UV fluorescence method is 220 nm. we acquire the linear relation between the concentration of CH4 and relative intensity and the linear relation between the concentration of SO2 and output voltage after conducting the experiment of spectral absorption method, and the linearity are 98.7%, 99.2% respectively. Through the experiment of UV fluorescence method we acquire that the relation between the concentration of SO2 and the voltage is linear, and the linearity is 99.5%. Research shows that the system is able to be applied to detect the polluted gas by absorption spectrum method and UV fluorescence method. Combing these two measurement methods decreases the costing and the volume, and this system can also be used to measure the other gases. Such system has a certain value of application.

  9. Two-color two-photon excited fluorescence of indole: Determination of wavelength-dependent molecular parameters

    NASA Astrophysics Data System (ADS)

    Herbrich, Sebastian; Al-Hadhuri, Tawfik; Gericke, Karl-Heinz; Shternin, Peter S.; Smolin, Andrey G.; Vasyutinskii, Oleg S.

    2015-01-01

    We present a detailed study of two-color two-photon excited fluorescence in indole dissolved in propylene glycol. Femtosecond excitation pulses at effective wavelengths from 268 to 293.33 nm were used to populate the two lowest indole excited states 1La and 1Lb and polarized fluorescence was then detected. All seven molecular parameters and the two-photon polarization ratio Ω containing information on two-photon absorption dynamics, molecular lifetime τf, and rotation correlation time τrot have been determined from experiment and analyzed as a function of the excitation wavelength. The analysis of the experimental data has shown that 1Lb-1La inversion occurred under the conditions of our experiment. The two-photon absorption predominantly populated the 1La state at all excitation wavelengths but in the 287-289 nm area which contained an absorption hump of the 1Lb state 0-0 origin. The components of the two-photon excitation tensor S were analyzed giving important information on the principal tensor axes and absorption symmetry. The results obtained are in a good agreement with the results reported by other groups. The lifetime τf and the rotation correlation time τrot showed no explicit dependence on the effective excitation wavelength. Their calculated weighted average values were found to be τf = 3.83 ± 0.14 ns and τrot = 0.74 ± 0.06 ns.

  10. Hyperspectral fluorescence imaging with multi wavelength LED excitation

    NASA Astrophysics Data System (ADS)

    Luthman, A. Siri; Dumitru, Sebastian; Quirós-Gonzalez, Isabel; Bohndiek, Sarah E.

    2016-04-01

    Hyperspectral imaging (HSI) can combine morphological and molecular information, yielding potential for real-time and high throughput multiplexed fluorescent contrast agent imaging. Multiplexed readout from targets, such as cell surface receptors overexpressed in cancer cells, could improve both sensitivity and specificity of tumor identification. There remains, however, a need for compact and cost effective implementations of the technology. We have implemented a low-cost wide-field multiplexed fluorescence imaging system, which combines LED excitation at 590, 655 and 740 nm with a compact commercial solid state HSI system operating in the range 600 - 1000 nm. A key challenge for using reflectance-based HSI is the separation of contrast agent fluorescence from the reflectance of the excitation light. Here, we illustrate how it is possible to address this challenge in software, using two offline reflectance removal methods, prior to least-squares spectral unmixing. We made a quantitative comparison of the methods using data acquired from dilutions of contrast agents prepared in well-plates. We then established the capability of our HSI system for non-invasive in vivo fluorescence imaging in small animals using the optimal reflectance removal method. The HSI presented here enables quantitative unmixing of at least four fluorescent contrast agents (Alexa Fluor 610, 647, 700 and 750) simultaneously in living mice. A successful unmixing of the four fluorescent contrast agents was possible both using the pure contrast agents and with mixtures. The system could in principle also be applied to imaging of ex vivo tissue or intraoperative imaging in a clinical setting. These data suggest a promising approach for developing clinical applications of HSI based on multiplexed fluorescence contrast agent imaging.

  11. Excitation of emission lines by fluorescence and recombination in IC 418

    NASA Astrophysics Data System (ADS)

    Escalante, Vladimir; Morisset, Cristophe; Georgiev, Leonid

    2012-08-01

    We predict intensities of lines of CII, NI, NII, OI and OII and compare them with a deep spectroscopic survey of IC 418 to test the effect of excitation of nebular emission lines by continuum fluorescence of starlight. Our calculations use a nebular model and a synthetic spectrum of its central star to take into account excitation of the lines by continuum fluorescence and recombination. The NII spectrum is mostly produced by fluorescence due to the low excitation conditions of the nebula, but many CII and OII lines have more excitation by fluorescence than recombination. In the neutral envelope, the NI permitted lines are excited by fluorescence, and almost all the OI lines are excited by recombination. Electron excitation produces the forbidden optical lines of OI, but continuum fluorescence excites most of the NI forbidden line intensities. Lines excited by fluorescence of light below the Lyman limit thus suggest a new diagnostic to explore the photodissociation region of a nebula.

  12. Small fluorescence-activating and absorption-shifting tag for tunable protein imaging in vivo

    PubMed Central

    Plamont, Marie-Aude; Billon-Denis, Emmanuelle; Maurin, Sylvie; Gauron, Carole; Pimenta, Frederico M.; Specht, Christian G.; Shi, Jian; Quérard, Jérôme; Pan, Buyan; Rossignol, Julien; Moncoq, Karine; Morellet, Nelly; Volovitch, Michel; Lescop, Ewen; Chen, Yong; Triller, Antoine; Vriz, Sophie; Le Saux, Thomas; Jullien, Ludovic; Gautier, Arnaud

    2016-01-01

    This paper presents Yellow Fluorescence-Activating and absorption-Shifting Tag (Y-FAST), a small monomeric protein tag, half as large as the green fluorescent protein, enabling fluorescent labeling of proteins in a reversible and specific manner through the reversible binding and activation of a cell-permeant and nontoxic fluorogenic ligand (a so-called fluorogen). A unique fluorogen activation mechanism based on two spectroscopic changes, increase of fluorescence quantum yield and absorption red shift, provides high labeling selectivity. Y-FAST was engineered from the 14-kDa photoactive yellow protein by directed evolution using yeast display and fluorescence-activated cell sorting. Y-FAST is as bright as common fluorescent proteins, exhibits good photostability, and allows the efficient labeling of proteins in various organelles and hosts. Upon fluorogen binding, fluorescence appears instantaneously, allowing monitoring of rapid processes in near real time. Y-FAST distinguishes itself from other tagging systems because the fluorogen binding is highly dynamic and fully reversible, which enables rapid labeling and unlabeling of proteins by addition and withdrawal of the fluorogen, opening new exciting prospects for the development of multiplexing imaging protocols based on sequential labeling. PMID:26711992

  13. Small fluorescence-activating and absorption-shifting tag for tunable protein imaging in vivo.

    PubMed

    Plamont, Marie-Aude; Billon-Denis, Emmanuelle; Maurin, Sylvie; Gauron, Carole; Pimenta, Frederico M; Specht, Christian G; Shi, Jian; Quérard, Jérôme; Pan, Buyan; Rossignol, Julien; Moncoq, Karine; Morellet, Nelly; Volovitch, Michel; Lescop, Ewen; Chen, Yong; Triller, Antoine; Vriz, Sophie; Le Saux, Thomas; Jullien, Ludovic; Gautier, Arnaud

    2016-01-19

    This paper presents Yellow Fluorescence-Activating and absorption-Shifting Tag (Y-FAST), a small monomeric protein tag, half as large as the green fluorescent protein, enabling fluorescent labeling of proteins in a reversible and specific manner through the reversible binding and activation of a cell-permeant and nontoxic fluorogenic ligand (a so-called fluorogen). A unique fluorogen activation mechanism based on two spectroscopic changes, increase of fluorescence quantum yield and absorption red shift, provides high labeling selectivity. Y-FAST was engineered from the 14-kDa photoactive yellow protein by directed evolution using yeast display and fluorescence-activated cell sorting. Y-FAST is as bright as common fluorescent proteins, exhibits good photostability, and allows the efficient labeling of proteins in various organelles and hosts. Upon fluorogen binding, fluorescence appears instantaneously, allowing monitoring of rapid processes in near real time. Y-FAST distinguishes itself from other tagging systems because the fluorogen binding is highly dynamic and fully reversible, which enables rapid labeling and unlabeling of proteins by addition and withdrawal of the fluorogen, opening new exciting prospects for the development of multiplexing imaging protocols based on sequential labeling. PMID:26711992

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

    NASA Astrophysics Data System (ADS)

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

    1991-10-01

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

  15. Laser-excited fluorescence spectra of atomic uranium

    SciTech Connect

    Wang Songyue; Jin Changtai; Shen Mingtao; Wang Xiulan

    1987-05-01

    Using a dc-supply hollow-cathode lamp as a source of uranium vapor and a rhodamine 6G dye laser to excite the vapor optically, it was simple and convenient to detect fluorescence from uranium atoms at 753.393, 763.175, and 763.954 nm. We give a detailed discussion of how we eliminated the intense background emissions, which were principally due to the lamp.

  16. Resonant excited state absorption and relaxation mechanisms in Tb3+-doped calcium aluminosilicate glasses: an investigation by thermal mirror spectroscopy.

    PubMed

    Bianchi, G S; Zanuto, V S; Astrath, F B G; Malacarne, L C; Terra, I A A; Catunda, T; Nunes, L A O; Jacinto, C; Andrade, L H C; Lima, S M; Baesso, M L; Astrath, N G C

    2013-11-15

    Resonant excited state absorption (ESA) and relaxation processes in Tb(3+)-doped aluminosilicate glasses are quantitatively evaluated. A model describing the excitation steps and upconversion emission is developed and applied to interpret the results from laser-induced surface deformation using thermal mirror spectroscopy. The fluorescence quantum efficiency of level (5)D(4) was found to be close to unity and concentration independent while, for the level (5)D(3), it decreases with Tb(3+) concentration. Emission spectroscopy measurements supported these results. ESA cross sections are found to be more than three orders of magnitude higher than the ground state absorption cross section. PMID:24322101

  17. Excitation volumetric effects (EVE) in metal-enhanced fluorescence.

    PubMed

    Dragan, A I; Geddes, C D

    2011-03-01

    Metal-Enhanced Fluorescence (MEF) effects from different density silver island films (SiFs) and the effects of far-field excitation irradiance on the observed enhancement of fluorescence were studied. It is shown that MEF non-linearly depends on silver nanoparticle (NP) size/density, reaching a maximum value for SiFs made at a deposition time (DT) of ∼5 minutes, i.e. just before SiFs become continuous. Numerical simulations of the silver-islands growing on glass revealed that the near-field magnitude depends non-linearly on size and interparticle distance exhibiting dramatic enhancement at ∼10 nm distance between the NPs. In addition, a remarkable effect of modulation in MEF efficiency by far-field excitation irradiance has been observed, which can be correlated well with numerical simulations that show an excitation power volume dependence. The near-field volume changes non-linearly with far-field power. This unique observation has profound implications in MEF, which has rapidly emerged as a powerful tool in the biosciences and ultimately allows for tunable fluorescence enhancement factors.

  18. Real-time absorption reduced surface fluorescence imaging

    PubMed Central

    Yang, Bin; Tunnell, James W.

    2014-01-01

    Abstract. We introduce a technique that limits absorption effects in fluorescence imaging and does not require extensive imaging processing, thus allowing for video rate imaging. The absorption minimization is achieved using spatial frequency domain imaging at a single high spatial frequency with standard three-phase demodulation. At a spatial frequency f=0.5  mm−1, we demonstrated in both in-vitro phantoms and ex-vivo tissue that the absorption can be significantly reduced. In the real-time implementation, we achieved a video rate of 19  frames/s. This technique has potential in cancer visualization and tumor margin detection. PMID:25250826

  19. Real-time absorption reduced surface fluorescence imaging.

    PubMed

    Yang, Bin; Tunnell, James W

    2014-09-01

    We introduce a technique that limits absorption effects in fluorescence imaging and does not require extensive imaging processing, thus allowing for video rate imaging. The absorption minimization is achieved using spatial frequency domain imaging at a single high spatial frequency with standard three-phase demodulation. At a spatial frequency f ¼ 0.5 mm−1, we demonstrated in both in-vitro phantoms and ex-vivo tissue that the absorption can be significantly reduced. In the real-time implementation, we achieved a video rate of 19 frames∕s. This technique has potential in cancer visualization and tumor margin detection. PMID:25250826

  20. Topographic mapping of subsurface fluorescent structures in tissue using multiwavelength excitation

    PubMed Central

    Kim, Anthony; Roy, Mathieu; Dadani, Farhan N.; Wilson, Brian C.

    2010-01-01

    Different colors of visible light penetrate to varying depths in tissue due to the wavelength dependence of tissue optical absorption and elastic scattering. We exploit this to map the contour of the closest surface of a buried fluorescent object. This uses a novel algorithm based on the diffusion theory description of light propagation in tissue at each excitation wavelength to derive metrics that define the depth of the top surface of the object. The algorithm was validated using a tissue-simulating phantom. It was then demonstrated in vivo by subsurface brain tumor topography in a rodent model, using the fluorescence signal from protoporphyrin IX that is preferentially synthesized within malignant cells following systemic application of aminolevulinic acid. Comparisons to histomorphometry in the brain post mortem show the spatial accuracy of the technique. This method has potential for fluorescence image-guided tumor surgery, as well as other biomedical and nonbiological applications in subsurface sensing. PMID:21198200

  1. Multi-color femtosecond source for simultaneous excitation of multiple fluorescent proteins in two-photon fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Wang, Ke; Liu, Tzu-Ming; Wu, Juwell; Horton, Nicholas G.; Lin, Charles P.; Xu, Chris

    2013-02-01

    Simultaneous imaging of cells expressing multiple fluorescent proteins (FPs) is of particular interest in applications such as mapping neural circuits, tracking multiple immune cell populations, etc. To visualize both in vivo and ex vivo tissue morphology and physiology at a cellular level deep within scattering tissues, two-photon fluorescence microscopy (2PM) is a powerful tool that has found wide applications. However, simultaneous imaging of multiple FPs with 2PM is greatly hampered by the lack of proper ultrafast lasers offering multi-color femtosecond pulses, each targeting the two-photon absorption peak of a different FP. Here we demonstrate simultaneous two-photon fluorescence excitation of RFP, YFP, and CFP in human melanoma cells engineered to express a "rainbow" pallet of colors, using a novel fiber-based source with energetic, three-color femtosecond pulses. The three-color pulses, centered at 775 nm, 864 nm and 950 nm, are obtained through second harmonic generation of the 1550 nm pump laser and SHG of the solitons at 1728 nm and 1900 nm generated through soliton self-frequency shift (SSFS) of the pump laser in a large-mode-area (LMA) fiber. The resulting wavelengths are well matched to the two-photon absorption peaks of the three FPs for efficient excitation. Our results demonstrate that multi-color femtosecond pulse generation using SSFS and a turn-key, fiber-based femtosecond laser can fulfill the requirements for simultaneous imaging of multiple FPs in 2PM, opening new opportunities for a wide range of biological applications where non-invasive, high-resolution imaging of multiple fluorescent indicators is required.

  2. Atomic Absorption, Atomic Fluorescence, and Flame Emission Spectrometry.

    ERIC Educational Resources Information Center

    Horlick, Gary

    1984-01-01

    This review is presented in six sections. Sections focus on literature related to: (1) developments in instrumentation, measurement techniques, and procedures; (2) performance studies of flames and electrothermal atomizers; (3) applications of atomic absorption spectrometry; (4) analytical comparisons; (5) atomic fluorescence spectrometry; and (6)…

  3. Laser line illumination scheme allowing the reduction of background signal and the correction of absorption heterogeneities effects for fluorescence reflectance imaging.

    PubMed

    Fantoni, Frédéric; Hervé, Lionel; Poher, Vincent; Gioux, Sylvain; Mars, Jérôme I; Dinten, Jean-Marc

    2015-10-01

    Intraoperative fluorescence imaging in reflectance geometry is an attractive imaging modality as it allows to noninvasively monitor the fluorescence targeted tumors located below the tissue surface. Some drawbacks of this technique are the background fluorescence decreasing the contrast and absorption heterogeneities leading to misinterpretations concerning fluorescence concentrations. We propose a correction technique based on a laser line scanning illumination scheme. We scan the medium with the laser line and acquire, at each position of the line, both fluorescence and excitation images. We then use the finding that there is a relationship between the excitation intensity profile and the background fluorescence one to predict the amount of signal to subtract from the fluorescence images to get a better contrast. As the light absorption information is contained both in fluorescence and excitation images, this method also permits us to correct the effects of absorption heterogeneities. This technique has been validated on simulations and experimentally. Fluorescent inclusions are observed in several configurations at depths ranging from 1 mm to 1 cm. Results obtained with this technique are compared with those obtained with a classical wide-field detection scheme for contrast enhancement and with the fluorescence by an excitation ratio approach for absorption correction. PMID:26442963

  4. Remote excitation fluorescence correlation spectroscopy using silver nanowires

    NASA Astrophysics Data System (ADS)

    Su, Liang; Yuan, Haifeng; Lu, Gang; Hofkens, Johan; Roeffaers, Maarten; Uji-i, Hiroshi

    2014-11-01

    Fluorescence correlation spectroscopy (FCS), a powerful tool to resolve local properties, dynamical process of molecules, rotational and translational diffusion motions, relies on the fluctuations of florescence observables in the observation volume. In the case of rare transition events or small dynamical fluctuations, FCS requires few molecules or even single molecules in the observation volume at a time to minimize the background signals. Metal nanoparticle which possess unique localized surface plasmon resonance (LSPR) have been used to reduce the observation volume down to sub-diffraction limited scale while maintain at high analyst concentration up to tens of micromolar. Nevertheless, the applications of functionalized nanoparticles in living cell are limited due to the continuous diffusion after cell uptake, which makes it difficult to target the region of interests in the cell. In this work, we demonstrate the use of silver nanowires for remote excitation FCS on fluorescent molecules in solution. By using propagation surface plasmon polaritons (SPPs) which supported by the silver nanowire to excite the fluorescence, both illumination and observation volume can be reduced simultaneously. In such a way, less perturbation is induced to the target region, and this will broaden the application scope of silver nanowire as tip in single cell endoscopy.

  5. Local excitation and collection in polymeric fluorescent microstructures

    NASA Astrophysics Data System (ADS)

    Henrique, Franciele Renata; Mendonca, Cleber Renato

    2016-04-01

    Integrated photonics has gained attention in recent years due to its wide range of applications which span from biology to optical communications. The use of polymer-based platforms for photonic devices is of great interest because organic compounds can be easily incorporated to polymers, enabling modifications to the system physical properties. The two-photon polymerization technique has emerged as an interesting tool for the production of three-dimensional polymeric microstructures. However, for their further incorporation in photonic devices it is necessary to develop methods to perform optical excitation and signal collection on such microstructures. With such purpose, we demonstrate approaches to perform local excitation and collection in polymeric microstructures doped with fluorescent dyes, employing tapered fibers. The obtained results indicate that fiber tapers are suitable to couple light in and out of fluorescent polymeric microstructures, paving the way for their incorporation in photonic devices. We also show that microstructures doped with more than one dye can be used as built-in broadband light sources to photonic circuits and their emission spectrum can be tuned by the right choice of the excitation position.

  6. The origin of radiationless conversion of the excited state in the kindling fluorescent protein (KFP): femtosecond studies and quantum modeling

    NASA Astrophysics Data System (ADS)

    Shelaev, I.; Mironov, V.; Rusanov, A.; Gostev, F.; Bochenkova, A.; Sarkisov, O.; Nemukhin, A.; Savitsky, A.

    2011-06-01

    The Ala143Gly variant of the chromoprotein asCP from the sea anemony Anemonia sulcata, called the kindling fluorescent protein (KFP), is a promising candidate for the development of novel subdiffraction method of fluorescent microscopy. The pump-probe method with the delay times between the pump and probe pulses up to 5 ps was applied to study dynamics of the primary processes upon excitation of KFP. The differential absorption spectra at 80 fs delay showed the absorption peak in the range 450-510 nm with the maximum wavelength at 490 nm, which diminished almost twice by intensity by 400 fs and practically disappeared by 1.5 ps. The quantum calculations showed that upon photo-excitation of KFP to the first excited state S1, the fast radiationless relaxation occurred to the ground state S0 due to rotation of the phenolic fragment of the chromophore.

  7. Fluorescence excitation spectrum and solvent-assisted conformational isomerization (SACI) of jet-cooled acetaminophen

    NASA Astrophysics Data System (ADS)

    Sohn, Woon Yong; Kang, Jeong Seok; Lee, So Young; Kang, Hyuk

    2013-08-01

    Fluorescence excitation spectrum of jet-cooled acetaminophen was obtained. When AAP was expanded with a buffer gas containing 0.3-1.1% of water, absorption peaks of the less stable trans conformer was significantly reduced by solvent-assisted conformational isomerization (SACI), which is confirmed by a separately measured UV-UV hole burning spectroscopy. It is also confirmed by quantum mechanical calculation and RRKM calculation that it is energetically and kinetically possible to induce SACI in AAP with water. The SACI mechanism suggests a possible pathway that acetaminophen can adopt an active conformation in vivo, which is need for molecular recognition and drug activity.

  8. Simulations of fluorescence solvatochromism in substituted PPV oligomers from excited state molecular dynamics with implicit solvent

    SciTech Connect

    Bjorgaard, J. A.; Nelson, T.; Kalinin, K.; Kuzmenko, V.; Velizhanin, K. A.; Tretiak, S.

    2015-04-28

    In this study, an efficient method of treating solvent effects in excited state molecular dynamics (ESMD) is implemented and tested by exploring the solvatochromic effects in substituted p-phenylene vinylene oligomers. A continuum solvent model is used which has very little computational overhead. This allows simulations of ESMD with solvent effects on the scale of hundreds of picoseconds for systems of up to hundreds of atoms. At these time scales, solvatochromic shifts in fluoresence spectra can be described. Solvatochromic shifts in absorption and fluorescence spectra from ESMD are compared with time-dependent density functional theory calculations and experiments.

  9. Laser-excited confocal-fluorescence gel scanner

    SciTech Connect

    Mathies, R.A.; Scherer, J.R.; Quesada, M.A. ); Rye, H.S.; Glazer, A.N. )

    1994-04-01

    A high-sensitivity, laser-excited, confocal-fluorescence scanner has been developed for the detection of fluorescently labeled nucleic acids separated on slab gels. The gel is placed on a motor-driven, two-dimensional scan stage and raster scanned past the optical detection system. The 488-nm argon ion laser beam is introduced into the confocal optical system at a long-pass dichroic beam splitter and focused within the gel to an [similar to]2 [mu]m diameter spot by a high-numerical aperture microscope objective. The resulting fluorescence is gathered by the objective, passed back through the first long-pass beam splitter, and relayed to a second dichroic beam splitter that separates the red and green emissions. The fluorescence is then focused on confocal spatial filters to reduce stray and scattered light, passed through spectral filters, and detected with photomultipliers. The resulting signals are amplified, filtered, and digitized for display on a computer. This system can detect as little as 5[times]10[sup [minus]12] M fluorescein, the resolution as operated is 160 [mu]m, and it can scan a 6 cm[times]6 cm gel using a scan rate of 4 cm/s in 12 min. The detection of DNA on slab gels, two-color DNA fragment sizing, and microtiter plate scanning are presented to illustrate some of the possible applications of this apparatus.

  10. Fluorescent vibration-rotation excitation of cometary C2

    NASA Technical Reports Server (NTRS)

    Gredel, Roland; Van Dishoeck, Ewine F.; Black, John H.

    1989-01-01

    The statistical equilibrium equations that determine the population densities of the energy levels in cometary C2 molecules due to fluorescent excitation are examined in detail. The adopted model and molecular parameters are discussed, and a theoretical estimate is made of the two intercombination transition moments. From the theoretical population densities in the various rotational levels, flux ratios and synthetic emission profiles are calculated as functions of the a 3Pi(u) - X 1Sigma(g)+ and the c 3Sigma(u)+ - X 3Sigma(g)+ intercombination transition moments. The influence of each of these two transitions separately on the vibrational and rotational excitation temperatures is investigated. The observed emission spectra of the (0,0) Swan band in Comet Halley are presented and compared to the synthetic profiles.

  11. The photophysics of phenylenevinylene oligomers and self-absorption of their fluorescence in polymer films.

    PubMed

    Winch, Nicola M; Smith, Gerald J; Breukers, Robert D; Bhuiyan, Delower H; Kay, Andrew J; Smith, Trevor A; Ghiggino, Kenneth P; Raymond, Sebastiampillai G

    The fluorescence spectra, quantum yields and lifetimes of a series of alkoxy-substituted phenylenevinylene molecules, which serve as short chain oligomer models for poly(p-phenylenevinylene), have been determined in fluid solvents and in a high viscosity polymer matrix. The effects of solvent polarity and a high viscosity molecular environment on the fluorescence yields and spectral shapes have been established. Alkoxy group substitution on the phenyl ring moieties of the molecules has an important effect on the vibronic structures and profiles of the absorption spectra. This was interpreted in terms of hot-band, ground to excited singlet state transitions from energetically closely-spaced torsional vibrational levels of the vinylene double bond in the ground state. The shapes of the absorption bands affect the overlaps of the absorption and fluorescence spectra. This has been quantified as the probability of fluorescence reabsorption in solid polymer films as a function of pathlength. This is an important determinant of the efficacies of these compounds for "harvesting" solar energy in luminescent solar concentrator systems. The reabsorption probabilities of these compounds are lower for all pathlengths than those determined in the same polymer film for the fluorophores, perylene and perylene diimide, which have been considered for concentrating spatially diffuse sunlight. PMID:27480331

  12. Ratiometric fluorescent/colorimetric cyanide-selective sensor based on excited-state intramolecular charge transfer-excited-state intramolecular proton transfer switching.

    PubMed

    Lin, Wei-Chi; Fang, Sin-Kai; Hu, Jiun-Wei; Tsai, Hsing-Yang; Chen, Kew-Yu

    2014-05-20

    A novel salicylideneaniline-based fluorescent sensor, SB1, with a unique excited-state intramolecular charge transfer-excited-state intramolecular proton transfer (ESICT-ESIPT) coupled system was synthesized and demonstrated to fluorescently sense CN(-) with specific selectivity and high sensitivity in aqueous media based on ESICT-ESIPT switching. A large blue shift (96 nm) was also observed in the absorption spectra in response to CN(-). The bleaching of the color could be clearly observed by the naked eye. Moreover, SB1-based test strips were easily fabricated and low-cost, and could be used in practical and efficient CN(-) test kits. Density functional theory (DFT) and time-dependent DFT (TD-DFT) calculations further support the cyanide-induced ESICT-ESIPT switching mechanism. The results provide the proof of concept that the colorimetric and ratiometric fluorescent cyanide-selective chemodosimeter can be created based on an ESICT-ESIPT coupled system. PMID:24809868

  13. Fluorescence Excitation-Emission Matrix Regional Integration to Quantify Spectra for Dissolved Organic Matter

    USGS Publications Warehouse

    Chen, W.; Westerhoff, P.; Leenheer, J.A.; Booksh, K.

    2003-01-01

    Excitation-emission matrix (EEM) fluorescence spectroscopy has been widely used to characterize dissolved organic matter (DOM) in water and soil. However, interpreting the >10,000 wavelength-dependent fluorescence intensity data points represented in EEMs has posed a significant challenge. Fluorescence regional integration, a quantitative technique that integrates the volume beneath an EEM, was developed to analyze EEMs. EEMs were delineated into five excitation-emission regions based on fluorescence of model compounds, DOM fractions, and marine waters or freshwaters. Volumetric integration under the EEM within each region, normalized to the projected excitation-emission area within that region and dissolved organic carbon concentration, resulted in a normalized region-specific EEM volume (??i,n). Solid-state carbon nuclear magnetic resonance (13C NMR), Fourier transform infrared (FTIR) analysis, ultraviolet-visible absorption spectra, and EEMs were obtained for standard Suwannee River fulvic acid and 15 hydrophobic or hydrophilic acid, neutral, and base DOM fractions plus nonfractionated DOM from wastewater effluents and rivers in the southwestern United States. DOM fractions fluoresced in one or more EEM regions. The highest cumulative EEM volume (??T,n = ????i,n) was observed for hydrophobic neutral DOM fractions, followed by lower ??T,n values for hydrophobic acid, base, and hydrophilic acid DOM fractions, respectively. An extracted wastewater biomass DOM sample contained aromatic protein- and humic-like material and was characteristic of bacterial-soluble microbial products. Aromatic carbon and the presence of specific aromatic compounds (as indicated by solid-state 13C NMR and FTIR data) resulted in EEMs that aided in differentiating wastewater effluent DOM from drinking water DOM.

  14. [The Establishment of the Method of the Fiber Optic Chemical Sensor Synchronous Absorption-Fluorescence].

    PubMed

    Zhang Li-hua; Iburaim, Arkin

    2016-03-01

    A new method of simultaneously measuring fiber-optic chemical sensor absorption spectrum and fluorescence spectrum is established. Make synchronous absorption-fluorescence cuvette, establish synchronous absorption-fluorescence spectrometry instrumentation combined by fiber optic chemical sensor technology, measure the synchronous absorption-fluorescence spectrums of solutions of rhodamine B, vitamin B2 and vitamin B6, compared by absorption spectroscopy measured by traditional UV-Visible photometric method and fluorescence spectroscopy measured by traditional fluorescence method. Synchronous absorption-fluorescence method measure absorption spectrums and fluorescence spectrums the same to traditional photometric and fluorescence spectroscopy of rhodamine B, vitamin B2 and vitamin B6. The maximum wavelength of fluorescence intensity method has high accuracy relatively compared with fluorescence, but the maximum wavelength of absorption has a slight deviation. Synchronous absorption-fluorescence method means simultaneously measure the absorption spectrums and fluorescence spectrums of the fluorescent substance, making two spectrums to one. The method measured the maximum emission wavelength with high accuracy, though in measuring maximum absorption wavelength there is a slight deviation, but it is worth further studying. PMID:27400519

  15. Absorption and Transport of Fluorescent Brighteners by Microorganisms

    PubMed Central

    Darken, Marjorie A.

    1962-01-01

    The absorption of brighteners by living cells and their transport to subsequent growth is described. Brighteners are highly fluorescent, ultraviolet-absorbing compounds which appear to be essentially nontoxic, stable biological markers. They have been effectively absorbed by growing cultures of bacteria, yeasts, actinomycetes, and higher fungi, with active growth centers evidencing the greatest flourescence. Images FIG. 2 FIG. 3 FIG. 4 FIG. 5 FIG. 6 FIG. 7 FIG. 8 FIG. 9 FIG. 10-11 PMID:14025111

  16. Determination of optimal excitation and emission wavebands for detection of defect cherry tomato by using fluorescence emission and excitation matrix

    NASA Astrophysics Data System (ADS)

    Baek, In-Suck; Cho, Byoung-Kwan; Kim, Moon S.; Kim, Young-Sik

    2013-05-01

    Fluorescence imaging technique has been widely used for quality and safety measurements of agro-food materials. Fluorescence emission intensities of target materials are influenced by wavelengths of excitation sources. Hence, selection of a proper excitation wavelength is an important factor in differentiating target materials effectively. In this study, optimal fluorescence excitation wavelength was determined on the basis of fluorescence emission intensity of defect and sound areas of cherry tomatoes. The result showed that fluorescence responses of defect and sound surfaces of cherry tomatoes were most significantly separated with the excitation light wavelength range between 400 and 410 nm. Fluorescence images of defect cherry tomatoes were acquired with the LEDs with the central wavelength of 410 nm as the excitation source to verify the detection efficiency of cherry tomato defects. The resultant fluorescence images showed that the defects were discriminated from sound areas on cherry tomatoes with above 98% accuracy. This study shows that high power LEDs as the excitation source for fluorescence imaging are suitable for defect detection of cherry tomatoes.

  17. Effect of perfluorocarbons on the ultraviolet absorption and fluorescence characteristics of some saturated hydrocarbon liquids

    SciTech Connect

    Choi, H.T.; Lipsky, S.

    1981-12-24

    Perfluoro-n-alkanes and perfluorocycloalkanes are found to affect the photophysical properties of saturated hydrocarbons in markedly different ways.The addition of a low concentration (<0.2 M) of a perfluoro-n-alkane to a hydrocarbon liquid has no observable effect on the electronic absorption spectrum and only very slightly quenches the hydrocarbon fluorescence. In contrast, at the same concentration levels, the perfluorocycloalkane strongly perturbs the absorption spectrum and very effectively reduces the fluorescence quantum yield. The change in the absorption spectrum is attributed to a contact charge-transfer absorption with the hydrocarbon acting as electron donor. The efficiency with which the perfluorocycloalkane reduces the hydrocarbon fluorescence quantum yield increases with increasing perfluorocycloalkane concentration. The results are analyzed with a conventional diffusional model that includes transient terms. The model contains two parameters, an encounter distance, R, and the product of the relative diffusion coefficient, D, and the lifetime, tau/sub 0/, of the hydrocarbon excited state. The model is applied to perfluorodecalin quenching of cyclohexane, decalin, and 2,3-dimethylbutane at excitation wavelengths, lambda/sub ex/, ranging from 185 to 147 nm. An unrestricted, two-parameter, least-squares fit of the model to the data provides values of R and D(tau/sub 0/) for each solvent system at each lambda/sub ex/. Where values of D and tau/sub 0/ are known independently, their product agrees well with the D(tau/sub 0/) obtained from the fit. The value of R is found to be approx. = 14 angstrom for all solvents at all lambda/sub ex/. This value is estimated to be about 2 times larger than the ground-state hydrocarbon-perfluorodecalin contact distance.

  18. Excitation-Selectable Nanoprobe for Tumor Fluorescence Imaging and Near-Infrared Thermal Therapy.

    PubMed

    Wei, Yanchun; Chen, Qun; Wu, Baoyan; Xing, Da

    2016-01-01

    The combination of diagnostics and therapeutics is growing rapidly in cancer treatment. Here, using upconversion nanoparticles coated with chitosan conjugated with a targeting molecule and loaded with indocyanine green (ICG), we develop an excitation-selectable nanoprobe with highly integrated functionalities, including the emission of visible and near-infrared (NIR) light, strong optical absorption in the NIR region and high photostability. After intravenous injection in tumor bearing mice, the nanoprobes target to the tumor vascular system. NIR lasers (980 and 808 nm) are then selectively applied to the mice. The results show that the emitted upconversion fluorescence and NIR fluorescence can be used in a complementary manner for high signal/noise ratio and sensitive tumor imaging for more precise tumor localization. Highly effective photothermal therapy is realized using 808 nm laser irradiation, and the upconversion fluorescence at 654 nm can be used for monitoring treatment effect during the thermal therapy. In summary, using the nanoprobes, outstanding therapeutic efficacy could be realized through flexible excitation control, precise tumor localization, highly effective photothermal conversion and real-time treatment monitoring. The nanofabrication strategy highlights the promise of nanoparticles in cancer theranostics. PMID:27301175

  19. Spectral analysis on origination of the bands at 437 nm and 475.5 nm of chlorophyll fluorescence excitation spectrum in Arabidopsis chloroplasts.

    PubMed

    Zeng, Lizhang; Wang, Yongqiang; Zhou, Jun

    2016-05-01

    Chlorophyll fluorescence has been often used as an intrinsic optical molecular probe to study photosynthesis. In this study, the origin of bands at 437 and 475.5 nm in the chlorophyll fluorescence excitation spectrum for emission at 685 nm in Arabidopsis chloroplasts was investigated using various optical analysis methods. The results revealed that this fluorescence excitation spectrum was related to the absorption characteristics of pigment molecules in PSII complexes. Moreover, the excitation band centred at 475.5 nm had a blue shift, but the excitation band at 437 nm changed relatively less due to induction of non-photochemical quenching (NPQ). Furthermore, fluorescence emission spectra showed that this blue shift occurred when excitation energy transfer from both chlorophyll b (Chl b) and carotenoids (Cars) to chlorophyll a (Chl a) was blocked. These results demonstrate that the excitation band at 437 nm was mainly contributed by Chl a, while the excitation band at 475.5 nm was mainly contributed by Chl b and Cars. The chlorophyll fluorescence excitation spectrum, therefore, could serve as a useful tool to describe specific characteristics of light absorption and energy transfer between light-harvesting pigments.

  20. Absorption and Fluorescence Properties of Chromophoric Dissolved Organic Matter Produced by Algae.

    PubMed

    Peng, Tong; Lu, Xiao-lan; Su, Rong-guo; Zhang, Dong-mei

    2015-09-01

    Four kinds of diatom (Chaetoceros curvisetus, Phaeodactylum tricornutum, Nitzschia closterium f. minutissima and Navicula halophile) and two kinds of dinoflagellates (Prorocentrum donghaiense and Gymnodinium) were cultured under laboratory conditions. Variations of optical properties of chromophoric dissolved organic matter (CDOM) were studied with absorption and fluorescence excitation-emission matrix spectroscopy(EEM) during growth of marine microalgae in incubation experiment. Absorption spectrum revealed absorption coefficient a(355) (CDOM absorption coefficients at 355 nm) of 6 kinds of marine microalgae above increased by 64.8%, 242.3%, 535.1%, 903.2%, 836% and 196.4%, respectively. Simultaneously, the absorption spectral slope (Sg), determined between 270 and 350 nm, representing the size of molecular weight of CDOM and humic-like composition, decreased by 8.7%, 34.6%, 39.4%, 53.1%, 46.7%, and 35.7%, respectively. Applying parallel factor analysis (PARAFAC) together with EEM got four components of CDOM: C1(Ex/Em=350(260) nm/450 nm), C2 (Ex/Em=260(430) nm/525 nm), C3 (Ex/Em=325 nm/400 nm) and C4(Ex/Em=275 nm/325 nm), which were relative to three humic-like and one protein-like fluorescent components of Nitzschia closterium f. minutissima and Navicula halophile. In incubation experiment, fluorescence intensity of these four components during growth of Nitzschia closterium f. minutissima increased by, respectively, 8.68, 24.9, 7.19 and 39.8 times, and those of Navicula halophile increased by 2.64, 0.07, 4.39 and 12.4 times, respectively. Significant relationships were found between the fluorescence intensity of four components of CDOM, a(355) and Sg. All results demonstrated that both content and molecular weight of CDOM produced by diatom and dinoflagellate studied in incubation experiment increased, but these two parameters changed more obviously of the diatom than those of dinoflagellate; the proportion of humic-like components in the composition of CDOM

  1. Absorption and Fluorescence Properties of Chromophoric Dissolved Organic Matter Produced by Algae.

    PubMed

    Peng, Tong; Lu, Xiao-lan; Su, Rong-guo; Zhang, Dong-mei

    2015-09-01

    Four kinds of diatom (Chaetoceros curvisetus, Phaeodactylum tricornutum, Nitzschia closterium f. minutissima and Navicula halophile) and two kinds of dinoflagellates (Prorocentrum donghaiense and Gymnodinium) were cultured under laboratory conditions. Variations of optical properties of chromophoric dissolved organic matter (CDOM) were studied with absorption and fluorescence excitation-emission matrix spectroscopy(EEM) during growth of marine microalgae in incubation experiment. Absorption spectrum revealed absorption coefficient a(355) (CDOM absorption coefficients at 355 nm) of 6 kinds of marine microalgae above increased by 64.8%, 242.3%, 535.1%, 903.2%, 836% and 196.4%, respectively. Simultaneously, the absorption spectral slope (Sg), determined between 270 and 350 nm, representing the size of molecular weight of CDOM and humic-like composition, decreased by 8.7%, 34.6%, 39.4%, 53.1%, 46.7%, and 35.7%, respectively. Applying parallel factor analysis (PARAFAC) together with EEM got four components of CDOM: C1(Ex/Em=350(260) nm/450 nm), C2 (Ex/Em=260(430) nm/525 nm), C3 (Ex/Em=325 nm/400 nm) and C4(Ex/Em=275 nm/325 nm), which were relative to three humic-like and one protein-like fluorescent components of Nitzschia closterium f. minutissima and Navicula halophile. In incubation experiment, fluorescence intensity of these four components during growth of Nitzschia closterium f. minutissima increased by, respectively, 8.68, 24.9, 7.19 and 39.8 times, and those of Navicula halophile increased by 2.64, 0.07, 4.39 and 12.4 times, respectively. Significant relationships were found between the fluorescence intensity of four components of CDOM, a(355) and Sg. All results demonstrated that both content and molecular weight of CDOM produced by diatom and dinoflagellate studied in incubation experiment increased, but these two parameters changed more obviously of the diatom than those of dinoflagellate; the proportion of humic-like components in the composition of CDOM

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

    PubMed

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

    2016-08-22

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

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

    PubMed Central

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

    2016-01-01

    In green-sulfur bacteria sunlight is absorbed by antenna structures termed chlorosomes, and transferred to the RC via the Fenna-Matthews-Olson (FMO) complex. FMO consists of three monomers arranged in C3 symmetry where each monomer accommodates eight Bacteriochlorophyll a (BChl a) molecules. It was the first pigment-protein complex for which the structure has been determined with high resolution and since then this complex has been the subject of numerous studies both experimentally and theoretically. Here we report about fluorescence-excitation spectroscopy as well as emission spectroscopy from individual FMO complexes at low temperatures. The individual FMO complexes are subjected to very fast spectral fluctuations smearing out any possible different information from the ensemble data that were recorded under the same experimental conditions. In other words, on the time scales that are experimentally accessible by single-molecule techniques, the FMO complex exhibits ergodic behaviour. PMID:27545197

  4. Two-color two-photon excited fluorescence of indole: Determination of wavelength-dependent molecular parameters

    SciTech Connect

    Herbrich, Sebastian; Al-Hadhuri, Tawfik; Gericke, Karl-Heinz; Shternin, Peter S. Vasyutinskii, Oleg S.; Smolin, Andrey G.

    2015-01-14

    We present a detailed study of two-color two-photon excited fluorescence in indole dissolved in propylene glycol. Femtosecond excitation pulses at effective wavelengths from 268 to 293.33 nm were used to populate the two lowest indole excited states {sup 1}L{sub a} and {sup 1}L{sub b} and polarized fluorescence was then detected. All seven molecular parameters and the two-photon polarization ratio Ω containing information on two-photon absorption dynamics, molecular lifetime τ{sub f}, and rotation correlation time τ{sub rot} have been determined from experiment and analyzed as a function of the excitation wavelength. The analysis of the experimental data has shown that {sup 1}L{sub b}–{sup 1}L{sub a} inversion occurred under the conditions of our experiment. The two-photon absorption predominantly populated the {sup 1}L{sub a} state at all excitation wavelengths but in the 287–289 nm area which contained an absorption hump of the {sup 1}L{sub b} state 0-0 origin. The components of the two-photon excitation tensor S were analyzed giving important information on the principal tensor axes and absorption symmetry. The results obtained are in a good agreement with the results reported by other groups. The lifetime τ{sub f} and the rotation correlation time τ{sub rot} showed no explicit dependence on the effective excitation wavelength. Their calculated weighted average values were found to be τ{sub f} = 3.83 ± 0.14 ns and τ{sub rot} = 0.74 ± 0.06 ns.

  5. Fluorescent Ag nanoclusters prepared in aqueous poly(acrylic acid-co-maleic acid) solutions: a spectroscopic study of their excited state dynamics, size and local environment.

    PubMed

    Dandapat, Manika; Mandal, Debabrata

    2016-01-28

    Stable, fluorescent Ag nanoclusters were prepared in aqueous solutions of Na(+) salt of the carboxylate-rich polymer poly(acrylic acid-co-maleic acid) under brief spells of UV irradiation. The nanoclusters were nearly spherical, with diameters within 1.90 ± 0.50 nm, but displayed a prominent red edge excitation shift (REES) of fluorescence upon exciting within the visible absorption band, indicating heterogeneity of energy level distributions. Spectroscopic studies revealed that irrespective of whether the nanoclusters are excited in their UV or visible absorption bands, their fluorescence always ensues from the same manifold of emissive states, with a broad range of fluorescence lifetimes from ∼150 fs to 1 ns. PMID:26700465

  6. Excited state absorption of pump radiation as a loss mechanism in solid-state lasers

    SciTech Connect

    Kliewer, M.L.; Powell, R.C.

    1989-08-01

    The characteristics of optical pumping dynamics occurring in laser-pumped rare earth-doped, solid-state laser materials were investigated by using a tunable alexandrite laser to pump Y/sub 3/Al/sub 5/O/sub 12/:Nd/sup 3+/ in an optical cavity. It was found that the slope efficiency of the Nd laser operation depends strongly on the wavelength of the pump laser. For pump wavelengths resulting in low slope efficiencies, intense fluorescence emission is observed from the sample in the blue-green spectral region. This is attributed to the excited state absorption of pump photons which occurs during radiationless relaxation from the pump band to the metastable state. This type of process will be an important loss mechanism for monochromatic pumping of laser systems at specific pump wavelengths.

  7. Excited-state absorption of pump radiation as a loss mechanism in solid-state lasers

    SciTech Connect

    Kliewer, M.L.; Powell, R.C.

    1989-08-01

    The characteristics of optical pumping dynamics occuring in laser-pumped rare earth-doped, solid-state laser materials were investigated by using a tunable alexandrite laser to pump Y3Al5O12:Nd(3+) in an optical cavity. It was found that the slope efficiency of the Nd laser operation depends strongly on the wavelength of the pump laser. For pump wavelength resulting in low slope efficiencies, intense fluorescence emission is observed form the sample in the blue-green spectral region. This is attributed to the excited state absorption of pump photons which occurs during radiationless relaxation from the pump band to the metastable state. This type of process will be an important loss mechanism for monochromatic pumping of laser systems at specific pump wavelengths.

  8. Excited state absorption of pump radiation as a loss mechanism in solid-state lasers

    NASA Technical Reports Server (NTRS)

    Kliewer, Michael L.; Powell, Richard C.

    1989-01-01

    The characteristics of optical pumping dynamics in laser-pumped, rare-earth-doped, solid-state laser materials are investigated by using a tunable alexandrite laser to pump Y3Al5O12:Nd(3+) in an optical cavity. It is found that the slope efficiency of the Nd laser operation depends strongly on the wavelength of the pump laser. For pump wavelengths resulting in low slope efficiencies, intense fluorescence emission is observed from the sample in the blue-green spectral region. This is attributed to the excited-state absorption of pump photons which occurs during radiationless relaxation from the pump band to the metastable state. This type of process is an important loss mechanism for monochromatic pumping of laser systems at specific pump wavelengths.

  9. Solvent dependence of two-photon absorption spectra of the enhanced green fluorescent protein (eGFP) chromophore

    NASA Astrophysics Data System (ADS)

    Hosoi, Haruko; Tayama, Ryo; Takeuchi, Satoshi; Tahara, Tahei

    2015-06-01

    Two-photon absorption spectra of 4‧-hydroxybenzylidene-2,3-dimethylimidazolinone, a model chromophore of enhanced green fluorescent protein (eGFP), were measured in various solvents. The two-photon absorption band of its anionic form is markedly blue-shifted from the corresponding one-photon absorption band in all solvents. Moreover, the magnitude of the blue shift varies largely depending on the solvent, which does not accord with the assignment of the two-photon absorption band to the transitions to the vibrationally excited S1 state. Our finding is readily rationalized by considering overlapping contributions of the S1 ← S0 and S2 ← S0 transitions, suggesting the involvement of the S2 state also in two-photon fluorescence of eGFP.

  10. Two-color excitation system for fluorescence detection in DNA sequencing by capillary array electrophoresis.

    PubMed

    Xue, Gang; Yeung, Edward S

    2002-05-01

    Two computer-controlled galvanometer scanners are adapted for two-dimensional step scanning across a 96-capillary array for laser-induced fluorescence detection. 488 nm and 514 nm laser lines from the same Ar(+) laser were alternately coupled for two-color excitation in each capillary. The signal at a single photomultiplier tube is temporally sorted to distinguish among the capillaries and the excitation wavelengths. Based on the differences in absorption spectra for the dyes, the peak-height ratios in the 488 nm and 514 nm excitation electropherograms were used for peak identification for multiplexed capillary electrophoresis. Successful base calling for 24-capillary DNA sequencing was achieved to 450 bp with 99% accuracy. Advantages include the efficient utilization of light due to the high duty-cycle of step scan, good detection performance due to the reduction of stray light, ruggedness due to the small mass of the galvanometer mirror, low cost due to the simplicity of components and flexibility due to the independent paths for excitation and emission. PMID:12116160

  11. Two-photon excited fluorescence enhancement with broadband versus tunable femtosecond laser pulse excitation

    NASA Astrophysics Data System (ADS)

    Wang, Chao; Yeh, Alvin T.

    2012-02-01

    The inverse relationship between two-photon excited fluorescence (TPEF) and laser pulse duration suggests that two-photon microscopy (TPM) performance may be improved by decreasing pulse duration. However, for ultrashort pulses of sub-10 femtosecond (fs) in duration, its spectrum contains the effective gain bandwidth of Ti:Sapphire and its central wavelength is no longer tunable. An experimental study was performed to explore this apparent tradeoff between untuned sub-10 fs transform-limited pulse (TLP) and tunable 140 fs pulse for TPEF. Enhancement factors of 1.6, 6.7, and 5.2 are measured for Indo-1, FITC, and TRITC excited by sub-10 fs TLP compared with 140 fs pulse tuned to the two-photon excitation (TPE) maxima at 730 nm, 800 nm, and 840 nm, respectively. Both degenerate (v1=v2) and nondegenerate (v1≠v2) mixing of sub-10 fs TLP spectral components result in its broad second-harmonic (SH) power spectrum and high spectral density, which can effectively compensate for the lack of central wavelength tuning and lead to large overlap with dye TPE spectra for TPEF enhancements. These pulse properties were also exploited for demonstrating its potential applications in multicolor imaging with TPM.

  12. The fluorescence action spectra of some saturated hydrocarbon liquids for excitation energies above and below their ionization thresholds

    SciTech Connect

    Ostafin, A.E.; Lipsky, S. )

    1993-04-01

    Fluorescence action spectra have been obtained for the neat liquids, [ital cis]-decalin, [ital trans]-decalin, bicyclohexyl, cyclohexane, methylcyclohexane, isobutylcyclohexane, 2,3,4-trimethylpentane, 2,3-dimethylbutane, 3-methylhexane, 3-methylpentane, [ital n]-decane, [ital n]-dodecane, and [ital n]-pentadecane at excitation energies, [epsilon], ranging from their absorption onsets (at ca. 7 eV) to 10.3 eV. For all compounds, with the exception of [ital cis]-decalin, the fluorescence quantum yield is observed to monotonically decline with increasing [epsilon], reaching a minimum value at an energy, [epsilon][sub [ital m

  13. Two-photon fluorescence absorption and emission spectra of dyes relevant for cell imaging.

    PubMed

    Bestvater, F; Spiess, E; Stobrawa, G; Hacker, M; Feurer, T; Porwol, T; Berchner-Pfannschmidt, U; Wotzlaw, C; Acker, H

    2002-11-01

    Two-photon absorption and emission spectra for fluorophores relevant in cell imaging were measured using a 45 fs Ti:sapphire laser, a continuously tuneable optical parametric amplifier for the excitation range 580-1150 nm and an optical multichannel analyser. The measurements included DNA stains, fluorescent dyes coupled to antibodies as well as organelle trackers, e.g. Alexa and Bodipy dyes, Cy2, Cy3, DAPI, Hoechst 33342, propidium iodide, FITC and rhodamine. In accordance with the two-photon excitation theory, the majority of the investigated fluorochromes did not reveal significant discrepancies between the two-photon and the one-photon emission spectra. However, a blue-shift of the absorption maxima ranging from a few nanometres up to considerably differing courses of the spectrum was found for most fluorochromes. The potential of non-linear laser scanning fluorescence microscopy is demonstrated here by visualizing multiple intracellular structures in living cells. Combined with 3D reconstruction techniques, this approach gives a deeper insight into the spatial relationships of subcellular organelles. PMID:12423261

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

    SciTech Connect

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

    1986-05-22

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

  15. Energy level decay and excited state absorption processes in erbium-doped tellurite glass

    NASA Astrophysics Data System (ADS)

    Gomes, Laércio; Oermann, Michael; Ebendorff-Heidepriem, Heike; Ottaway, David; Monro, Tanya; Felipe Henriques Librantz, André; Jackson, Stuart D.

    2011-10-01

    The fundamental excited state decay processes relating to the 4I11/2 → 4I13/2 transition in singly Er3+-doped tellurite (TZNL) glass have been investigated in detail using time-resolved fluorescence spectroscopy. Selective laser excitation of the 4I11/2 energy level at 970 nm and selective laser excitation of the 4I13/2 energy level at 1485 nm has established that energy transfer upconversion by way of a dipole-dipole interaction between two excited erbium ions in the 4I13/2 level populates the 4I11/2 upper laser level of the 3 μm transition. This upconversion has been analyzed for Er2O3 concentrations between 0.5 mol. % and 2.2 mol. %. The 4I13/2 and 4I11/2 energy levels emit luminescence with peaks located at 1532 nm and 2734 nm, respectively, with radiative decay efficiencies of 65% and 6.8% for the higher (2.2 mol. %) concentration sample. The low 2.7 μm emission efficiency is due to the non-radiative decay bridging the 4I11/2 → 4I13/2 transition and energy transfer to the OH- groups in the glass. Excited state absorption was observed to occur from the 4I13/2 and 4I11/2 levels with peak absorptions occurring at 1550 nm and 971 nm, respectively. The decay time of the 4I11/2 excited state decreased with an increase in the Er3+ concentration, which related to energy transfer to OH- ions that had a measured concentration of 6.6 × 1018 cm-3. Results from numerical simulations showed that a population inversion is reached at a threshold pumping intensity of ˜80 kW cm-2 for a cw laser pump at 976 nm if [Er3+] ≥ 1.2 × 1021 cm-3 (or [Er2O3] ≥ 2.65 mol. %) without OH- impurities being present.

  16. Experimental determination and interpretation of the fluorescence and fluorescence excitation spectra of chrysene cooled in a supersonic jet

    NASA Astrophysics Data System (ADS)

    Borisevich, N. A.; Dyachenko, G. G.; Petukhov, V. A.; Semenov, M. A.

    2008-12-01

    The fluorescence and fluorescence excitation spectra of jet-cooled chrysene are measured. The frequencies of in-plane vibrations in the ground and first excited singlet electronic states, as well as the relative intensities of transitions between them, are calculated with the MO/M8ST method. Based on these data, experimental spectra are interpreted. In the fluorescence excitation spectrum, the position of the line of the 0-0 transition (28 195 ± 1 cm-1), which is the most intense, is determined. In the experimental fluorescence excitation spectrum, 21 lines correspond to fundamental vibrations (altogether, 37 lines are attributed). This supports our assignment and is consistent with the group-theoretical analysis of vibronic interactions. Upon excitation at the frequency of the 0-0 transition, 10 lines corresponding to the excitation of fundamental vibrations are detected, and all 17 lines observed are attributed. In the fluorescence excitation spectrum, the standard deviation between the calculated and measured frequencies of attributed fundamental vibrations is 19 cm-1, while that in the fluorescence spectrum is 15 cm-1.

  17. [Influence of the Experiment Energy Dispersive X-Ray Fluorescence Measurement of Uranium by Different Excitation Source].

    PubMed

    Xiong, Chao; Ge, Liang-quan; Liu, Duan; Zhang, Qing-xian; Gu, Yi; Luo, Yao-yao; Zhao, Jian-kun

    2016-03-01

    Aiming at the self-excitation effect on the interference of measurements which exist in the process of Energy dispersive X-ray fluorescence method for uranium measurement. To solve the problem of radioactive isotopes only used as excitation source in determination of uranium. Utilizing the micro X-ray tube to test Self-excitation effect to get a comparison of the results obtained by three different uranium ore samples--109 Cd, 241 Am and Mirco X-ray tube. The results showed that self-excitation effect produced the area measure of characteristic X-ray peak is less than 1% of active condition, also the interference of measurements can be negligible. Photoelectric effect cross-section excited by 109 Cd is higher, corresponding fluorescence yield is higher than excited by 241 Am as well due to characteristics X-ray energy of 109 Cd, 22.11 & 24.95 KeV adjacent to absorption edge energy of L(α), 21.75 KeV, based on the above, excitation efficiency by 109 Cd is higher than 241 Am; The fact that measurement error excited by 241 Am is significantly greater than by 109 Cd is mainly due to peak region overlap between L energy peaks of uranium and Scattering peak of 241 Am, 26.35 keV, These factors above caused the background of measured Spectrum higher; The error between the uranium content in ore samples which the X-ray tube as the excitation source and the chemical analysis results is within 10%. Conclusion: This paper come to the conclusion that the technical quality of uranium measurement used X-ray tube as excitation source is superior to that in radioactive source excitation mode. PMID:27400534

  18. [Influence of the Experiment Energy Dispersive X-Ray Fluorescence Measurement of Uranium by Different Excitation Source].

    PubMed

    Xiong, Chao; Ge, Liang-quan; Liu, Duan; Zhang, Qing-xian; Gu, Yi; Luo, Yao-yao; Zhao, Jian-kun

    2016-03-01

    Aiming at the self-excitation effect on the interference of measurements which exist in the process of Energy dispersive X-ray fluorescence method for uranium measurement. To solve the problem of radioactive isotopes only used as excitation source in determination of uranium. Utilizing the micro X-ray tube to test Self-excitation effect to get a comparison of the results obtained by three different uranium ore samples--109 Cd, 241 Am and Mirco X-ray tube. The results showed that self-excitation effect produced the area measure of characteristic X-ray peak is less than 1% of active condition, also the interference of measurements can be negligible. Photoelectric effect cross-section excited by 109 Cd is higher, corresponding fluorescence yield is higher than excited by 241 Am as well due to characteristics X-ray energy of 109 Cd, 22.11 & 24.95 KeV adjacent to absorption edge energy of L(α), 21.75 KeV, based on the above, excitation efficiency by 109 Cd is higher than 241 Am; The fact that measurement error excited by 241 Am is significantly greater than by 109 Cd is mainly due to peak region overlap between L energy peaks of uranium and Scattering peak of 241 Am, 26.35 keV, These factors above caused the background of measured Spectrum higher; The error between the uranium content in ore samples which the X-ray tube as the excitation source and the chemical analysis results is within 10%. Conclusion: This paper come to the conclusion that the technical quality of uranium measurement used X-ray tube as excitation source is superior to that in radioactive source excitation mode.

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

    SciTech Connect

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

    2009-08-20

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

  20. B{sub 2u} {yields} A{sub 1g} fluorescence excitation spectrum of liquid benzene from 250 to 150 nm

    SciTech Connect

    Saik, V.O.; Lipsky, S.

    1995-06-29

    The B{sub 2u} {yields} A{sub 1g} fluorescence quantum yield of benzene has been obtained for excitations from 250 to 150 nm in the solvents n-hexane, isooctane, n-perfluorohexane, n-pentane, cyclohexane, and cis-decalin. In n-hexane and isooctane, measurements have been made over a range of benzene concentrations from 0.11{sub 2} to 11.2 M (the neat liquid). In dilute solutions, the sudden losses in fluorescence as the excitation wavelength sweeps from the first into the second absorption system and then again as the excitation wavelength sweeps from the second into the third absorption system are correlated with the behavior of benzene vapor in the region of channel three. Increasing the benzene concentration causes abatements in the rate of loss of fluorescence with decreasing excitation wavelength in the regions of the second and third absorption systems and, above 2 M, causes a reversal of this rate so as to cause a recovery of the fluorescence in the region of the third absorption system. These changes are correlated with the effects of benzene concentration on the electronic absorption spectrum and with reported effects of clustering on the multiphoton photoionization efficiency. 59 refs., 13 figs.

  1. Ultrafast Laser Studies of Two-Photon Excited Fluorescence Intermittency in Single CdSe/ZnS Quantum Dots.

    PubMed

    Early, Kevin T; Nesbitt, David J

    2015-12-01

    Two-photon fluorescence microscopy of single quantum dots conditions has been reported by several groups, with contrasting observations regarding the kinetics and dynamics of fluorescence intermittency or "blinking". Here, we investigate the power dependence, kinetics, and statistics of two photon-excited fluorescence intermittency from single CdSe/ZnS quantum dots in a solid PMMA film as a function of sub-bandgap laser intensity at 800 nm. Fluorescence intermittency is observed at all excitation powers and a quadratic (n = 1.97(3)) dependence of the shot noise-limited fluorescence intensity on the incident laser power is verified, confirming essentially zero background contribution from one-photon excitation processes. Such analyses permit two photon absorption cross sections for single quantum dots to be extracted quantitatively from the data, which reveal good agreement with those obtained from previous two-photon FCS measurements. Strictly inverse power law-distributed off-state dwell times are observed for all excitation powers, with a mean power law exponent ⟨m(off)⟩ = 1.65(4) in excellent agreement with the behavior observed under one-photon excitation conditions. Finally, a superquadratic (n = 2.3(2)) rather than quartic (n = 4) power dependence is observed for the on-state blinking dwell times, which we kinetically analyze and interpret in terms of a novel 2 + 1 "hot" exciton ionization/blinking mechanism due to partially saturated 1-photon sub-bandgap excitation out of the two-photon single exciton state. The kinetic results are consistent with quantum dot photoionization quantum yields from "hot" exciton states (4(1) × 10(-6)) comparable with experimental estimates (10(-6)-10(-5)) of Auger ionization efficiencies out of the biexcitonic state.

  2. Influence of absorption and scattering on the quantification of fluorescence diffuse optical tomography using normalized data.

    PubMed

    Abascal, Juan Felipe Perez-Juste; Aguirre, Juan; Chamorro-Servent, Judit; Schweiger, Martin; Arridge, Simon; Ripoll, Jorge; Vaquero, Juan J; Desco, Manuel

    2012-03-01

    Reconstruction algorithms for imaging fluorescence in near infrared ranges usually normalize fluorescence light with respect to excitation light. Using this approach, we investigated the influence of absorption and scattering heterogeneities on quantification accuracy when assuming a homogeneous model and explored possible reconstruction improvements by using a heterogeneous model. To do so, we created several computer-simulated phantoms: a homogeneous slab phantom (P1), slab phantoms including a region with a two- to six-fold increase in scattering (P2) and in absorption (P3), and an atlas-based mouse phantom that modeled different liver and lung scattering (P4). For P1, reconstruction with the wrong optical properties yielded quantification errors that increased almost linearly with the scattering coefficient while they were mostly negligible regarding the absorption coefficient. This observation agreed with the theoretical results. Taking the quantification of a homogeneous phantom as a reference, relative quantification errors obtained when wrongly assuming homogeneous media were in the range +41 to +94% (P2), 0.1 to -7% (P3), and -39 to +44% (P4). Using a heterogeneous model, the overall error ranged from -7 to 7%. In conclusion, this work demonstrates that assuming homogeneous media leads to noticeable quantification errors that can be improved by adopting heterogeneous models.

  3. Influence of absorption and scattering on the quantification of fluorescence diffuse optical tomography using normalized data

    NASA Astrophysics Data System (ADS)

    Abascal, Juan Felipe Perez-Juste; Aguirre, Juan; Chamorro-Servent, Judit; Schweiger, Martin; Arridge, Simon; Ripoll, Jorge; Vaquero, Juan J.; Desco, Manuel

    2012-03-01

    Reconstruction algorithms for imaging fluorescence in near infrared ranges usually normalize fluorescence light with respect to excitation light. Using this approach, we investigated the influence of absorption and scattering heterogeneities on quantification accuracy when assuming a homogeneous model and explored possible reconstruction improvements by using a heterogeneous model. To do so, we created several computer-simulated phantoms: a homogeneous slab phantom (P1), slab phantoms including a region with a two- to six-fold increase in scattering (P2) and in absorption (P3), and an atlas-based mouse phantom that modeled different liver and lung scattering (P4). For P1, reconstruction with the wrong optical properties yielded quantification errors that increased almost linearly with the scattering coefficient while they were mostly negligible regarding the absorption coefficient. This observation agreed with the theoretical results. Taking the quantification of a homogeneous phantom as a reference, relative quantification errors obtained when wrongly assuming homogeneous media were in the range +41 to +94% (P2), 0.1 to -7% (P3), and -39 to +44% (P4). Using a heterogeneous model, the overall error ranged from -7 to 7%. In conclusion, this work demonstrates that assuming homogeneous media leads to noticeable quantification errors that can be improved by adopting heterogeneous models.

  4. Laser excited fluorescence in the cesium-xenon excimer and the cesium dimer

    NASA Technical Reports Server (NTRS)

    Exton, R. J.; Snow, W. L.; Hillard, M. E.

    1978-01-01

    Argon ion laser lines are used to excite fluorescence in a mixture of cesium and xenon. Excimer band fluorescence is observed at higher pressures (about 1 atm) while at lower pressures (several torr) a diffuse fluorescence due to the cesium dimer is observed whose character changes with exciting wavelength. The excimer fluorescence is shown to be directly related to the location of the exciting wavelength within previously measured Cs/Xe line shapes. This fact suggests that the excimer systems may be efficiently pumped through these line shapes. Qualitative energy-level schemes are proposed to explain the observations in both the excimer and dimer systems.

  5. Spectral fluorescent properties of tissues in vivo with excitation in the red wavelength range

    NASA Astrophysics Data System (ADS)

    Stratonnikov, Alexander A.; Loschenov, Victor B.; Klimov, D. V.; Edinac, N. E.; Wolnukhin, V. A.; Strashkevich, I. A.

    1997-12-01

    The spectral fluorescence analysis is a promising method for differential tissue diagnostic. Usually the UV and visible light is used for fluorescence excitation with emission registration in the visible wavelength range. The light penetration length in this wavelength range is very small allowing one to analyze only the surface region of the tissue. Here we present the tissue fluorescent spectra in vivo excited in the red wavelength region. As excitation light source we used compact He-Ne laser (632.8 nm) and observed the fluorescence in 650 - 800 nm spectral range. The various tissues including normal skin, psoriasis, tumors, necrosis as well as photosensitized tissues have been measured.

  6. Twisting in the excited state of an N-methylpyridinium fluorescent dye modulated by nano-heterogeneous micellar systems.

    PubMed

    Cesaretti, A; Carlotti, B; Gentili, P L; Germani, R; Spalletti, A; Elisei, F

    2016-04-01

    A push-pull N-methylpyridinium fluorescent dye with a pyrenyl group as the electron-donor portion was investigated within the nano-heterogeneous media provided by some micellar systems. The molecule was studied by stationary and time-resolved spectroscopic techniques in spherical micellar solutions and viscoelastic hydrogels, in order to throw light on the role played by twisting in its excited state deactivation. As proven by femtosecond fluorescence up-conversion and transient absorption experiments, the excited state dynamics of the molecule is ruled by charge transfer and twisting processes, which, from the locally excited (LE) state initially populated upon excitation, progressively lead to twisted (TICT) and planar (PICT) intramolecular charge transfer states. The inclusion within micellar aggregates was found to slow down and/or limit the rotation of the molecule with respect to what had previously been observed in water, while its confinement within the hydrophobic domains of the gel matrixes prevents any molecular torsion. The increasing viscosity of the medium, when passing from water to micellar systems, implies that the detected steady-state fluorescence comes from an excited state which is not fully relaxed, as is the case with the TICT state in micelles or the LE state in hydrogels, where the detected emission changes its usual orange colour to yellow.

  7. Laser excited confocal microscope fluorescence scanner and method

    DOEpatents

    Mathies, Richard A.; Peck, Konan

    1992-01-01

    A fluorescent scanner for scanning the fluorescence from a fluorescence labeled separated sample on a sample carrier including a confocal microscope for illuminating a predetermined volume of the sample carrier and/or receiving and processing fluorescence emissions from said volume to provide a display of the separated sample.

  8. Laser excited confocal microscope fluorescence scanner and method

    DOEpatents

    Mathies, R.A.; Peck, K.

    1992-02-25

    A fluorescent scanner is designed for scanning the fluorescence from a fluorescence labeled separated sample on a sample carrier. The scanner includes a confocal microscope for illuminating a predetermined volume of the sample carrier and/or receiving and processing fluorescence emissions from the volume to provide a display of the separated sample. 8 figs.

  9. Excited state absorption spectrum of chlorophyll a obtained with white-light continuum.

    PubMed

    De Boni, L; Correa, D S; Pavinatto, F J; dos Santos, D S; Mendonça, C R

    2007-04-28

    The study of excited state properties of chlorophyll a is a subject of foremost interest, given that it plays important roles in biological process and has also been proposed for applications in photonics. This work reports on the excited state absorption spectrum of chlorophyll a solution from 460 to 700 nm, obtained through the white-light continuum Z-scan technique. Saturation of absorption was observed due to the ground state depletion, induced by the white-light continuum region that is resonant with the Q band of chlorophyll a. The authors also observed reverse saturation of absorption related to the excitation from the first excited state to a higher energy level for wavelengths below 640 nm. An energy-level diagram, based on the electronic states of chlorophyll a, was employed to interpret their results, revealing that more states than the ones related to the Q and B bands participate in the excited state absorption of this molecule.

  10. Laser-induced fluorescence of formaldehyde in combustion using third harmonic Nd:YAG laser excitation.

    PubMed

    Brackmann, Christian; Nygren, Jenny; Bai, Xiao; Li, Zhongshan; Bladh, Henrik; Axelsson, Boman; Denbratt, Ingemar; Koopmans, Lucien; Bengtsson, Per-Erik; Aldén, Marcus

    2003-12-01

    Formaldehyde (CH2O) is an important intermediate species in combustion processes and it can through laser-induced fluorescence measurements be used for instantaneous flame front detection. The present study has focussed on the use of the third harmonic of a Nd:YAG laser at 355 nm as excitation wavelength for formaldehyde, and different dimethyl ether (C2H6O) flames were used as sources of formaldehyde in the experiments. The investigations included studies of the overlap between the laser profile and the absorption lines of formaldehyde, saturation effects and the potential occurrence of laser-induced photochemistry. The technique was applied for detection of formaldehyde in an internal combustion engine operated both as a spark ignition engine and as a homogenous charge compression ignition engine.

  11. Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood

    PubMed Central

    Saytashev, Ilyas; Glenn, Rachel; Murashova, Gabrielle A.; Osseiran, Sam; Spence, Dana; Evans, Conor L.; Dantus, Marcos

    2016-01-01

    Red blood cells (RBC) in two-photon excited fluorescence (TPEF) microscopy usually appear as dark disks because of their low fluorescent signal. Here we use 15fs 800nm pulses for TPEF, 45fs 1060nm pulses for three-photon excited fluorescence, and third harmonic generation (THG) imaging. We find sufficient fluorescent signal that we attribute to hemoglobin fluorescence after comparing time and wavelength resolved spectra of other expected RBC endogenous fluorophores: NADH, FAD, biliverdin, and bilirubin. We find that both TPEF and THG microscopy can be used to examine erythrocyte morphology non-invasively without breaching a blood storage bag. PMID:27699111

  12. Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood

    PubMed Central

    Saytashev, Ilyas; Glenn, Rachel; Murashova, Gabrielle A.; Osseiran, Sam; Spence, Dana; Evans, Conor L.; Dantus, Marcos

    2016-01-01

    Red blood cells (RBC) in two-photon excited fluorescence (TPEF) microscopy usually appear as dark disks because of their low fluorescent signal. Here we use 15fs 800nm pulses for TPEF, 45fs 1060nm pulses for three-photon excited fluorescence, and third harmonic generation (THG) imaging. We find sufficient fluorescent signal that we attribute to hemoglobin fluorescence after comparing time and wavelength resolved spectra of other expected RBC endogenous fluorophores: NADH, FAD, biliverdin, and bilirubin. We find that both TPEF and THG microscopy can be used to examine erythrocyte morphology non-invasively without breaching a blood storage bag.

  13. Large edge-excitation red-shift of the fluorescence of an ethanol solution of 4-amino-4'-nitrodiphenyl (and). The case of a solute with a dipole moment increase in the excited state

    NASA Astrophysics Data System (ADS)

    Al-Hassan, Khader A.; El-Bayoumi, M. Ashraf

    1987-08-01

    The observed large edge-excitation red-shift of the fluorescence of 4-amino-4'-nitrodiphenyl (AND) in a rigid ethanol medium and the red-shift of the absorption band as the temperature is lowered are explained in terms of a microscopic solvent heterogeneity. The AND results where the dipole moment is increased upon excitation are compared with the merocyanine dye results where the reverse is true.

  14. Excitation of emission lines by fluorescence and recombination in IC 418

    NASA Astrophysics Data System (ADS)

    Escalante, V.; Morisset, C.; Georgiev, L.

    2012-11-01

    We compare calculated intensities of lines of C II, N I, N II, O I and O II with a published deep spectroscopic survey of IC 418. Our calculations use a self-consistent nebular model and a synthetic spectrum of the central star atmosphere to take into account line excitation by continuum fluorescence and electron recombination. We found that the N II spectrum of the s, p and most d states is excited by fluorescence due to the low-excitation conditions of the nebula. Many C II and O II lines have significant amount of excitation by fluorescence. Recombination excites all the lines from the f and g states and most O II lines. In the neutral-ionized boundary, the N I quartet and O I triplet dipole-allowed lines are excited by fluorescence, while the quintet O I lines are excited by recombination. Electron excitation produces the forbidden optical lines of O I, and continuum fluorescence enhances the N I forbidden line intensities. Lines excited by fluorescence of light below the Lyman limit thus suggest a new diagnostic to explore the inner boundary of the photodissociation region of the nebula.

  15. Absorption, fluorescence, and Raman spectra of mass-selected rhenium dimers in argon matrices

    NASA Astrophysics Data System (ADS)

    Hu, Zhendong; Dong, Jian-Guo; Lombardi, John R.; Lindsay, D. M.; Harbich, W.

    1994-07-01

    We report absorption, laser fluorescence, and Raman spectra for Re2 in an argon matrix prepared by the mass-selected ion deposition technique. The dirhenium absorption spectrum consists of seven band systems (A-G) extending from the near infrared into the ultraviolet region. For the A system (a simple vibrational progression), we find T0=10 817(1) cm-1, ωe=317.1(5) cm-1 and ωexe=1.0(1) cm-1. A Franck-Condon analysis of the A system intensities predicts that this state has a smaller equilibrium internuclear distance than the ground state (Δre=-0.073 Å), in violation of Badger's rule. The B system starts at 13 250 cm-1 and consists of four overlapping (and possibly perturbed) subsystems, whose average vibrational spacing is 270(11) cm-1. The C, D, E, and F systems (vibrational spacings in parentheses) are centered at 22 300 cm-1 (210 cm-1), 24 500 cm-1 (195 cm-1), 29 150 cm-1 (175 cm-1), and 32 900 cm-1 (160 cm-1), respectively. Weak fluorescence spectra, obtained upon laser excitation into the A system, were characterized by vibrational progressions to the dimer ground (X) state and to a low lying (X') state for which T0=357.6(5) cm-1 and ωe=332.3(2) cm-1. Raman and fluorescence progressions to the ground state were observed when the B system was excited. These data give ωe=337.9(49) cm-1 for the dimer ground state in good agreement with measurements from photodetachment spectra [J. Am. Chem. Soc. 108, 178 (1986)]. We propose likely assignments for the low lying electronic states of Re2 and discuss our results in terms of the bonding in the other group VIIB dimers, Mn2 and Tc2.

  16. Excited State Absorption from Real-Time Time-Dependent Density Functional Theory.

    PubMed

    Fischer, Sean A; Cramer, Christopher J; Govind, Niranjan

    2015-09-01

    The optical response of excited states is a key property used to probe photophysical and photochemical dynamics. Additionally, materials with a large nonlinear absorption cross-section caused by two-photon (TPA) and excited state absorption (ESA) are desirable for optical limiting applications. The ability to predict the optical response of excited states would help in the interpretation of transient absorption experiments and aid in the search for and design of optical limiting materials. We have developed an approach to obtain excited state absorption spectra by combining real-time (RT) and linear-response (LR) time-dependent density functional theory (TDDFT). Being based on RT-TDDFT, our method is aimed at tackling larger molecular complexes and materials systems where excited state absorption is predominantly seen and many time-resolved experimental efforts are focused. To demonstrate our method, we have calculated the ground and excited state spectra of H₂⁺ and H₂ due to the simplicity in the interpretation of the spectra. We have validated our new approach by comparing our results for butadiene with previously published results based on quadratic response (QR). We also present results for oligofluorenes, where we compare our results with both QR-TDDFT and experimental measurements. Because our method directly measures the response of an excited state, stimulated emission features are also captured; although, these features are underestimated in energy which could be attributed to a change of the reference from the ground to the excited state.

  17. Metal-enhanced fluorescence and FRET on nanohole arrays excited at angled incidence.

    PubMed

    Poirier-Richard, H-P; Couture, M; Brule, T; Masson, J-F

    2015-07-21

    The influence of experimental parameters on the performance of plasmonic sensors is of great importance in analytical sciences. The plasmon coupling conditions (angle of incidence, metal composition, laser frequency and excitation/emission properties of fluorophores) were thus investigated for surface plasmon-enhanced fluorescence on metallic nanohole arrays. Optimal fluorescence enhancements were achieved when the plasmon resonance, the excitation laser and the fluorophore's excitation wavelengths were matched. The enhancement of the acceptor emission of a rhodamine 6G(Rh6G)-Quasar670™ FRET pair was achieved on the nanohole arrays by tuning the plasmon wavelength with the maximal overlap of the donor's emission and acceptor excitation. Silver nanohole arrays achieved larger fluorescence enhancement than gold nanohole arrays at 532 nm, while gold nanohole arrays led to larger fluorescence enhancement at 635 nm. These results demonstrate the importance of tuning the plasmon coupling conditions for surface plasmon-enhanced fluorescence sensing.

  18. Determination of the Residual Anthracene Concentration in Cultures of Haloalkalitolerant Actinomycetes by Excitation Fluorescence, Emission Fluorescence, and Synchronous Fluorescence: Comparative Study

    PubMed Central

    Lara-Severino, Reyna del Carmen; Camacho-López, Miguel Ángel; García-Macedo, Jessica Marlene; Gómez-Oliván, Leobardo M.; Sandoval-Trujillo, Ángel H.; Isaac-Olive, Keila; Ramírez-Durán, Ninfa

    2016-01-01

    Polycyclic aromatic hydrocarbons (PAHs) are compounds that can be quantified by fluorescence due to their high quantum yield. Haloalkalitolerant bacteria tolerate wide concentration ranges of NaCl and pH. They are potentially useful in the PAHs bioremediation of saline environments. However, it is known that salinity of the sample affects fluorescence signal regardless of the method. The objective of this work was to carry out a comparative study based on the sensitivity, linearity, and detection limits of the excitation, emission, and synchronous fluorescence methods, during the quantification of the residual anthracene concentration from the following haloalkalitolerant actinomycetes cultures Kocuria rosea, Kocuria palustris, Microbacterium testaceum, and 4 strains of Nocardia farcinica, in order to establish the proper fluorescence method to study the PAHs biodegrading capacity of haloalkalitolerant actinobacteria. The study demonstrated statistical differences among the strains and among the fluorescence methods regarding the anthracene residual concentration. The results showed that excitation and emission fluorescence methods performed very similarly but sensitivity in excitation fluorescence is slightly higher. Synchronous fluorescence using Δλ = 150 nm is not the most convenient method. Therefore we propose the excitation fluorescence as the fluorescence method to be used in the study of the PAHs biodegrading capacity of haloalkalitolerant actinomycetes. PMID:26925294

  19. Multiphoton excitation fluorescence microscopy and spectroscopy of in vivo human skin.

    PubMed Central

    Masters, B R; So, P T; Gratton, E

    1997-01-01

    Multiphoton excitation microscopy at 730 nm and 960 nm was used to image in vivo human skin autofluorescence from the surface to a depth of approximately 200 microm. The emission spectra and fluorescence lifetime images were obtained at selected locations near the surface (0-50 microm) and at deeper depths (100-150 microm) for both excitation wavelengths. Cell borders and cell nuclei were the prominent structures observed. The spectroscopic data suggest that reduced pyridine nucleotides, NAD(P)H, are the primary source of the skin autofluorescence at 730 nm excitation. With 960 nm excitation, a two-photon fluorescence emission at 520 nm indicates the presence of a variable, position-dependent intensity component of flavoprotein. A second fluorescence emission component, which starts at 425 nm, is observed with 960-nm excitation. Such fluorescence emission at wavelengths less than half the excitation wavelength suggests an excitation process involving three or more photons. This conjecture is further confirmed by the observation of the super-quadratic dependence of the fluorescence intensity on the excitation power. Further work is required to spectroscopically identify these emitting species. This study demonstrates the use of multiphoton excitation microscopy for functional imaging of the metabolic states of in vivo human skin cells. Images FIGURE 2 FIGURE 3 FIGURE 4 FIGURE 5 PMID:9168018

  20. Optimal fluorescence excitation wavelengths for detection of squamous intra-epithelial neoplasia: results from an animal model

    NASA Astrophysics Data System (ADS)

    Coghlan, Lezlee; Utzinger, Urs; Drezek, Rebekah A.; Heintzelmann, Doug; Zuluaga, Andres F.; Brookner, Carrie; Richards-Kortum, Rebecca R.; Gimenez-Conti, Irma; Follen, Michele

    2000-12-01

    Using the hamster cheek pouch carcinogenesis model, we explore which fluorescence excitation wavelengths are useful for the detection of neoplasia. 42 hamsters were treated with DMBA to induce carcinogenesis, and 20 control animals were treated only with mineral oil. Fluorescence excitation emission matrices were measured from the cheek pouches of the hamsters weekly. Results showed increased fluorescence near 350-370 nm and 410 nm excitation and decreased fluorescence near 450-470 nm excitation with neoplasia. The optimal diagnostic excitation wavelengths identified using this model - 350-370 nm excitation and 400-450 nm excitation - are similar to those identified for detection of human oral cavity neoplasia.

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

    NASA Astrophysics Data System (ADS)

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

    2012-11-01

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

  2. Water-Soluble Triarylborane Chromophores for One- and Two-Photon Excited Fluorescence Imaging of Mitochondria in Cells.

    PubMed

    Griesbeck, Stefanie; Zhang, Zuolun; Gutmann, Marcus; Lühmann, Tessa; Edkins, Robert M; Clermont, Guillaume; Lazar, Adina N; Haehnel, Martin; Edkins, Katharina; Eichhorn, Antonius; Blanchard-Desce, Mireille; Meinel, Lorenz; Marder, Todd B

    2016-10-01

    Three water-soluble tetracationic quadrupolar chromophores comprising two three-coordinate boron π-acceptor groups bridged by thiophene-containing moieties were synthesised for biological imaging applications. Compound 3 containing the bulkier 5-(3,5-Me2 C6 H2 )-2,2'-(C4 H2 S)2 -5'-(3,5-Me2 C6 H2 ) bridge is stable over a long period of time, exhibits a high fluorescence quantum yield and strong one- and two-photon absorption (TPA), and has a TPA cross section of 268 GM at 800 nm in water. Confocal laser scanning fluorescence microscopy studies in live cells indicated localisation of the chromophore at the mitochondria; moreover, cytotoxicity measurements proved biocompatibility. Thus, chromophore 3 has excellent potential for one- and two-photon-excited fluorescence imaging of mitochondrial function in cells.

  3. Water-Soluble Triarylborane Chromophores for One- and Two-Photon Excited Fluorescence Imaging of Mitochondria in Cells.

    PubMed

    Griesbeck, Stefanie; Zhang, Zuolun; Gutmann, Marcus; Lühmann, Tessa; Edkins, Robert M; Clermont, Guillaume; Lazar, Adina N; Haehnel, Martin; Edkins, Katharina; Eichhorn, Antonius; Blanchard-Desce, Mireille; Meinel, Lorenz; Marder, Todd B

    2016-10-01

    Three water-soluble tetracationic quadrupolar chromophores comprising two three-coordinate boron π-acceptor groups bridged by thiophene-containing moieties were synthesised for biological imaging applications. Compound 3 containing the bulkier 5-(3,5-Me2 C6 H2 )-2,2'-(C4 H2 S)2 -5'-(3,5-Me2 C6 H2 ) bridge is stable over a long period of time, exhibits a high fluorescence quantum yield and strong one- and two-photon absorption (TPA), and has a TPA cross section of 268 GM at 800 nm in water. Confocal laser scanning fluorescence microscopy studies in live cells indicated localisation of the chromophore at the mitochondria; moreover, cytotoxicity measurements proved biocompatibility. Thus, chromophore 3 has excellent potential for one- and two-photon-excited fluorescence imaging of mitochondrial function in cells. PMID:27627995

  4. Dipolar relaxation within the protein matrix of the green fluorescent protein: a red edge excitation shift study.

    PubMed

    Haldar, Sourav; Chattopadhyay, Amitabha

    2007-12-27

    The fluorophore in green fluorescent protein (GFP) is localized in a highly constrained environment, protected from the bulk solvent by the barrel-shaped protein matrix. We have used the wavelength-selective fluorescence approach (red edge excitation shift, REES) to monitor solvent (environment) dynamics around the fluorophore in enhanced green fluorescent protein (EGFP) under various conditions. Our results show that EGFP displays REES in buffer and glycerol, i.e., the fluorescence emission maxima exhibit a progressive shift toward the red edge, as the excitation wavelength is shifted toward the red edge of the absorption spectrum. Interestingly, EGFP displays REES when incorporated in reverse micelles of sodium bis(2-ethylhexyl)sulfosuccinate (AOT), independent of the hydration state. We interpret the observed REES to the constrained environment experienced by the EGFP fluorophore in the rigid protein matrix, rather than to the dynamics of the bulk solvent. These results are supported by the temperature dependence of REES and characteristic wavelength-dependent changes in fluorescence anisotropy.

  5. [Study on the characters of phytoplankton chlorophyll fluorescence excitation spectra based on fourth-derivative].

    PubMed

    Lu, Lu; Su, Rong-Guo; Wang, Xiu-Lin; Zhu, Chen-Jian

    2007-11-01

    Chlorophyll fluorescence excitation spectra of six phytoplankton species, belonging to Bacillariophyta and Dinophyta, were dealt by fourth-derivative analysis with the Matlab program. The results show that between 350 nm and 550 nm six fluorescence peaks were found in the fourth-derivative spectra, which are representatives of non-pigments, chlorophylls and carotenoides respectively. The method makes Bacillariophyta and Dinophyta more distinguishable when the fourth-derivative spectra are compared with the chlorophyll fluorescence excitation spectra. It can be used not only to discriminate the two groups of algaes, but also to reduce the effect of noise. The fluorescence peaks in the fourth-derivative spectra are proved to be stable.

  6. Enhancing the depth of tissue microscope imaging using two-photon excitation of the second singlet state of fluorescent agents

    NASA Astrophysics Data System (ADS)

    Pu, Yang; Shi, Lingyan; Pratavieira, Sebastião.; Alfano, R. R.

    2014-03-01

    Increasing the depth to image inside tissue is critical in biomedicine. Two-photon (2P) excitation of the second singlet (S2) state of a group of fluorescent agents with near infrared emission, Chlorophyll a (Chl a) and Indocyanine green (ICG), is used to extend the optical imaging regime of 2PM into "tissue optical window" for deep tissue penetration. The fast nonradiative from S2 to S1 yields both emission and absorption wavelengths in the therapeutic window. The salient feature is to place both the 2P excitation and emission wavelengths of the imaging agents falling into the "tissue optical window". As a first step to achieve deeper optical imaging, Chl a and ICG are investigated and demonstrated as imaging agents for 2P S2 excitation microscope image.

  7. Time-resolved multicolor two-photon excitation fluorescence microscopy of cells and tissues

    NASA Astrophysics Data System (ADS)

    Zheng, Wei

    2014-11-01

    Multilabeling which maps the distribution of different targets is an indispensable technique in many biochemical and biophysical studies. Two-photon excitation fluorescence (TPEF) microscopy of endogenous fluorophores combining with conventional fluorescence labeling techniques such as genetically encoded fluorescent protein (FP) and fluorescent dyes staining could be a powerful tool for imaging living cells. However, the challenge is that the excitation and emission wavelength of these endogenous fluorophores and fluorescent labels are very different. A multi-color ultrafast source is required for the excitation of multiple fluorescence molecules. In this study, we developed a two-photon imaging system with excitations from the pump femtosecond laser and the selected supercontinuum generated from a photonic crystal fiber (PCF). Multiple endogenous fluorophores, fluorescent proteins and fluorescent dyes were excited in their optimal wavelengths simultaneously. A time- and spectral-resolved detection system was used to record the TPEF signals. This detection technique separated the TPEF signals from multiple sources in time and wavelength domains. Cellular organelles such as nucleus, mitochondria, microtubule and endoplasmic reticulum, were clearly revealed in the TPEF images. The simultaneous imaging of multiple fluorophores of cells will greatly aid the study of sub-cellular compartments and protein localization.

  8. Invited Review Article: Imaging techniques for harmonic and multiphoton absorption fluorescence microscopy

    PubMed Central

    Carriles, Ramón; Schafer, Dawn N.; Sheetz, Kraig E.; Field, Jeffrey J.; Cisek, Richard; Barzda, Virginijus; Sylvester, Anne W.; Squier, Jeffrey A.

    2009-01-01

    We review the current state of multiphoton microscopy. In particular, the requirements and limitations associated with high-speed multiphoton imaging are considered. A description of the different scanning technologies such as line scan, multifoci approaches, multidepth microscopy, and novel detection techniques is given. The main nonlinear optical contrast mechanisms employed in microscopy are reviewed, namely, multiphoton excitation fluorescence, second harmonic generation, and third harmonic generation. Techniques for optimizing these nonlinear mechanisms through a careful measurement of the spatial and temporal characteristics of the focal volume are discussed, and a brief summary of photobleaching effects is provided. Finally, we consider three new applications of multiphoton microscopy: nonlinear imaging in microfluidics as applied to chemical analysis and the use of two-photon absorption and self-phase modulation as contrast mechanisms applied to imaging problems in the medical sciences. PMID:19725639

  9. Resolution of heterogeneous fluorescence into component decay-associated excitation spectra. Application to subtilisins.

    PubMed

    Willis, K J; Szabo, A G; Drew, J; Zuker, M; Ridgeway, J M

    1990-02-01

    Direct and indirect methods are described to combine steady-state and picosecond time-resolved fluorescence decay data to generate decay-associated excitation spectra. The heterogeneous fluorescence from a fluorophore mixture that models protein fluorescence was resolved into individual component excitation spectra. The two methods were also used to determine the excitation spectra associated with each of the decay time components for the proteins subtilisin Carlsberg and BPN'. On the basis of associated spectra, the decay components of both proteins were assigned to individual (or groups of) emitting species. The two approaches used to generate the decay-associated excitation spectra are compared and their general application to protein fluorescence studies is discussed. PMID:2180489

  10. Solvents effect on the absorption and fluorescence spectra of 7-diethylamino-3-thenoylcoumarin: Evaluation and correlation between solvatochromism and solvent polarity parameters

    NASA Astrophysics Data System (ADS)

    Basavaraja, Jana; Inamdar, S. R.; Suresh Kumar, H. M.

    2015-02-01

    Effect of solvents of varying polarities on absorption and fluorescence spectra and dipole moment of laser dye: 7-diethylamino-3-thenoylcoumarin (DETC) has been investigated. A small band shift is obtained in the absorption spectra compared to emission spectra. The spectral shifts were correlated with Catalan's parameters using linear solvation energy relationship. It reveals that non-specific interaction measured by solvent polarity has more influence on absorption and solvent dipolarity contribution is significant in case of fluorescence. A bathochromic shift observed in absorption and emission spectra with increasing solvent polarity, which implied that the transition involved is π → π∗. The solvatochromic correlations were used to estimate the excited state dipole moment using experimentally determined ground state dipole moment. The observed single-state excited state dipole moment is found to be greater than the ground state.

  11. Strong excited state absorption (ESA) in Yb-doped fiber lasers

    NASA Astrophysics Data System (ADS)

    Engholm, Magnus; Rydberg, Sara; Hammarling, Krister

    2013-03-01

    Excited state absorption (ESA) measurements performed on Yb-doped silica bers show the onset of a strong absorption band in the visible range. In this work, we perform experiments to investigate the possibility for ESA to be part of the induced optical losses (photodarkening) observed in Yb-doped ber lasers. Our results indicate that an ESA process, from the 2F5/2 excited state manifold in the Yb3+ ion to the charge-transfer state with absorption bands in the UV range, may constitute a transfer route for pump- and laser photons in the near-infrared range.

  12. o-nitrobenzyl photolabile protecting groups with red-shifted absorption: syntheses and uncaging cross-sections for one- and two-photon excitation.

    PubMed

    Aujard, Isabelle; Benbrahim, Chouaha; Gouget, Marine; Ruel, Odile; Baudin, Jean-Bernard; Neveu, Pierre; Jullien, Ludovic

    2006-09-01

    We evaluated the o-nitrobenzyl platform for designing photolabile protecting groups with red-shifted absorption that could be photolyzed upon one- and two-photon excitation. Several synthetic pathways to build different conjugated o-nitrobenzyl backbones, as well as to vary the benzylic position, are reported. Relative to the reference 4,5-dimethoxy-2-nitrobenzyl group, several o-nitrobenzyl derivatives exhibit a large and red-shifted one-photon absorption within the near-UV range. Uncaging after one-photon excitation was studied by measuring UV-visible absorption and steady-state fluorescence emission on model caged ethers and esters. In the whole series investigated, the caged substrates were released cleanly upon photolysis. Quantum yields of uncaging after one-photon absorption lie within the 0.1-1 % range. We observed that these drop as the maximum wavelength absorption of the o-nitrobenzyl protecting group is increased. A new method based on fluorescence correlation spectroscopy (FCS) after two-photon excitation was used to measure the action uncaging cross section for two-photon excitation. The series of o-nitrobenzyl caged fluorescent coumarins investigated exhibit values within the 0.1-0.01 Goeppert-Mayer (GM) range. Such results are in line with the low quantum yields of uncaging associated with cross-sections of 1-50 GM for two-photon absorption. Although the cross-sections for one- and two-photon absorption of o-nitrobenzyl photolabile protecting groups can be readily improved, we emphasize the difficulty in enlarging the corresponding action uncaging cross-sections in view of the observed trend of their quantum yield of uncaging.

  13. o-nitrobenzyl photolabile protecting groups with red-shifted absorption: syntheses and uncaging cross-sections for one- and two-photon excitation.

    PubMed

    Aujard, Isabelle; Benbrahim, Chouaha; Gouget, Marine; Ruel, Odile; Baudin, Jean-Bernard; Neveu, Pierre; Jullien, Ludovic

    2006-09-01

    We evaluated the o-nitrobenzyl platform for designing photolabile protecting groups with red-shifted absorption that could be photolyzed upon one- and two-photon excitation. Several synthetic pathways to build different conjugated o-nitrobenzyl backbones, as well as to vary the benzylic position, are reported. Relative to the reference 4,5-dimethoxy-2-nitrobenzyl group, several o-nitrobenzyl derivatives exhibit a large and red-shifted one-photon absorption within the near-UV range. Uncaging after one-photon excitation was studied by measuring UV-visible absorption and steady-state fluorescence emission on model caged ethers and esters. In the whole series investigated, the caged substrates were released cleanly upon photolysis. Quantum yields of uncaging after one-photon absorption lie within the 0.1-1 % range. We observed that these drop as the maximum wavelength absorption of the o-nitrobenzyl protecting group is increased. A new method based on fluorescence correlation spectroscopy (FCS) after two-photon excitation was used to measure the action uncaging cross section for two-photon excitation. The series of o-nitrobenzyl caged fluorescent coumarins investigated exhibit values within the 0.1-0.01 Goeppert-Mayer (GM) range. Such results are in line with the low quantum yields of uncaging associated with cross-sections of 1-50 GM for two-photon absorption. Although the cross-sections for one- and two-photon absorption of o-nitrobenzyl photolabile protecting groups can be readily improved, we emphasize the difficulty in enlarging the corresponding action uncaging cross-sections in view of the observed trend of their quantum yield of uncaging. PMID:16763952

  14. Deeper Insight into Fluorescence-Excitation of Molecules by Light

    ERIC Educational Resources Information Center

    Wahab, M. Farooq; Gore, Gordon R.

    2013-01-01

    In a recent issue of "TPT," Gordon Gore made interesting observations about the red or yellow fluorescence when laser beams are passed through olive oil. With the excellent visuals contained in that article, we present a pictorial explanation of the questions that were raised in Ref 1 ("Fun with Fluorescence in Olive Oil,"…

  15. Noninvasive fluorescence excitation spectroscopy for the diagnosis of oral neoplasia in vivo

    NASA Astrophysics Data System (ADS)

    Ebenezar, Jeyasingh; Ganesan, Singaravelu; Aruna, Prakasarao; Muralinaidu, Radhakrishnan; Renganathan, Kannan; Saraswathy, Thillai Rajasekaran

    2012-09-01

    Fluorescence excitation spectroscopy (FES) is an emerging approach to cancer detection. The goal of this pilot study is to evaluate the diagnostic potential of FES technique for the detection and characterization of normal and cancerous oral lesions in vivo. Fluorescence excitation (FE) spectra from oral mucosa were recorded in the spectral range of 340 to 600 nm at 635 nm emission using a fiberoptic probe spectrofluorometer to obtain spectra from the buccal mucosa of 30 sites of 15 healthy volunteers and 15 sites of 10 cancerous patients. Significant FE spectral differences were observed between normal and well differentiated squamous cell carcinoma (WDSCC) oral lesions. The FE spectra of healthy volunteers consists of a broad emission band around 440 to 470 nm, whereas in WDSCC lesions, a new primary peak was seen at 410 nm with secondary peaks observed at 505, 540, and 580 nm due to the accumulation of porphyrins in oral lesions. The FE spectral bands of the WDSCC lesions resemble the typical absorption spectra of a porphyrin. Three potential ratios (I410/I505, I410/I540, and I410/I580) were calculated from the FE spectra and used as input variables for a stepwise linear discriminant analysis (SLDA) for normal and WDSCC groups. Leave-one-out (LOO) method of cross-validation was performed to check the reliability on spectral data for tissue characterization. The diagnostic sensitivity and specificity were determined for normal and WDSCC lesions from the scatter plot of the discriminant function scores. It was observed that diagnostic algorithm based on discriminant function scores obtained by SLDA-LOO method was able to distinguish WDSCC from normal lesions with a sensitivity of 100% and specificity of 100%. Results of the pilot study demonstrate that the FE spectral changes due to porphyrin have a good diagnostic potential; therefore, porphyrin can be used as a native tumor marker.

  16. Hyperspectral imaging fluorescence excitation scanning for detecting colorectal cancer: pilot study

    NASA Astrophysics Data System (ADS)

    Leavesley, Silas J.; Wheeler, Mikayla; Lopez, Carmen; Baker, Thomas; Favreau, Peter F.; Rich, Thomas C.; Rider, Paul F.; Boudreaux, Carole W.

    2016-03-01

    Optical spectroscopy and hyperspectral imaging have shown the theoretical potential to discriminate between cancerous and non-cancerous tissue with high sensitivity and specificity. To date, these techniques have not been able to be effectively translated to endoscope platforms. Hyperspectral imaging of the fluorescence excitation spectrum represents a new technology that may be well-suited for endoscopic implementation. However, the feasibility of detecting differences between normal and cancerous mucosa using fluorescence excitation-scanning hyperspectral imaging has not been evaluated. The objective of this pilot study was to evaluate the changes in the fluorescence excitation spectrum of resected specimen pairs of colorectal adenocarcinoma and normal colorectal mucosa. Patients being treated for colorectal adenocarcinoma were enrolled. Representative adenocarcinoma and normal colonic mucosa specimens were collected from each case. Specimens were flash frozen in liquid nitrogen. Adenocarcinoma was confirmed by histologic evaluation of H&E permanent sections. Hyperspectral image data of the fluorescence excitation of adenocarcinoma and surrounding normal tissue were acquired using a custom microscope configuration previously developed in our lab. Results demonstrated consistent spectral differences between normal and cancerous tissues over the fluorescence excitation spectral range of 390-450 nm. We conclude that fluorescence excitation-scanning hyperspectral imaging may offer an alternative approach for differentiating adenocarcinoma and surrounding normal mucosa of the colon. Future work will focus on expanding the number of specimen pairs analyzed and will utilize fresh tissues where possible, as flash freezing and reconstituting tissues may have altered the autofluorescence properties.

  17. Enhanced energy transfer in respiratory-deficient endothelial cells probed by microscopic fluorescence excitation spectroscopy

    NASA Astrophysics Data System (ADS)

    Schneckenburger, Herbert; Gschwend, Michael H.; Bauer, Manfred; Strauss, Wolfgang S. L.; Steiner, Rudolf W.

    1996-12-01

    Mitochondrial malfunction may be concomitant with changes of the redox states of the coenzymes nicotinamide adenine dinucleotide (NAD+/NADH), as well as flavin.mononucleotide or dinucleotide. The intrinsic fluorescence of these coenzymes was therefore proposed to be a measure of malfunction. Since mitochondrial fluorescence is strongly superposed by autofluorescence from various cytoplasmatic fluorophores, cultivated endothelial cells were incubated with the mitochondrial marker rhodamine 123 (R123), and after excitation of flavin molecules, energy transfer to R123 was investigated. Due to spectral overlap of flavin and R123 fluorescence, energy transfer flavin yields R123 could not be detected from their emission spectra. Therefore, the method of microscopic fluorescence excitation spectroscopy was established. When detecting R123 fluorescence, excitation maxima at 370 - 390 nm and 420-460 nm were assigned to flavins, whereas a pronounced excitation band at 465 - 490 nm was attributed to R123. Therefore, excitation at 475 nm reflected the intracellular concentration of R123, whereas excitation at 385 nm reflected flavin excitation with a subsequent energy transfer to R123 molecules. An enhanced energy transfer after inhibition of specific enzyme complexes of the respiratory chain is discussed in the present article.

  18. Vibrationally Resolved Absorption and Fluorescence Spectra of Firefly Luciferin: A Theoretical Simulation in the Gas Phase and in Solution.

    PubMed

    Cheng, Yuan-Yuan; Liu, Ya-Jun

    2016-07-01

    Firefly bioluminescence has been applied in several fields. However, the absorption and fluorescence spectra of the substrate, luciferin, have not been observed at the vibrational level. In this study, the vibrationally resolved absorption and fluorescence spectra of firefly luciferin (neutral form LH2 , phenolate ion form LH(-) and dianion form L(2-) ) are simulated using the density functional method and convoluted by a Gaussian function, with displacement, distortion and Duschinsky effects in the framework of the Franck-Condon approximation. Both neutral and anionic forms of the luciferin are considered in the gas phase and in solution. The simulated spectra have desired band maxima with the experimental ones. The vibronic structure analysis reveals that the features of the most contributive vibrational modes coincide with the key geometry-changing region during transition between the ground state and the first singlet excited state. PMID:27165852

  19. Multi-Channel Hyperspectral Fluorescence Detection Excited by Coupled Plasmon-Waveguide Resonance

    PubMed Central

    Du, Chan; Liu, Le; Zhang, Lin; Guo, Jun; Guo, Jihua; Ma, Hui; He, Yonghong

    2013-01-01

    We propose in this paper a biosensor scheme based on coupled plasmon-waveguide resonance (CPWR) excited fluorescence spectroscopy. A symmetrical structure that offers higher surface electric field strengths, longer surface propagation lengths and depths is developed to support guided waveguide modes for the efficient excitation of fluorescence. The optimal parameters for the sensor films are theoretically and experimentally investigated, leading to a detection limit of 0.1 nM (for a Cy5 solution). Multiplex analysis possible with the fluorescence detection is further advanced by employing the hyperspectral fluorescence technique to record the full spectra for every pixel on the sample plane. We demonstrate experimentally that highly overlapping fluorescence (Cy5 and Dylight680) can be distinguished and ratios of different emission sources can be determined accurately. This biosensor shows great potential for multiplex detections of fluorescence analytes. PMID:24129023

  20. Excitation-emission matrices and synchronous fluorescence spectroscopy for the diagnosis of gastrointestinal cancers

    NASA Astrophysics Data System (ADS)

    Genova, Ts; Borisova, E.; Penkov, N.; Vladimirov, B.; Zhelyazkova, A.; Avramov, L.

    2016-06-01

    We report the development of an improved fluorescence technique for cancer diagnostics in the gastrointestinal tract. We investigate the fluorescence of ex vivo colorectal (cancerous and healthy) tissue samples using excitation-emission matrix (EEM) and synchronous fluorescence spectroscopy (SFS) steady-state approaches. The obtained results are processed for revealing characteristic fluorescence spectral features with a valuable diagnostic meaning. The main tissue fluorophores, contributing to the observed fluorescence, are tyrosine, tryptophan, NADH, FAD, collagen and elastin. Based on the results of the Mann-Whitney test as useful parameters for differentiation of gastrointestinal cancer from normal mucosa, we suggest using excitation wavelengths in the range 300 - 360 nm for fluorescence spectroscopy and wavelengths intervals of 60 nm and 90 nm for SFS.

  1. Reconstruction of an excited-state molecular wave packet with attosecond transient absorption spectroscopy

    NASA Astrophysics Data System (ADS)

    Cheng, Yan; Chini, Michael; Wang, Xiaowei; González-Castrillo, Alberto; Palacios, Alicia; Argenti, Luca; Martín, Fernando; Chang, Zenghu

    2016-08-01

    Attosecond science promises to allow new forms of quantum control in which a broadband isolated attosecond pulse excites a molecular wave packet consisting of a coherent superposition of multiple excited electronic states. This electronic excitation triggers nuclear motion on the molecular manifold of potential energy surfaces and can result in permanent rearrangement of the constituent atoms. Here, we demonstrate attosecond transient absorption spectroscopy (ATAS) as a viable probe of the electronic and nuclear dynamics initiated in excited states of a neutral molecule by a broadband vacuum ultraviolet pulse. Owing to the high spectral and temporal resolution of ATAS, we are able to reconstruct the time evolution of a vibrational wave packet within the excited B'Σ1u+ electronic state of H2 via the laser-perturbed transient absorption spectrum.

  2. Moving Towards a Technical Specification for Fluorescence Excitation-Emission Mapping and Absorbance Analysis of Colored Dissolved Organic Matter

    NASA Astrophysics Data System (ADS)

    Gilmore, A. M.

    2010-12-01

    Colored dissolved organic matter (CDOM) measurements with fluorescence and absorbance are important for evaluating a wide variety natural and industrial water sources. However, uncertainties and ambiguities continue to be propagated regarding interpretation of CDOM spectral data due to the variety of instruments, sampling chemistry conditions and types of analysis algorithms. Recent efforts have focused on standardization and interlaboratory comparisons of CDOM samples with respect to preparation, spectroscopic evaluation and mathematical analysis. This study deals with correlating absorbance and fluorescence data measured with the same sample to minimize interlaboratory variation. The theoretical significance of true simultaneous acquisition of the corrected (NIST Traceable) absorbance spectrum and fluorescence excitation spectral profile and excitation emission map is discussed as a means to provide the least ambiguous spectral data. Key issues considered are the variations introduced by ‘serial’ acquisitions of absorbance and fluorescence data. Variation can be caused by the different light-exposure history (especially UV) in the instruments, dissolved oxygen content associated with temperature changes and oxidation kinetics of the CDOM and in many cases concentration- and pH-related changes associated with diluting and pH buffering of the CDOM sample, respectively. Concentration changes in CDOM can be associated with optical anomalies including self-quenching and -absorption which systematically alter the fluorescence spectrum. Clearly, monitoring the absorbance and fluorescence simultaneously would deal with the above sampling variations and facilitate correcting the absorbance based fluorescence anomalies. The proposed method(s) described will be discussed in view of their potential to serve as the basis for an international technical specification in terms of the optical instrument and sampling conditions for CDOM analysis and reporting.

  3. Further evidence for charge transfer complexes in brown carbon aerosols from excitation-emission matrix fluorescence spectroscopy.

    PubMed

    Phillips, Sabrina M; Smith, Geoffrey D

    2015-05-14

    The light-absorbing fraction of organic molecules in ambient aerosols, known as "brown carbon," is an important yet poorly characterized component. Despite the fact that brown carbon could alter the radiative forcing of aerosols significantly, identification of specific chromophores has remained challenging. We recently demonstrated that charge transfer (CT) complexes formed in organic molecules could be responsible for a large fraction of absorption observed in water-extracted ambient particulate matter.1 In the present study, we use excitation-emission matrix fluorescence spectroscopy to further corroborate the importance of CT complexes in defining aerosol optical properties. Monotonically increasing and decreasing quantum yields, decreasing Stokes shifts, and red-shifting emission maxima are observed from ambient particulate matter collected in Athens, Georgia, strongly suggesting that a superposition of independent chromophores is not sufficient to explain brown carbon absorption and fluorescence. Instead, we show that a model in which such chromophores are energetically coupled to a dense manifold of CT complexes is consistent with all of the observations. Further, we suggest that a significant fraction of the observed fluorescence originates from CT complexes and that their contribution to brown carbon absorption is likely greater than we reported previously.

  4. Low temperature absorption, fluorescence, and hole-burning spectroscopy of photosystem II reaction center complex containing 1 and 2 carotenoides

    NASA Astrophysics Data System (ADS)

    Dědic, R.; Lovčinský, M.; Pšenčík, J.; Vácha, M.; Vácha, F.; Hála, J.

    1999-05-01

    Well defined photosystem II reaction centers from Pisum sativum containing 5 or 6 chlorophyll a (Chl a), 2 pheophytine a (Pheo a), and 1 or 2 β-carotene ( β Car) molecules were prepared by using immobilized metal affinity chromatography. Samples containing 6 Chl a and 1 or 2 β-Car and containing 5 Chl a and 1 β-Car were measured using low temperature absorption, fluorescence and hole-burning spectroscopy. Absorption bands of the β Car (462, 490, and 508 nm) can be clearly distinguished next to the Soret absorption band of Chl a at low temperature. Their relative intensities strongly depend on Chl/Car ratio. The shapes of fluorescence bands are the same for all samples. Persistent spectral holes were burnt into both absorption and fluorescence spectra. This technique provides lifetime of excited state τ1 and Huang-Rhys factor S. Values of τ1 correspond to two picoseconds energy transfer in reaction centers. Huang-Rhys factor S=0.4 appears to be the same for all studied samples.

  5. Near-unity broadband absorption designs for semiconducting nanowire arrays via localized radial mode excitation.

    PubMed

    Fountaine, Katherine T; Kendall, Christian G; Atwater, Harry A

    2014-05-01

    We report design methods for achieving near-unity broadband light absorption in sparse nanowire arrays, illustrated by results for visible absorption in GaAs nanowires on Si substrates. Sparse (<5% fill fraction) nanowire arrays achieve near unity absorption at wire resonant wavelengths due to coupling into 'leaky' radial waveguide modes of individual wires and wire-wire scattering processes. From a detailed conceptual development of radial mode resonant absorption, we demonstrate two specific geometric design approaches to achieve near unity broadband light absorption in sparse nanowire arrays: (i) introducing multiple wire radii within a small unit cell array to increase the number of resonant wavelengths, yielding a 15% absorption enhancement relative to a uniform nanowire array and (ii) tapering of nanowires to introduce a continuum of diameters and thus resonant wavelengths excited within a single wire, yielding an 18% absorption enhancement over a uniform nanowire array.

  6. Conjugated Polymer-Based Hybrid Nanoparticles with Two-Photon Excitation and Near-Infrared Emission Features for Fluorescence Bioimaging within the Biological Window.

    PubMed

    Lv, Yanlin; Liu, Peng; Ding, Hui; Wu, Yishi; Yan, Yongli; Liu, Heng; Wang, Xuefei; Huang, Fei; Zhao, Yongsheng; Tian, Zhiyuan

    2015-09-23

    Hybrid fluorescent nanoparticles (NPs) capable of fluorescing near-infrared (NIR) light (centered ∼730 nm) upon excitation of 800 nm laser light were constructed. A new type of conjugated polymer with two-photon excited fluorescence (TPEF) feature, P-F8-DPSB, was used as the NIR-light harvesting component and the energy donor while a NIR fluorescent dye, DPA-PR-PDI, was used as the energy acceptor and the NIR-light emitting component for the construction of the fluorescent NPs. The hybrid NPs possess δ value up to 2.3 × 10(6) GM per particle upon excitation of 800 nm pulse laser. The excellent two-photon absorption (TPA) property of the conjugated polymer component, together with its high fluorescence quantum yield (ϕ) up to 45% and the efficient energy transfer from the conjugated polymer to NIR-emitting fluorophore with efficiency up to 90%, imparted the hybrid NPs with TPEF-based NIR-input-NIR-output fluorescence imaging ability with penetration depth up to 1200 μm. The practicability of the hybrid NPs for fluorescence imaging in Hela cells was validated. PMID:26340609

  7. Fluorescent detection and imaging of Hg(2+) using a novel phenanthroline derivative based single- and two-photon excitation.

    PubMed

    Zhang, Xian; Li, Long-long; Liu, Ying-kai

    2016-02-01

    A novel phenanthroline derivative, 4-[4-(N-methyl)styrene]-imidazo[4,5-f][1,10]phenanthroline-benzene iodated salt (MSIPBI), was synthesized, and the linear absorption and fluorescent spectra of MSIPBI in different solvents were investigated. The photophysical properties in unbound and in ligand-metal complexes were evaluated by UV absorption and one- and two-photon fluorescent spectra, and the quantum yields, two-photon active cross-sections and the binding constant of dye-metal were calculated. The results indicated that MSIPBI has a large Stokes shift (more than 167nm), and the dye was selective and sensitive for the detection of Hg(2+) with a two-photon active cross-section of 55.5GM in tris-HCl buffer solution at 800nm. Furthermore, the results of the fluorescence microscopy imaging indicated that MSIPBI is an efficient fluorescent probe for the detection of Hg(2+) in living cells by one- and two-photon excitation. Moreover, the experiments of determination Hg(2+) in river water and tap water were finished.

  8. Short-range ordered photonic structures of lamellae-forming diblock copolymers for excitation-regulated fluorescence enhancement

    NASA Astrophysics Data System (ADS)

    Kim, Se Hee; Kim, Ki-Se; Char, Kookheon; Yoo, Seong Il; Sohn, Byeong-Hyeok

    2016-05-01

    Photonic crystals can be represented by periodic nanostructures with alternating refractive indices, which create artificial stop bands with the appearance of colors. In this regard, nanodomains of block copolymers and the corresponding structural colors have been intensively studied in the past. However, the practical application of photonic crystals of block copolymers has been limited to a large degree because of the presence of large defects and grain boundaries in the nanodomains of block copolymers. The present study focuses on the alternative opportunity of short-range ordered nanodomains of block copolymers for fluorescence enhancement, which also has a direct relevance to the development of fluorescence sensors or detectors. The enhancement mechanism was found to be interconnected with the excitation process rather than the alternation of the decay kinetics. In particular, we demonstrate that randomly oriented, but regular grains of lamellae of polystyrene-block-polyisoprene, PS-b-PI, diblock copolymers and their blend with PS homopolymers can behave as Bragg mirrors to induce multiple reflections of the excitation source inside the photonic structures. This process in turn significantly increases the effective absorption of the given fluorophores inside the polymeric photonic structures to amplify the fluorescence signal.Photonic crystals can be represented by periodic nanostructures with alternating refractive indices, which create artificial stop bands with the appearance of colors. In this regard, nanodomains of block copolymers and the corresponding structural colors have been intensively studied in the past. However, the practical application of photonic crystals of block copolymers has been limited to a large degree because of the presence of large defects and grain boundaries in the nanodomains of block copolymers. The present study focuses on the alternative opportunity of short-range ordered nanodomains of block copolymers for fluorescence

  9. Ultrafast Excited-State Dynamics in the Green Fluorescent Protein Variant S65T/H148D 1. Mutagenesis and Structural Studies†

    PubMed Central

    Shu, Xiaokun; Kallio, Karen; Shi, Xinghua; Abbyad, Paul; Kanchanawong, Pakorn; Childs, William; Boxer, Steven G.; Remington, S. James

    2008-01-01

    Wild type green fluorescent protein (wt-GFP) and the variant S65T/H148D each exhibit two absorption bands, A and B, which are associated with the protonated and deprotonated chromophores respectively. Excitation of either band leads to green emission. In wt-GFP, excitation of band A (~390 nm) leads to green emission with a rise time of 10–15 picoseconds, due to excited state proton transfer (ESPT) from the chromophore hydroxyl group to an acceptor. This process produces an anionic excited state intermediate I* that subsequently emits a green photon. In the variant S65T/H148D, the A band absorbance maximum is red-shifted to ~415 nm and as detailed in the accompanying papers (1, 2), when the A band is excited, green fluorescence appears with rise time shorter than the instrument time resolution (~170 fs). Based on steady state spectroscopy and high resolution crystal structures of several variants described herein, we propose that in S65T/H148D, the red shift of absorption band A and the ultrafast appearance of green fluorescence upon excitation of band A is due to a very short (≤ 2.4 Å), and possibly low barrier, hydrogen bond between the chromophore hydroxyl and introduced Asp148. PMID:17918959

  10. Plant Cell Imaging Based on Nanodiamonds with Excitation-Dependent Fluorescence.

    PubMed

    Su, Li-Xia; Lou, Qing; Jiao, Zhen; Shan, Chong-Xin

    2016-12-01

    Despite extensive work on fluorescence behavior stemming from color centers of diamond, reports on the excitation-dependent fluorescence of nanodiamonds (NDs) with a large-scale redshift from 400 to 620 nm under different excitation wavelengths are so far much fewer, especially in biological applications. The fluorescence can be attributed to the combined effects of the fraction of sp(2)-hybridized carbon atoms among the surface of the fine diamond nanoparticles and the defect energy trapping states on the surface of the diamond. The excitation-dependent fluorescent NDs have been applied in plant cell imaging for the first time. The results reported in this paper may provide a promising route to multiple-color bioimaging using NDs. PMID:27664016

  11. Plant Cell Imaging Based on Nanodiamonds with Excitation-Dependent Fluorescence

    NASA Astrophysics Data System (ADS)

    Su, Li-Xia; Lou, Qing; Jiao, Zhen; Shan, Chong-Xin

    2016-09-01

    Despite extensive work on fluorescence behavior stemming from color centers of diamond, reports on the excitation-dependent fluorescence of nanodiamonds (NDs) with a large-scale redshift from 400 to 620 nm under different excitation wavelengths are so far much fewer, especially in biological applications. The fluorescence can be attributed to the combined effects of the fraction of sp2-hybridized carbon atoms among the surface of the fine diamond nanoparticles and the defect energy trapping states on the surface of the diamond. The excitation-dependent fluorescent NDs have been applied in plant cell imaging for the first time. The results reported in this paper may provide a promising route to multiple-color bioimaging using NDs.

  12. Excited state X-ray absorption spectroscopy: Probing both electronic and structural dynamics

    NASA Astrophysics Data System (ADS)

    Neville, Simon P.; Averbukh, Vitali; Ruberti, Marco; Yun, Renjie; Patchkovskii, Serguei; Chergui, Majed; Stolow, Albert; Schuurman, Michael S.

    2016-10-01

    We investigate the sensitivity of X-ray absorption spectra, simulated using a general method, to properties of molecular excited states. Recently, Averbukh and co-workers [M. Ruberti et al., J. Chem. Phys. 140, 184107 (2014)] introduced an efficient and accurate L 2 method for the calculation of excited state valence photoionization cross-sections based on the application of Stieltjes imaging to the Lanczos pseudo-spectrum of the algebraic diagrammatic construction (ADC) representation of the electronic Hamiltonian. In this paper, we report an extension of this method to the calculation of excited state core photoionization cross-sections. We demonstrate that, at the ADC(2)x level of theory, ground state X-ray absorption spectra may be accurately reproduced, validating the method. Significantly, the calculated X-ray absorption spectra of the excited states are found to be sensitive to both geometric distortions (structural dynamics) and the electronic character (electronic dynamics) of the initial state, suggesting that core excitation spectroscopies will be useful probes of excited state non-adiabatic dynamics. We anticipate that the method presented here can be combined with ab initio molecular dynamics calculations to simulate the time-resolved X-ray spectroscopy of excited state molecular wavepacket dynamics.

  13. Excited state absorption properties of Pt(II) terpyridyl complexes bearing π-conjugated arylacetylides.

    PubMed

    Wang, Xianghuai; Goeb, Sébastien; Ji, Zhiqiang; Castellano, Felix N

    2010-11-18

    The synthesis, photophysics, and excited state absorption properties of three platinum(II) terpyridyl acetylide charge transfer (CT) complexes possessing a lone ancillary ligand systematically varied in phenylacetylide (PA) π-conjugation length, [Pt((t)Bu(3)tpy)([C≡C-C(6)H(4)](n)-H)]ClO(4) (n = 1, 2, 3), are described. Density functional theory (DFT) calculations performed on the ground states of complexes 1, 2, and 3 reveal that their HOMOs reside mainly on the ancillary π-conjugated PA moiety, ranging from 86 to 97%, with LUMOs predominantly centered on the terpyridyl acceptor ligand (91-92%). This electronic structure leads to the production of a triplet ligand-to-ligand CT ((3)LLCT) excited state upon visible light excitation with minor contributions from the corresponding triplet metal-to-ligand CT ((3)MLCT) excited state. Unusually strong red-to-near-IR transient absorptions are produced in the excited states of these molecules following selective long wavelength visible excitation of the low energy CT bands that do not emanate from the terpyridyl radical anion produced in the CT excited state or from an arylacetylide-based triplet intraligand ((3)IL) excited state. The extinction coefficients of these low energy absorption transients were determined using the energy transfer method with anthracene serving as the triplet acceptor. A detailed theoretical investigation using DFT and TDDFT methods reveals that these intense near-IR transient absorptions involve transitions resulting from transient oxidation of the PA subunit. In essence, the production of the (3)LLCT excited state transiently oxidizes the PA moiety by one electron, producing the corresponding highly absorbing radical cation-like species, analogous to that experienced in related intramolecular photoinduced electron transfer reactions. The computational work successfully predicts the oscillator strength and peak wavelength of the measured excited state absorption transients across this series

  14. The Fe XI-excited fluorescent cascade in Ne IV

    NASA Technical Reports Server (NTRS)

    Bhatia, A. K.; Kastner, S. O.

    1990-01-01

    A theoretical spectroscopic investigation of the fluorescent cascade to be expected in nitrogen-like Ne IV, when it is resonantly photoexcited by Fe XI in the solar atmosphere, symbiotic stars, or novae, is described. Primary and secondary cascade intensities are obtained as functions of photoexcitation rate, and expected absolute intensities in the solar atmosphere are derived on the basis of observed Fe XI and Ne EUV emission. Comparisons between the spatially resolved solar situation and spatially unresolved stellar cases of cataclysmic variables, in which these ions have been found to coexist, are made. An interesting possibility of periodic time-dependent fluorescence exists for the binary cataclysmics.

  15. Statistical image segmentation for the detection of skin lesion borders in UV fluorescence excitation

    NASA Astrophysics Data System (ADS)

    Ortega-Martinez, Antonio; Padilla-Martinez, Juan Pablo; Franco, Walfre

    2016-04-01

    The skin contains several fluorescent molecules or fluorophores that serve as markers of structure, function and composition. UV fluorescence excitation photography is a simple and effective way to image specific intrinsic fluorophores, such as the one ascribed to tryptophan which emits at a wavelength of 345 nm upon excitation at 295 nm, and is a marker of cellular proliferation. Earlier, we built a clinical UV photography system to image cellular proliferation. In some samples, the naturally low intensity of the fluorescence can make it difficult to separate the fluorescence of cells in higher proliferation states from background fluorescence and other imaging artifacts -- like electronic noise. In this work, we describe a statistical image segmentation method to separate the fluorescence of interest. Statistical image segmentation is based on image averaging, background subtraction and pixel statistics. This method allows to better quantify the intensity and surface distributions of fluorescence, which in turn simplify the detection of borders. Using this method we delineated the borders of highly-proliferative skin conditions and diseases, in particular, allergic contact dermatitis, psoriatic lesions and basal cell carcinoma. Segmented images clearly define lesion borders. UV fluorescence excitation photography along with statistical image segmentation may serve as a quick and simple diagnostic tool for clinicians.

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

    NASA Astrophysics Data System (ADS)

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

    1999-05-01

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

  17. Intrinsic fluorescence spectroscopy in turbid media: disentangling effects of scattering and absorption.

    PubMed

    Müller, M G; Georgakoudi, I; Zhang, Q; Wu, J; Feld, M S

    2001-09-01

    The fluorescence from a turbid medium such as biologic tissue contains information about scattering and absorption, as well as the intrinsic fluorescence, i.e., the fluorescence from an optically thin sample of pure fluorophores. The interplay of scattering and absorption can result in severe distortion of the intrinsic spectral features. These distortions can be removed by use of a photon-migration-based picture and information from simultaneously acquired fluorescence and reflectance spectra. We present experimental evidence demonstrating the validity of such an approach for extracting the intrinsic fluorescence for a wide range of scatterer and absorber concentrations in tissue models, ex vivo and in vivo tissues. We show that variations in line shape and intensity in intrinsic tissue fluorescence are significantly reduced compared with the corresponding measured fluorescence.

  18. Ultrafast Dynamics of a Green Fluorescent Protein Chromophore Analogue: Competition between Excited-State Proton Transfer and Torsional Relaxation.

    PubMed

    Chatterjee, Tanmay; Lacombat, Fabien; Yadav, Dheerendra; Mandal, Mrinal; Plaza, Pascal; Espagne, Agathe; Mandal, Prasun K

    2016-09-15

    The competition between excited-state proton transfer (ESPT) and torsion plays a central role in the photophysics of fluorescent proteins of the green fluorescent protein (GFP) family and their chromophores. Here, it was investigated in a single GFP chromophore analogue bearing o-hydroxy and p-diethylamino substituents, OHIM. The light-induced dynamics of OHIM was studied by femtosecond transient absorption spectroscopy, at different pH. We found that the photophysics of OHIM is determined by the electron-donating character of the diethylamino group: torsional relaxation dominates when the diethylamino group is neutral, whereas ultrafast ESPT followed by cis/trans isomerization and ground-state reprotonation are observed when the diethylamino group is protonated and therefore inactive as an electron donor. PMID:27548114

  19. Describing Two-Photon Absorptivity of Fluorescent Proteins with a New Vibronic Coupling Mechanism

    PubMed Central

    Makarov, N. S.; Tillo, S. E.; Hughes, T. E.; Rebane, A.

    2012-01-01

    Fluorescent proteins (FPs) are widely used in two-photon microscopy as genetically-encoded probes. Understanding the physical basics of their two-photon absorption (2PA) properties is therefore crucial for creation of two-photon brighter mutants. On the other hand, it can give us better insight into molecular interactions of the FP chromophore with complex protein environment. It is known that, compared to one-photon absorption spectrum, where the pure electronic transition is the strongest, the 2PA spectrum of a number of FPs is dominated by a vibronic transition. The physical mechanism of such intensity redistribution is not understood. Here we present a new physical model that explains this effect through the “Herzberg-Teller”-type vibronic coupling of the difference between the permanent dipole moments in the ground and excited states (Δμ) to the bond-length-alternating coordinate. This model also enables us to quantitatively describe a large variability of the 2PA peak intensity in a series of red FPs with the same chromophore through the interference between the “Herzberg-Teller” and Franck-Condon terms. PMID:22224830

  20. Deeper Insight into Fluorescence--Excitation of Molecules by Light

    NASA Astrophysics Data System (ADS)

    Wahab, M. Farooq; Gore, Gordon R.

    2013-05-01

    In a recent issue of TPT, Gordon Gore made interesting observations about the red or yellow fluorescence when laser beams are passed through olive oil. With the excellent visuals contained in that article, we present a pictorial explanation of the questions that were raised in Ref 1.

  1. Correction method for the self-absorption effects in fluorescence extended X-ray absorption fine structure on multilayer samples.

    PubMed

    Li, Wen Bin; Yang, Xiao Yue; Zhu, Jing Tao; Tu, Yu Chun; Mu, Bao Zhong; Yu, Hai Sheng; Wei, Xiang Jun; Huang, Yu Ying; Wang, Zhan Shan

    2014-05-01

    A novel correction method for self-absorption effects is proposed for extended X-ray absorption fine structure (EXAFS) detected in the fluorescence mode on multilayer samples. The effects of refraction and multiple reflection at the interfaces are fully considered in this correction method. The correction is performed in k-space before any further data analysis, and it can be applied to single-layer or multilayer samples with flat surfaces and without thickness limit when the model parameters for the samples are known. The validity of this method is verified by the fluorescence EXAFS data collected for a Cr/C multilayer sample measured at different experimental geometries. PMID:24763646

  2. UV-Excited Fluorescence of Rocks in Gale Crater, Mars

    NASA Astrophysics Data System (ADS)

    Fisk, M. R.; Pommerenck, J.; Watkins-Brandt, K.; Edgett, K. S.; Minitti, M. E.; Hardgrove, C. J.; Popa, R.; Goetz, W.; Cloutis, E.; Nixon, B.; Kennedy, M. R.; Treiman, A. H.

    2015-12-01

    Curiosity, the Mars Science Laboratory rover, landed in Gale crater in 2012. The Mars Hand Lens Imager (MAHLI), located on Curiosity's robotic arm, can be placed as close as 0.2 cm from targets. At this distance it has a pixel scale of ~13 µm/pixel. MAHLI usually images in daylight, but light emitting diodes (LEDs) located around the lens make nighttime imaging possible. Two of the six LEDs produce 365 nm long wave UV and near-UV light and small amounts of green and red light. On Mars MAHLI has taken images of UV-illuminated rocks, drill tailings, and a fluorescent calibration standard. In a lab test bed we took images of UV-illuminated terrestrial minerals. The test bed UV LEDs are equivalent to those on MAHLI. Fluorescence in the test bed was inferred when the color of the mineral in UV light was shifted from the color of the reflected incident UV light on non-fluorescing targets. We demonstrate this shift with terrestrial minerals: willemite (ZnSiO4) is green, fluorite (CaF2) is blue, fluorapatite (Ca5(PO4)3F) is yellow, and (some) calcites are red (Figure). Bassanite (CaSO4•½H2O) has been identified in sedimentary rock in Gale, and under long wave UV illumination, terrestrial bassanite fluoresces blue. In addition to bassanite, fluorite and apatite are candidate minerals in Gale crater. Portions of a bassanite-bearing target in Gale appear to fluoresce blue under MAHLI UV illumination. Adjacent to the bassanite is a dark mineral that yields a signal with more blue and less red and green than the bassanite suggesting a second fluorescing mineral. If these findings are supported by additional MAHLI and test bed measurements, then this will be the first fluorescence detection of its kind on Mars. This will be a step toward using UV light for the detection of organic compounds as has been proposed for the Mars 2020 mission.

  3. Energy-Looping Nanoparticles: Harnessing Excited-State Absorption for Deep-Tissue Imaging.

    PubMed

    Levy, Elizabeth S; Tajon, Cheryl A; Bischof, Thomas S; Iafrati, Jillian; Fernandez-Bravo, Angel; Garfield, David J; Chamanzar, Maysamreza; Maharbiz, Michel M; Sohal, Vikaas S; Schuck, P James; Cohen, Bruce E; Chan, Emory M

    2016-09-27

    Near infrared (NIR) microscopy enables noninvasive imaging in tissue, particularly in the NIR-II spectral range (1000-1400 nm) where attenuation due to tissue scattering and absorption is minimized. Lanthanide-doped upconverting nanocrystals are promising deep-tissue imaging probes due to their photostable emission in the visible and NIR, but these materials are not efficiently excited at NIR-II wavelengths due to the dearth of lanthanide ground-state absorption transitions in this window. Here, we develop a class of lanthanide-doped imaging probes that harness an energy-looping mechanism that facilitates excitation at NIR-II wavelengths, such as 1064 nm, that are resonant with excited-state absorption transitions but not ground-state absorption. Using computational methods and combinatorial screening, we have identified Tm(3+)-doped NaYF4 nanoparticles as efficient looping systems that emit at 800 nm under continuous-wave excitation at 1064 nm. Using this benign excitation with standard confocal microscopy, energy-looping nanoparticles (ELNPs) are imaged in cultured mammalian cells and through brain tissue without autofluorescence. The 1 mm imaging depths and 2 μm feature sizes are comparable to those demonstrated by state-of-the-art multiphoton techniques, illustrating that ELNPs are a promising class of NIR probes for high-fidelity visualization in cells and tissue. PMID:27603228

  4. Photoacoustic-fluorescence in vitro flow cytometry for quantification of absorption, scattering and fluorescence properties of the cells

    NASA Astrophysics Data System (ADS)

    Nedosekin, D. A.; Sarimollaoglu, M.; Foster, S.; Galanzha, E. I.; Zharov, V. P.

    2013-03-01

    Fluorescence flow cytometry is a well-established analytical tool that provides quantification of multiple biological parameters of cells at molecular levels, including their functional states, morphology, composition, proliferation, and protein expression. However, only the fluorescence and scattering parameters of the cells or labels are available for detection. Cell pigmentation, presence of non-fluorescent dyes or nanoparticles cannot be reliably quantified. Herewith, we present a novel photoacoustic (PA) flow cytometry design for simple integration of absorbance measurements into schematics of conventional in vitro flow cytometers. The integrated system allow simultaneous measurements of light absorbance, scattering and of multicolor fluorescence from single cells in the flow at rates up to 2 m/s. We compared various combinations of excitation laser sources for multicolor detection, including simultaneous excitation of PA and fluorescence using a single 500 kHz pulsed nanosecond laser. Multichannel detection scheme allows simultaneous detection of up to 8 labels, including 4 fluorescent tags and 4 PA colors. In vitro PA-fluorescence flow cytometer was used for studies of nanoparticles uptake and for the analysis of cell line pigmentation, including genetically encoded melanin expression in breast cancer cell line. We demonstrate that this system can be used for direct nanotoxicity studies with simultaneous quantification of nanoparticles content and assessment of cell viability using a conventional fluorescent apoptosis assays.

  5. Strategic emission color tuning of highly fluorescent imidazole-based excited-state intramolecular proton transfer molecules.

    PubMed

    Park, Sanghyuk; Kwon, Ji Eon; Park, Soo Young

    2012-07-01

    Highly fluorescent molecules harnessing the excited state intramolecular proton transfer (ESIPT) process are promising for a new generation of displays and light sources because they can offer very unique and novel optoelectronic properties which are different from those of conventional fluorescent dyes. To realize innovative ESIPT devices comprising full emission colors over the whole visible region, a molecular design strategy for predictable emission color tuning should be established. Here, we have developed a general strategy for a wide-range spectral tuning of imidazole-based ESIPT materials based on three different strategies--introduction of a nodal plane model, extension of effective conjugation length, and modification of heterocyclic rings. A series of nine ESIPT molecules were designed, synthesized and comprehensively investigated for their characteristic emission properties. All these molecules commonly showed no clear and transparent visible range absorption with no absorption color, but showed different colors of intense photoluminescence over broad visible regions from 450 nm (HPI) to 630 nm (HPNO) depending on their molecular structure. With the aid of density functional theory and time-dependent DFT calculations using M06, wB97XD, and B3LYP parameters with the 6-31G(d,p) basis set, these tuned emission bands of nine emitters were assigned from the stabilized excited state conformations that were derived from modified molecular structures.

  6. Broadband transient absorption spectroscopy with 1- and 2-photon excitations: Relaxation paths and cross sections of a triphenylamine dye in solution

    SciTech Connect

    Moreno, J.; Dobryakov, A. L.; Hecht, S. E-mail: skovale@chemie.hu-berlin.de; Kovalenko, S. A. E-mail: skovale@chemie.hu-berlin.de; Ioffe, I. N.; Granovsky, A. A.

    2015-07-14

    1-photon (382 nm) and 2-photon (752 nm) excitations to the S{sub 1} state are applied to record and compare transient absorption spectra of a push-pull triphenylamine (TrP) dye in solution. After 1-photon excitation, ultrafast vibrational and structural molecular relaxations are detected on a 0.1 ps time scale in nonpolar hexane, while in polar acetonitrile, the spectral evolution is dominated by dipolar solvation. Upon 2-photon excitation, transient spectra in hexane reveal an unexpected growth of stimulated emission (SE) and excited-state absorption (ESA) bands. The behavior is explained by strong population transfer S{sub 1} → S{sub n} due to resonant absorption of a third pump photon. Subsequent S{sub n} → S{sub 1} internal conversion (with τ{sub 1} = 1 ps) prepares a very hot S{sub 1} state which cools down with τ{sub 2} = 13 ps. The pump pulse energy dependence proves the 2-photon origin of the bleach signal. At the same time, SE and ESA are strongly affected by higher-order pump absorptions that should be taken into account in nonlinear fluorescence applications. The 2-photon excitation cross sections σ{sup (2)} = 32 ⋅ 10{sup −50} cm{sup 4} s at 752 nm are evaluated from the bleach signal.

  7. Saturable absorption and two-photon absorption of 1,2,5-thiadiazolo[3,4-g]quinoxaline based derivatives with near-infrared fluorescence

    NASA Astrophysics Data System (ADS)

    Du, Yabing; Lin, Xiaodong; Jia, Tingjian; Dong, Jun

    2015-03-01

    Organic molecules with near-infrared (NIR) fluorescence are extremely interesting for the applications in nonlinear optical devices and bioimaging. However, such kind of materials have been relatively rarely studied. In this work, the nonlinear optical properties of 1,2,5-thiadiazolo[3,4-g]quinoxaline based derivatives with NIR fluorescence emission have been investigated for the first time. Under the excitation of femtosecond pulses at 532 nm, the chromophore with dithienyl as donor (TQ2) presents saturable absorption (SA) behavior, while no SA has been observed in the derivative with biphenyl (TQ1) as donor. Moreover, TQ2 exhibits much larger two-photon absorption (TPA) cross-sections with strong NIR fluorescence in the second biological window. The larger nonlinear optical properties of TQ2 is due to the introduction of stronger electron-donating group (dithienyl) and the resultant enhanced intramolecular charge transfer properties. At the end, TPA based optical limiting behaviors of the molecules are demonstrated in THF solutions, thanks to their large solubility and strong TPA.

  8. Hybridization assay based on evanescent fluorescence excitation and collection

    NASA Astrophysics Data System (ADS)

    Sumner, James J.; Mmerole, Robert U.; Stratis-Cullum, Dimitra N.; Yi, Hyunmin; Bentley, William E.; Gillespie, James B.

    2003-08-01

    There is a great need for high throughput and sensitive sensors for genetic analysis. These sensors can be used for varied purposes from monitoring gene expression in organims to speciation of possible pathogens. Consequently, an instrument capable of these tasks would be a great benefit for food and water safety, medical diagnostics and defense of military and civilian populations from biological threats. This work examines the development of a hybridization-based biosensor using a novel tapered fiber optic rpobe. The immobilization of single-stranded, synthetic ologinucleotides utilizing aminoproplytriethoxysilane and glutaraldehyde was implemented on the fiber optic sensor. Hybridization takes place with a complementary analyte sequence followed by a fluorescent, labeled signaling probe to form a sandwich assay. Following hybridization, the fiber is interrogated with a diode laser source and the resulting fluorescence signal is detected using a miniature spectrometer.

  9. Characterization of one- and two-photon excitation fluorescence resonance energy transfer microscopy.

    PubMed

    Elangovan, Masilamani; Wallrabe, Horst; Chen, Ye; Day, Richard N; Barroso, Margarida; Periasamy, Ammasi

    2003-01-01

    Advances in molecular biology provide various methods to define the structure and function of the individual proteins that form the component parts of subcellular structures. The ability to see the dynamic behavior of a specific protein inside the living cell became possible through the application of advanced fluorescence resonance energy transfer (FRET) microscope techniques. The fluorophore molecule used for FRET imaging has a characteristic absorption and emission spectrum that should be considered for characterizing the FRET signal. In this article we describe the system development for the image acquisition for one- and two-photon excitation FRET microscopy. We also describe the precision FRET (PFRET) data analysis algorithm that we developed to remove spectral bleed-through and variation in the fluorophore expression level (or concentration) for the donor and acceptor molecules. The acquired images have been processed using a PFRET algorithm to calculate the energy transfer efficiency and the distance between donor and acceptor molecules. We implemented the software correction to study the organization of the apical endosome in epithelial polarized MDCK cells and dimerization of the CAATT/enhancer binding protein alpha (C/EBPalpha). For these proteins, the results revealed that the extent of correction affects the conventionally calculated energy transfer efficiency (E) and the distance (r) between donor and acceptor molecules by 38 and 9%, respectively. PMID:12543072

  10. Short-range ordered photonic structures of lamellae-forming diblock copolymers for excitation-regulated fluorescence enhancement.

    PubMed

    Kim, Se Hee; Kim, Ki-Se; Char, Kookheon; Yoo, Seong Il; Sohn, Byeong-Hyeok

    2016-05-19

    Photonic crystals can be represented by periodic nanostructures with alternating refractive indices, which create artificial stop bands with the appearance of colors. In this regard, nanodomains of block copolymers and the corresponding structural colors have been intensively studied in the past. However, the practical application of photonic crystals of block copolymers has been limited to a large degree because of the presence of large defects and grain boundaries in the nanodomains of block copolymers. The present study focuses on the alternative opportunity of short-range ordered nanodomains of block copolymers for fluorescence enhancement, which also has a direct relevance to the development of fluorescence sensors or detectors. The enhancement mechanism was found to be interconnected with the excitation process rather than the alternation of the decay kinetics. In particular, we demonstrate that randomly oriented, but regular grains of lamellae of polystyrene-block-polyisoprene, PS-b-PI, diblock copolymers and their blend with PS homopolymers can behave as Bragg mirrors to induce multiple reflections of the excitation source inside the photonic structures. This process in turn significantly increases the effective absorption of the given fluorophores inside the polymeric photonic structures to amplify the fluorescence signal.

  11. Photodynamic tumor therapy and on-line fluorescence spectroscopy after ALA administration using 633-nm light as therapeutic and fluorescence excitation radiation

    NASA Astrophysics Data System (ADS)

    Koenig, Karsten; Kienle, Alwin; Boehncke, Wolf-Henning; Kaufmann, Roland; Rueck, Angelika C.; Meier, Thomas H.; Steiner, Rudolf W.

    1994-09-01

    Photodynamic therapy (PDT) and on-line fluorescence spectroscopy were carried out on human tumors after 5-aminolevulinic acid (ALA) administration using 633-nm light of a dye laser as therapeutic radiation and as fluorescence excitation radiation. This has the advantages of (1) enabling use of one laser for PDT and fluorescence diagnosis only, (2) enabling the possibility of on-line fluorescence measurements, and (3) exciting protoporphyrin molecules in deep tissue layers. Monte Carlo calculations were carried out to determine excitation and fluorescence phonon distribution in case of red and violet excitation radiation. The results show the possibility of depth-resolved measurements on the fluorophore distribution by variation of excitation wavelength. The high penetration depth of 633-nm radiation results in a higher ratio of the 700-nm protoporphyrin fluorescence of the xenotransplanted tumor It to Is compared with 407-nm excitation. No values greater than 1 for the ratio I/Is were found, however, in case of intravenous ALA injection even for red excitation. Therefore, a large amount of ALA will be metabolized in the skin and can cause photosensitivity of the patient when applied systematically. In contrast, protoporphyrin fluorescence limited to the pretreated skin area was detected in case of topically applied ALA to patients with mycosis funcoides and erythroplasy of Queyrat. The influence of remitted excitation light and of the spontaneous radiation from the laser as well as the possible excitation of foodbased degradation products of chlorophyll has to be considered in high-sensitivity fluorescence measurements.

  12. Boron Difluoride Curcuminoid Fluorophores with Enhanced Two-Photon Excited Fluorescence Emission and Versatile Living-Cell Imaging Properties.

    PubMed

    Kamada, Kenji; Namikawa, Tomotaka; Senatore, Sébastien; Matthews, Cédric; Lenne, Pierre-François; Maury, Olivier; Andraud, Chantal; Ponce-Vargas, Miguel; Le Guennic, Boris; Jacquemin, Denis; Agbo, Peter; An, Dahlia D; Gauny, Stacey S; Liu, Xin; Abergel, Rebecca J; Fages, Frédéric; D'Aléo, Anthony

    2016-04-01

    The synthesis of boron difluoride complexes of a series of curcuminoid derivatives containing various donor end groups is described. Time-dependent (TD)-DFT calculations confirm the charge-transfer character of the second lowest-energy transition band and ascribe the lowest energy band to a "cyanine-like" transition. Photophysical studies reveal that tuning the donor strength of the end groups allows covering a broad spectral range, from the visible to the NIR region, of the UV-visible absorption and fluorescence spectra. Two-photon-excited fluorescence and Z-scan techniques prove that an increase in the donor strength or in the rigidity of the backbone results in a considerable increase in the two-photon cross section, reaching 5000 GM, with predominant two-photon absorption from the S0-S2 charge-transfer transition. Direct comparisons with the hemicurcuminoid derivatives show that the two-photon active band for the curcuminoid derivatives has the same intramolecular charge-transfer character and therefore arises from a dipolar structure. Overall, this structure-relationship study allows the optimization of the two-photon brightness (i.e., 400-900 GM) with one dye that emits in the NIR region of the spectrum. In addition, these dyes demonstrate high intracellular uptake efficiency in Cos7 cells with emission in the visible region, which is further improved by using porous silica nanoparticles as dye vehicles for the imaging of two mammalian carcinoma cells type based on NIR fluorescence emission. PMID:26919627

  13. Boron Difluoride Curcuminoid Fluorophores with Enhanced Two-Photon Excited Fluorescence Emission and Versatile Living-Cell Imaging Properties.

    PubMed

    Kamada, Kenji; Namikawa, Tomotaka; Senatore, Sébastien; Matthews, Cédric; Lenne, Pierre-François; Maury, Olivier; Andraud, Chantal; Ponce-Vargas, Miguel; Le Guennic, Boris; Jacquemin, Denis; Agbo, Peter; An, Dahlia D; Gauny, Stacey S; Liu, Xin; Abergel, Rebecca J; Fages, Frédéric; D'Aléo, Anthony

    2016-04-01

    The synthesis of boron difluoride complexes of a series of curcuminoid derivatives containing various donor end groups is described. Time-dependent (TD)-DFT calculations confirm the charge-transfer character of the second lowest-energy transition band and ascribe the lowest energy band to a "cyanine-like" transition. Photophysical studies reveal that tuning the donor strength of the end groups allows covering a broad spectral range, from the visible to the NIR region, of the UV-visible absorption and fluorescence spectra. Two-photon-excited fluorescence and Z-scan techniques prove that an increase in the donor strength or in the rigidity of the backbone results in a considerable increase in the two-photon cross section, reaching 5000 GM, with predominant two-photon absorption from the S0-S2 charge-transfer transition. Direct comparisons with the hemicurcuminoid derivatives show that the two-photon active band for the curcuminoid derivatives has the same intramolecular charge-transfer character and therefore arises from a dipolar structure. Overall, this structure-relationship study allows the optimization of the two-photon brightness (i.e., 400-900 GM) with one dye that emits in the NIR region of the spectrum. In addition, these dyes demonstrate high intracellular uptake efficiency in Cos7 cells with emission in the visible region, which is further improved by using porous silica nanoparticles as dye vehicles for the imaging of two mammalian carcinoma cells type based on NIR fluorescence emission.

  14. Investigating the 3.3 micron infrared fluorescence from naphthalene following ultraviolet excitation

    NASA Technical Reports Server (NTRS)

    Williams, Richard M.; Leone, Stephen R.

    1994-01-01

    Polycyclic aromatic hydrocarbon (PAH) type molecules are proposed as the carriers of the unidentified infrared (UIR) bands. Detailed studies of the 3.3 micrometer infrared emission features from naphthalene, the simplest PAH, following ultraviolet laser excitation are used in the interpretation of the 3.29 micrometer (3040 cm(sup -1)) UIR band. A time-resolved Fourier transform spectrometer is used to record the infrared emission spectrum of gas-phase naphthalene subsequent to ultraviolet excitation facilitated by an excimer laser operated at either 193 nm or 248 nm. The emission spectra differ significantly from the absorption spectrum in the same spectral region. Following 193 nm excitation the maximum in the emission profile is red-shifted 45 cm(sup -1) relative to the absorption maximum; a 25 cm(sup -1) red-shift is observed after 248 nm excitation. The red-shifting of the emission spectrum is reduced as collisional and radiative relaxation removes energy from the highly vibrationally excited molecules. Coupling between the various vibrational modes is thought to account for the differences between absorption and emission spectra. Strong visible emission is also observed following ultraviolet excitation. Visible emission may play an important role in the rate of radiative relaxation, which according to the interstellar PAH hypothesis occurs only by the slow emission of infrared photons. Studying the visible emission properties of PAH type molecules may be useful in the interpretation of the DIB's observed in absorption.

  15. Excitation emission and time-resolved fluorescence spectroscopy of selected varnishes used in historical musical instruments.

    PubMed

    Nevin, Austin; Echard, Jean-Philippe; Thoury, Mathieu; Comelli, Daniela; Valentini, Gianluca; Cubeddu, Rinaldo

    2009-11-15

    The analysis of various varnishes from different origins, which are commonly found on historical musical instruments was carried out for the first time with both fluorescence excitation emission spectroscopy and laser-induced time-resolved fluorescence spectroscopy. Samples studied include varnishes prepared using shellac, and selected diterpenoid and triterpenoid resins from plants, and mixtures of these materials. Fluorescence excitation emission spectra have been collected from films of naturally aged varnishes. In parallel, time-resolved fluorescence spectroscopy of varnishes provides means for discriminating between short- (less than 2.0 ns) and long-lived (greater than 7.5 ns) fluorescence emissions in each of these complex materials. Results suggest that complementary use of the two non destructive techniques allows a better understanding of the main fluorophores responsible for the emission in shellac, and further provides means for distinguishing the main classes of other varnishes based on differences in fluorescence lifetime behaviour. Spectrofluorimetric data and time resolved spectra presented here may form the basis for the interpretation of results from future in situ fluorescence examination and time resolved fluorescence imaging of varnished musical instruments.

  16. Fluorescence excitation by enhanced plasmon upconversion under continuous wave illumination

    NASA Astrophysics Data System (ADS)

    Tasgin, Mehmet Emre; Salakhutdinov, Ildar; Kendziora, Dania; Abak, Musa Kurtulus; Turkpence, Deniz; Piantanida, Luca; Fruk, Ljiljana; Lazzarino, Marco; Bek, Alpan

    2016-09-01

    We demonstrate effective background-free continuous wave nonlinear optical excitation of molecules that are sandwiched between asymmetrically constructed plasmonic gold nanoparticle clusters. We observe that near infrared photons are converted to visible photons through efficient plasmonic second harmonic generation. Our theoretical model and simulations demonstrate that Fano resonances may be responsible for being able to observe nonlinear conversion using a continuous wave light source. We show that nonlinearity enhancement of plasmonic nanostructures via coupled quantum mechanical oscillators such as molecules can be several orders larger as compared to their classical counterparts.

  17. Fluorescence excitation enhancement by Bloch surface wave in all-polymer one-dimensional photonic structure

    SciTech Connect

    Fornasari, L.; Floris, F.; Patrini, M.; Guizzetti, G.; Marabelli, F.; Canazza, G.; Comoretto, D.

    2014-08-04

    We demonstrate photoluminescence excitation enhancement in an all-polymer flexible one-dimensional photonic crystal structure capped with a fluorescent organic ultrathin film. When optical matching conditions between the excitation beam and the Bloch Surface Wave mode supported by the photonic structure are achieved, a ten times enhancement of the photoluminescence is observed. We notice that in these systems luminescence signal reinforcement is achieved by increasing the pump efficiency with no need of spectral resonance to the emission of the chosen fluorophore. All these features make these systems suitable candidates for easy, flexible, and cheap fluorescent sensing.

  18. Optimal fluorescence excitation and emission bands for detection of fecal contamination.

    PubMed

    Kim, Moon S; Lefcourt, Alan M; Chen, Yud-Ren

    2003-07-01

    Fecal contamination of food products is a critical health issue. To test the feasibility of the use fluorescent techniques to detect fecal contamination, fluorescence excitation and emission characteristics of fecal matter from cows, deer, swine, chickens, and turkeys in the UV to far-red regions of the spectrum were evaluated. To allow the optimization of the detection of fecal contamination on animal carcasses and cut meats, emission-excitation spectra of the feces were compared with spectra for animal meats. The feedstuffs for the swine, chickens, and turkeys were also analyzed. Excitation at approximately 410 to 420 nm yielded the highest level of fluorescence for both feces and feedstuffs. Emission maxima were in the red region (at 632 nm for chicken feces and at 675 nm for the feces of the other species). The major constituent responsible for emission at 632 nm was tentatively identified as protoporphyrin IX; emission at 675 nm most likely emanates from chlorophyll a or its metabolites. Animal meats emitted strong fluorescence in the blue-green regions, but no emission peaks were observed in the red region for these meats. These results suggest that fluorescence emissions from naturally occurring chlorophyll a and its metabolites are good markers for fecal contamination and that with excitation at 410 to 420 nm, the responses of fecal matter can easily be differentiated from the responses of animal meats. We suggest that the detection of fecal contamination can be enhanced by requiring a minimum chlorophyll a content in the finishing diets of all farm animals.

  19. Construction, figures of merit, and testing of a single-cell fluorescence excitation spectroscopy system

    PubMed Central

    Hill, Laura S.; Richardson, Tammi L.; Profeta, Luisa T. M.; Shaw, Timothy J.; Hintz, Christopher J.; Twining, Benjamin S.; Lawrenz, Evelyn; Myrick, Michael L.

    2010-01-01

    Characterization of phytoplankton community composition is critical to understanding the ecology and biogeochemistry of the oceans. One approach to taxonomic characterization takes advantage of differing pigmentation between algal taxa and thus differences in fluorescence excitation spectra. Analyses of bulk water samples, however, may be confounded by interference from chromophoric dissolved organic matter or suspended particulate matter. Here, we describe an instrument that uses a laser trap based on a Nikon TE2000-U microscope to position individual phytoplankton cells for confocal fluorescence excitation spectroscopy, thus avoiding interference from the surrounding medium. Quantitative measurements of optical power give data in the form of photons emitted per photon of exposure for an individual phytoplankton cell. Residence times for individual phytoplankton in the instrument can be as long as several minutes with no substantial change in their fluorescence excitation spectra. The laser trap was found to generate two-photon fluorescence from the organisms so a modification was made to release the trap momentarily during data acquisition. Typical signal levels for an individual cell are in the range of 106 photons∕s of fluorescence using a monochromated 75 W Xe arc lamp excitation source with a 2% transmission neutral density filter. PMID:20113077

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

    NASA Technical Reports Server (NTRS)

    Lin, C.; Dienes, A.

    1973-01-01

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

  1. Magnetic field effects on yields and decays of fluorescence of pyrimidine vapor: Excited rovibronic level dependence

    NASA Astrophysics Data System (ADS)

    Ohta, Nobuhiro; Takemura, Takeshi; Fujita, Masahisa; Baba, Hiroaki

    1988-04-01

    External magnetic field effects on yields and decays of fluorescence of pyrimidine vapor on excitation into various rotational levels belonging to the vibrationless level or the 6a1 level of S1 have been studied in a supersonic jet or in a bulk gas at room temperature with a field strength of 0-150 G. The fast component of fluorescence is not affected by an external magnetic field, whereas the slow fluorescence is quenched by a field except for excitation at the R(0) line belonging to the 0-0 transition. The fluorescence quenching is more effective at the 6a1 level than that at 00, indicating that the level density of the triplet state coupled to the singlet state plays an important role in the magnetic mixing of the triplet spin sublevels, in terms of which the fluorescence quenching by a magnetic field is interpreted. The excited rotational level dependence of the fluorescence quenching by a magnetic field is attributed to K scrambling in the triplet manifold following intersystem crossing.

  2. A novel non-fluorescent excited state intramolecular proton transfer phenomenon induced by intramolecular hydrogen bonds: an experimental and theoretical investigation

    PubMed Central

    Yin, Hang; Li, Hui; Xia, Guomin; Ruan, Chengyan; Shi, Ying; Wang, Hongming; Jin, Mingxing; Ding, Dajun

    2016-01-01

    Two molecules, 1-hydroxypyrene-2-carbaldehyde (HP) and 1-methoxypyrene-2-carbaldehyde (MP) were explored. We investigated their photophysical properties, using experimental transient absorption and theoretical density functional theory/time-dependent density functional theory (DFT/TDDFT). HP and MP have similar geometric conformations but exhibit entirely different photophysical properties upon excitation into the S1 state. In contrast to traditional excited state intramolecular proton transfer (ESIPT) in molecules that exhibit either single or dual fluorescence, HP has an unusual non-fluorescent property. Specifically, the ultrafast ESIPT process occurs in 158 fs and is followed by an intersystem crossing (ISC) component of 11.38 ps. In contrast to HP, MP undergoes only an 8 ps timescale process, which was attributed to interactions between solute and solvent. We concluded that this difference arises from intramolecular hydrogen bonds (IMHBs), which induce drastic structural alterntion upon excitation. PMID:26790961

  3. Two-photon-excited fluorescence resonance energy transfer in an aqueous system of CdTe quantum dots and Rhodamine B

    SciTech Connect

    Li, Muye; Lu, Peixiang; Li, Fang He, Zhicong; Zhang, Junpei; Han, Junbo

    2014-12-21

    Two-photon excited fluorescence resonance energy transfer (FRET) between CdTe quantum dots with different emission peaks and Rhodamine B in aqueous solution are investigated both experimentally and theoretically. The photoluminescence and lifetime are measured using a time-resolved fluorescence test system. The two-photon excited FRET efficiency is found to increase as the degree of spectral overlap of the emission spectrum of CdTe and the absorption spectrum of Rhodamine B increases, which is due to the increase of Forster radius of the sample. Moreover, FRET efficiency increases when the ratio of acceptor/donor concentration increases. The two-photon excited FRET efficiency was found to reach 40%.

  4. Delayed fluorescence during the deactivation of highly excited triplet states

    SciTech Connect

    Skvortsov, V.I.; Alfimov, M.V.

    1987-06-01

    It has been suggested that the T state may be not only an electron donor but an acceptor, i.e., it may give a charge transfer state in a photoreduction reaction: A(T) + M ..-->.. /sup 3/(A/sup .-/M/sup +./). In this connection, it may be assumed that the quenching may also be connected with the nonradiative deactivation of excitation energy in charge transfer states (A/sup .-/M/sup +./). Aromatic molecules are characterized by an extremely low quantum yield for intramolecular intersystem crossing. However, in charge-transfer states the efficiency of intersystem crossing may be enhanced. The authors have investigated the laws governing DF in conditions where the deactivation of T states occurs. The systems studied were frozen (77 K) solutions of the aromatic additives naphthalene, diphenyl, and chrysene in toluene, MCH, and ethanol.

  5. Diagnostics of a see-through hollow cathode discharge by emission, absorption, and fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Taylor, Nicholas

    Atomic line filters have been suggested to be attractive in areas of Doppler velocimetry, resonance fluorescence detection, and resonance ionization detection. They are based on the resonant absorption of photons by an atomic vapor, and allow all other radiation to pass. This allows the detection of very low levels of light superimposed on a large optical background. Several elements have been studied for use as atomic line filters, such as the alkali metals, alkaline earths, and thallium. As previously recognized, thallium is especially attractive since the 535.046 nm metastable transition overlaps with the second harmonic output of an Nd:La2Be2O 5 (BEL) laser (1070 nm). This makes thallium ideal for certain applications as an atomic line filter. Recently a see-through hollow cathode lamp, or galvatron (Hamamatsu), was made commercially available. The galvatron geometry is unique compared to traditional hollow cathode lamps since the cathode and cell are oriented in a T-shape, with the cathode bored completely through to allow the propagation of a light source through the cathode. This allows multi-step excitation of the atomic vapor, not easily accomplished with a traditional hollow cathode lamp. The advantages that a galvatron offers over conventional atomic reservoirs make it an attractive candidate for the application as an atomic line filter; however, little spectroscopic data have been found in the literature. For this reason, Doppler temperatures, number densities, quantum efficiencies, and lifetimes have been determined in order to characterize this atomic reservoir as a potential atomic line filter. These parameters are determined by use of various spectroscopic techniques which include emission, absorption, time-resolved fluorescence, and time-resolved laser-induced saturated fluorescence spectroscopy. From these measurements, it has been demonstrated that a galvatron is an attractive atomic reservoir for applications as an atomic line filter. The

  6. An experimental study of the electronic absorption and fluorescence spectral properties of new p-substituted-N-phenylpyrroles and their electrosynthesized polymers.

    PubMed

    Diaw, A K D; Gningue-Sall, D; Yassar, A; Brochon, J-C; Henry, E; Aaron, J-J

    2015-01-25

    Electronic absorption and fluorescence spectral properties of new p-substituted-N-phenylpyrroles (N-PhPys), including HOPhPy, MeOPhPy, ThPhPy, PhDPy, DPhDPy, PyPhThThPhPy, and their available, electrosynthesized polymers were investigated. Electronic absorption spectra, fluorescence excitation and emission spectra, fluorescence quantum yields (ΦF) and lifetimes (τF), and other photophysical parameters of these N-PhPy derivatives and their polymers were measured in DMF, DMSO diluted solutions and/or solid state at room temperature. The electronic absorption spectra of N-PhPy derivatives and their polymers included one to several bands, located in the 270-395 nm region, according to the p-phenyl substituent electron-donating effect and conjugated heteroaromatic system length. The fluorescence excitation spectra were characterized by one broad main peak, with, in most cases, one (or more) poorly resolved shoulder (s), appearing in the 270-405 nm region, and their emission spectra were generally constituted of several bands located in the 330-480 nm region. No significant shift of the absorption, fluorescence excitation and emission spectra wavelengths was found upon going from the monomers to the corresponding polymers. ΦF values were high, varying between 0.11 and 0.63, according to the nature of substituents(s) and to the conjugated system extension. Fluorescence decays were mono-exponential for the monomers and poly-exponential for PyPhThThPhPy and for polymers. τF values were relatively short (0.35-5.17 ns), and markedly decreased with the electron-donor character of the phenyl group p-substituent and the conjugated system extension.

  7. Multispectral fluorescence lifetime imaging of feces-contaminated apples by time-resolved laser-induced fluorescence imaging system with tunable excitation wavelengths

    NASA Astrophysics Data System (ADS)

    Kim, Moon S.; Cho, Byoung-Kwan; Lefcourt, Alan M.; Chen, Yud-Ren; Kang, Sukwon

    2008-04-01

    We recently developed a time-resolved multispectral laser-induced fluorescence (LIF) imaging system capable of tunable wavelengths in the visible region for sample excitation and nanosecond-scale characterizations of fluorescence responses (lifetime imaging). Time-dependent fluorescence decay characteristics and fluorescence lifetime imaging of apples artificially contaminated with a range of diluted cow feces were investigated at 670 and 685 nm emission bands obtained by 418, 530, and 630 nm excitations. The results demonstrated that a 670 nm emission with a 418 nm excitation provided the greatest difference in time-dependent fluorescence responses between the apples and feces-treated spots. The versatilities of the time-resolved LIF imaging system, including fluorescence lifetime imaging of a relatively large biological object in a multispectral excitation-emission wavelength domain, were demonstrated.

  8. Dual fluorescence excitation spectra of methyl salicylate in a free jet

    NASA Astrophysics Data System (ADS)

    Heimbrook, Lou Ann; Kenny, Jonathan E.; Kohler, Bryan E.; Scott, Gary W.

    1981-11-01

    Separate fluorescence excitation spectra of the blue- and UV-emitting forms of methyl salicylate cooled in a free-jet expansion are reported. This study represents the first observation of the detailed vibrational structure of these transitions. The two excitation spectra have no features in common, and their intensity patterns are very different. Many individual lines are ˜2 cm-1 wide (nearly laser limited), although in the excitation spectrum of the UV emission, spectral congestion persists at high energies despite the high degree of cooling. (AIP)

  9. Effect of anthocyanins, carotenoids, and flavonols on chlorophyll fluorescence excitation spectra in apple fruit: signature analysis, assessment, modelling, and relevance to photoprotection.

    PubMed

    Merzlyak, Mark N; Melø, Thor Bernt; Naqvi, K Razi

    2008-01-01

    Whole apple fruit (Malus domestica Borkh.) widely differing in pigment content and composition has been examined by recording its chlorophyll fluorescence excitation and diffuse reflection spectra in the visible and near UV regions. Spectral bands sensitive to the pigment concentration have been identified, and linear models for non-destructive assessment of anthocyanins, carotenoids, and flavonols via chlorophyll fluorescence measurements are put forward. The adaptation of apple fruit to high light stress involves accumulation of these protective pigments, which absorb solar radiation in broad spectral ranges extending from UV to the green and, in anthocyanin-containing cultivars, to the red regions of the spectrum. In ripening apples the protective effect in the blue region could be attributed to extrathylakoid carotenoids. A simple model, which allows the simulation of chlorophyll fluorescence excitation spectra in the visible range and a quantitative evaluation of competitive absorption by anthocyanins, carotenoids, and flavonols, is described. Evidence is presented to support the view that anthocyanins, carotenoids, and flavonols play, in fruit with low-to-moderate pigment content, the role of internal traps (insofar as they compete with chlorophylls for the absorption of incident light in specific spectral bands), affecting thereby the shape of the chlorophyll fluorescence excitation spectrum.

  10. Effect of gold nanoparticles on the fluorescence excitation spectrum of α-fetoprotein: Local environment dependent fluorescence quenching

    NASA Astrophysics Data System (ADS)

    Li, Jian-jun; Chen, Yu; Wang, A.-qing; Zhu, Jian; Zhao, Jun-wu

    2011-01-01

    The effect of colloid gold nanoparticles (AuNPs) on the fluorescence excitation spectrum of α-fetoprotein (AFP) has been investigated experimentally. The excitation spectral peaks of AFP with low concentration from 0.01 ng ml -1 to 12 ng ml -1 increase monotonically with increasing of AFP concentration. When some gold colloids were added to the AFP solution, the excitation peak at 285 nm decreases distinctly. By comparing the excitation peak intensity of AFP solution with gold colloids and without gold colloids at different AFP concentrations, the quenching effect from gold nanoparticle was more effective at lower AFP concentration. So the range of concentration from 0.01 ng ml -1 to 0.09 ng ml -1 will be the potential range of applications because of the higher sensitivity. The physical origin based on local field effect was investigated to illuminate this local environment dependent fluorescence quenching. The changing extent of quenching with different AFP concentrations can be attributed to the nonlinear decreasing of the local field factor of gold nanoparticles as a function of environmental dielectric constant.

  11. Ultrafast Excited State Relaxation of a Metalloporphyrin Revealed by Femtosecond X-ray Absorption Spectroscopy.

    PubMed

    Shelby, Megan L; Lestrange, Patrick J; Jackson, Nicholas E; Haldrup, Kristoffer; Mara, Michael W; Stickrath, Andrew B; Zhu, Diling; Lemke, Henrik T; Chollet, Matthieu; Hoffman, Brian M; Li, Xiaosong; Chen, Lin X

    2016-07-20

    Photoexcited Nickel(II) tetramesitylporphyrin (NiTMP), like many open-shell metalloporphyrins, relaxes rapidly through multiple electronic states following an initial porphyrin-based excitation, some involving metal centered electronic configuration changes that could be harnessed catalytically before excited state relaxation. While a NiTMP excited state present at 100 ps was previously identified by X-ray transient absorption (XTA) spectroscopy at a synchrotron source as a relaxed (d,d) state, the lowest energy excited state (J. Am. Chem. Soc., 2007, 129, 9616 and Chem. Sci., 2010, 1, 642), structural dynamics before thermalization were not resolved due to the ∼100 ps duration of the available X-ray probe pulse. Using the femtosecond (fs) X-ray pulses of the Linac Coherent Light Source (LCLS), the Ni center electronic configuration from the initial excited state to the relaxed (d,d) state has been obtained via ultrafast Ni K-edge XANES (X-ray absorption near edge structure) on a time scale from hundreds of femtoseconds to 100 ps. This enabled the identification of a short-lived Ni(I) species aided by time-dependent density functional theory (TDDFT) methods. Computed electronic and nuclear structure for critical excited electronic states in the relaxation pathway characterize the dependence of the complex's geometry on the electron occupation of the 3d orbitals. Calculated XANES transitions for these excited states assign a short-lived transient signal to the spectroscopic signature of the Ni(I) species, resulting from intramolecular charge transfer on a time scale that has eluded previous synchrotron studies. These combined results enable us to examine the excited state structural dynamics of NiTMP prior to thermal relaxation and to capture intermediates of potential photocatalytic significance.

  12. Decomposing the First Absorption Band of OCS Using Photofragment Excitation Spectroscopy.

    PubMed

    Toulson, Benjamin W; Murray, Craig

    2016-09-01

    Photofragment excitation spectra of carbonyl sulfide (OCS) have been recorded from 212-260 nm by state-selectively probing either electronically excited S((1)D) or ground state S((3)P) photolysis products via 2 + 1 resonance-enhanced multiphoton ionization. Probing the major S((1)D) product results in a broad, unstructured action spectrum that reproduces the overall shape of the first absorption band. In contrast, spectra obtained probing S((3)P) products display prominent resonances superimposed on a broad continuum; the resonances correspond to the diffuse vibrational structure observed in the conventional absorption spectrum. The vibrational structure is assigned to four progressions, each dominated by the C-S stretch, ν1, following direct excitation to quasi-bound singlet and triplet states. The S((3)PJ) products are formed with a near-statistical population distribution over the J = 2, 1, and 0 spin-orbit levels across the wavelength range investigated. Although a minor contributor to the S atom yield near the peak of the absorption cross section, the relative yield of S((3)P) increases significantly at longer wavelengths. The experimental measurements validate recent theoretical work characterizing the electronic states responsible for the first absorption band by Schmidt and co-workers. PMID:27552402

  13. Optimal optical filters of fluorescence excitation and emission for poultry fecal detection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Purpose: An analytic method to design excitation and emission filters of a multispectral fluorescence imaging system is proposed and was demonstrated in an application to poultry fecal inspection. Methods: A mathematical model of a multispectral imaging system is proposed and its system parameters, ...

  14. The Effective of Different Excitation Wavelengths on the Identification of Plant Species Based on Fluorescence LIDAR

    NASA Astrophysics Data System (ADS)

    Yang, Jian; Gong, Wei; Shi, Shuo; Du, Lin; Sun, Jia; Song, Shalei

    2016-06-01

    Laser-induced fluorescence (LIF) served as an active technology has been widely used in many field, and it is closely related to excitation wavelength (EW). The objective of this investigation is to discuss the performance of different EWs of LIF LiDAR in identifying plant species. In this study, the 355, 460 and 556 nm lasers were utilized to excite the leaf fluorescence and the fluorescence spectra were measured by using the LIF LiDAR system built in the laboratory. Subsequently, the principal component analysis (PCA) with the help of support vector machine (SVM) was utilized to analyse fluorescence spectra. For the three EWs, the overall identification rates of the six plant species were 80 %, 83.3 % and 90 %. Experimental results demonstrated that 556 nm excitation light source is superior to 355 and 460 nm for the classification of the plant species for the same genus in this study. Thus, an appropriate excitation wavelength should be considered when the LIF LiDAR was utilized in the field of remote sensing based on the LIF technology.

  15. Fluorescence excitation and multiphoton ionization spectroscopy of 3-methylindole in a supersonic jet

    NASA Astrophysics Data System (ADS)

    Hays, T. R.; Henke, W. E.; Selzle, H. L.; Schlag, E. W.

    1983-05-01

    The fluorescence excitation and multiphoton ionization spectroscopy of 3-methylindole (skatole) is reported. One electronic origin ( 1L b) is assigned at 34875 cm -1, the second ( 1L a) suspected at 35483 cm -1. Some 1L b vibrational assignments are also made. Complex formation between skatole and some small molecules is indicated but not directly observed.

  16. Shape of the absorption and fluorescence spectra of condensed phases and transition energies.

    PubMed

    Lagos, Miguel; Paredes, Rodrigo

    2014-11-13

    General integral expressions for the temperature-dependent profile of the spectral lines of photon absorption and emission by atomic or molecular species in a condensed environment are derived with no other hypothesis than: (a) The acoustic vibrational modes of the condensed host medium constitute the thermodynamic energy reservoir at a given constant temperature, and local electronic transitions modifying the equilibrium configuration of the surroundings are multiphonon events, regardless of the magnitude of the transition energy. (b) Electron-phonon coupling is linear in the variations of the bond length. The purpose is to develop a theoretical tool for the analysis of the spectra, allowing us to grasp highly accurate information from fitting the theoretical line shape function to experiment, including those spectra displaying wide features. The method is illustrated by applying it to two dyes, Lucifer Yellow CH and Coumarin 1, which display fluorescence maxima of 0.41 and 0.51 eV fwhm. Fitting the theoretical curves to the spectra indicates that the neat excitation energies are 2.58 eV ± 2.5% and 3.00 eV ± 2.0%, respectively.

  17. Fluorescence excitation-emission matrix spectroscopy as a tool for determining quality of sparkling wines.

    PubMed

    Elcoroaristizabal, Saioa; Callejón, Raquel M; Amigo, Jose M; Ocaña-González, Juan A; Morales, M Lourdes; Ubeda, Cristina

    2016-09-01

    Browning in sparkling wines was assessed by the use of excitation-emission fluorescence spectroscopy combined with PARAllel FACtor analysis (PARAFAC). Four different cava sparkling wines were monitored during an accelerated browning process and subsequently storage. Fluorescence changes observed during the accelerated browning process were monitored and compared with other conventional parameters: absorbance at 420nm (A420) and the content of 5-hydroxymethyl-2-furfural (5-HMF). A high similarity of the spectral profiles for all sparkling wines analyzed was observed, being explained by a four component PARAFAC model. A high correlation between the third PARAFAC factor (465/530nm) and the commonly used non-enzymatic browning indicators was observed. The fourth PARAFAC factor (280/380nm) gives us also information about the browning process following a first order kinetic reaction. Hence, excitation-emission fluorescence spectroscopy, together with PARAFAC, provides a faster alternative for browning monitoring to conventional methods, as well as useful key indicators for quality control.

  18. Multifocal multiphoton excitation and time correlated single photon counting detection for 3-D fluorescence lifetime imaging.

    PubMed

    Kumar, S; Dunsby, C; De Beule, P A A; Owen, D M; Anand, U; Lanigan, P M P; Benninger, R K P; Davis, D M; Neil, M A A; Anand, P; Benham, C; Naylor, A; French, P M W

    2007-10-01

    We report a multifocal multiphoton time-correlated single photon counting (TCSPC) fluorescence lifetime imaging (FLIM) microscope system that uses a 16 channel multi-anode PMT detector. Multiphoton excitation minimizes out-of-focus photobleaching, multifocal excitation reduces non-linear in-plane photobleaching effects and TCSPC electronics provide photon-efficient detection of the fluorescence decay profile. TCSPC detection is less prone to bleaching- and movement-induced artefacts compared to wide-field time-gated or frequency-domain FLIM. This microscope is therefore capable of acquiring 3-D FLIM images at significantly increased speeds compared to single beam multiphoton microscopy and we demonstrate this with live cells expressing a GFP tagged protein. We also apply this system to time-lapse FLIM of NAD(P)H autofluorescence in single live cells and report measurements on the change in the fluorescence decay profile following the application of a known metabolic inhibitor. PMID:19550524

  19. Red-edge excitation fluorescence measurements of several two-tryptophan-containing proteins.

    PubMed

    Wasylewski, Z; Kołoczek, H; Waśniowska, A; Slizowska, K

    1992-05-15

    The dependence of the fluorescence emission maximum of the tryptophan residues in several two-tryptophan-containing proteins (horse liver alcohol dehydrogenase, yeast 3-phosphoglycerate kinase, Staphylococcus aureus metalloprotease and bee venom phospholipase A2) on the excitation wavelengths has been studied. Using fluorescence-resolved spectroscopy, we have dissected the contributions of particular tryptophan residues located in different parts of the protein molecule. The results demonstrate that dipolar structural relaxation can occur in the environment of tryptophan residues buried within protein molecules. The observed spectral shifts upon red-edge excitation of these residues can depend on temperature or ligand binding, as demonstrated in case of metalloprotease and alcohol dehydrogenase. No spectral shifts upon red-edge excitation have been observed for tryptophan residues totally exposed to the rapidly relaxing aqueous solvent.

  20. 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

  1. Determination of the in vivo redox potential using roGFP and fluorescence spectra obtained from one-wavelength excitation

    NASA Astrophysics Data System (ADS)

    Wierer, S.; Elgass, K.; Bieker, S.; Zentgraf, U.; Meixner, A. J.; Schleifenbaum, F.

    2011-02-01

    The analysis of molecular processes in living (plant) cells such as signal transduction, DNA replication, carbon metabolism and senescence has been revolutionized by the use of green fluorescent protein (GFP) and its variants as specific cellular markers. Many cell biological processes are accompanied by changes in the intracellular redox potential. To monitor the redox potential, a redox-sensitive mutant of GFP (roGFP) was created, which shows changes in its optical properties in response to changes in the redox state of its surrounding medium. For a quantitative analysis in living systems, it is essential to know the optical properties of roGFP in vitro. Therefore, we applied spectrally resolved fluorescence spectroscopy on purified roGFP exposed to different redox potentials to determine shifts in both the absorption and the emission spectra of roGFP. Based on these in vitro findings, we introduce a new approach using one-wavelength excitation to use roGFP for the in vivo analysis of cell biological processes. We demonstrate the ability this technique by investigating chloroplast-located Grx1-roGFP2 expressing Arabidopsis thaliana cells as example for dynamically moving intracellular compartments. This is not possible with the two-wavelength excitation technique established so far, which hampers a quantitative analysis of highly mobile samples due to the time delay between the two measurements and the consequential displacement of the investigated area.

  2. Photothermal measurement of absorption and scattering losses in thin films excited by surface plasmons.

    PubMed

    Domené, Esteban A; Balzarotti, Francisco; Bragas, Andrea V; Martínez, Oscar E

    2009-12-15

    We present a novel noncontact, photothermal technique, based on the focus error signal of a commercial CD pickup head that allows direct determination of absorption in thin films. Combined with extinction methods, this technique yields the scattering contribution to the losses. Surface plasmon polaritons are excited using the Kretschmann configuration in thin Au films of varying thickness. By measuring the extinction and absorption simultaneously, it is shown that dielectric constants and thickness retrieval leads to inconsistencies if the model does not account for scattering. PMID:20016617

  3. UV absorption study of collisional energy transfer in vibrationally highly excited SO/sub 2/ molecules

    SciTech Connect

    Heymann, M.; Hippler, H.; Nahr, D.; Plach, H.J.; Troe, J.

    1988-09-22

    Transient UV absorption spectra after UV laser excitation of SO/sub 2/ were recorded and analyzed with respect to collisional energy transfer. Byuse of previously determined calibration curves, the absorption-time signals were converted into average energy-number of collision profiles. Energy-dependent average energies transferred per collision (..delta..E) were derived for 22 different collision partners. The temperature dependence of (..delta..E) was determined over the range 300-1500 K by experiments in a CO/sub 2/ CW laser-heated reactor and in shock waves.

  4. Real-time intraoperative fluorescence imaging system using light-absorption correction

    NASA Astrophysics Data System (ADS)

    Themelis, George; Yoo, Jung Sun; Soh, Kwang-Sup; Schulz, Ralf; Ntziachristos, Vasilis

    2009-11-01

    We present a novel fluorescence imaging system developed for real-time interventional imaging applications. The system implements a correction scheme that improves the accuracy of epi-illumination fluorescence images for light intensity variation in tissues. The implementation is based on the use of three cameras operating in parallel, utilizing a common lens, which allows for the concurrent collection of color, fluorescence, and light attenuation images at the excitation wavelength from the same field of view. The correction is based on a ratio approach of fluorescence over light attenuation images. Color images and video is used for surgical guidance and for registration with the corrected fluorescence images. We showcase the performance metrics of this system on phantoms and animals, and discuss the advantages over conventional epi-illumination systems developed for real-time applications and the limits of validity of corrected epi-illumination fluorescence imaging.

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

    NASA Astrophysics Data System (ADS)

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

    2015-05-01

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

  6. Study of the Photodegradation Process of Vitamin E Acetate by Optical Absorption, Fluorescence, and Thermal Lens Spectroscopy

    NASA Astrophysics Data System (ADS)

    Tiburcio-Moreno, J. A.; Marcelín-Jiménez, G.; Leanos-Castaneda, O. L.; Yanez-Limon, J. M.; Alvarado-Gil, J. J.

    2012-11-01

    The stability of vitamin E acetate exposed to ultraviolet (UV) light was studied using three spectroscopic methods. An ethanol solution of vitamin E acetate was treated with either UVC light (254 nm) or UVA light (366 nm) during a period of 10 min followed by a study of UV-Vis optical absorption, then by fluorescence spectroscopy excitation by UV radiation at either 290 nm or 368 nm and, finally the solution was studied by thermal lens spectroscopy. Immediately, the same solution of vitamin E acetate was subjected to the UV irradiation process until completion of six periods of irradiation and measurements. UVC light treatment induced the appearance of a broad absorption band in the range of 310 nm to 440 nm with maximum absorbance at 368 nm, which progressively grew as the time of the exposure to UVC light increases. In contrast, UVA light treatment did not affect the absorption spectra of vitamin E acetate. Fluorescence spectra of the vitamin E acetate (without UV light treatment) showed no fluorescence when excited with 368 nm while exciting with 290 nm, an intense and broad emission band (300 nm to 440 nm) with a maximum at 340 nm appeared. When vitamin E acetate was treated with UVC light, this emission band progressively decreased as the time of the UVC light irradiation grew. No signal from UV-untreated vitamin E acetate could be detected by the thermal lens method. Interestingly, as the time of the UVC light treatment increased, the thermal lens signal progressively grew. Additional experiments performed to monitor the time evolution of the process during continuous UVC treatment of the vitamin E acetate using thermal lens spectroscopy exhibited a progressive increase of the thermal lens signal reaching a plateau at about 8000 s. This study shows that the vitamin E acetate is stable when it is irradiated with UVA light, while the irradiation with UVC light induces the formation of photodegradation products. Interestingly, this photodegradation process using

  7. Magnetic fluorescent lamp having reduced ultraviolet self-absorption

    DOEpatents

    Berman, Samuel M.; Richardson, Robert W.

    1985-01-01

    The radiant emission of a mercury-argon discharge in a fluorescent lamp assembly (10) is enhanced by providing means (30) for establishing a magnetic field with lines of force along the path of electron flow through the bulb (12) of the lamp assembly, to provide Zeeman splitting of the ultraviolet spectral line. Optimum results are obtained when the magnetic field strength causes a Zeeman splitting of approximately 1.7 times the thermal line width.

  8. Determination of K shell absorption jump factors and jump ratios of 3d transition metals by measuring K shell fluorescence parameters.

    PubMed

    Kaçal, Mustafa Recep; Han, Ibrahim; Akman, Ferdi

    2014-10-29

    Energy dispersive X-ray fluorescence technique (EDXRF) has been employed for measuring K-shell absorption jump factors and jump ratios for Ti, Cr, Fe, Co, Ni and Cu elements. The jump factors and jump ratios for these elements were determined by measuring K shell fluorescence parameters such as the Kα X-ray production cross-sections, K shell fluorescence yields, Kβ-to-Kα X-rays intensity ratios, total atomic absorption cross sections and mass attenuation coefficients. The measurements were performed using a Cd-109 radioactive point source and an Si(Li) detector in direct excitation and transmission experimental geometry. The measured values for jump factors and jump ratios were compared with theoretically calculated and the ones available in the literature. PMID:25464198

  9. Environment-sensitive quinolone demonstrating long-lived fluorescence and unusually slow excited-state intramolecular proton transfer kinetics

    NASA Astrophysics Data System (ADS)

    Zamotaiev, O. M.; Shvadchak, V.; Sych, T. P.; Melnychuk, N. A.; Yushchenko, D.; Mely, Y.; Pivovarenko, V. G.

    2016-09-01

    A new small fluorescent dye based on 3-hydroxybenzo[g]quinolone, a benzo-analogue of Pseudomonas quinolone signal species, has been synthesized. The dye demonstrates interesting optical properties, with absorption in the visible region, two band emission due to an excited-state intramolecular proton transfer (ESIPT) reaction and high fluorescence quantum yield in both protic and aprotic media. Time-resolved fluorescence spectroscopy shows that the ESIPT reaction time is unusually long (up to 8 ns), indicating that both forward and backward ESIPT reactions are very slow in comparison to other 3-hydroxyquinolones. In spite of these slow rate constants, the ESIPT reaction was found to show a reversible character as a result of the very long lifetimes of both N* and T* forms (up to 16 ns). The ESIPT reaction rate is mainly controlled by the hydrogen bond donor ability in protic solvents and the polarity in aprotic solvents. Using large unilamellar vesicles and giant unilamellar vesicles of different lipid compositions, the probe was shown to preferentially label liquid disordered phases.

  10. Role of excited state solvent fluctuations on time-dependent fluorescence Stokes shift

    SciTech Connect

    Li, Tanping E-mail: revatik@lsu.edu; Kumar, Revati E-mail: revatik@lsu.edu

    2015-11-07

    We explore the connection between the solvation dynamics of a chromophore upon photon excitation and equilibrium fluctuations of the solvent. Using molecular dynamics simulations, fluorescence Stokes shift for the tryptophan in Staphylococcus nuclease was examined using both nonequilibrium calculations and linear response theory. When the perturbed and unperturbed surfaces exhibit different solvent equilibrium fluctuations, the linear response approach on the former surface shows agreement with the nonequilibrium process. This agreement is excellent when the perturbed surface exhibits Gaussian statistics and qualitative in the case of an isomerization induced non-Gaussian statistics. However, the linear response theory on the unperturbed surface breaks down even in the presence of Gaussian fluctuations. Experiments also provide evidence of the connection between the excited state solvent fluctuations and the total fluorescence shift. These observations indicate that the equilibrium statistics on the excited state surface characterize the relaxation dynamics of the fluorescence Stokes shift. Our studies specifically analyze the Gaussian fluctuations of the solvent in the complex protein environment and further confirm the role of solvent fluctuations on the excited state surface. The results are consistent with previous investigations, found in the literature, of solutes dissolved in liquids.

  11. Toward Fourier interferometry fluorescence excitation/emission imaging of malignant cells combined with photoacoustic microscopy

    NASA Astrophysics Data System (ADS)

    Kohen, Elli; Hirschberg, Joseph G.; Berry, John P.; Ozkutuk, Nuri; Ornek, Ceren; Monti, Marco; Leblanc, Roger M.; Schachtschabel, Dietrich O.; Haroon, Sumaira

    2003-10-01

    Dual excitation fluorescence imaging has been used as a first step towards multi-wavelength excitation/emission fluorescence spectral imaging. Target cells are transformed keratinocytes, and other osteosarcoma, human breast and color cancer cells. Mitochondrial membrane potential probes, e.g. TMRM (tetramethylrhodamine methyl ester), Mitotracker Green (Molecular Probes, Inc., Eugene OR,USA; a recently synthesized mitochondrial oxygen probe, [PRE,P1"- pyrene butyl)-2-rhodamine ester] allow dual excitation in the UV plus in teh blue-green spectral regions. Also, using the natural endogenous probe NAD(P)H, preliminary results indicate mitochondrial responses to metabolic challenges (e.g. glucose addition), plus changes in mitochonrial distribution and morphology. In terms of application to biomedicine (for diagnostiscs, prognostsics and drug trials) three parameters have been selected in addition to the natural probe NAD(P)H, i.e. vital fluorescence probing of mitochondria, lysosomes and Golgi apparatus. It is hoped that such a multiparameter approach will allow malignant cell characterization and grading. A new area being introduced is the use of similar methodology for biotechnical applications such as the study of the hydrogen-producing alga Chlamydomonas Reinhardtii, and possible agricultural applications, such as Saccharomyces yeast for oenology. Complementation by Photoacoustic Microscopy is also contemplated, to study the internal conversion component which follows the excitation by photons.

  12. Excitation wavelength dependence of fluorescence intermittency in CdSe/ZnS core/shell quantum dots.

    PubMed

    Knappenberger, Kenneth L; Wong, Daryl B; Romanyuk, Yaroslav E; Leone, Stephen R

    2007-12-01

    Core/shell CdSe/ZnS quantum dot fluorescence-blinking statistics depend strongly on excitation wavelength. Excitation on the band gap (575 nm) results in inverse-power law "on" time distributions. However, distributions resulting from excitation above the band gap (525 nm) require a truncated power law and are 100 times less likely to display 10-s fluorescence. "Off" time statistics are insensitive to the excitation wavelength. The results may be explained by nonemissive trap states accessed with the higher-photon excitation energies.

  13. Absorption-ablation-excitation mechanism of laser-cluster interactions in a nanoaerosol system.

    PubMed

    Ren, Yihua; Li, Shuiqing; Zhang, Yiyang; Tse, Stephen D; Long, Marshall B

    2015-03-01

    The absorption-ablation-excitation mechanism in laser-cluster interactions is investigated by measuring Rayleigh scattering of aerosol clusters along with atomic emission from phase-selective laser-induced breakdown spectroscopy. For 532 nm excitation, as the laser intensity increases beyond 0.16  GW/cm^{2}, the scattering cross section of TiO_{2} clusters begins to decrease, concurrent with the onset of atomic emission of Ti, indicating a scattering-to-ablation transition and the formation of nanoplasmas. With 1064 nm laser excitation, the atomic emissions are more than one order of magnitude weaker than that at 532 nm, indicating that the thermal effect is not the main mechanism. To better clarify the process, time-resolved measurements of scattering signals are examined for different excitation laser intensities. For increasing laser intensity, the cross section of clusters decreases during a single pulse, evincing the shorter ablation delay time and larger ratios of ablation clusters. Assessment of the electron energy distribution during the ablation process is conducted by nondimensionalizing the Fokker-Planck equation, with analogous Strouhal Sl_{E}, Peclet Pe_{E}, and Damköhler Da_{E} numbers defined to characterize the laser-induced aerothermochemical environment. For conditions where Sl_{E}≫1, Pe_{E}≫1, and Da_{E}≪1, the electrons are excited to the conduction band by two-photon absorption, then relax to the bottom of the conduction band by electron energy loss to the lattice, and finally serve as the energy transfer media between laser field and lattice. The relationship between delay time and excitation intensity is well correlated by this simplified model with quasisteady assumption.

  14. Absorption-Ablation-Excitation Mechanism of Laser-Cluster Interactions in a Nanoaerosol System

    NASA Astrophysics Data System (ADS)

    Ren, Yihua; Li, Shuiqing; Zhang, Yiyang; Tse, Stephen D.; Long, Marshall B.

    2015-03-01

    The absorption-ablation-excitation mechanism in laser-cluster interactions is investigated by measuring Rayleigh scattering of aerosol clusters along with atomic emission from phase-selective laser-induced breakdown spectroscopy. For 532 nm excitation, as the laser intensity increases beyond 0.16 GW /cm2 , the scattering cross section of TiO2 clusters begins to decrease, concurrent with the onset of atomic emission of Ti, indicating a scattering-to-ablation transition and the formation of nanoplasmas. With 1064 nm laser excitation, the atomic emissions are more than one order of magnitude weaker than that at 532 nm, indicating that the thermal effect is not the main mechanism. To better clarify the process, time-resolved measurements of scattering signals are examined for different excitation laser intensities. For increasing laser intensity, the cross section of clusters decreases during a single pulse, evincing the shorter ablation delay time and larger ratios of ablation clusters. Assessment of the electron energy distribution during the ablation process is conducted by nondimensionalizing the Fokker-Planck equation, with analogous Strouhal SlE , Peclet PeE , and Damköhler DaE numbers defined to characterize the laser-induced aerothermochemical environment. For conditions where SlE≫1 , PeE≫1 , and DaE≪1 , the electrons are excited to the conduction band by two-photon absorption, then relax to the bottom of the conduction band by electron energy loss to the lattice, and finally serve as the energy transfer media between laser field and lattice. The relationship between delay time and excitation intensity is well correlated by this simplified model with quasisteady assumption.

  15. Absorption-ablation-excitation mechanism of laser-cluster interactions in a nanoaerosol system.

    PubMed

    Ren, Yihua; Li, Shuiqing; Zhang, Yiyang; Tse, Stephen D; Long, Marshall B

    2015-03-01

    The absorption-ablation-excitation mechanism in laser-cluster interactions is investigated by measuring Rayleigh scattering of aerosol clusters along with atomic emission from phase-selective laser-induced breakdown spectroscopy. For 532 nm excitation, as the laser intensity increases beyond 0.16  GW/cm^{2}, the scattering cross section of TiO_{2} clusters begins to decrease, concurrent with the onset of atomic emission of Ti, indicating a scattering-to-ablation transition and the formation of nanoplasmas. With 1064 nm laser excitation, the atomic emissions are more than one order of magnitude weaker than that at 532 nm, indicating that the thermal effect is not the main mechanism. To better clarify the process, time-resolved measurements of scattering signals are examined for different excitation laser intensities. For increasing laser intensity, the cross section of clusters decreases during a single pulse, evincing the shorter ablation delay time and larger ratios of ablation clusters. Assessment of the electron energy distribution during the ablation process is conducted by nondimensionalizing the Fokker-Planck equation, with analogous Strouhal Sl_{E}, Peclet Pe_{E}, and Damköhler Da_{E} numbers defined to characterize the laser-induced aerothermochemical environment. For conditions where Sl_{E}≫1, Pe_{E}≫1, and Da_{E}≪1, the electrons are excited to the conduction band by two-photon absorption, then relax to the bottom of the conduction band by electron energy loss to the lattice, and finally serve as the energy transfer media between laser field and lattice. The relationship between delay time and excitation intensity is well correlated by this simplified model with quasisteady assumption. PMID:25793812

  16. Excitation-emission matrices (EEMs) and synchronous fluorescence spectroscopy (SFS) investigations of gastrointestinal tissues

    NASA Astrophysics Data System (ADS)

    Genova, Ts.; Borisova, E.; Zhelyazkova, Al.; Semyachkina-Glushkovskaya, O.; Penkov, N.; Keremedchiev, M.; Vladimirov, B.; Avramov, L.

    2015-01-01

    In this report we will present our recent investigations of the fluorescence properties of lower part gastrointestinal tissues using excitation-emission matrix and synchronous fluorescence spectroscopy measurement modalities. The spectral peculiarities observed will be discussed and the endogenous sources of the fluorescence signal will be addressed. For these fluorescence spectroscopy measurements the FluoroLog 3 system (HORIBA Jobin Yvon, France) was used. It consists of a Xe lamp (300 W, 200-650 nm), a double mono-chromators, and a PMT detector with a work region at 220- 850 nm. Autofluorescence signals were detected in the form of excitation-emission matrices for the samples of normal mucosa, dysphasia and colon carcinoma and specific spectral features for each tissue were found. Autofluorescence signals from the same samples are observed through synchronous fluorescence spectroscopy, which is a novel promising modality for fluorescence spectroscopy measurements of bio-samples. It is one of the most powerful techniques for multicomponent analysis, because of its sensitivity. In the SFS regime, the fluorescence signal is recorded while both excitation λexc and emission wavelengths λem are simultaneously scanned. A constant wavelength interval is maintained between the λexc and λem wavelengths throughout the spectrum. The resulted fluorescence spectrum shows narrower peak widths, in comparison with EEMs, which are easier for identification and minimizes the chance for false determinations or pretermission of specific spectral feature. This modality is also faster, than EEMs, a much smaller number of data points are required.1 In our measurements we use constant wavelength interval Δλ in the region of 10-200 nm. Measurements are carried out in the terms of finding Δλ, which results in a spectrum with most specific spectral features for comparison with spectral characteristics observed in EEMs. Implementing synchronous fluorescence spectroscopy in optical

  17. Novel xenon calibration scheme for two-photon absorption laser induced fluorescence of hydrogen

    NASA Astrophysics Data System (ADS)

    Elliott, Drew; Scime, Earl; Short, Zachary

    2016-11-01

    Two photon absorption laser induced fluorescence (TALIF) measurements of neutral hydrogen and its isotopes are typically calibrated by performing TALIF measurements on krypton with the same diagnostic system and using the known ratio of the absorption cross sections [K. Niemi et al., J. Phys. D 34, 2330 (2001)]. Here we present the measurements of a new calibration method based on a ground state xenon scheme for which the fluorescent emission wavelength is nearly identical to that of hydrogen, thereby eliminating chromatic effects in the collection optics and simplifying detector calibration. We determine that the ratio of the TALIF cross sections of xenon and hydrogen is 0.024 ± 0.001.

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

    PubMed Central

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

    2016-01-01

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

  19. Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein

    PubMed Central

    Chu, Jun; Haynes, Russell D; Corbel, Stéphane Y; Li, Pengpeng; González-González, Emilio; Burg, John S; Ataie, Niloufar J; Lam, Amy J; Cranfill, Paula J; Baird, Michelle A; Davidson, Michael W; Ng, Ho-Leung; Garcia, K Christopher; Contag, Christopher H; Shen, Kang; Blau, Helen M; Lin, Michael Z

    2014-01-01

    A method for non-invasive visualization of genetically labelled cells in animal disease models with micron-level resolution would greatly facilitate development of cell-based therapies. Imaging of fluorescent proteins (FPs) using red excitation light in the “optical window” above 600 nm is one potential method for visualizing implanted cells. However, previous efforts to engineer FPs with peak excitation beyond 600 nm have resulted in undesirable reductions in brightness. Here we report three new red-excitable monomeric FPs obtained by structure-guided mutagenesis of mNeptune, previously the brightest monomeric FP when excited beyond 600 nm. Two of these, mNeptune2 and mNeptune2.5, demonstrate improved maturation and brighter fluorescence, while the third, mCardinal, has a red-shifted excitation spectrum without reduction in brightness. We show that mCardinal can be used to non-invasively and longitudinally visualize the differentiation of myoblasts and stem cells into myocytes in living mice with high anatomical detail. PMID:24633408

  20. Spatial distribution of fluorescent light emitted from neon and nitrogen excited by low energy electron beams

    SciTech Connect

    Morozov, A.; Kruecken, R.; Ulrich, A.; Wieser, J.

    2006-11-01

    Side-view intensity profiles of fluorescent light were measured for neon and nitrogen excited with 12 keV electron beams at gas pressures from 250 to 1400 hPa. The intensity profiles were compared with theoretical profiles calculated using the CASINO program which performs Monte Carlo simulations of electron scattering. It was assumed that the spatial distribution of fluorescent intensity is directly proportional to the spatial distribution of energy loss by primary electrons. The comparison shows good correlation of experimental data and the results of numeric simulations.

  1. Steady state fluorescence spectroscopic characterization of normal and diabetic urine at selective excitation wavelength 280 nm

    NASA Astrophysics Data System (ADS)

    Kesavan, Anjana; Pachaiappan, Rekha; Aruna, Prakasa Rao; Ganesan, Singaravelu

    2016-03-01

    Urine is considered diagnostically important for tits native fluorophores and they vary in their distribution, concentration and physiochemical properties, depending upon the metabolic condition of the subject. In this study, we have made an attempt, to characterize the urine of normal subject and diabetic patients under medication by native fluorescence spectroscopy at 280 nm excitation. Further, the fluorescence data were analyzed employing the multivariate statistical method linear discriminant analysis (LDA) using leave one out cross validation method. The results were promising in discriminating diabetic urine from that of normal urine. This study in future may be extended to check the feasibility in ruling out the coexisting disorders such as cancer.

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

    SciTech Connect

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

    2010-03-15

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

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

    PubMed

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

    2006-03-20

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

  4. Evidence for an excited-state reaction contributing to NADH fluorescence.

    PubMed

    Ladokhin, A S; Brand, L

    1995-03-01

    The fluorescence of reduced β-nicotinamide adenine dinucleotide (NADH) was monitored as a function of the excitation and emission wavelengths. In aqueous and organic solvents the intensity decay was found to be more heterogeneous than reported earlier. When the ternary complex of NADH with the enzyme (horse liver alcohol dehydrogenase) and substrate analog (iso-butyramide) is formed, three exponents are required to fit the data. The decay-associated spectrum for the shortest lifetime undergoes a sign change from positive at the blue edge of emission to negative at the red edge. This phenomenon is interpreted as an outcome of reversible excited-state reaction leading to the appearance of at least one fluorescent product.

  5. A Study on the Excitation and Resonant Absorption of Coronal Loop Kink Oscillations

    NASA Astrophysics Data System (ADS)

    Yu, Dae Jung; Van Doorsselaere, Tom

    2016-11-01

    We study theoretically the issue of externally driven excitations of standing kink waves and their resonant absorption into torsionally polarized m = 1 waves in the coronal loops in pressureless plasmas. We use the ideal MHD equations, for which we develop an invariant imbedding method available in cylindrical geometry. We assume a sinusoidal density profile at the loop boundary where the density inside the loop is lower than the outside and vice versa. We present field distributions for these two cases and find that they have similar behaviors. We compare the results for the overdense loops, which describe the usual coronal loops, with the analytical solutions of Soler et al. obtained using the Frobenius method. Our results show some similarity for thin nonuniform layers but deviate a lot for thick nonuniform layers. For the first case, which describes the wave train propagation in funnels, we find that resonant absorption depends crucially on the thickness of the nonuniform boundary, loop length, and density contrast. The resonant absorption of the kink mode is dominant when the loop length is sufficiently larger compared with its radius (thin loop). The behavior of the far-field pattern of the scattered wave by the coronal loop is closely related to that of the resonant absorption. For the mode conversion phenomena in inhomogeneous plasmas, a certain universal behavior of the resonant absorption is found for the first time. We expect that the main feature may also apply to the overdense loops and discuss its relation to the damping rate.

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

    PubMed

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

    2003-12-01

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

  7. Two-photon absorption of fluorescent protein chromophores incorporating non-canonical amino acids: TD-DFT screening and classical dynamics.

    PubMed

    Alaraby Salem, M; Brown, Alex

    2015-10-14

    Two-photon spectroscopy of fluorescent proteins is a powerful bio-imaging tool characterized by deep tissue penetration and little damage. However, two-photon spectroscopy has lower sensitivity than one-photon microscopy alternatives and hence a protein with a large two-photon absorption cross-section is needed. We use time-dependent density functional theory (TD-DFT) at the B3LYP/6-31+G(d,p) level of theory to screen twenty-two possible chromophores that can be formed upon replacing the amino-acid Tyr66 that forms the green fluorescent protein (GFP) chromophore with a non-canonical amino acid. A proposed chromophore with a nitro substituent was found to have a large two-photon absorption cross-section (29 GM) compared to other fluorescent protein chromophores as determined at the same level of theory. Classical molecular dynamics are then performed on a nitro-modified fluorescent protein to test its stability and study the effect of the conformational flexibility of the chromophore on its two-photon absorption cross-section. The theoretical results show that the large cross-section is primarily due to the difference between the permanent dipole moments of the excited and ground states of the nitro-modified chromophore. This large difference is maintained through the various conformations assumed by the chromophore in the protein cavity. The nitro-derived protein appears to be very promising as a two-photon absorption probe.

  8. [Lake algae chemotaxonomy technology based on fluorescence excitation emission matrix and parallel factor analysis].

    PubMed

    Chen, Xiao-Na; Han, Xiu-Rong; Su, Rong-Guo; Shi, Xiao-Yong

    2014-03-01

    An in vivo three-dimensional fluorescence method for the determination of algae community structure was developed by parallel factor (PARAFAC) analysis and CHEMTAX. The PARAFAC model was applied to fluorescence excitation-emission matrix (EEM) of 23 algae species and 12 fluorescent components were identified according to the residual sum of squares and specificity of the composition profiles of fluorescent. Based on the 12 fluorescent components, the algae species at different growth stages were correctly classified at the division level using Bayesian discriminant analysis (BDA). Then the reference fluorescent component ratio matrix was constructed for CHEMTAX, and the EEM-PARAFAC-CHEMTAX method was developed to differentiate taxonomic groups of algae. When the fluorometric method was used for 531 single-species samples, the average correct discrimination ratio (CDR) was 99.1% and the correct discrimination ratios (CDRs) were 100% at the division level except Chlorophyta, the CDR of which was 97.5%. The CDRs for 95 mixtures were above 98.5% for the dominant algae species and above 90.5% for the subdominant algae species, with average relative contents of 69.7% and 26.4%, respectively. This technique would be of great aid when low-cost and rapid analysis is needed for samples in a large batch.

  9. Correlating Fluorescence and Absorption Properties of Dissolved Organic Matter in Northern Peatland Soil Porewaters with Molecular Composition Information

    NASA Astrophysics Data System (ADS)

    Tfaily, M. M.; D'Andrilli, J.; Corbett, J.; Chanton, J.; Cooper, W. T.

    2009-12-01

    Northern peatlands store roughly half as much carbon as is stored as CO2 in the atmosphere in the form of complex biogeopolymers. As climate change proceeds, increasing carbon release from these historically carbon-sequestering environments may represent an unidentified forcing stress on global temperatures. In this work, we have applied absorption spectroscopy, Excitation/Emission Matrix (EEM) spectroscopy and ultrahigh resolution Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) to study the differences in composition of dissolved organic matter (DOM) in soil porewaters at varying depths from different sites (fens and bogs) within the Glacial Lake Agassiz Peatlands (GLAP) of northern Minnesota. UV/Vis absorbance and Excitation Emission Matrix Fluorescence Spectroscopy (EEMS) were used to identify changes in the optical properties associated with the chromophoric fractions of DOM (CDOM). Higher specific UV absorbance (SUVA) at 254 nm indicated relatively more aromatic content in the DOM in surface bog and deep fen horizons. EEMS results were also found to be in agreement with the absorption spectra and molecular characterization as determined by FT-ICR-MS. The strong correlations we have observed suggest that optical spectroscopy techniques represent an effective surrogate approach to characterizing the composition of DOM provided some detailed molecular information is available for calibrating the observed correlations.

  10. Absorption and fluorescence anisotropies of monoclinic crystals: the case of Nd:YCOB.

    PubMed

    Petit, Yannick; Boulanger, Benoît; Segonds, Patricia; Félix, Corinne; Ménaert, Bertrand; Zaccaro, Julien; Aka, Gérard

    2008-05-26

    We report for the first time measurements and modelization of the angular distributions of absorption and fluorescence in a monoclinic crystal. Studies on Nd:YCOB revealed specific topologies with ombilics. These new data upgrade the knowledge on low symmetry crystal optics.

  11. Optical imaging of non-fluorescent nanodiamonds in live cells using transient absorption microscopy.

    PubMed

    Chen, Tao; Lu, Feng; Streets, Aaron M; Fei, Peng; Quan, Junmin; Huang, Yanyi

    2013-06-01

    We directly observe non-fluorescent nanodiamonds in living cells using transient absorption microscopy. This label-free technology provides a novel modality to study the dynamic behavior of nanodiamonds inside the cells with intrinsic three-dimensional imaging capability. We apply this method to capture the cellular uptake of nanodiamonds under various conditions, confirming the endocytosis mechanism.

  12. Combined two-photon excited fluorescence and second-harmonic generation backscattering microscopy of turbid tissues

    NASA Astrophysics Data System (ADS)

    Zoumi, Aikaterini; Yeh, Alvin T.; Tromberg, Bruce J.

    2002-06-01

    A broad range of excitation wavelengths (730-880nm) was used to demonstrate the co-registration of two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) in unstained turbid tissues in reflection geometry. The composite TPEF/SHG microscopic technique was applied to imaging an organotypic tissue model (RAFT). The origin of the image-forming signal from the various RAFT constituents was determined by spectral measurements. It was shown that at shorter excitation wavelengths the signal emitted from the extracellular matrix (ECM) is a combination of SHG and TPEF from collagen, whereas at longer excitation wavelengths the ECM signal is exclusively due to SHG. The cellular signal is due to TPEF at all excitation wavelengths. The reflected SHG intensity followed a quadratic dependence on the excitation power and exhibited a spectral dependence in accordance with previous theoretical studies. Understanding the structural origin of signal provided a stratagem for enhancing contrast between cellular structures, and components of the extracellular matrix. The use of SHG and TPEF in combination provides complementary information that allows non-invasive, spatially localized in vivo characterization of cell-ECM interactions and pathology.

  13. Photosystem II does not possess a simple excitation energy funnel: time-resolved fluorescence spectroscopy meets theory.

    PubMed

    Shibata, Yutaka; Nishi, Shunsuke; Kawakami, Keisuke; Shen, Jian-Ren; Renger, Thomas

    2013-05-01

    The experimentally obtained time-resolved fluorescence spectra of photosystem II (PS II) core complexes, purified from a thermophilic cyanobacterium Thermosynechococcus vulcanus, at 5-180 K are compared with simulations. Dynamic localization effects of excitons are treated implicitly by introducing exciton domains of strongly coupled pigments. Exciton relaxations within a domain and exciton transfers between domains are treated on the basis of Redfield theory and generalized Förster theory, respectively. The excitonic couplings between the pigments are calculated by a quantum chemical/electrostatic method (Poisson-TrEsp). Starting with previously published values, a refined set of site energies of the pigments is obtained through optimization cycles of the fits of stationary optical spectra of PS II. Satisfactorily agreement between the experimental and simulated spectra is obtained for the absorption spectrum including its temperature dependence and the linear dichroism spectrum of PS II core complexes (PS II-CC). Furthermore, the refined site energies well reproduce the temperature dependence of the time-resolved fluorescence spectrum of PS II-CC, which is characterized by the emergence of a 695 nm fluorescence peak upon cooling down to 77 K and the decrease of its relative intensity upon further cooling below 77 K. The blue shift of the fluorescence band upon cooling below 77 K is explained by the existence of two red-shifted chlorophyll pools emitting at around 685 and 695 nm. The former pool is assigned to Chl45 or Chl43 in CP43 (Chl numbering according to the nomenclature of Loll et al. Nature2005, 438, 1040) while the latter is assigned to Chl29 in CP47. The 695 nm emitting chlorophyll is suggested to attract excitations from the peripheral light-harvesting complexes and might also be involved in photoprotection.

  14. Calculation of vibrational and electronic excited state absorption spectra of arsenic-water complexes using density functional theory

    NASA Astrophysics Data System (ADS)

    Huang, L.; Lambrakos, S. G.; Shabaev, A.; Massa, L.

    2016-05-01

    Calculations are presented of vibrational and electronic excited-state absorption spectra for As-H2O complexes using density function theory (DFT) and time-dependent density functional theory (TD-DFT). DFT and TD-DFT can provide interpretation of absorption spectra with respect to molecular structure for excitation by electromagnetic waves at frequencies within the IR and UV-visible ranges. The absorption spectrum corresponding to excitation states of As-H2O complexes consisting of relatively small numbers of water molecules should be associated with response features that are intermediate between that of isolated molecules and that of a bulk system. DFT and TD-DFT calculated absorption spectra represent quantitative estimates that can be correlated with additional information obtained from laboratory measurements and other types of theory based calculations. The DFT software GAUSSIAN was used for the calculations of excitation states presented here.

  15. Spoilage of foods monitored by native fluorescence spectroscopy with selective excitation wavelength

    NASA Astrophysics Data System (ADS)

    Pu, Yang; Wang, Wubao; Alfano, Robert R.

    2015-03-01

    The modern food processing and storage environments require the real-time monitoring and rapid microbiological testing. Optical spectroscopy with selective excitation wavelengths can be the basis of a novel, rapid, reagent less, noncontact and non-destructive technique for monitoring the food spoilage. The native fluorescence spectra of muscle foods stored at 2-4°C (in refrigerator) and 20-24°C (in room temperature) were measured as a function of time with a selective excitation wavelength of 340nm. The contributions of the principal molecular components to the native fluorescence spectra of meat were measured spectra of each fluorophore: collagen, reduced nicotinamide adenine dinucleotide (NADH), and flavin. The responsible components were extracted using a method namely Multivariate Curve Resolution with Alternating Least-Squares (MCR-ALS). The native fluorescence combined with MCR-ALS can be used directly on the surface of meat to produce biochemically interpretable "fingerprints", which reflects the microbial spoilage of foods involved with the metabolic processes. The results show that with time elapse, the emission from NADH in meat stored at 24°C increases much faster than that at 4°C. This is because multiplying of microorganisms and catabolism are accompanied by the generation of NADH. This study presents changes of relative content of NADH may be used as criterion for detection of spoilage degree of meat using native fluorescence spectroscopy.

  16. Excitation-emission matrix fluorescence coupled to chemometrics for the exploration of essential oils.

    PubMed

    Mbogning Feudjio, William; Ghalila, Hassen; Nsangou, Mama; Mbesse Kongbonga, Yvon G; Majdi, Youssef

    2014-12-01

    Excitation-emission matrix fluorescence (EEMF) coupled to chemometrics was used to explore essential oils (EOs). The spectrofluorometer was designed with basic and inexpensive materials and was accompanied by appropriate tools for data pre-treatment. Excitation wavelengths varied between 320 nm and 600 nm while emission wavelengths were from 340 nm to 700 nm. Excitation-emission matrix (EEM) spectra of EOs presented different features, revealing the presence of varying fluorophores. EOs from the same species but from different origins presented almost the same spectra, showing the possibility that EEM spectra could be used as additional parameters in the standardisation of EOs. With the aid of unfold principal component analysis (UPCA), resemblances obtained by spectral analysis of EOs were confirmed. A five components parallel factor analysis (PARAFAC) model was used to find the profiles of fluorophores in EOs. One of those components was associated to chlorophyll a.

  17. Characterizing the Structures, Spectra, and Energy Landscapes Involved in the Excited-State Proton Transfer Process of Red Fluorescent Protein LSSmKate1.

    PubMed

    Chen, Fasheng; Zeng, Qiao; Zhuang, Wei; Liang, WanZhen

    2016-09-22

    By applying molecular dynamics (MD) simulations and quantum chemical calculations, we have characterized the states and processes involved in the excited-state proton transfer (ESPT) of LSSmKate1. MD simulations identify two stable structures in the electronic ground state of LSSmKate1, one with a protonated chromophore and the other with a deprotonated chromophore, thus leading to two separate low-energy absorption maxima with a large energy spacing, as observed in the calculated and experimentally measured absorption spectra. Proton transfer is induced by electronic excitation. When LSSmKate1 is excited, the electrons in the chromophore are transferred from the phenol ring to the N-acylimine moiety; the acidity of a phenolic hydroxyl group is thus enhanced. The calculated potential energy curves (PECs) exhibit energetic feasibility in the generation of the fluorescent species in LSSmKate1, and the exact agreement between the calculated and experimentally measured values of the large Stokes shift further provides solid theoretical evidence for the ESPT process taking place in photoexcited LSSmKate1. The molecular environments play a significant role in the geometries and absorption/emission energies of the chromophores. Overall, TD-ωB97X-D/molecular mechanics (MM) provides a better description of the optical properties of LSSmKate1 than TD-B3LYP/MM, although it always overestimates the excitation energies. PMID:27581731

  18. Characterizing the Structures, Spectra, and Energy Landscapes Involved in the Excited-State Proton Transfer Process of Red Fluorescent Protein LSSmKate1.

    PubMed

    Chen, Fasheng; Zeng, Qiao; Zhuang, Wei; Liang, WanZhen

    2016-09-22

    By applying molecular dynamics (MD) simulations and quantum chemical calculations, we have characterized the states and processes involved in the excited-state proton transfer (ESPT) of LSSmKate1. MD simulations identify two stable structures in the electronic ground state of LSSmKate1, one with a protonated chromophore and the other with a deprotonated chromophore, thus leading to two separate low-energy absorption maxima with a large energy spacing, as observed in the calculated and experimentally measured absorption spectra. Proton transfer is induced by electronic excitation. When LSSmKate1 is excited, the electrons in the chromophore are transferred from the phenol ring to the N-acylimine moiety; the acidity of a phenolic hydroxyl group is thus enhanced. The calculated potential energy curves (PECs) exhibit energetic feasibility in the generation of the fluorescent species in LSSmKate1, and the exact agreement between the calculated and experimentally measured values of the large Stokes shift further provides solid theoretical evidence for the ESPT process taking place in photoexcited LSSmKate1. The molecular environments play a significant role in the geometries and absorption/emission energies of the chromophores. Overall, TD-ωB97X-D/molecular mechanics (MM) provides a better description of the optical properties of LSSmKate1 than TD-B3LYP/MM, although it always overestimates the excitation energies.

  19. Excited-State Intramolecular Proton Transfer in a Blue Fluorescence Chromophore Induces Dual Emission.

    PubMed

    Wu, Dan; Guo, Wei-Wei; Liu, Xiang-Yang; Cui, Ganglong

    2016-08-01

    Compared with green fluorescence protein (GFP) chromophores, the recently synthesized blue fluorescence protein (BFP) chromophore variant presents intriguing photochemical properties, for example, dual fluorescence emission, enhanced fluorescence quantum yield, and ultra-slow excited-state intramolecular proton transfer (ESIPT; J. Phys. Chem. Lett., 2014, 5, 92); however, its photochemical mechanism is still elusive. Herein we have employed the CASSCF and CASPT2 methods to study the mechanistic photochemistry of a truncated BFP chromophore variant in the S0 and S1 states. Based on the optimized minima, conical intersections, and minimum-energy paths (ESIPT, photoisomerization, and deactivation), we have found that the system has two competitive S1 relaxation pathways from the Franck-Condon point of the BFP chromophore variant. One is the ESIPT path to generate an S1 tautomer that exhibits a large Stokes shift in experiments. The generated S1 tautomer can further evolve toward the nearby S1 /S0 conical intersection and then jumps down to the S0 state. The other is the photoisomerization path along the rotation of the central double bond. Along this path, the S1 system runs into an S1 /S0 conical intersection region and eventually hops to the S0 state. The two energetically allowed S1 excited-state deactivation pathways are responsible for the in-part loss of fluorescence quantum yield. The considerable S1 ESIPT barrier and the sizable barriers that separate the S1 tautomers from the S1 /S0 conical intersections make these two tautomers establish a kinetic equilibrium in the S1 state, which thus results in dual fluorescence emission. PMID:27128380

  20. Excited-State Intramolecular Proton Transfer in a Blue Fluorescence Chromophore Induces Dual Emission.

    PubMed

    Wu, Dan; Guo, Wei-Wei; Liu, Xiang-Yang; Cui, Ganglong

    2016-08-01

    Compared with green fluorescence protein (GFP) chromophores, the recently synthesized blue fluorescence protein (BFP) chromophore variant presents intriguing photochemical properties, for example, dual fluorescence emission, enhanced fluorescence quantum yield, and ultra-slow excited-state intramolecular proton transfer (ESIPT; J. Phys. Chem. Lett., 2014, 5, 92); however, its photochemical mechanism is still elusive. Herein we have employed the CASSCF and CASPT2 methods to study the mechanistic photochemistry of a truncated BFP chromophore variant in the S0 and S1 states. Based on the optimized minima, conical intersections, and minimum-energy paths (ESIPT, photoisomerization, and deactivation), we have found that the system has two competitive S1 relaxation pathways from the Franck-Condon point of the BFP chromophore variant. One is the ESIPT path to generate an S1 tautomer that exhibits a large Stokes shift in experiments. The generated S1 tautomer can further evolve toward the nearby S1 /S0 conical intersection and then jumps down to the S0 state. The other is the photoisomerization path along the rotation of the central double bond. Along this path, the S1 system runs into an S1 /S0 conical intersection region and eventually hops to the S0 state. The two energetically allowed S1 excited-state deactivation pathways are responsible for the in-part loss of fluorescence quantum yield. The considerable S1 ESIPT barrier and the sizable barriers that separate the S1 tautomers from the S1 /S0 conical intersections make these two tautomers establish a kinetic equilibrium in the S1 state, which thus results in dual fluorescence emission.

  1. Two-photon excited fluorescence from a pseudoisocyanine-attached gold-coated tip via a thin tapered fiber under a weak continuous wave excitation.

    PubMed

    Ren, Fang; Takashima, Hideaki; Tanaka, Yoshito; Fujiwara, Hideki; Sasaki, Keiji

    2013-11-18

    A simple tapered fiber based photonic-plasmonic hybrid nanostructure composed of a thin tapered fiber and a pseudoisocyanine (PIC)-attached Au-coated tip was demonstrated. Using this simple hybrid nanostructure, we succeeded in observing two-photon excited fluorescence from the PIC dye molecules under a weak continuous wave excitation condition. From the results of the tip-fiber distance dependence and excitation polarization dependence, we found that using a thin tapered fiber and an Au-coated tip realized efficient coupling of the incident light (~95%) and LSP excitation at the Au-coated tip, suggesting the possibility of efficiently inducing two-photon excited fluorescence from the PIC dye molecules attached on the Au-coated tip. This simple photonic-plasmonic hybrid system is one of the promising tools for single photon sources, highly efficient plasmonic sensors, and integrated nonlinear plasmonic devices.

  2. O 1s excitation and ionization processes in the CO2 molecule studied via detection of low-energy fluorescence emission

    NASA Astrophysics Data System (ADS)

    Kivimäki, A.; Alvarez-Ruiz, J.; Wasowicz, T. J.; Callegari, C.; de Simone, M.; Alagia, M.; Richter, R.; Coreno, M.

    2011-08-01

    Oxygen 1s excitation and ionization processes in the CO2 molecule have been studied with dispersed and non-dispersed fluorescence spectroscopy as well as with the vacuum ultraviolet (VUV) photon-photoion coincidence technique. The intensity of the neutral O emission line at 845 nm shows particular sensitivity to core-to-Rydberg excitations and core-valence double excitations, while shape resonances are suppressed. In contrast, the partial fluorescence yield in the wavelength window 300-650 nm and the excitation functions of selected O+ and C+ emission lines in the wavelength range 400-500 nm display all of the absorption features. The relative intensity of ionic emission in the visible range increases towards higher photon energies, which is attributed to O 1s shake-off photoionization. VUV photon-photoion coincidence spectra reveal major contributions from the C+ and O+ ions and a minor contribution from C2 +. No conclusive changes in the intensity ratios among the different ions are observed above the O 1s threshold. The line shape of the VUV-O+ coincidence peak in the mass spectrum carries some information on the initial core excitation.

  3. Experimental recovery of intrinsic fluorescence and fluorophore concentration in the presence of hemoglobin: spectral effect of scattering and absorption on fluorescence

    NASA Astrophysics Data System (ADS)

    Du Le, Vinh Nguyen; Patterson, Michael S.; Farrell, Thomas J.; Hayward, Joseph E.; Fang, Qiyin

    2015-12-01

    The ability to recover the intrinsic fluorescence of biological fluorophores is crucial to accurately identify the fluorophores and quantify their concentrations in the media. Although some studies have successfully retrieved the fluorescence spectral shape of known fluorophores, the techniques usually came with heavy computation costs and did not apply for strongly absorptive media, and the intrinsic fluorescence intensity and fluorophore concentration were not recovered. In this communication, an experimental approach was presented to recover intrinsic fluorescence and concentration of fluorescein in the presence of hemoglobin (Hb). The results indicated that the method was efficient in recovering the intrinsic fluorescence peak and fluorophore concentration with an error of 3% and 10%, respectively. The results also suggested that chromophores with irregular absorption spectra (e.g., Hb) have more profound effects on fluorescence spectral shape than chromophores with monotonic absorption and scattering spectra (e.g., black India ink and polystyrene microspheres).

  4. Strong UV absorption and visible luminescence in ytterbium-doped aluminosilicate glass under UV excitation.

    PubMed

    Engholm, M; Norin, L; Aberg, D

    2007-11-15

    A broad visible luminescence band and characteristic IR luminescence of Yb(3+) ions are observed under UV excitation in ytterbium-doped aluminosilicate glass. Samples made under both oxidizing and reducing conditions are analyzed. A strong charge-transfer absorption band in the UV range is observed for glass samples containing ytterbium. Additional absorption bands are observed for the sample made under reducing conditions, which are associated with f-d transitions of divalent ytterbium. The visible luminescence band is attributed to 5d-4f emission from Yb(2+) ions, and the IR luminescence is concluded to originate from a relaxed charge-transfer transition. The findings are important to explain induced optical losses (photodarkening) in high-power fiber lasers. PMID:18026305

  5. Edge physics of the quantum spin Hall insulator from a quantum dot excited by optical absorption.

    PubMed

    Vasseur, Romain; Moore, Joel E

    2014-04-11

    The gapless edge modes of the quantum spin Hall insulator form a helical liquid in which the direction of motion along the edge is determined by the spin orientation of the electrons. In order to probe the Luttinger liquid physics of these edge states and their interaction with a magnetic (Kondo) impurity, we consider a setup where the helical liquid is tunnel coupled to a semiconductor quantum dot that is excited by optical absorption, thereby inducing an effective quantum quench of the tunneling. At low energy, the absorption spectrum is dominated by a power-law singularity. The corresponding exponent is directly related to the interaction strength (Luttinger parameter) and can be computed exactly using boundary conformal field theory thanks to the unique nature of the quantum spin Hall edge.

  6. Excited-state absorption in the lasing wavelength region of Alexandrite

    SciTech Connect

    Shand, M.L.; Walling, J.C.

    1982-07-01

    The excited-state absorption cross section sigma/sub 2/ /sub a/ (E) in the gain wavelength region of alexandrite has been determined and is shown to limit the vibronic laser range at both high and low energy. The maximum in vibronic laser emission is due to a minimum in sigma/sub 2/ /sub a/ (E) near 13 000 cm/sup -1/. sigma/sub 2/ /sub a/ (E) is less than 10/sup -20/ cm/sup 2/ between 12 000 and 14 000 cm/sup -1/.

  7. Absorption and fluorescence emission spectroscopic characters of naphtho-homologated yy-DNA bases and effect of methanol solution and base pairing.

    PubMed

    Zhang, Laibin; Li, Huifang; Li, Jilai; Chen, Xiaohua; Bu, Yuxiang

    2010-03-01

    A comprehensive theoretical study of electronic transitions of naphtho-homologated base analogs, namely, yy-T, yy-C, yy-A, and yy-G, was performed. The nature of the low-lying excited states is discussed, and the results are compared with those from experiment and also with those of y-bases. Geometrical characteristics of the lowest excited singlet pipi* and npi* states were explored using the CIS method, and the effects of methanol solution and paring with their complementary natural bases on the relevant absorption and emission spectra of these modified bases were examined. The calculated excitation and emission energies agree well with the measured data, where experimental results are available. In methanol solution, the fluorescence from yy-A and yy-G would be expected to occur around 539 and 562 nm, respectively, suggesting that yy-A is a green-colored fluorophore, whereas yy-G is a yellow-colored fluorophore. The methanol solution was found to red-shift both the absorption and emission maxima of yy-A, yy-T, and yy-C, but blue-shift those for yy-G. Generally, though base pairing has no significant effects on the absorption and fluorescence maxima of yy-A, yy-C, and yy-T, it blue-shifts those for yy-G.

  8. Determination of tellurium and antimony in nickel alloys by laser excited atomic fluorescence spectrometry in a graphite furnace

    NASA Astrophysics Data System (ADS)

    Liang, Zhongwen; Lonardo, Robert F.; Michel, Robert G.

    1993-01-01

    Analytical laser excited atomic fluorescence of the metalloids tellurium and antimony in an electrothermal atomizer was studied. The detection limits were 20 fg and 10 fg for tellurium and antimony respectively, equivalent to about 0.01 ng g -1 in nickel based alloys by direct solid sample analysis, for a 1 mg solid sample, or 1 ng g -1 by the dissolution method, for a 100 mg solid sample in 100 ml solution. The detection limits were three orders of magnitude better than those obtained by graphite furnace atomic absorption spectrometry. They were also comparable to, or better than, those by inductively coupled plasma mass spectrometry. The linear dynamic ranges of the calibration curves were found to be six and seven orders of magnitude for antimony and tellurium respectively. By use of aqueous calibration, tellurium was accurately determined in NIST nickel alloy Standard Reference Materials by both a solid sample method, with a relative standard deviation (RSD) of about 13%, and a dissolution method with an RSD of about 9%. Antimony in Pratt and Whitney "A" series nickel alloy standards was successfully determined by the dissolution method, with an RSD of about 7%, but by solid sampling the antimony method gave incomplete recovery. Molecular fluorescence backgrounds from nitric oxide and silicon monoxide were observed and discussed.

  9. Intrinsic fluorescence excitation-emission matrix spectral features of cottonseed protein fractions and the effects of denaturants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To better understand the functional and physicochemical properties of cottonseed protein, we investigated the intrinsic fluorescence excitation-emission matrix (EEM) spectral features of cottonseed protein isolate (CSPI) and sequentially extracted water (CSPw) and alkali (CSPa) protein fractions, an...

  10. Highly-sensitive Eu3+ ratiometric thermometers based on excited state absorption with predictable calibration

    NASA Astrophysics Data System (ADS)

    Souza, Adelmo S.; Nunes, Luiz A. O.; Silva, Ivan G. N.; Oliveira, Fernando A. M.; da Luz, Leonis L.; Brito, Hermi F.; Felinto, Maria C. F. C.; Ferreira, Rute A. S.; Júnior, Severino A.; Carlos, Luís D.; Malta, Oscar L.

    2016-02-01

    Temperature measurements ranging from a few degrees to a few hundreds of Kelvin are of great interest in the fields of nanomedicine and nanotechnology. Here, we report a new ratiometric luminescent thermometer using thermally excited state absorption of the Eu3+ ion. The thermometer is based on the simple Eu3+ energy level structure and can operate between 180 and 323 K with a relative sensitivity ranging from 0.7 to 1.7% K-1. The thermometric parameter is defined as the ratio between the emission intensities of the 5D0 --> 7F4 transition when the 5D0 emitting level is excited through the 7F2 (physiological range) or 7F1 (down to 180 K) level. Nano and microcrystals of Y2O3:Eu3+ were chosen as a proof of concept of the operational principles in which both excitation and detection are within the first biological transparent window. A novel and of paramount importance aspect is that the calibration factor can be calculated from the Eu3+ emission spectrum avoiding the need for new calibration procedures whenever the thermometer operates in different media.Temperature measurements ranging from a few degrees to a few hundreds of Kelvin are of great interest in the fields of nanomedicine and nanotechnology. Here, we report a new ratiometric luminescent thermometer using thermally excited state absorption of the Eu3+ ion. The thermometer is based on the simple Eu3+ energy level structure and can operate between 180 and 323 K with a relative sensitivity ranging from 0.7 to 1.7% K-1. The thermometric parameter is defined as the ratio between the emission intensities of the 5D0 --> 7F4 transition when the 5D0 emitting level is excited through the 7F2 (physiological range) or 7F1 (down to 180 K) level. Nano and microcrystals of Y2O3:Eu3+ were chosen as a proof of concept of the operational principles in which both excitation and detection are within the first biological transparent window. A novel and of paramount importance aspect is that the calibration factor can be

  11. Characterization of dissolved organic matter in fogwater by excitation-emission matrix fluorescence spectroscopy

    USGS Publications Warehouse

    Birdwell, J.E.; Valsaraj, K.T.

    2010-01-01

    Dissolved organic matter (DOM) present in fogwater samples collected in southeastern Louisiana and central-eastern China has been characterized using excitation-emission matrix fluorescence spectroscopy. The goal of the study was to illustrate the utility of fluorescence for obtaining information on the large fraction of organic carbon in fogwaters (typically >40% by weight) that defies characterization in terms of specific chemical compounds without the difficulty inherent in obtaining sufficient fogwater volume to isolate DOM for assessment using other spectroscopic and chemical analyses. Based on the findings of previous studies using other characterization methods, it was anticipated that the unidentified organic carbon fraction would have characteristic peaks associated with humic substances and fluorescent amino acids. Both humic- and protein-like fluorophores were observed in the fogwater spectra and fluorescence-derived indices for the fogwater had similar values to those of soil and sediment porewater. Greater biological character was observed in samples with higher organic carbon concentrations. Fogwaters are shown to contain a mixture of terrestrially- and microbially-derived fluorescent organic material, which is expected to be derived from an array of different sources, such as suspended soil and dust particles, biogenic emissions and organic substances generated by atmospheric processes. The fluorescence results indicate that much of the unidentified organic carbon present in fogwater can be represented by humic-like and biologically-derived substances similar to those present in other aquatic systems, though it should be noted that fluorescent signatures representative of DOM produced by atmospheric processing of organic aerosols may be contributing to or masked by humic-like fluorophores. ?? 2010.

  12. High-sensitivity DNA detection with a laser-excited confocal fluorescence gel scanner.

    PubMed

    Quesada, M A; Rye, H S; Gingrich, J C; Glazer, A N; Mathies, R A

    1991-05-01

    A high-sensitivity, laser-excited confocal fluorescence gel scanner has been developed and applied to the detection of fluorescently labeled DNA. An argon ion laser (1-10 mW at 488 nm) is focused in the gel with a high-numerical aperture microscope objective. The laser-excited fluorescence is gathered by the objective and focused on a confocal spatial filter, followed by a spectral filter and photodetector. The gel is placed on a computer-controlled scan stage, and the scanned image of the gel fluorescence is stored and analyzed in a computer. This scanner has been used to detect DNA separated on sequencing gels, agarose mapping gels and pulsed field gels. Sanger sequencing gels were run on M13mp18 DNA using a fluoresceinated primer. The 400-microns-thick gels, loaded with 30 fmol of DNA fragments in 3-mm lanes, were scanned at 78-microns resolution. The high resolution of our scanner coupled with image processing allows us to read up to approximately 300 bases in four adjacent sequencing lanes. The minimum band size that could be detected and read was approximately 200 microns. This instrument has a limiting detection sensitivity of approximately 10 amol of fluorescein-labeled DNA in a 1 x 3-mm band. In applications to agarose mapping gels, we have exploited the fact that DNA can be prestained with ethidium homodimer, followed by electrophoresis and fluorescence detection to achieve picogram sensitivity. We have also developed methods using both ethidium homodimer and thiazole orange staining which permit two-color detection of DNA in one lane.(ABSTRACT TRUNCATED AT 250 WORDS)

  13. Comparison of Cherenkov excited fluorescence and phosphorescence molecular sensing from tissue with external beam irradiation.

    PubMed

    Lin, Huiyun; Zhang, Rongxiao; Gunn, Jason R; Esipova, Tatiana V; Vinogradov, Sergei; Gladstone, David J; Jarvis, Lesley A; Pogue, Brian W

    2016-05-21

    Ionizing radiation delivered by a medical linear accelerator (LINAC) generates Cherenkov emission within the treated tissue. A fraction of this light, in the 600-900 nm wavelength region, propagates through centimeters of tissue and can be used to excite optical probes in vivo, enabling molecular sensing of tissue analytes. The success of isolating the emission signal from this Cherenkov excitation background is dependent on key factors such as: (i) the Stokes shift of the probe spectra; (ii) the excited state lifetime; (iii) the probe concentration; (iv) the depth below the tissue surface; and (v) the radiation dose used. Previous studies have exclusively focused on imaging phosphorescent dyes, rather than fluorescent dyes. However there are only a few biologically important phosphorescent dyes and yet in comparison there are thousands of biologically relevant fluorescent dyes. So in this study the focus was a study of efficacy of Cherenkov-excited luminescence using fluorescent commercial near-infrared probes, IRDye 680RD, IRDye 700DX, and IRDye 800CW, and comparing them to the well characterized phosphorescent probe Oxyphor PtG4, an oxygen sensitive dye. Each probe was excited by Cherenkov light from a 6 MV external radiation beam, and measured in continuous wave or time-gated modes. The detection was performed by spectrally resolving the luminescence signals, and measuring them with spectrometer-based separation on an ICCD detector. The results demonstrate that IRDye 700DX and PtG4 allowed for the maximal signal to noise ratio. In the case of the phosphorescent probe, PtG4, with emission decays on the microsecond (μs) time scale, time-gated acquisition was possible, and it allowed for higher efficacy in terms of the probe concentration and detection depth. Phantoms containing the probe at 5 mm depth could be detected at concentrations down to the nanoMolar range, and at depths into the tissue simulating phantom near 3 cm. In vivo studies showed that 5

  14. Comparison of Cherenkov excited fluorescence and phosphorescence molecular sensing from tissue with external beam irradiation.

    PubMed

    Lin, Huiyun; Zhang, Rongxiao; Gunn, Jason R; Esipova, Tatiana V; Vinogradov, Sergei; Gladstone, David J; Jarvis, Lesley A; Pogue, Brian W

    2016-05-21

    Ionizing radiation delivered by a medical linear accelerator (LINAC) generates Cherenkov emission within the treated tissue. A fraction of this light, in the 600-900 nm wavelength region, propagates through centimeters of tissue and can be used to excite optical probes in vivo, enabling molecular sensing of tissue analytes. The success of isolating the emission signal from this Cherenkov excitation background is dependent on key factors such as: (i) the Stokes shift of the probe spectra; (ii) the excited state lifetime; (iii) the probe concentration; (iv) the depth below the tissue surface; and (v) the radiation dose used. Previous studies have exclusively focused on imaging phosphorescent dyes, rather than fluorescent dyes. However there are only a few biologically important phosphorescent dyes and yet in comparison there are thousands of biologically relevant fluorescent dyes. So in this study the focus was a study of efficacy of Cherenkov-excited luminescence using fluorescent commercial near-infrared probes, IRDye 680RD, IRDye 700DX, and IRDye 800CW, and comparing them to the well characterized phosphorescent probe Oxyphor PtG4, an oxygen sensitive dye. Each probe was excited by Cherenkov light from a 6 MV external radiation beam, and measured in continuous wave or time-gated modes. The detection was performed by spectrally resolving the luminescence signals, and measuring them with spectrometer-based separation on an ICCD detector. The results demonstrate that IRDye 700DX and PtG4 allowed for the maximal signal to noise ratio. In the case of the phosphorescent probe, PtG4, with emission decays on the microsecond (μs) time scale, time-gated acquisition was possible, and it allowed for higher efficacy in terms of the probe concentration and detection depth. Phantoms containing the probe at 5 mm depth could be detected at concentrations down to the nanoMolar range, and at depths into the tissue simulating phantom near 3 cm. In vivo studies showed that 5

  15. Comparison of Cherenkov excited fluorescence and phosphorescence molecular sensing from tissue with external beam irradiation

    NASA Astrophysics Data System (ADS)

    Lin, Huiyun; Zhang, Rongxiao; Gunn, Jason R.; Esipova, Tatiana V.; Vinogradov, Sergei; Gladstone, David J.; Jarvis, Lesley A.; Pogue, Brian W.

    2016-05-01

    Ionizing radiation delivered by a medical linear accelerator (LINAC) generates Cherenkov emission within the treated tissue. A fraction of this light, in the 600-900 nm wavelength region, propagates through centimeters of tissue and can be used to excite optical probes in vivo, enabling molecular sensing of tissue analytes. The success of isolating the emission signal from this Cherenkov excitation background is dependent on key factors such as: (i) the Stokes shift of the probe spectra; (ii) the excited state lifetime; (iii) the probe concentration; (iv) the depth below the tissue surface; and (v) the radiation dose used. Previous studies have exclusively focused on imaging phosphorescent dyes, rather than fluorescent dyes. However there are only a few biologically important phosphorescent dyes and yet in comparison there are thousands of biologically relevant fluorescent dyes. So in this study the focus was a study of efficacy of Cherenkov-excited luminescence using fluorescent commercial near-infrared probes, IRDye 680RD, IRDye 700DX, and IRDye 800CW, and comparing them to the well characterized phosphorescent probe Oxyphor PtG4, an oxygen sensitive dye. Each probe was excited by Cherenkov light from a 6 MV external radiation beam, and measured in continuous wave or time-gated modes. The detection was performed by spectrally resolving the luminescence signals, and measuring them with spectrometer-based separation on an ICCD detector. The results demonstrate that IRDye 700DX and PtG4 allowed for the maximal signal to noise ratio. In the case of the phosphorescent probe, PtG4, with emission decays on the microsecond (μs) time scale, time-gated acquisition was possible, and it allowed for higher efficacy in terms of the probe concentration and detection depth. Phantoms containing the probe at 5 mm depth could be detected at concentrations down to the nanoMolar range, and at depths into the tissue simulating phantom near 3 cm. In vivo studies showed that 5

  16. [Fluorescence excitation-emission matrix spectroscopy of CDOM from Yundang Lagoon and its indication for organic pollution].

    PubMed

    Zhuo, Jian-Fu; Guo, Wei-Dong; Deng, Xun; Zhang, Zhi-Ying; Xu, Jing; Huang, Ling-Feng

    2010-06-01

    Fluorescence excitation-emission matrix spectroscopy (EEMs) combined with absorption spectroscopy were applied to study the optical properties of CDOM samples from highly-polluted Yundang Lagoon in Xiamen in order to demonstrate the feasibility of using these spectral properties as a tracer of the degree of organic pollution in similar polluted coastal waters. Surface water samples were collected from 13 stations 4 times during April and May, 2008. Parallel factor analysis (PARAFAC) model was used to resolve the EEMs of CDOM. Five separate fluorescent components were identified, including two humic-like components (C1: 240, 325/422 nm; C5: 260, 380/474 nm), two protein-like components (C2: 225, 275/350 nm; C4: 240, 300/354 nm) and one xenobiotic-like component (C3: 225/342 nm), which could be used as a good tracer for the input of the anthropogenic organic, pollutants. The concentrations of component C3 and dissolved organic carbon (DOC) are much higher near the inlet of sewage discharge, demonstrating that the discharge of surrounding sewage is a major source of organic pollutants in Yundang Lagoon. CDOM absorption coefficient alpha (280) and the score of humic-like component C1 showed significant linear relationships with COD(Mn), and a strong positive correlation was also found between the score of protein-like component C2 and BOD5. This suggested that the optical properties of CDOM may provide a fast in-situ way to monitor the variation of the water quality in Yundang Lagoon and that of similar polluted coastal waters. PMID:20707146

  17. [Fluorescence excitation-emission matrix spectroscopy of CDOM from Yundang Lagoon and its indication for organic pollution].

    PubMed

    Zhuo, Jian-Fu; Guo, Wei-Dong; Deng, Xun; Zhang, Zhi-Ying; Xu, Jing; Huang, Ling-Feng

    2010-06-01

    Fluorescence excitation-emission matrix spectroscopy (EEMs) combined with absorption spectroscopy were applied to study the optical properties of CDOM samples from highly-polluted Yundang Lagoon in Xiamen in order to demonstrate the feasibility of using these spectral properties as a tracer of the degree of organic pollution in similar polluted coastal waters. Surface water samples were collected from 13 stations 4 times during April and May, 2008. Parallel factor analysis (PARAFAC) model was used to resolve the EEMs of CDOM. Five separate fluorescent components were identified, including two humic-like components (C1: 240, 325/422 nm; C5: 260, 380/474 nm), two protein-like components (C2: 225, 275/350 nm; C4: 240, 300/354 nm) and one xenobiotic-like component (C3: 225/342 nm), which could be used as a good tracer for the input of the anthropogenic organic, pollutants. The concentrations of component C3 and dissolved organic carbon (DOC) are much higher near the inlet of sewage discharge, demonstrating that the discharge of surrounding sewage is a major source of organic pollutants in Yundang Lagoon. CDOM absorption coefficient alpha (280) and the score of humic-like component C1 showed significant linear relationships with COD(Mn), and a strong positive correlation was also found between the score of protein-like component C2 and BOD5. This suggested that the optical properties of CDOM may provide a fast in-situ way to monitor the variation of the water quality in Yundang Lagoon and that of similar polluted coastal waters.

  18. A Step Beyond BRET: Fluorescence by Unbound Excitation from Luminescence (FUEL)

    PubMed Central

    Dragavon, Joseph; Sinow, Carolyn; Holland, Alexandra D.; Rekiki, Abdessalem; Theodorou, Ioanna; Samson, Chelsea; Blazquez, Samantha; Rogers, Kelly L.; Tournebize, Régis; Shorte, Spencer L.

    2014-01-01

    Fluorescence by Unbound Excitation from Luminescence (FUEL) is a radiative excitation-emission process that produces increased signal and contrast enhancement in vitro and in vivo. FUEL shares many of the same underlying principles as Bioluminescence Resonance Energy Transfer (BRET), yet greatly differs in the acceptable working distances between the luminescent source and the fluorescent entity. While BRET is effectively limited to a maximum of 2 times the Förster radius, commonly less than 14 nm, FUEL can occur at distances up to µm or even cm in the absence of an optical absorber. Here we expand upon the foundation and applicability of FUEL by reviewing the relevant principles behind the phenomenon and demonstrate its compatibility with a wide variety of fluorophores and fluorescent nanoparticles. Further, the utility of antibody-targeted FUEL is explored. The examples shown here provide evidence that FUEL can be utilized for applications where BRET is not possible, filling the spatial void that exists between BRET and traditional whole animal imaging. PMID:24894759

  19. Fluorescence excitation-emission matrix spectroscopy as a tool for determining quality of sparkling wines.

    PubMed

    Elcoroaristizabal, Saioa; Callejón, Raquel M; Amigo, Jose M; Ocaña-González, Juan A; Morales, M Lourdes; Ubeda, Cristina

    2016-09-01

    Browning in sparkling wines was assessed by the use of excitation-emission fluorescence spectroscopy combined with PARAllel FACtor analysis (PARAFAC). Four different cava sparkling wines were monitored during an accelerated browning process and subsequently storage. Fluorescence changes observed during the accelerated browning process were monitored and compared with other conventional parameters: absorbance at 420nm (A420) and the content of 5-hydroxymethyl-2-furfural (5-HMF). A high similarity of the spectral profiles for all sparkling wines analyzed was observed, being explained by a four component PARAFAC model. A high correlation between the third PARAFAC factor (465/530nm) and the commonly used non-enzymatic browning indicators was observed. The fourth PARAFAC factor (280/380nm) gives us also information about the browning process following a first order kinetic reaction. Hence, excitation-emission fluorescence spectroscopy, together with PARAFAC, provides a faster alternative for browning monitoring to conventional methods, as well as useful key indicators for quality control. PMID:27041327

  20. Site-selective excitation and polarized absorption and emission spectra of trivalent thulium and erbium in strontium fluorapatite

    NASA Astrophysics Data System (ADS)

    Gruber, John B.; Wright, Andrew O.; Seltzer, Michael D.; Zandi, Bahram; Merkle, Larry D.; Hutchinson, J. Andrew; Morrison, Clyde A.; Allik, Toomas H.; Chai, Bruce H. T.

    1997-05-01

    Polarized fluorescence spectra produced by site-selective excitation, and conventional polarized absorption spectra were obtained for Tm3+ and Er3+ ions individually incorporated into single crystals of strontium fluorapatite, Sr5(PO4)3F, also known as SFAP. Substitution of the trivalent rare earth ion for divalent strontium was achieved by passive charge compensation during Czochralski growth of the fluorapatite crystals. Spectra were obtained between 1780 and 345 nm at temperatures from 4 K to room temperature on crystals having the hexagonal structure [P63/m(C6h2)]. The polarized fluorescence spectra due to transitions from multiplet manifolds of Tm3+(4f12), including 1D2, 1G4, and 3H4 to manifolds 3H6 (the ground-state manifold), 3F4, 3H5, 3H4, and 3F3 were analyzed for the details of the crystal-field splitting of the manifolds. Fluorescence lifetimes were measured for Tm3+ transitions from 1D2, 1G4, and 3H4 at room temperature and from 1G4 at 16 K. Results of the analysis indicate that the majority of Tm3+ ions occupy sites having Cs symmetry. A point-charge lattice-sum calculation was made in which the crystal-field components, Anm, were determined assuming that trivalent thulium replaces divalent strontium in the metal site having Cs symmetry. Results support the conclusion that the nearest-neighbor fluoride (F-) is replaced by divalent oxygen (O2-), thus preserving overall charge neutrality and local symmetry. Crystal-field splitting calculations predict energy levels in agreement with results obtained from an analysis of the experimental data. By varying the crystal-field parameters, Bnm, we obtained a rms difference of 7 cm-1 between 43 calculated and experimental Stark levels for Tm3+(4f12) in Tm:SFAP. Absorption and fluorescence spectra are also reported for Er3+ ions in Er:SFAP. Measurement of the temporal decay of the room temperature fluorescence from the 4I11/2 and 4I13/2 manifolds yielded fluorescence lifetimes of 230±20 μs and 8.9±0.1 ms

  1. Two-photon excited fluorescence in the LYB:Eu monoclinic crystal: towards a new scheme of single-beam dual-voxel direct laser writing in crystals.

    PubMed

    Petit, Y; Royon, A; Marquestaut, N; Dussauze, M; Fargues, A; Veber, P; Jubera, V; Cardinal, T; Canioni, L

    2013-01-14

    We report on two-photon excited fluorescence in the oriented Eu(3+)doped LYB monoclinic crystal under femtosecond laser tight focusing. Due to spatial walk-off, the two polarization modes of the incident femtosecond beam simultaneously provide the independent excitation of two distinct focuses, leading to a single-beam dual-voxel nonlinear excitation of fluorescence below material modification threshold. These observations emphasize on the anisotropy of both two-photon absorption as well as fluorescence emission. They demonstrate the localized control of the nonlinear energy deposit, thanks to the adjustment of both the input power and polarization, by properly balancing the injected energy in each voxel. Such approach should be considered for future direct laser writing of waveguides in propagation directions out of the dielectric axes, so as to optimally cope with the highly probable anisotropy of laser-induced material modification thresholds in these crystals. These results open new ways for further potential developments in direct laser writing as the simultaneous inscription of double-line structures for original waveguides processes.

  2. Two-photon excited fluorescence in the LYB:Eu monoclinic crystal: new scheme for single-beam dual-voxel direct laser writing in crystals

    NASA Astrophysics Data System (ADS)

    Petit, Y.; Royon, A.; Marquestaut, N.; Dussauze, M.; Fargues, A.; Veber, P.; Jubera, V.; Cardinal, T.; Canioni, L.

    2013-03-01

    We report on two-photon excited fluorescence in the oriented Eu3+-doped LYB monoclinic crystal under femtosecond laser tight focusing. Due to spatial walk-off, the two polarization modes of the incident femtosecond beam simultaneously provide the independent excitation of two distinct focuses, leading to a single-beam dual-voxel nonlinear excitation of fluorescence below material modification threshold. These observations emphasize on the anisotropy of both two-photon absorption as well as fluorescence emission. They demonstrate the localized control of the nonlinear energy deposit, thanks to the adjustment of both the input power and polarization, by properly balancing the injected energy in each voxel. Such approach should be considered for future direct laser writing of waveguides in propagation directions out of the dielectric axes, so as to optimally cope with the highly probable anisotropy of laser-induced material modification thresholds in these crystals. These results open new ways for further potential developments in direct laser writing as the simultaneous inscription of double-line structures for original waveguides processes.

  3. Near infrared excited micro-Raman spectra of 4:1 methanol-ethanol mixture and ruby fluorescence at high pressure

    NASA Astrophysics Data System (ADS)

    Wang, X. B.; Shen, Z. X.; Tang, S. H.; Kuok, M. H.

    1999-06-01

    Near infrared (NIR) lasers, as a new excitation source for Raman spectroscopy, has shown its unique advantages and is being increasingly used for some special samples, such as those emitting strong fluorescence in the visible region. This article focuses on some issues related to high-pressure micro-Raman spectroscopy using NIR excitation source. The Raman spectra of 4:1 methanol-ethanol mixture (4:1 M-E) show a linear variation in both Raman shifts and linewidths under pressure up to 18 GPa. This result is useful in distinguishing Raman scattering of samples from that of the alcohol mixture, an extensively used pressure-transmitting medium. The R1 fluorescence in the red region induced by two-photon absorption of the NIR laser is strong enough to be used as pressure scale. The frequency and line width of the R1 lines are very sensitive to pressure change and the glass transition of the pressure medium. Our results manifest that it is reliable and convenient to use NIR induced two-photon excited fluorescence of ruby for both pressure calibration and distribution of pressure in the 4:1 M-E pressure transmitting medium.

  4. Truly Fluorescent Excitation-Dependent Carbon Dots and Their Applications in Multicolor Cellular Imaging and Multidimensional Sensing.

    PubMed

    Pan, Lulu; Sun, Shan; Zhang, Aidi; Jiang, Kai; Zhang, Ling; Dong, Chaoqing; Huang, Qing; Wu, Aiguo; Lin, Hengwei

    2015-12-16

    Truly fluorescent excitation-dependent carbon dots are prepared, and the relationship between their chemical composition and fluorescent emission is discussed. Furthermore, potential applications of the as-prepared carbon dots to multicolor bio-labeling and multidimodal sensing are demonstrated.

  5. Eliminating Unwanted Far-Field Excitation in Objective-Type TIRF. Part II. Combined Evanescent-Wave Excitation and Supercritical-Angle Fluorescence Detection Improves Optical Sectioning

    PubMed Central

    Brunstein, Maia; Hérault, Karine; Oheim, Martin

    2014-01-01

    Azimuthal beam scanning makes evanescent-wave (EW) excitation isotropic, thereby producing total internal reflection fluorescence (TIRF) images that are evenly lit. However, beam spinning does not fundamentally address the problem of propagating excitation light that is contaminating objective-type TIRF. Far-field excitation depends more on the specific objective than on cell scattering. As a consequence, the excitation impurities in objective-type TIRF are only weakly affected by changes of azimuthal or polar beam angle. These are the main results of the first part of this study (Eliminating unwanted far-field excitation in objective-type TIRF. Pt.1. Identifying sources of nonevanescent excitation light). This second part focuses on exactly where up beam in the illumination system stray light is generated that gives rise to nonevanescent components in TIRF. Using dark-field imaging of scattered excitation light we pinpoint the objective, intermediate lenses and, particularly, the beam scanner as the major sources of stray excitation. We study how adhesion-molecule coating and astrocytes or BON cells grown on the coverslip surface modify the dark-field signal. On flat and weakly scattering cells, most background comes from stray reflections produced far from the sample plane, in the beam scanner and the objective lens. On thick, optically dense cells roughly half of the scatter is generated by the sample itself. We finally show that combining objective-type EW excitation with supercritical-angle fluorescence (SAF) detection efficiently rejects the fluorescence originating from deeper sample regions. We demonstrate that SAF improves the surface selectivity of TIRF, even at shallow penetration depths. The coplanar microscopy scheme presented here merges the benefits of beam spinning EW excitation and SAF detection and provides the conditions for quantitative wide-field imaging of fluorophore dynamics at or near the plasma membrane. PMID:24606929

  6. Cellular characterization of adenylate kinase and its isoform: two-photon excitation fluorescence imaging and fluorescence correlation spectroscopy.

    PubMed Central

    Ruan, Qiaoqiao; Chen, Yan; Gratton, Enrico; Glaser, Michael; Mantulin, William W

    2002-01-01

    Adenylate kinase (AK) is a ubiquitous enzyme that regulates the homeostasis of adenine nucleotides in the cell. AK1beta (long form) from murine cells shares the same protein sequence as AK1 (short form) except for the addition of 18 amino acid residues at its N-terminus. It is hypothesized that these residues serve as a signal for protein lipid modification and targeting of the protein to the plasma membrane. To better understand the cellular function of these AK isoforms, we have used several modern fluorescence techniques to characterize these two isoforms of AK enzyme. We fused cytosolic adenylate kinase (AK1) and its isoform (AK1beta) with enhanced green fluorescence protein (EGFP) and expressed the chimera proteins in HeLa cells. Using two-photon excitation scanning fluorescence imaging, we were able to directly visualize the localization of AK1-EGFP and AK1beta-EGFP in live cells. AK1beta-EGFP mainly localized on the plasma membrane, whereas AK1-EGFP distributed throughout the cell except for trace amounts in the nuclear membrane and some vesicles. We performed fluorescence correlation spectroscopy measurements and photon-counting histogram analysis in specific domains of live cells. For AK1-EGFP, we observed only one diffusion component in the cytoplasm. For AK1beta-EGFP, we observed two distinct diffusion components on the plasma membrane. One corresponded to the free diffusing protein, whereas the other represented the membrane-bound AK1beta-EGFP. The diffusion rate of AK1-EGFP was slowed by a factor of 1.8 with respect to that of EGFP, which was 50% more than what we would expect for a free diffusing AK1-EGFP. To rule out the possibility of oligomer formation, we performed photon-counting histogram analysis to direct analyze the brightness difference between AK1-EGFP and EGFP. From our analysis, we concluded that cytoplasmic AK1-EGFP is monomeric. fluorescence correlation spectroscopy proved to be a powerful technique for quantitatively studying the

  7. Highly-sensitive Eu(3+) ratiometric thermometers based on excited state absorption with predictable calibration.

    PubMed

    Souza, Adelmo S; Nunes, Luiz A O; Silva, Ivan G N; Oliveira, Fernando A M; da Luz, Leonis L; Brito, Hermi F; Felinto, Maria C F C; Ferreira, Rute A S; Júnior, Severino A; Carlos, Luís D; Malta, Oscar L

    2016-03-01

    Temperature measurements ranging from a few degrees to a few hundreds of Kelvin are of great interest in the fields of nanomedicine and nanotechnology. Here, we report a new ratiometric luminescent thermometer using thermally excited state absorption of the Eu(3+) ion. The thermometer is based on the simple Eu(3+) energy level structure and can operate between 180 and 323 K with a relative sensitivity ranging from 0.7 to 1.7% K(-1). The thermometric parameter is defined as the ratio between the emission intensities of the (5)D0 → (7)F4 transition when the (5)D0 emitting level is excited through the (7)F2 (physiological range) or (7)F1 (down to 180 K) level. Nano and microcrystals of Y2O3:Eu(3+) were chosen as a proof of concept of the operational principles in which both excitation and detection are within the first biological transparent window. A novel and of paramount importance aspect is that the calibration factor can be calculated from the Eu(3+) emission spectrum avoiding the need for new calibration procedures whenever the thermometer operates in different media.

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

    PubMed Central

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

    2016-01-01

    Multimodal imaging probes a variety of tissue properties in a single image acquisition by merging complimentary imaging technologies. Exploiting synergies amongst the data, algorithms can be developed that lead to better tissue characterization than could be accomplished by the constituent imaging modalities taken alone. The combination of optical coherence tomography (OCT) with fluorescence lifetime imaging microscopy (FLIM) provides access to detailed tissue morphology and local biochemistry. The optical system described here merges 1310 nm swept-source OCT with time-domain FLIM having excitation at 355 and 532 nm. The pulses from 355 and 532 nm lasers have been interleaved to enable simultaneous acquisition of endogenous and exogenous fluorescence signals, respectively. The multimodal imaging system was validated using tissue phantoms. Nonspecific tagging with Alexa Flour 532 in a Watanbe rabbit aorta and active tagging of the LOX-1 receptor in human coronary artery, demonstrate the capacity of the system for simultaneous acquisition of OCT, endogenous FLIM, and exogenous FLIM in tissues. PMID:27699091

  9. Absorption spectrophotometric, fluorescence, transient absorption and quantum chemical investigations on fullerene/phthalocyanine supramolecular complexes.

    PubMed

    Ray, Anamika; Santhosh, Kotni; Bhattacharya, Sumanta

    2011-05-01

    The present paper reports the photophysical investigations on supramolecular interaction of a phthalocyanine derivative, namely, 2,9,16,23-tetra-tert-butyl-29H,31H-Pc (1) with C(60) and C(70) in toluene. The binding constants of the C(60) and C(70) complexes of 1 are estimated to be 27,360 and 25,205 dm(3), respectively. Transient absorption measurements in the visible region establishes that energy transfer from C60*T (and C70*T) to 1 occurs predominantly in toluene which is subsequently confirmed by the consecutive appearance of the triplet states of 1. Quantum chemical calculations at DFT level of theory explore the geometry and electronic structure of the supramolecules and testify the significant redistribution of charge between fullerenes and 1.

  10. Investigation of Fluorescence Characteristic in Transversely Excited Dye and Nanoparticle-Doped PMMA Fibre

    NASA Astrophysics Data System (ADS)

    NG, C. S.; Yap, S. S.; Chin, O. H.; Wong, H. Y.; Tou, T. Y.

    2011-03-01

    A simple extruder was used to fabricate poly (methyl methacrylate) (PMMA) fibers with optimum diameter 0.5 mm from the modified, low-softening (50-60° C) PMMA that was prepared using the reactive polymerization. The laser dye Rhodamine 6G (R6G) and Zinc Oxide (ZnO) nanoparticles of nominal diameter 30nm were added to the monomer MMA prior to the polymerization process, hence producing a random media. The nanoparticle embedded dye-doped PMMA fibers were transversely excited by TEA Nitrogen (N2) laser for fluorescence studies. It was observed that ZnO nanoparticles embedded in the dye-doped PMMA fibre significantly improved the fluorescence characteristic.

  11. Photocatalytic degradation-excitation-emission matrix fluorescence for increasing the selectivity of polycyclic aromatic hydrocarbon analyses.

    PubMed

    Kim, Yoon-Chang; Jordan, James A; Nahorniak, Michelle L; Booksh, Karl S

    2005-12-01

    The application of photocatalysis enhancement to calibration of fluorescence excitation-emission matrixes (EEMs) with parallel factor (PARAFAC) analysis is described. In this study, three- and four-way PARAFAC analysis was employed to extract the fluorescent species' spectra from overlapping EEMs. Time-dependent photocatalysis degradation of the polycyclic aromatic hydrocarbons (PAHs) was employed to create an additional dimension for analysis. The consequent four-dimension degradation-EEM data cubes have greater selectivity for each PAH than do three-dimension EEM data cubes alone. On a scale of 0 to 1, with 0 being completely collinear spectra and 1 being orthogonal spectra, including the time-dependent measurements increased the selectivity an average of 21%, from 0.73 to 0.87.

  12. Excited-State Proton Transfer Can Tune the Color of Protein Fluorescent Markers.

    PubMed

    Mancini, Daiana T; Sen, Kakali; Barbatti, Mario; Thiel, Walter; Ramalho, Teodorico C

    2015-11-16

    We show by quantum mechanical/molecular mechanical (QM/MM) simulations that phenylbenzothiazoles undergoing an excited-state proton transfer (ESPT) can be used to probe protein binding sites. For 2-(2'-hydroxy-4'-aminophenyl)benzothiazole (HABT) bound to a tyrosine kinase, the absolute and relative intensities of the fluorescence bands arising from the enol and keto forms are found to be strongly dependent on the active-site conformation. The emission properties are tuned by hydrogen-bonding interactions of HABT with the neighboring amino acid T766 and with active-site water. The use of ESPT tuners opens the possibility of creating two-color fluorescent markers for protein binding sites, with potential applications in the detection of mutations in cancer cell lines. PMID:26333875

  13. Continuous flow real-time PCR device using multi-channel fluorescence excitation and detection.

    PubMed

    Hatch, Andrew C; Ray, Tathagata; Lintecum, Kelly; Youngbull, Cody

    2014-02-01

    High throughput automation is greatly enhanced using techniques that employ conveyor belt strategies with un-interrupted streams of flow. We have developed a 'conveyor belt' analog for high throughput real-time quantitative Polymerase Chain Reaction (qPCR) using droplet emulsion technology. We developed a low power, portable device that employs LED and fiber optic fluorescence excitation in conjunction with a continuous flow thermal cycler to achieve multi-channel fluorescence detection for real-time fluorescence measurements. Continuously streaming fluid plugs or droplets pass through tubing wrapped around a two-temperature zone thermal block with each wrap of tubing fluorescently coupled to a 64-channel multi-anode PMT. This work demonstrates real-time qPCR of 0.1-10 μL droplets or fluid plugs over a range of 7 orders of magnitude concentration from 1 × 10(1) to 1 × 10(7). The real-time qPCR analysis allows dynamic range quantification as high as 1 × 10(7) copies per 10 μL reaction, with PCR efficiencies within the range of 90-110% based on serial dilution assays and a limit of detection of 10 copies per rxn. The combined functionality of continuous flow, low power thermal cycling, high throughput sample processing, and real-time qPCR improves the rates at which biological or environmental samples can be continuously sampled and analyzed. PMID:24297040

  14. In vivo Diagnosis of Cervical Intraepithelial Neoplasia Using 337-nm- Excited Laser-Induced Fluorescence

    NASA Astrophysics Data System (ADS)

    Ramanujam, N.; Mitchell, M. F.; Mahadevan, A.; Warren, S.; Thomsen, S.; Silva, E.; Richards-Kortum, R.

    1994-10-01

    Laser-induced fluorescence at 337-nm excitation was used in vivo to differentiate neoplastic [cervical intraepithelial neoplasia (CIN)], nonneoplastic abnormal (inflammation and human papilloma viral infection), and normal cervical tissues. A colposcope (low-magnification microscope used to view the cervix with reflected light) was used to identify 66 normal and 49 abnormal (5 inflammation, 21 human papilloma virus infection, and 23 CIN) sites on the cervix in 28 patients. These sites were then interrogated spectroscopically. A two-stage algorithm was developed to diagnose CIN. The first stage differentiated histologically abnormal tissues from colposcopically normal tissues with a sensitivity, specificity, and positive predictive value of 92%, 90%, and 88%, respectively. The second stage differentiated preneoplastic and neoplastic tissues from nonneoplastic abnormal tissues with a sensitivity, specificity, and positive predictive value of 87%, 73%, and 74%, respectively. Spectroscopic differences were consistent with a decrease in the absolute contribution of collagen fluorescence, an increase in the absolute contribution of oxyhemoglobin attenuation, and an increase in the relative contribution of reduced nicotinamide dinucleotide phosphate [NAD(P)H] fluorescence as tissue progresses from normal to abnormal in the same patient. These results suggest that in vivo fluorescence spectroscopy of the cervix can be used to diagnose CIN at colposcopy.

  15. Two-photon excited fluorescence lifetime imaging and spectroscopy of melanins in vitro and in vivo

    NASA Astrophysics Data System (ADS)

    Krasieva, Tatiana B.; Stringari, Chiara; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L.; Gratton, Enrico; Tromberg, Bruce J.

    2013-03-01

    Changes in the amounts of cellular eumelanin and pheomelanin have been associated with carcinogenesis. The goal of this work is to develop methods based on two-photon-excited-fluorescence (TPEF) for measuring relative concentrations of these compounds. We acquire TPEF emission spectra (λex=1000 nm) of melanin in vitro from melanoma cells, hair specimens, and in vivo from healthy volunteers. We find that the pheomelanin emission peaks at approximately 615 to 625 nm and eumelanin exhibits a broad maximum at 640 to 680 nm. Based on these data we define an optical melanin index (OMI) as the ratio of fluorescence intensities at 645 and 615 nm. The measured OMI for the MNT-1 melanoma cell line is 1.6±0.22 while the Mc1R gene knockdown lines MNT-46 and MNT-62 show substantially greater pheomelanin production (OMI=0.5±0.05 and 0.17±0.03, respectively). The measured values are in good agreement with chemistry-based melanin extraction methods. In order to better separate melanin fluorescence from other intrinsic fluorophores, we perform fluorescence lifetime imaging microscopy of in vitro specimens. The relative concentrations of keratin, eumelanin, and pheomelanin components are resolved using a phasor approach for analyzing lifetime data. Our results suggest that a noninvasive TPEF index based on spectra and lifetime could potentially be used for rapid melanin ratio characterization both in vitro and in vivo.

  16. Label-free in vivo imaging of human leukocytes using two-photon excited endogenous fluorescence

    NASA Astrophysics Data System (ADS)

    Zeng, Yan; Yan, Bo; Sun, Qiqi; Teh, Seng Khoon; Zhang, Wei; Wen, Zilong; Qu, Jianan Y.

    2013-04-01

    We demonstrate that two-photon excited endogenous fluorescence enables label-free morphological and functional imaging of various human blood cells. Specifically, we achieved distinctive morphological contrast to visualize morphology of important leukocytes, such as polymorphonuclear structure of granulocyte and mononuclear feature of agranulocyte, through the employment of the reduced nicotinamide adenine dinucleotide (NADH) fluorescence signals. In addition, NADH fluorescence images clearly reveal the morphological transformation process of neutrophils during disease-causing bacterial infection. Our findings also show that time-resolved NADH fluorescence can be potentially used for functional imaging of the phagocytosis of pathogens by leukocytes (neutrophils) in vivo. In particular, we found that free-to-bound NADH ratios measured in infected neutrophils increased significantly, which is consistent with a previous study that the energy consumed in the phagocytosis of neutrophils is mainly generated through the glycolysis pathway that leads to the accumulation of free NADH. Future work will focus on further developing and applying label-free imaging technology to investigate leukocyte-related diseases and disorders.

  17. Two-photon excited fluorescence lifetime imaging and spectroscopy of melanins in vitro and in vivo.

    PubMed

    Krasieva, Tatiana B; Stringari, Chiara; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L; Gratton, Enrico; Tromberg, Bruce J

    2013-03-01

    Changes in the amounts of cellular eumelanin and pheomelanin have been associated with carcinogenesis. The goal of this work is to develop methods based on two-photon-excited-fluorescence (TPEF) for measuring relative concentrations of these compounds. We acquire TPEF emission spectra (λ(ex)=1000  nm) of melanin in vitro from melanoma cells, hair specimens, and in vivo from healthy volunteers. We find that the pheomelanin emission peaks at approximately 615 to 625 nm and eumelanin exhibits a broad maximum at 640 to 680 nm. Based on these data we define an optical melanin index (OMI) as the ratio of fluorescence intensities at 645 and 615 nm. The measured OMI for the MNT-1 melanoma cell line is 1.6 ± 0.22 while the Mc1R gene knockdown lines MNT-46 and MNT-62 show substantially greater pheomelanin production (OMI=0.5 ± 0.05 and 0.17 ± 0.03, respectively). The measured values are in good agreement with chemistry-based melanin extraction methods. In order to better separate melanin fluorescence from other intrinsic fluorophores, we perform fluorescence lifetime imaging microscopy of in vitro specimens. The relative concentrations of keratin, eumelanin, and pheomelanin components are resolved using a phasor approach for analyzing lifetime data. Our results suggest that a noninvasive TPEF index based on spectra and lifetime could potentially be used for rapid melanin ratio characterization both in vitro and in vivo.

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

    NASA Astrophysics Data System (ADS)

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

    2004-11-01

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

  19. Ultrafast Excited-State Dynamics of 6-Azauracil Studied by Femtosecond Transient Absorption Spectroscopy.

    PubMed

    Hua, XinZhong; Hua, LinQiang; Liu, XiaoJun

    2015-12-31

    The excited-state dynamics of 6-azauracil in different solvents have been studied using femtosecond transient absorption spectroscopy. The molecule is populated to the S2 state with a pump pulse at 264 nm. Broad-band white light continuum which covers from 320 to 600 nm is used as the probe. With a global fitting analysis of the measured transient spectra, three decay time constants, i.e., <0.3, 5.2 ± 0.1, and >1000 ps, are directly obtained in the solvent of acetonitrile. These newly observed lifetime constants are important in clarifying its decay dynamics as well as in providing a criterion for the ultrafast dynamics simulations in 6-azauracil using quantum chemical theories. In combination with previous theoretical works, the main decay channel is proposed: the initially populated S2 decays to S1 through internal conversion in <0.3 ps, followed by an intersystem crossing from S1 to T1 in 5.2 ± 0.1 ps. The >1000 ps component is due to the decay of the T1 state. A comparison of the excited-state dynamics in different solvents reveals that the decay from S1 to T1 shows a clear dependence on the polarity of the solvents. With higher polarity, the S1 excited state decays faster. This observation is in line with the prediction by Etinski et al. [ Phys. Chem. Chem. Phys. 2010 , 12 , 15665 - 15671 ], where a blue-shift of the T1 state potential energy surface leading to an increase of the intersystem crossing rate was proposed. With the new information obtained in the present measurement, a clearer picture of the decay dynamics of 6-azauracil on the S2 excited state is provided.

  20. Biomolecular imaging based on far-red fluorescent protein with a high two-photon excitation action cross section

    NASA Astrophysics Data System (ADS)

    Tsai, Tsung-Han; Lin, Cheng-Yung; Tsai, Huai-Jen; Chen, Szu-Yu; Tai, Shih-Peng; Lin, Kung-Hsuan; Sun, Chi-Kuang

    2006-04-01

    The two-photon excitation action cross section of Hc-Red fluorescent proteins (Hc-RFPs) is measured and found to be of the same order as that of enhanced green fluorescent proteins. With a 618 nm emission wavelength in the far-red region and with an excitation wavelength around 1200 nm, Hc-RPF-based two-photon fluorescence microscopy (2PFM) can offer deep penetration capability inside live samples and is ideal for in vivo gene expression study and biomolecular imaging in live objects. In vivo 2PFM of the developing heart deep inside a transgenic zebrafish embryo tagged by Hc-RFP is also successfully demonstrated.

  1. Comments on laser-excited fluorescence of the hydroxyl radical: Relaxation coefficients at atmospheric pressure, appendix 5

    NASA Technical Reports Server (NTRS)

    Wang, C. C.

    1983-01-01

    The lifetime of the excited state of a atom or molecule is often determined from the rate of fluorescence decay originating as a function of buffer gas pressure, an accurate determination is made of the rates of collision induced transitions away from the excited state. Deconvolution can in principle be employed to resolve fluorescence times shorter than the response times of the system. However, attainable reproducibility and accuracy in actual experiments usually set a limit beyond which no meaningful results are expected. Prudence thus dictates that the results of deconvolution be viewed with extreme caution whenever fluorescence time much shorter than the response of times of the system are indicated.

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

    SciTech Connect

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

    1994-03-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2002-05-01

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

  4. Quantitative absorption and fluorescence study of CO from 1060 to 1550 A

    NASA Technical Reports Server (NTRS)

    Lee, L. C.; Guest, J. A.

    1980-01-01

    Measurement of the photoabsorption cross section of CO in the 1060-1550 A region using synchrotron radiation is described. The oscillator strengths for the transitions from CO (Chi1Sigma+ to various excited states are obtained from these data. Fluorescence from excited CO was observed in the 1150 to 3000 A and 3000 to 8000 A regions. The quantum yields for the production of fluroescence from the Alpha(1)P and B(1)Sigma(+) states are unity because their excitation energies are below the dissociation limit. The C(1)Sigma(+) , v = O level has significant fluorescence quantum yields both in the UV and visible region, but the yields for the E(1)Pi, v = O and C(1)Sigma(+), v = 1 levels are very small. The C(1)Sigma(+), v = 1 level is presumably predissociated. The cross sections for the production of fluroescence from the a'(3)Sigma(+), d(3)Delta sub 1, e(3)Sigma(-) yields a(3)Pi, and B(1)Sigma(+), C(1)Sigma(+) yields A(1)Pi transitions upon excitation from the X(1)Sigma(+) were measured.

  5. Ratiometric fluorescent chemosensor for fluoride ion based on inhibition of excited state intramolecular proton transfer

    NASA Astrophysics Data System (ADS)

    Gupta, Akul Sen; Paul, Kamaldeep; Luxami, Vijay

    2015-03-01

    ESIPT based benzimidazole derivative has been synthesized and investigated their photophysical behavior towards various anions. The probe 2 has been used for selective estimation of F- ions as compared to other anions and signaled the binding event through formation of new absorption band at 360 nm and emission band at 420 nm. The probe 2 showed fluorescence behavior towards fluoride ions through hydrogen bonding interactions and restricted the ESIPT emission at 540 nm from OH to nitrogen of benzimidazole moiety to release its enol emission at 420 nm.

  6. X-ray microprobe for micro x-ray fluorescence and absorption spectroscopies at GSECARS

    NASA Astrophysics Data System (ADS)

    Newville, M.; Sutton, S.; Rivers, M.

    2002-12-01

    The hard x-ray microprobe for x-ray fluorescence and absorption spectroscopy at GeoSoilEnviroCARS is presented. Using focused synchrotron radiation from an undulator beamline at the Advanced Photon Source at Argonne National Lab, the x-ray microprobe provides bright, monochromatic x-rays with typical spot sizes down to 1x1 μm for x-ray fluorescence and absorption spectroscopies. Quantitative x-ray fluorescence (XRF) analysis gives precise elemental composition and correlations, while x-ray absorption spectroscopy (XAS) gives the chemical state and local atomic coordination for a selected atomic species. These two techniques can be used in conjunction with one another on a wide range of samples, including minerals, glasses, fluid inclusions, soils, sediments, and plant tissue. This x-ray microprobe is part of the GeoSoilEnviroCARS user facility, available for use in all areas geological, soil, and environmental sciences, and selected examples from these fields will be given.

  7. Excitation laser energy dependence of surface-enhanced fluorescence showing plasmon-induced ultrafast electronic dynamics in dye molecules

    NASA Astrophysics Data System (ADS)

    Itoh, Tamitake; Yamamoto, Yuko S.; Tamaru, Hiroharu; Biju, Vasudevanpillai; Murase, Norio; Ozaki, Yukihiro

    2013-06-01

    We find unique properties accompanying surface-enhanced fluorescence (SEF) from dye molecules adsorbed on Ag nanoparticle aggregates, which generate surface-enhanced Raman scattering. The properties are observed in excitation laser energy dependence of SEF after excluding plasmonic spectral modulation in SEF. The unique properties are large blue shifts of fluorescence spectra, deviation of ratios between anti-Stokes SEF intensity and Stokes from those of normal fluorescence, super-broadening of Stokes spectra, and returning to original fluorescence by lower energy excitation. We elucidate that these properties are induced by electromagnetic enhancement of radiative decay rates exceeding the vibrational relaxation rates within an electronic excited state, which suggests that molecular electronic dynamics in strong plasmonic fields can be largely deviated from that in free space.

  8. Substituent and Solvent Effects on Excited State Charge Transfer Behavior of Highly Fluorescent Dyes Containing Thiophenylimidazole-Based Aldehydes

    NASA Technical Reports Server (NTRS)

    Santos, Javier; Bu, Xiu R.; Mintz, Eric A.

    2001-01-01

    The excited state charge transfer for a series of highly fluorescent dyes containing thiophenylimidazole moiety was investigated. These systems follow the Twisted Intramolecular Charge Transfer (TICT) model. Dual fluorescence was observed for each substituted dye. X-ray structures analysis reveals a twisted ground state geometry for the donor substituted aryl on the 4 and 5 position at the imidazole ring. The excited state charge transfer was modeled by a linear solvation energy relationship using Taft's pi and Dimroth's E(sub T)(30) as solvent parameters. There is linear relation between the energy of the fluorescence transition and solvent polarity. The degree of stabilization of the excited state charge transfer was found to be consistent with the intramolecular molecular charge transfer. Excited dipole moment was studied by utilizing the solvatochromic shift method.

  9. 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.

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

    NASA Astrophysics Data System (ADS)

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

    1995-10-01

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

  11. Excited State Proton Transfer in the Red Fluorescent Protein mKeima

    PubMed Central

    Henderson, J. Nathan; Osborn, Maire F.; Koon, Nayden; Gepshtein, Rinat; Huppert, Dan; Remington, S. James

    2009-01-01

    mKeima is an unusual monomeric red fluorescent protein (λemmax ~620 nm) that is maximally excited in the blue (λexmax ~440 nm). The large Stokes shift suggests that the chromophore is normally protonated. A 1.63 Å resolution structure of mKeima reveals the chromophore to be imbedded in a novel hydrogen bond network, different than in GFP, which could support proton transfer from the chromophore hydroxyl, via Ser142, to Asp157. At low temperatures the emission contains a green component (λemmax ~535 nm), enhanced by deuterium substitution, presumably resulting from reduced proton transfer efficiency. Ultrafast pump/probe studies reveal a rising component in the 610 nm emission with lifetime ~4 ps, characterizing the rate of proton transfer. Mutation of Asp157 to neutral Asn changes the chromophore resting charge state to anionic (λexmax ~565 nm, λemmax ~620 nm). Thus, excited state proton transfer (ESPT) explains the large Stokes shift. This work unambiguously characterizes green emission from the protonated acylimine chromophore of red fluorescent proteins. PMID:19708654

  12. Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with cameleons.

    PubMed

    Fan, G Y; Fujisaki, H; Miyawaki, A; Tsay, R K; Tsien, R Y; Ellisman, M H

    1999-05-01

    A video-rate (30 frames/s) scanning two-photon excitation microscope has been successfully tested. The microscope, based on a Nikon RCM 8000, incorporates a femtosecond pulsed laser with wavelength tunable from 690 to 1050 nm, prechirper optics for laser pulse-width compression, resonant galvanometer for video-rate point scanning, and a pair of nonconfocal detectors for fast emission ratioing. An increase in fluorescent emission of 1.75-fold is consistently obtained with the use of the prechirper optics. The nonconfocal detectors provide another 2.25-fold increase in detection efficiency. Ratio imaging and optical sectioning can therefore be performed more efficiently without confocal optics. Faster frame rates, at 60, 120, and 240 frames/s, can be achieved with proportionally reduced scan lines per frame. Useful two-photon images can be acquired at video rate with a laser power as low as 2.7 mW at specimen with the genetically modified green fluorescent proteins. Preliminary results obtained using this system confirm that the yellow "cameleons" exhibit similar optical properties as under one-photon excitation conditions. Dynamic two-photon images of cardiac myocytes and ratio images of yellow cameleon-2.1, -3.1, and -3.1nu are also presented. PMID:10233058

  13. Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with cameleons.

    PubMed Central

    Fan, G Y; Fujisaki, H; Miyawaki, A; Tsay, R K; Tsien, R Y; Ellisman, M H

    1999-01-01

    A video-rate (30 frames/s) scanning two-photon excitation microscope has been successfully tested. The microscope, based on a Nikon RCM 8000, incorporates a femtosecond pulsed laser with wavelength tunable from 690 to 1050 nm, prechirper optics for laser pulse-width compression, resonant galvanometer for video-rate point scanning, and a pair of nonconfocal detectors for fast emission ratioing. An increase in fluorescent emission of 1.75-fold is consistently obtained with the use of the prechirper optics. The nonconfocal detectors provide another 2.25-fold increase in detection efficiency. Ratio imaging and optical sectioning can therefore be performed more efficiently without confocal optics. Faster frame rates, at 60, 120, and 240 frames/s, can be achieved with proportionally reduced scan lines per frame. Useful two-photon images can be acquired at video rate with a laser power as low as 2.7 mW at specimen with the genetically modified green fluorescent proteins. Preliminary results obtained using this system confirm that the yellow "cameleons" exhibit similar optical properties as under one-photon excitation conditions. Dynamic two-photon images of cardiac myocytes and ratio images of yellow cameleon-2.1, -3.1, and -3.1nu are also presented. PMID:10233058

  14. Selective two-photon absorptive resonance femtosecond-laser electronic-excitation tagging velocimetry.

    PubMed

    Jiang, Naibo; Halls, Benjamin R; Stauffer, Hans U; Danehy, Paul M; Gord, James R; Roy, Sukesh

    2016-05-15

    Selective two-photon absorptive resonance femtosecond-laser electronic-excitation tagging (STARFLEET), a nonseeded ultrafast-laser-based velocimetry technique, is demonstrated in reactive and nonreactive flows. STARFLEET is pumped via a two-photon resonance in N2 using 202.25 nm 100 fs light. STARFLEET greatly reduces the per-pulse energy required (30 μJ/pulse) to generate the signature FLEET emission compared to the conventional FLEET technique (1.1 mJ/pulse). This reduction in laser energy results in less energy deposited in the flow, which allows for reduced flow perturbations (reactive and nonreactive), increased thermometric accuracy, and less severe damage to materials. Velocity measurements conducted in a free jet of N2 and in a premixed flame show good agreement with theoretical velocities, and further demonstrate the significantly less intrusive nature of STARFLEET. PMID:27176968

  15. Excited electronic structure of methylcyanoacetylene probed by VUV Fourier-transform absorption spectroscopy

    NASA Astrophysics Data System (ADS)

    Lamarre, N.; Gans, B.; Vieira Mendes, L. A.; Gronowski, M.; Guillemin, J.-C.; De Oliveira, N.; Douin, S.; Chevalier, M.; Crépin, C.; Kołos, R.; Boyé-Péronne, S.

    2016-10-01

    High resolution photoabsorption spectrum of gas-phase methylcyanoacetylene (CH3C3 N) has been recorded from 44 500 to 130 000 cm-1 at room temperature with a vacuum ultraviolet Fourier-transform spectrometer on the DESIRS synchrotron beamline (SOLEIL). The absolute photoabsorption cross section in this range is reported for the first time. Valence shell transitions and Rydberg series converging to the ground state X˜+2E of the cation as well as series converging to electronically excited states (A˜+A21 and C˜+) are observed and assigned. Time-dependent density-functional-theory calculations have been performed to support the assignment of the experimental spectrum in the low energy range. A tentative scaling of the previously measured CH3C3N+ ion yield by Lamarre et al. [17] is proposed, based on the comparison of the absorption data above the first ionization potential with the observed autoionization structures.

  16. Fluorescence detection of camptothecin anticancer drugs by two-photon excitation

    NASA Astrophysics Data System (ADS)

    Burke, Thomas G.; Malak, Magda; Bom, David; Curran, Dennis P.; Malak, Henryk M.; Gryczynski, Ignacy; Lakowicz, Joseph R.

    1998-04-01

    Hycamtin is a camptothecin anticancer analogue containing a dimethylaminomethyl substituent at position 9 and a hydroxy functionality at position 10. Using an excitation wavelength of 800 nm we have compared the two-photon cross sections and excited-state lifetimes from several camptothecins in phosphate buffered saline solution with and without the presence of human serum albumin (HSA). Drug and HSA concentrations of 10 (mu) M and 46 (mu) M were employed in our studies. In phosphate buffered saline solution containing HSA the following excited-state lifetimes (ns) and two- photon cross-sections (10-50 cm4 s/photon), respectively, were determined: hycamtin (4.3 nm, 36); camptothecin (1.3 ns, 1); 7-t-butyldimethylsilyl-10- hydroxycamptothecin (1.7 ns, 3.7); 7-t-butyldimethylsilyl- camptothecin (1.9 ns, 1.9); 7-trimethylsilyl-10- aminocamptothecin (6.3 ns; 35); and 7-trimethylsilyl-10- hydroxycamptothecin (1.8 ns; 2.2). Our results indicate that Hycamtin exhibits a high cross-section relative to the parent camptothecin molecule and represents one of the best camptothecin analogues to detect using two-photon excitation. Hycamtin was detected at concentrations as low as 0.05 (mu) M and 1 (mu) M in plasma and whole blood, respectively. The newly synthesized analogue 7- trimethylsilyl-10-aminocamptothecin was found to display similar lifetime and two-photon cross section values relative to Hycamtin. Thus, fluorescence detection with two- photon excitation may prove to be of advantage in the development of this promising new experimental therapeutic.

  17. Observation of ultralow-level Al impurities on a silicon surface by high-resolution grazing emission x-ray fluorescence excited by synchrotron radiation

    SciTech Connect

    Kubala-Kukus, A.; Banas, D.; Pajek, M.; Cao, W.; Dousse, J.-Cl.; Hoszowska, J.; Kayser, Y.; Szlachetko, M.; Salome, M.; Susini, J.; Szlachetko, J.

    2009-09-15

    We demonstrate that ultralow-level Al impurities on a silicon surface can be measured by using the high-resolution grazing emission x-ray fluorescence (GEXRF) technique combined with synchrotron-radiation excitation. An Al-impurity level of about 10{sup 12} atoms/cm{sup 2} was reached by observing the Al K{alpha} x-ray fluorescence in the resonant Raman-scattering background-''free'' regime by choosing an appropriate beam energy below the Si K absorption edge. Present results show that by combining the GEXRF method with the vapor phase decomposition technique the 10{sup 7} atoms/cm{sup 2} level can be reached for Al detection on silicon. Finally, we found that the high-resolution GEXRF technique is a sensitive tool to study the morphology of surface nanostructures.

  18. Excited-State Absorption from Real-Time Time-Dependent Density Functional Theory: Optical Limiting in Zinc Phthalocyanine.

    PubMed

    Fischer, Sean A; Cramer, Christopher J; Govind, Niranjan

    2016-04-01

    Optical-limiting materials are capable of attenuating light to protect delicate equipment from high-intensity light sources. Phthalocyanines have attracted a lot of attention for optical-limiting applications due to their versatility and large nonlinear absorption. With excited-state absorption (ESA) being the primary mechanism for optical limiting behavior in phthalocyanines, the ability to tune the optical absorption of ground and excited states in phthalocyanines would allow for the development of advanced optical limiters. We recently developed a method for the calculation of ESA based on real-time time-dependent density functional theory propagation of an excited-state density. In this work, we apply the approach to zinc phthalocyanine, demonstrating the ability of our method to efficiently identify the optical limiting potential of a molecular complex.

  19. Differences in excitation energy transfer of Arthrospira platensis cells grown in seawater medium and freshwater medium, probed by time-resolved fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Arba, Muhammad; Aikawa, Shimpei; Niki, Kenta; Yokono, Makio; Kondo, Akihiko; Akimoto, Seiji

    2013-11-01

    Excitation energy transfer of Arthrospira platensis cells grown in f/2 medium (a high salinity medium) and SOT medium (a control) was investigated by steady-state and time-resolved spectroscopies. Growth in f/2 medium induced changes in absorption and fluorescence spectra as well as in the energy transfer pathways. Excitation energy captured by phycobilisome (PBS) was transferred directly to photosystem (PS) I, instead of being first transferred to an intermediate (PBS → PSII → PSI), as observed in SOT medium. The respiration rate increased while photosynthetic rate reduced in f/2 medium. Possible causes of the differences in light-harvesting and energy-transfer processes between the two media are discussed.

  20. Three-dimensional tracking of a single fluorescent nanoparticle using four-focus excitation in a confocal microscope.

    PubMed

    Germann, James A; Davis, Lloyd M

    2014-03-10

    We report high sensitivity detection and tracking of a single fluorescent nanoparticle in solution by use of four alternately pulsed laser diodes for fluorescence excitation in a confocal microscope. Slight offsets between the centers of the overlapping laser foci together with time-resolved photon counting enable sub-micron precision position measurements. Real-time correction for diffusional motion with a xyz-piezo stage then enables tracking of a nanoparticle with diffusivity up to ~12 μm(2) s(-1). Fluorescence correlation spectroscopy and calibration measurements indicate a net fluorescence photon detection efficiency of ~6-9%, comparable to that of an optimized single-molecule microscope. PMID:24663905

  1. Source-corrected two-photon excited fluorescence measurements between 700 and 880 nm

    SciTech Connect

    Fisher, W.G.; Wachter, E.A.; Lytle, F.E.; Armas, M.; Seaton, C.

    1998-04-01

    Passively mode-locked titanium:sapphire (Ti:S) lasers are capable of generating a high-frequency train of transform-limited subpico-second pulses, producing peak powers near 10{sup 5}thinspW at moderate average powers. The low energy per pulse ({lt}20 nJ) permits low fluence levels to be maintained in tightly focused beams, reducing the possibility of saturating fluorescence transitions. These properties, combined with a wavelength tunability from approximately 700 nm to 1 {mu}m, provide excellent opportunities for studying simultaneous two-photon excitation (TPE). However, pulse formation is very sensitive to a variety of intracavity parameters, including group velocity dispersion compensation, which leads to wavelength-dependent pulse profiles as the wavelength is scanned. This wavelength dependence can seriously distort band shapes and apparent peak heights during collection of two-photon spectral data. Since two-photon excited fluorescence is proportional to the product of the peak and average powers, it is not possible to obtain source-independent spectra by using average power correction schemes alone. Continuous-wave, single-mode lasers can be used to generate source-independent two-photon data, but these sources are four to five orders of magnitude less efficient than the mode-locked Ti:S laser and are not practical for general two-photon measurements. Hence, a continuous-wave, single-mode Ti:S laser has been used to collect a source-independent excitation spectrum for the laser dye Coumarin 480. This spectrum may be used to correct data collected with multimode sources; this possibility is demonstrated by using a simple ratiometric method to collect accurate TPE spectra with the mode-locked Ti:S laser. An approximate value of the two-photon cross section for Coumarin 480 is also given. {copyright} {ital 1998} {ital Society for Applied Spectroscopy}

  2. Integrated OLED as excitation light source in fluorescent lateral flow immunoassays.

    PubMed

    Venkatraman, Vishak; Steckl, Andrew J

    2015-12-15

    The integration of organic light emitting diodes (OLEDs) as excitation light sources for quantum dot-based fluorescent lateral flow immunoassay systems (LFIA) was investigated. This approach has the potential to deliver a sensitive visible detection scheme for low-cost, disposable lab-on-chip point-of-care (POC) diagnosis system. Thin film phosphorescent green OLEDs fabricated on plastic substrates were integrated on-chip to excite the test line of a quantum dot-based LFIA (QD-LFIA). OLEDs were fabricated by sequential deposition of organic thin films (total of ~100 nm) onto ITO-coated PET substrates. CdSe/ZnS QDs emitting at 655 nm and Au nanoparticles (NP - 10 nm size) conjugated antibodies were used for the fluorescence QD-LFIA and conventional reflection-mode Au NP-LFIA, respectively. Thin plastic color light filters were integrated for filtering the excitation light source and, thereby, increasing the contrast of the emitted light for optimized visual detection. Integration of the OLED and color filters with the analytical membrane was achieved using adhesive techniques facilitated by the planar nature of the layers, which suggests possible large scale manufacturing using roll-to-roll processing. Gray scale analysis from digital images captured with a digital camera was used to quantify the visual sensitivity. The signal intensity, signal-to-noise ratio (SNR) and the limit of detection (LOD) of OLED integrated QD-LFIAs were compared to Au NP LFIAs. OLED QD-LFIA exhibited superior performance in all signal aspects: 7-8× higher signal intensity and SNR, and a 7× lower LOD of 3 nM (measured at S/N=3). These results demonstrate the potential of OLED-integrated in LFIA devices for obtaining sensitive, fast and low-cost POC diagnostics.

  3. A theoretical study of the isotope effects on the fluorescence excitation spectrum of 5-aminotropolone

    NASA Astrophysics Data System (ADS)

    Paz, Juan J.; Moreno, Miquel; Lluch, José M.

    1998-05-01

    A combined ab initio+nuclear dynamics study is performed to theoretically analyze the intramolecular H-atom transfer process in 5-aminotropolone in both the ground (S0) and first excited (S1) singlet electronic states. A complete active space self-consistent field (CASSCF) method is used to optimize the geometries. Energies are then corrected through the second order Møller-Plesset perturbation theory. These results are used to build up reduced bidimensional energy surfaces so that the nuclear wave functions for the nuclear motions in both electronic states are obtained. In particular we have analyzed the six isotopomers that result from deuteration of the amino and hydroxy groups of 5-aminotropolone. It is found that for symmetric structures (-OH/-NH2, -OH/-ND2, -OD/-NH2, and -OD/-ND2), the two lowest vibrational levels in both S0 and S1 appear as a quasidegenerated tunneling doublet. The tunneling splitting in S0 is much lower so that the doublet at the origin, seen in the fluorescence excitation spectra of 5-aminotropolone, can be entirely assigned to the S1 state. In agreement with the experimental findings, this splitting greatly diminishes when the transferring hydrogen is substituted by a deuterium, whereas deuteration of the amino group produces only a modest decrease of such a splitting. A quite different result is found for the nonsymmetric isotopically substituted structures (-OH/-NHD and -OD/-NHD), as the isotope induced asymmetry, combined with the high energy barrier in the S0 potential energy surface, leads to a complete localization of the two lowest vibrational wave functions in S0. On the other hand, for S1 the asymmetry and energy barriers are low enough so that an important degree of delocalization of the two lowest vibrational wave functions is found. These results are again in agreement with the presence of an isotope induced quartet in the fluorescence excitation spectra of these species.

  4. Integrated OLED as excitation light source in fluorescent lateral flow immunoassays.

    PubMed

    Venkatraman, Vishak; Steckl, Andrew J

    2015-12-15

    The integration of organic light emitting diodes (OLEDs) as excitation light sources for quantum dot-based fluorescent lateral flow immunoassay systems (LFIA) was investigated. This approach has the potential to deliver a sensitive visible detection scheme for low-cost, disposable lab-on-chip point-of-care (POC) diagnosis system. Thin film phosphorescent green OLEDs fabricated on plastic substrates were integrated on-chip to excite the test line of a quantum dot-based LFIA (QD-LFIA). OLEDs were fabricated by sequential deposition of organic thin films (total of ~100 nm) onto ITO-coated PET substrates. CdSe/ZnS QDs emitting at 655 nm and Au nanoparticles (NP - 10 nm size) conjugated antibodies were used for the fluorescence QD-LFIA and conventional reflection-mode Au NP-LFIA, respectively. Thin plastic color light filters were integrated for filtering the excitation light source and, thereby, increasing the contrast of the emitted light for optimized visual detection. Integration of the OLED and color filters with the analytical membrane was achieved using adhesive techniques facilitated by the planar nature of the layers, which suggests possible large scale manufacturing using roll-to-roll processing. Gray scale analysis from digital images captured with a digital camera was used to quantify the visual sensitivity. The signal intensity, signal-to-noise ratio (SNR) and the limit of detection (LOD) of OLED integrated QD-LFIAs were compared to Au NP LFIAs. OLED QD-LFIA exhibited superior performance in all signal aspects: 7-8× higher signal intensity and SNR, and a 7× lower LOD of 3 nM (measured at S/N=3). These results demonstrate the potential of OLED-integrated in LFIA devices for obtaining sensitive, fast and low-cost POC diagnostics. PMID:26134292

  5. Functional imaging of living Paramecium by means of confocal and two-photon excitation fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Diaspro, Alberto; Fronte, Paola; Raimondo, Marco; Fato, Marco; DeLeo, Gianluca; Beltrame, Francesco; Cannone, Fabio; Chirico, Giberto; Ramoino, Paola

    2002-05-01

    Confocal and Two-photon excitation laser scanning microscopy allow gathering three-dimensional and temporal information from biological systems exploiting fluorescence labeling and autofluorescence properties. In this work we study biological events linked to functionality in Paramecium primaurelia. The internalization of material in ciliated one-celled organisms (protozoa) occurs via different mechanisms, even if most of nutrients, particulate or not, is taken up by food vacuoles formed at the bottom of the oral cavity. The endocytosis of small-sized molecules occurs at the parasomal sacs, located next the ciliar basal bodies. Vital fluorescent dyes (BSA-FITC, WGA-FITC, dextran-Texas Red, cholesteryl-Bodipy) and autofluorescence were used to study formation, movement, and fusion of vesicles during endocytosis and phagocytosis of Paramecium primaurelia. By immobilizing living cells pulsed with food vacuole and endosome markers at successive times after chasing in unlabeled medium, the intracellular movement and fusion of food vacuoles and of endosomes were visualized. A temporal analysis of fluorescence images and the false-color technique were used. Starting from time series or 3D data sets composite images were generated by associating with each originally acquired image a different color corresponding to each sampling point in time and along the z-axis. Second Harmonic Generation Imaging attempts are also outlined.

  6. Intracavity laser excitation of NCO fluorescence in an atmospheric pressure flame

    NASA Astrophysics Data System (ADS)

    Anderson, William R.; Vanderhoff, John A.; Kotlar, Anthony J.; Dewilde, Mark A.; Beyer, Richard A.

    1982-08-01

    Laser excited fluorescence of the NCO radical has been obtained using discrete prism selected lines of an argon ion laser pump source. To our knowledge this is the first time NCO fluorescence has been obtained in a flame environment. NCO was formed in a slightly rich atmospheric pressure CH4/N2O flame. This flame was placed inside the extended cavity of the argon laser to take advantage of the much higher light intensity levels. All of the available laser lines pump vibrational hot bands of the NCO A 2Σ+ ← X 2Π system. The 4658 Å line appears to be the most useful for probing NCO densities. This line pumps in the A 2Σ+(0,00,0) ← X 2Π (1,01,0) vibrational band. NCO is pumped to N' = 31 by this line, probably via the Q231 transition although the R230 and P232 transitions could not be ruled out in the present analysis. The 4658 Å line was used to determine a relative NCO density profile through the reaction zone of a CH4/N2O flame. Profiles of C2, CN, and temperature were also obtained in this flame and are compared with the NCO profile. A lower limit of approximately 3×1014 cm-3 was placed on the peak NCO density in the flame. Attempts to find NCO or CN fluorescence in a CH4/air flame failed indicating probable differences in nitrogen chemistry for the two flames.

  7. Temperature and bath gas composition dependence of effective fluorescence lifetimes of toluene excited at 266 nm

    NASA Astrophysics Data System (ADS)

    Faust, S.; Dreier, T.; Schulz, C.

    2011-05-01

    Time-resolved fluorescence spectra of gas-phase toluene upon picosecond excitation at 266 nm were investigated as a function of temperature (296-1074 K) and bath gas composition (varying amounts of N 2, O 2, and CO 2) at 1 bar total pressure with a temporal resolution of 50 ps. In the investigated temperature range the effective fluorescence lifetime drops with increasing temperature from 46 ± 3 ns to 0.05 ± 0.01 ns in N 2 and CO 2. In the presence of O 2 at constant temperature the lifetimes also decrease significantly (e.g., from 46 ± 3 ns without O 2 to 0.63 ± 0.05 ns in air at room temperature), whereas lifetimes are independent on the CO 2 concentration. The implications of the results for the existing phenomenological model of predicting temporally integrated fluorescence intensities in toluene [W. Koban, J.D. Koch, R.K. Hanson, C. Schulz, Appl. Phys. B 80 (2005) 777] are discussed.

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

    NASA Astrophysics Data System (ADS)

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

    2014-02-01

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

  9. Imaging single fluorescent molecules at the interface of an optical fiber probe by evanescent wave excitation.

    PubMed

    Fang, X; Tan, W

    1999-08-01

    We have developed a new fluorescent method for single-molecule detection (SMD) and imaging using an optical fiber probe. The fluorophores were excited by the evanescent wave field produced on the core surface of the optical fiber. This was achieved by exposing a section of the core of the optical fiber probe to the fluorophore solution. Both cylindrical and square optical fiber probes were used for SMD. The fluorescent signals were detected by an intensified charge-coupled device. Single rhodamine 6G molecules have been detected. The number of rhodamine 6G molecules imaged by the optical fiber probe showed an excellent linear relationship with the concentrations of the fluorophores. The SMD scheme was also applied to the imaging of biomolecules, such as molecular beacon DNA molecules, labeled with tetramethylrhodamine. Our results have shown that using an optical fiber is an easy yet effective approach to SMD. It represents a simpler fluorescent method for the detection of single-molecules in solution and at an interface.

  10. Production and excitation-emission fluorescence properties of colored dissolved organic matter from marine tropical species

    NASA Astrophysics Data System (ADS)

    Mendoza, W. G.; Zika, R. G.

    2009-12-01

    Colored dissolved organic matter (CDOM) plays an important key role in the photochemistry and biogeochemical cycling of carbon in the coastal region. Their distribution can vary in space and time due to supply of CDOM from different sources. To determine properties of fluorescence-CDOM produced by various marine tropical species, two species from each of the different marine communities were examined after incubation in the dark for forty-nine (49) days: seagrasses-Enhalus acoroides (EA), Thalassia testudinium (TT); corals-Pocillopora cylindrical (PC), Seriatopora hystrix (SH) ; mangroves- Avicennia marina (AM), Sonneratia alba (SA); brown algae-Hormophysa cuneiformis (HC), Sargassum sp.(SS). Average CDOM production is highest from mangrove species (218 QSU/g-sample/day), followed by seagrass (42 QSU/g-sample/day), brown alga (26 QSU/g-sample/day) then corals (19 QSU/g-sample/day).The fluorescence maximum at 312; 380-420 nm emission-excitation pair appears to be present in all species that is an identified humic-like signature. These results suggest that the production of the fluorescent CDOM fraction is a common phenomenon of tropical marine species and as such constitutes a major part of the marine CDOM pool in coastal regions.

  11. Fluorescence and UV/VIS absorption spectroscopy studies on polymer blend films for photovoltaics

    NASA Astrophysics Data System (ADS)

    van Stam, Jan; Lindqvist, Camilla; Hansson, Rickard; Ericsson, Leif; Moons, Ellen

    2015-08-01

    The quinoxaline-based polymer TQ1 (poly[2,3-bis-(3-octyloxyphenyl)quinoxaline-5,8-diyl-alt-thiophene-2,5- diyl]) is a promising candidate as electron donor in organic solar cells. In combination with the electron acceptor [6,6]- phenyl-C71- butyric acid methyl ester (PC70BM), TQ1 has resulted in solar cells with power conversion efficiencies of 7 %. We have studied TQ1 films, with and without PC70BM, spin-casted from different solvents, by fluorescence spectroscopy and UV/VIS absorption spectroscopy. We used chloroform (CF), chlorobenzene (CB), and odichlorobenzene (o-DCB) as solvents for the coating solutions and 1-chloronaphthalene (CN) as solvent additive. CN addition has been shown to enhance photo-conversion efficiency of these solar cells. Phase-separation causes lateral domain formation in the films and the domain size depends on the solvent . These morphological differences coincide with changes in the spectroscopic patterns of the films. From a spectroscopic point of view, TQ1 acts as fluorescent probe and PC70BM as quencher. The degree of fluorescence quenching is coupled to the morphology through the distance between TQ1 and PC70BM. Furthermore, if using a bad solvent for PC70BM, morphological regions rich in the fullerene yield emission characteristic for aggregated PC70BM. Clear differences were found, comparing the TQ1:PC70BM blend films casted from different solvents and at different ratios between the donor and acceptor. The morphology also influences the UV/VIS absorption spectra, yielding further information on the composition. The results show that fluorescence and UV/VIS absorption spectroscopy can be used to detect aggregation in blended films and that these methods extend the morphological information beyond the scale accessible with microscopy.

  12. Intermediate Coupling For Core-Level Excited States: Consequences For X-Ray Absorption Spectroscopy

    SciTech Connect

    Bagus, Paul S.; Sassi, Michel JPC; Rosso, Kevin M.

    2015-04-15

    The origin of the complex NEXAFS features of X-Ray Absorption, XAS, spectra in transition metal complexes is analyzed and interpreted in terms of the angular momentum coupling of the open shell electrons. Especially for excited configurations where a core-electron is promoted to an open valence shell, the angular momentum coupling is intermediate between the two limits of Russell- Saunders, RS, coupling where spin-orbit splitting of the electron shells is neglected and j-j coupling where this splitting is taken as dominant. The XAS intensities can be understood in terms of two factors: (1) The dipole selection rules that give the allowed excited RS multiplets and (2) The contributions of these allowed multiplets to the wavefunctions of the intermediate coupled levels. It is shown that the origin of the complex XAS spectra is due to the distribution of the RS allowed multiplets over several different intermediate coupled excited levels. The specific case that is analyzed is the L2,3 edge XAS of an Fe3+ cation, because this cation allows a focus on the angular momentum coupling to the exclusion of other effects; e.g., chemical bonding. Arguments are made that the properties identified for this atomic case are relevant for more complex materials. The analysis is based on the properties of fully relativistic, ab initio, many-body wavefunctions for the initial and final states of the XAS process. The wavefunction properties considered include the composition of the wavefunctions in terms of RS multiplets and the occupations of the spin-orbit split open shells; the latter vividly show whether the coupling is j-j or not.

  13. Absorption spectra of e-beam-excited Ne, Ar, and Kr, pure and in binary mixtures.

    PubMed

    Levchenko, A O; Ustinovskii, N N; Zvorykin, V D

    2010-10-21

    A technique using the broadband emission of a laser plume as probe radiation is applied to record UV-visible (190-510 nm) absorption spectra of Ne, Ar, and Kr, pure and in binary mixtures under moderate e-beam excitation up to 1 MW/cm(3). In all the rare gases and mixtures, the absorption spectra show continuum related to Rg(2) (+) homonuclear ions [peaking at λ∼285, 295, and 320 nm in Ne, Ar, and Kr(Ar/Kr), respectively] and a number of atomic lines related mainly to Rg(∗)(ms) levels, where m is the lowest principal quantum number of the valence electron. In argon, a continuum related to Ar(2) (∗) (λ∼325 nm) is also recorded. There are also trains of narrow bands corresponding to Rg(2) (∗)(npπ (3)Π(g))←Rg(2) (∗)(msσ (3)Σ(u) (+)) transitions. All the spectral features mentioned above were reported in literature but have never been observed simultaneously. Although charge transfer to a homonuclear ion of the heavier additive is commonly believed to dominate in binary rare-gas mixtures, it is found in this study that in Ne/Kr mixture, the charge is finally transferred from the buffer gas Ne(2) (+) ion not to Kr(2) (+) but to heteronuclear NeKr(+) ion.

  14. Two-photon excitation microscopy using the second singlet state of fluorescent agents within the ``tissue optical window''

    NASA Astrophysics Data System (ADS)

    Pu, Yang; Shi, Lingyan; Pratavieira, Sebastião; Alfano, R. R.

    2013-10-01

    Two-photon (2P) excitation of the second singlet (S2) state of a group of fluorescent agents with near infrared emission was used to extend the optical excitation and imaging regime of 2P microscope into "tissue optical window" (650-1100 nm). As the first step to achieve deeper optical imaging, Chlorophyll a and Indocyanine green are investigated and demonstrated as imaging agents using 2P S2 excitation at 800 nm for microscope imaging at 685 nm. The salient feature is to drive both the 2P excitation and emission wavelengths of the imaging agents to fall into the "tissue optical window."

  15. Enhancement of light absorption in polyazomethines due to plasmon excitation on randomly distributed metal nanoparticles

    NASA Astrophysics Data System (ADS)

    Wróbel, P.; Antosiewicz, T. J.; Stefaniuk, T.; Ciesielski, A.; Iwan, A.; Wronkowska, A. A.; Wronkowski, A.; Szoplik, T.

    2015-05-01

    In photovoltaic devices, metal nanoparticles embedded in a semiconductor layer allow the enhancement of solar-toelectric energy conversion efficiency due to enhanced light absorption via a prolonged optical path, enhanced electric fields near the metallic inclusions, direct injection of hot electrons, or local heating. Here we pursue the first two avenues. In the first, light scattered at an angle beyond the critical angle for reflection is coupled into the semiconductor layer and confined within such planar waveguide up to possible exciton generation. In the second, light is trapped by the excitation of localized surface plasmons on metal nanoparticles leading to enhanced near-field plasmon-exciton coupling at the peak of the plasmon resonance. We report on results of a numerical experiment on light absorption in polymer- (fullerene derivative) blends, using the 3D FDTD method, where exact optical parameters of the materials involved are taken from our recent measurements. In simulations we investigate light absorption in randomly distributed metal nanoparticles dispersed in polyazomethine-(fullerene derivative) blends, which serve as active layers in bulkheterojunction polymer solar cells. In the study Ag and Al nanoparticles of different diameters and fill factors are diffused in two air-stable aromatic polyazomethines with different chemical structures (abbreviated S9POF and S15POF) mixed with phenyl-C61-butyric acid methyl ester (PCBM) or [6,6]-phenyl-C71-butyric acid methyl ester (PC71BM). The mixtures are spin coated on a 100 nm thick Al layer deposited on a fused silica substrate. Optical constants of the active layers are taken from spectroscopic ellipsometry and reflectance measurements using a rotating analyzer type ellipsometer with auto-retarder performed in the wavelength range from 225 nm to 2200 nm. The permittivities of Ag and Al particles of diameters from 20 to 60 nm are assumed to be equal to those measured on 100 to 200 nm thick metal films.

  16. Excitations, optical absorption spectra, and optical excitonic gaps of heterofullerenes. I. C60, C59N+, and C48N12: theory and experiment.

    PubMed

    Xie, Rui-Hua; Bryant, Garnett W; Sun, Guangyu; Nicklaus, Marc C; Heringer, David; Frauenheim, Th; Manaa, M Riad; Smith, Vedene H; Araki, Yasuyuki; Ito, Osamu

    2004-03-15

    Low-energy excitations and optical absorption spectrum of C(60) are computed by using time-dependent (TD) Hartree-Fock, TD-density functional theory (TD-DFT), TD DFT-based tight-binding (TD-DFT-TB), and a semiempirical Zerner intermediate neglect of diatomic differential overlap method. A detailed comparison of experiment and theory for the excitation energies, optical gap, and absorption spectrum of C(60) is presented. It is found that electron correlations and correlation of excitations play important roles in accurately assigning the spectral features of C(60), and that the TD-DFT method with nonhybrid functionals or a local spin density approximation leads to more accurate excitation energies than with hybrid functionals. The level of agreement between theory and experiment for C(60) justifies similar calculations of the excitations and optical absorption spectrum of a monomeric azafullerene cation C(59)N(+), to serve as a spectroscopy reference for the characterization of carborane anion salts. Although it is an isoelectronic analogue to C(60), C(59)N(+) exhibits distinguishing spectral features different from C(60): (1) the first singlet is dipole-allowed and the optical gap is redshifted by 1.44 eV; (2) several weaker absorption maxima occur in the visible region; (3) the transient triplet-triplet absorption at 1.60 eV (775 nm) is much broader and the decay of the triplet state is much faster. The calculated spectra of C(59)N(+) characterize and explain well the measured ultraviolet-visible (UV-vis) and transient absorption spectra of the carborane anion salt [C(59)N][Ag(CB(11)H(6)Cl(6))(2)] [Kim et al., J. Am. Chem. Soc. 125, 4024 (2003)]. For the most stable isomer of C(48)N(12), we predict that the first singlet is dipole-allowed, the optical gap is redshifted by 1.22 eV relative to that of C(60), and optical absorption maxima occur at 585, 528, 443, 363, 340, 314, and 303 nm. We point out that the characterization of the UV-vis and transient absorption

  17. Excitations, optical absorption spectra, and optical excitonic gaps of heterofullerenes. I. C60, C59N+, and C48N12: Theory and experiment

    NASA Astrophysics Data System (ADS)

    Xie, Rui-Hua; Bryant, Garnett W.; Sun, Guangyu; Nicklaus, Marc C.; Heringer, David; Frauenheim, Th.; Manaa, M. Riad; Smith, Vedene H.; Araki, Yasuyuki; Ito, Osamu

    2004-03-01

    Low-energy excitations and optical absorption spectrum of C60 are computed by using time-dependent (TD) Hartree-Fock, TD-density functional theory (TD-DFT), TD DFT-based tight-binding (TD-DFT-TB), and a semiempirical Zerner intermediate neglect of diatomic differential overlap method. A detailed comparison of experiment and theory for the excitation energies, optical gap, and absorption spectrum of C60 is presented. It is found that electron correlations and correlation of excitations play important roles in accurately assigning the spectral features of C60, and that the TD-DFT method with nonhybrid functionals or a local spin density approximation leads to more accurate excitation energies than with hybrid functionals. The level of agreement between theory and experiment for C60 justifies similar calculations of the excitations and optical absorption spectrum of a monomeric azafullerene cation C59N+, to serve as a spectroscopy reference for the characterization of carborane anion salts. Although it is an isoelectronic analogue to C60, C59N+ exhibits distinguishing spectral features different from C60: (1) the first singlet is dipole-allowed and the optical gap is redshifted by 1.44 eV; (2) several weaker absorption maxima occur in the visible region; (3) the transient triplet-triplet absorption at 1.60 eV (775 nm) is much broader and the decay of the triplet state is much faster. The calculated spectra of C59N+ characterize and explain well the measured ultraviolet-visible (UV-vis) and transient absorption spectra of the carborane anion salt [C59N][Ag(CB11H6Cl6)2] [Kim et al., J. Am. Chem. Soc. 125, 4024 (2003)]. For the most stable isomer of C48N12, we predict that the first singlet is dipole-allowed, the optical gap is redshifted by 1.22 eV relative to that of C60, and optical absorption maxima occur at 585, 528, 443, 363, 340, 314, and 303 nm. We point out that the characterization of the UV-vis and transient absorption spectra of C48N12 isomers is helpful in

  18. Absorption and fluorescence of alexandrite and of titanium in sapphire and glass

    NASA Technical Reports Server (NTRS)

    Byvik, C. E.; Hess, R. V.; Buoncristiani, A. M.

    1985-01-01

    The fluorescence and absorption data for titanium in crystalline sapphire and titanium doped into two silicate and one phosphate glass structures are analyzed. It is observed that the Ti-doped silicate glass sample exhibits no absorption related to the Ti(III) ion, the Ti-doped phosphate glass is deep blue, the absorption line width of the glass samples are a factor of two larger than that of sapphire, and the absorption peak for the Ti in the glass shifted about 100 nm to the red from the Ti:sapphire absorption peak. This shift reveals that the Ti(III) ion is sensitive to the crystalline environment and not to the glass environment. The photoluminescence spectra for Ti-doped sapphire and alexandrite are compared. It is detected that the Ti:sapphire exhibits a broader spectrum than that for alexandrite with a peak at 750 nm. The three zero phonon transitions of Ti:Al2O3 at liquid nitrogen temperatures are studied.

  19. Fluorescence excitation involving multiple electron transition states of N{sub 2} and CO{sub 2}

    SciTech Connect

    Wu, C.Y.R.; Chen, F.Z.; Hung, T.; Judge, D.L.

    1997-04-01

    The electronic states and electronic structures of N{sub 2} and CO{sub 2} in the 8-50 eV energy region have been studied extensively both experimentally and theoretically. In the energy region higher than 25 eV there exists many electronic states including multiple electron transition (MET) states which are responsible for producing most of the dissociative photoionization products. The electronic states at energies higher than 50 eV have been mainly determined by Auger spectroscopy, double charge transfer, photofragment spectroscopy and ion-ion coincidence spectroscopy. The absorption and ionization spectra of these molecules at energies higher than 50 eV mainly show a monotonic decrease in cross section values and exhibit structureless features. The decay channels of MET and Rydberg (or superexcited) states include autoionization, ionization, dissociative ionization, predissociation, and dissociation while those of single ion and multiple ion states may involve predissociation. and dissociation processes. The study of fluorescence specifically probes electronically excited species resulting from the above-mentioned decay channels and provides information for understanding the competition among these channels.

  20. Modeling of fluorescence line-narrowed spectra in weakly coupled dimers in the presence of excitation energy transfer

    SciTech Connect

    Lin, Chen; Reppert, Mike; Feng, Ximao; Jankowiak, Ryszard

    2014-07-21

    This work describes simple analytical formulas to describe the fluorescence line-narrowed (FLN) spectra of weakly coupled chromophores in the presence of excitation energy transfer (EET). Modeling studies for dimer systems (assuming low fluence and weak coupling) show that the FLN spectra (including absorption and emission spectra) calculated for various dimers using our model are in good agreement with spectra calculated by: (i) the simple convolution method and (ii) the more rigorous treatment using the Redfield approach [T. Renger and R. A. Marcus, J. Chem. Phys. 116, 9997 (2002)]. The calculated FLN spectra in the presence of EET of all three approaches are very similar. We argue that our approach provides a simplified and computationally more efficient description of FLN spectra in the presence of EET. This method also has been applied to FLN spectra obtained for the CP47 antenna complex of Photosystem II reported by Neupane et al. [J. Am. Chem. Soc. 132, 4214 (2010)], which indicated the presence of uncorrelated EET between pigments contributing to the two lowest energy (overlapping) exciton states, each mostly localized on a single chromophore. Calculated and experimental FLN spectra for CP47 complex show very good qualitative agreement.

  1. Excited state electron distribution and role of the terminal amine in acidic and basic tryptophan dipeptide fluorescence

    NASA Astrophysics Data System (ADS)

    Eisenberg, Azaria S.; Nathan, Moshe; Juszczak, Laura J.

    2016-08-01

    The results of quantum yield (QY) study of tryptophanyl glutamate (Trp-Glu), tryptophanyl lysine (Trp-Lys) and lysinyl tryptophan (Lys-Trp) dipeptides over the pH range, 1.5-13, show that the charge state of the N-terminal amine, and not the nominal molecular charge determines the QY. When the terminal amine is protonated, QY is low (10-2) for all three dipeptides. As the terminal amine cation is found proximal to the indole ring in Trp-Glu and Trp-Lys conformers but not in those for Lys-Trp, its effect may lie only in the partitioning of energy between nonradiative processes, not on QY reduction. QY is also low when both the N-terminal amine and indole amine are deprotonated. These two low QY states can be distinguished by fluorescence lifetime measurement. Molecular dynamics simulation shows that the Chi 1 conformers persist for tens of nanoseconds such that 100-101 ns lifetimes may be associated with individual Chi 1 conformers. The ground state electron density or isosurface of high QY (0.30) 3-methyindole has a uniform electron density over the indole ring as do the higher QY Trp dipeptide conformers. This validates the association of ground state isosurfaces with QY. Excited state orbitals from calculated high intensity, low energy absorption transitions are typically centered over the indole ring for higher QY dipeptide species and off the ring in lower QY species. Thus excited state orbitals substantiate the earlier finding that the ground state isosurface charge density pattern on the indole ring can be predictive of QY.

  2. Simulating One-Photon Absorption and Resonance Raman Scattering Spectra Using Analytical Excited State Energy Gradients within Time-Dependent Density Functional Theory

    SciTech Connect

    Silverstein, Daniel W.; Govind, Niranjan; van Dam, Hubertus J. J.; Jensen, Lasse

    2013-12-10

    A parallel implementation of analytical time-dependent density functional theory gradients is presented for the quantum chemistry program NWChem. The implementation is based on the Lagrangian approach developed by Furche and Ahlrichs. To validate our implementation, we first calculate the Stokes shifts for a range of organic dye molecules using a diverse set of exchange-correlation functionals (traditional density functionals, global hybrids, and range-separated hybrids) followed by simulations of the one-photon absorption and resonance Raman scattering spectrum of the phenoxyl radical, the well-studied dye molecule rhodamine 6G, and a molecular host–guest complex (TTFcCBPQT4+). The study of organic dye molecules illustrates that B3LYP and CAM-B3LYP generally give the best agreement with experimentally determined Stokes shifts unless the excited state is a charge transfer state. Absorption, resonance Raman, and fluorescence simulations for the phenoxyl radical indicate that explicit solvation may be required for accurate characterization. For the host–guest complex and rhodamine 6G, it is demonstrated that absorption spectra can be simulated in good agreement with experimental data for most exchange-correlation functionals. Finally, however, because one-photon absorption spectra generally lack well-resolved vibrational features, resonance Raman simulations are necessary to evaluate the accuracy of the exchange-correlation functional for describing a potential energy surface.

  3. A method for tuning the excitation wavelength of an LED light source during fluorescence-based cystoscopy (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Lindvold, Lars R.; Hermannn, Gregers G.

    2016-02-01

    In clinical applications of fluorescence-guided endoscopy of the bladder (cystoscopy) it can be observed that the contrast in light from autofluorescence and from photodynamic diagnosis (PDD) varies from patient to patient. To compensate for this effect, a new method is presented for tuning the wavelength of a LED-based light source during fluorescence guided endoscopy of the bladder i.e. photodynamic diagnosis of bladder tumours. In the present embodiment, the wavelength of the LED source, developed in our laboratory, can be tuned to vary the excitation wavelength of both the sensitised fluorescence in the tumours (PDD) as well as the native fluorescence of the bladder mucosa and blood vessels. The contrast of the image observed through the CCD-camera attached to the cystoscope is thereby increased. In this way, patient to patient variations in autofluorescence and in sensitised fluorescence of tumours can be compensated for during fluorescence-guided cystoscopy in the clinic.

  4. Standing-wave-excited multiplanar fluorescence in a laser scanning microscope reveals 3D information on red blood cells

    NASA Astrophysics Data System (ADS)

    Amor, Rumelo; Mahajan, Sumeet; Amos, William Bradshaw; McConnell, Gail

    2014-12-01

    Standing-wave excitation of fluorescence is highly desirable in optical microscopy because it improves the axial resolution. We demonstrate here that multiplanar excitation of fluorescence by a standing wave can be produced in a single-spot laser scanning microscope by placing a plane reflector close to the specimen. We report here a variation in the intensity of fluorescence of successive planes related to the Stokes shift of the dye. We show by the use of dyes specific for the cell membrane how standing-wave excitation can be exploited to generate precise contour maps of the surface membrane of red blood cells, with an axial resolution of ~90 nm. The method, which requires only the addition of a plane mirror to an existing confocal laser scanning microscope, may well prove useful in studying diseases which involve the red cell membrane, such as malaria.

  5. Standing-wave-excited multiplanar fluorescence in a laser scanning microscope reveals 3D information on red blood cells.

    PubMed

    Amor, Rumelo; Mahajan, Sumeet; Amos, William Bradshaw; McConnell, Gail

    2014-12-08

    Standing-wave excitation of fluorescence is highly desirable in optical microscopy because it improves the axial resolution. We demonstrate here that multiplanar excitation of fluorescence by a standing wave can be produced in a single-spot laser scanning microscope by placing a plane reflector close to the specimen. We report here a variation in the intensity of fluorescence of successive planes related to the Stokes shift of the dye. We show by the use of dyes specific for the cell membrane how standing-wave excitation can be exploited to generate precise contour maps of the surface membrane of red blood cells, with an axial resolution of ≈90 nm. The method, which requires only the addition of a plane mirror to an existing confocal laser scanning microscope, may well prove useful in studying diseases which involve the red cell membrane, such as malaria.

  6. Study on discrimination of oral cancer from normal using blood plasma based on fluorescence steady and excited state at excitation wavelength 280 nm

    NASA Astrophysics Data System (ADS)

    Rekha, Pachaiappan; Aruna, Prakasa Rao; Ganesan, Singaravelu

    2016-03-01

    Many research works based on fluorescence spectroscopy have proven its potential in the diagnosis of various diseases using the spectral signatures of the native key fluorophores such as tryptophan, tyrosine, collagen, NADH, FAD and porphyrin. These fluorophores distribution, concentration and their conformation may be changed depending upon the pathological and metabolic conditions of cells and tissues. In this study, we have made an attempt to characterize the blood plasma of normal subject and oral cancer patients by native fluorescence spectroscopy at 280 nm excitation. Further, the fluorescence data were analyzed by employing the multivariate statistical method - linear discriminant analyses (LDA) using leaves one out cross validation method. The results illustrate the potential of fluorescence spectroscopy technique in the diagnosis of oral cancer using blood plasma.

  7. Excited-State Proton-Transfer-Induced Trapping Enhances the Fluorescence Emission of a Locked GFP Chromophore

    PubMed Central

    2016-01-01

    The chemical locking of the central single bond in core chromophores of green fluorescent proteins (GFPs) influences their excited-state behavior in a distinct manner. Experimentally, it significantly enhances the fluorescence quantum yield of GFP chromophores with an ortho-hydroxyl group, while it has almost no effect on the photophysics of GFP chromophores with a para-hydroxyl group. To unravel the underlying physical reasons for this different behavior, we report static electronic structure calculations and nonadiabatic dynamics simulations on excited-state intramolecular proton transfer, cis–trans isomerization, and excited-state deactivation in a locked ortho-substituted GFP model chromophore (o-LHBI). On the basis of our previous and present results, we find that the S1 keto species is responsible for the fluorescence emission of the unlocked o-HBI and the locked o-LHBI species. Chemical locking does not change the parts of the S1 and S0 potential energy surfaces relevant to enol–keto tautomerization; hence, in both chromophores, there is an ultrafast excited-state intramolecular proton transfer that takes only 35 fs on average. However, the locking effectively hinders the S1 keto species from approaching the keto S1/S0 conical intersections so that most of trajectories are trapped in the S1 keto region for the entire 2 ps simulation time. Therefore, the fluorescence quantum yield of o-LHBI is enhanced compared with that of unlocked o-HBI, in which the S1 excited-state decay is efficient and ultrafast. In the case of the para-substituted GFP model chromophores p-HBI and p-LHBI, chemical locking hardly affects their efficient excited-state deactivation via cis–trans isomerization; thus, the fluorescence quantum yields in these chromophores remain very low. The insights gained from the present work may help to guide the design of new GFP chromophores with improved fluorescence emission and brightness. PMID:26744782

  8. Excited-State Proton-Transfer-Induced Trapping Enhances the Fluorescence Emission of a Locked GFP Chromophore.

    PubMed

    Liu, Xiang-Yang; Chang, Xue-Ping; Xia, Shu-Hua; Cui, Ganglong; Thiel, Walter

    2016-02-01

    The chemical locking of the central single bond in core chromophores of green fluorescent proteins (GFPs) influences their excited-state behavior in a distinct manner. Experimentally, it significantly enhances the fluorescence quantum yield of GFP chromophores with an ortho-hydroxyl group, while it has almost no effect on the photophysics of GFP chromophores with a para-hydroxyl group. To unravel the underlying physical reasons for this different behavior, we report static electronic structure calculations and nonadiabatic dynamics simulations on excited-state intramolecular proton transfer, cis-trans isomerization, and excited-state deactivation in a locked ortho-substituted GFP model chromophore (o-LHBI). On the basis of our previous and present results, we find that the S1 keto species is responsible for the fluorescence emission of the unlocked o-HBI and the locked o-LHBI species. Chemical locking does not change the parts of the S1 and S0 potential energy surfaces relevant to enol-keto tautomerization; hence, in both chromophores, there is an ultrafast excited-state intramolecular proton transfer that takes only 35 fs on average. However, the locking effectively hinders the S1 keto species from approaching the keto S1/S0 conical intersections so that most of trajectories are trapped in the S1 keto region for the entire 2 ps simulation time. Therefore, the fluorescence quantum yield of o-LHBI is enhanced compared with that of unlocked o-HBI, in which the S1 excited-state decay is efficient and ultrafast. In the case of the para-substituted GFP model chromophores p-HBI and p-LHBI, chemical locking hardly affects their efficient excited-state deactivation via cis-trans isomerization; thus, the fluorescence quantum yields in these chromophores remain very low. The insights gained from the present work may help to guide the design of new GFP chromophores with improved fluorescence emission and brightness. PMID:26744782

  9. Excited-State Proton-Transfer-Induced Trapping Enhances the Fluorescence Emission of a Locked GFP Chromophore.

    PubMed

    Liu, Xiang-Yang; Chang, Xue-Ping; Xia, Shu-Hua; Cui, Ganglong; Thiel, Walter

    2016-02-01

    The chemical locking of the central single bond in core chromophores of green fluorescent proteins (GFPs) influences their excited-state behavior in a distinct manner. Experimentally, it significantly enhances the fluorescence quantum yield of GFP chromophores with an ortho-hydroxyl group, while it has almost no effect on the photophysics of GFP chromophores with a para-hydroxyl group. To unravel the underlying physical reasons for this different behavior, we report static electronic structure calculations and nonadiabatic dynamics simulations on excited-state intramolecular proton transfer, cis-trans isomerization, and excited-state deactivation in a locked ortho-substituted GFP model chromophore (o-LHBI). On the basis of our previous and present results, we find that the S1 keto species is responsible for the fluorescence emission of the unlocked o-HBI and the locked o-LHBI species. Chemical locking does not change the parts of the S1 and S0 potential energy surfaces relevant to enol-keto tautomerization; hence, in both chromophores, there is an ultrafast excited-state intramolecular proton transfer that takes only 35 fs on average. However, the locking effectively hinders the S1 keto species from approaching the keto S1/S0 conical intersections so that most of trajectories are trapped in the S1 keto region for the entire 2 ps simulation time. Therefore, the fluorescence quantum yield of o-LHBI is enhanced compared with that of unlocked o-HBI, in which the S1 excited-state decay is efficient and ultrafast. In the case of the para-substituted GFP model chromophores p-HBI and p-LHBI, chemical locking hardly affects their efficient excited-state deactivation via cis-trans isomerization; thus, the fluorescence quantum yields in these chromophores remain very low. The insights gained from the present work may help to guide the design of new GFP chromophores with improved fluorescence emission and brightness.

  10. Linkage of oxygen deficiency defects and rare earth concentrations in silica glass optical fiber probed by ultraviolet absorption and laser excitation spectroscopy.

    PubMed

    Liu, Y-S; Galvin, T C; Hawkins, T; Ballato, J; Dong, L; Foy, P R; Dragic, P D; Eden, J G

    2012-06-18

    Ultraviolet absorption measurements and laser excitation spectroscopy in the vicinity of 248 nm provide compelling evidence for linkages between the oxygen deficiency center (ODC) and rare earth concentrations in Yb and Er-doped glass optical fibers. Investigations of YAG-derived and solution-doped glass fibers are described. For both Yb and Er-doped fibers, the dependence of Type II ODC absorption on the rare earth number density is approximately linear, but the magnitude of the effect is greater for Yb-doped fibers. Furthermore, laser excitation spectra demonstrate unambiguously the existence of an energy transfer mechanism coupling an ODC with Yb(3+). Photopumping glass fibers with a Ti:sapphire laser/optical parametric amplifier system, tunable over the 225-265 nm region, or with a KrF laser at 248.4 nm show: 1) emission features in the 200-1100 nm interval attributable only to the ODC (Type II) defect or Yb(3+), and 2) the excitation spectra for ODC (II) emission at ~280 nm and Yb(3+) fluorescence (λ ~1.03 μm) to be, within experimental uncertainty, identical. The latter demonstrates that, when irradiating Yb-doped silica fibers between ~240 and 255 nm, the ODC (II) defect is at least the primary precursor to Yb(3+) emission. Consistent with previous reports in the literature, the data show the ODC (II) absorption spectrum to have a peak wavelength and breadth of ~246 nm and ~19 nm (FWHM). Experiments also reveal that, in the absence of Yb, incorporating either Al(2)O(3) or Y(2)O(3) into glass fibers has a negligible impact on the ODC concentration. Not only do the data reported here demonstrate the relationship between the ODC (II) number density and the Yb doping concentration, but they also suggest that the appearance of ODC defects in the fiber is associated with the introduction of Yb and the process by which the fiber is formed.

  11. Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation

    NASA Astrophysics Data System (ADS)

    Zipfel, Warren R.; Williams, Rebecca M.; Christie, Richard; Nikitin, Alexander Yu; Hyman, Bradley T.; Webb, Watt W.

    2003-06-01

    Multicolor nonlinear microscopy of living tissue using two- and three-photon-excited intrinsic fluorescence combined with second harmonic generation by supermolecular structures produces images with the resolution and detail of standard histology without the use of exogenous stains. Imaging of intrinsic indicators within tissue, such as nicotinamide adenine dinucleotide, retinol, indoleamines, and collagen provides crucial information for physiology and pathology. The efficient application of multiphoton microscopy to intrinsic imaging requires knowledge of the nonlinear optical properties of specific cell and tissue components. Here we compile and demonstrate applications involving a range of intrinsic molecules and molecular assemblies that enable direct visualization of tissue morphology, cell metabolism, and disease states such as Alzheimer's disease and cancer.

  12. Coherence gated wavefront sensorless adaptive optics for two photon excited fluorescence retinal imaging (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Jian, Yifan; Cua, Michelle; Bonora, Stefano; Pugh, Edward N.; Zawadzki, Robert J.; Sarunic, Marinko V.

    2016-03-01

    We present a novel system for adaptive optics two photon imaging. We utilize the bandwidth of the femtosecond excitation beam to perform coherence gated imaging (OCT) of the sample. The location of the focus is directly observable in the cross sectional OCT images, and adjusted to the desired depth plane. Next, using real time volumetric OCT, we perform Wavefront Sensorless Adaptive Optics (WSAO) aberration correction using a multi-element adaptive lens capable of correcting up to 4th order Zernike polynomials. The aberration correction is performed based on an image quality metric, for example intensity. The optimization time is limited only by the OCT acquisition rate, and takes ~30s. Following aberration correction, two photon fluorescence images are acquired, and compared to results without adaptive optics correction. This technique is promising for multiphoton imaging in multi-layered, scattering samples such as eye and brain, in which traditional wavefront sensing and guide-star sensorless adaptive optics approaches may not be suitable.

  13. Raman Spectroscopy of Lithium Hydride Corrosion: Selection of an Appropriate Excitation Wavelength to Minimize Fluorescence

    SciTech Connect

    Stowe, A. C.; Smyrl, N. R.

    2011-05-26

    The recent interest in a hydrogen-based fuel economy has renewed research into metal hydride chemistry. Many of these compounds react readily with water to release hydrogen gas and form a caustic. Diffuse Reflectance Infrared Fourier Transform Spectroscopy (DRIFT) has been used to study the hydrolysis reaction. The LiOH stretch appears at 3670 cm{sup -1}. Raman spectroscopy is a complementary technique that employs monochromatic excitation (laser) allowing access to the low energy region of the vibrational spectrum (<600 cm{sup -1}). Weak scattering and fluorescence typically prevent Raman from being used for many compounds. The role of Li{sub 2}O in the moisture reaction has not been fully studied for LiH. Li{sub 2}O can be observed by Raman while being hidden in the Infrared spectrum.

  14. Technical Note: Dissolved organic matter fluorescence - a finite mixture approach to deconvolve excitation-emission matrices

    NASA Astrophysics Data System (ADS)

    Butturini, A.; Ejarque, E.

    2013-09-01

    The analysis of the shape of excitation-emission matrices (EEMs) is a relevant tool for exploring the origin, transport and fate of dissolved organic matter (DOM) in aquatic ecosystems. Within this context, the decomposition of EEMs is acquiring a notable relevance. A simple mathematical algorithm that automatically deconvolves individual EEMs is described, creating new possibilities for the comparison of DOM fluorescence properties and EEMs that are very different from each other. A mixture model approach is adopted to decompose complex surfaces into sub-peaks. The laplacian operator and the Nelder-Mead optimisation algorithm are implemented to individuate and automatically locate potential peaks in the EEM landscape. The EEMs of a simple artificial mixture of fluorophores and DOM samples collected in a Mediterranean river are used to describe the model application and to illustrate a strategy that optimises the search for the optimal output.

  15. Technical Note: Dissolved organic matter fluorescence - a finite mixture approach to deconvolve excitation-emission matrices

    NASA Astrophysics Data System (ADS)

    Butturini, A.; Ejarque, E.

    2013-03-01

    The analysis of the shape of excitation-emission matrices (EEMs) is a relevant tool for exploring the origin, transport and fate of dissolved organic matter (DOM) in aquatic ecosystems. Within this context, the decomposition of EEMs is acquiring a notable relevance. A simple mathematical algorithm that automatically deconvolves single EEM is described, creating new possibilities for the comparison of DOM fluorescence properties and EEMs that are very different from each other. A mixture model approach is adopted to decompose complex surfaces into sub-peaks. The laplacian operator and the Nelder-Mead optimization algorithm are implemented to individuate and automatically locate potential peaks in the EEM landscape. A small heterogeneous data set of 21 EEMs from a human-impacted Mediterranean river is used to describe the model application and to illustrate a strategy that optimises the search for the optimal output.

  16. Imaging of surgical margin in pancreatic metastasis using two-photon excited fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Chen, Jing; Hong, Zhipeng; Chen, Hong; Chen, Youting; Xu, Yahao; Zhu, Xiaoqin; Zhuo, Shuangmu; Shi, Zheng; Chen, Jianxin

    2014-09-01

    Two-photon excited fluorescence (TPEF) microscopy, has become a powerful tool for imaging unstained tissue samples at subcellular level in biomedical research. The purpose of this study was to determine whether TPEF imaging of histological sections without H-E staining can be used to identify the boundary between normal pancreas and pancreatic metastasis from renal cell carcinoma (RCC). The typical features such as the significant increase of cancerous nests, the absence of pancreatic ductal, the appearance of cancer cells were observed to present the boundary between normal pancreas and pancreatic metastasis from RCC. These results correlated well with the corresponding histological outcomes. With the advent of clinically miniaturized TPEF microscopy and integrative endoscopy, TPEF microscopy has the potential application on surgical location of pancreatic metastasis from RCC in the near future.

  17. Fluorescence from excitation of CH4, CH3OH and CH3SH by extreme vacuum ultraviolet radiation

    NASA Technical Reports Server (NTRS)

    Ma, Guang; Suto, Masako; Lee, L. C.

    1990-01-01

    The photoabsorption and fluorescence cross sections of CH4, CH3OH, and CH3SH were measured in the wavelength regions of 52-106, 48-106, and 48-106 nm, respectively. The fluorescence spectra were dispersed to identify the emitting species. Emissions from the excited species of H(asterisk) and CH(asterisk) are commonly observed for all three molecules. Emission from the excited CH2(asterisk) is observed from CH4, OH(asterisk) from CH3OH and CS(asterisk) from CH3SH. The photoexcitation processes that may produce the observed emission bands are discussed.

  18. Integrated fluorescence detection of labeled biomolecules using a prism-like PDMS microfluidic chip and lateral light excitation.

    PubMed

    Novo, Pedro; Chu, Virginia; Conde, João Pedro

    2014-06-21

    Microfabricated amorphous silicon photodiodes were integrated with prism-like PDMS microfluidics for the detection and quantification of fluorescence signals. The PDMS device was fabricated with optical quality surfaces and beveled sides. A 405 nm laser beam perpendicular to the lateral sides of the microfluidic device excites the fluorophores in the microchannel at an angle of 70° to the normal to the microchannel/photodiode surface. This configuration, which makes use of the total internal reflection of the excitation beam and the isotropy of the fluorescence emission, minimizes the intensity of excitation light that reaches the integrated photodetector. A difference of two orders of magnitude was achieved in the reduction of the detection noise level as compared with a normally incident excitation configuration. A limit-of-detection of 5.6 × 10(10) antibodies per square centimeter was achieved using antibodies labeled with a model organic fluorophore. Furthermore, the results using the lateral excitation scheme are in good proportionality agreement with those by fluorescence quantification using wide-field fluorescence microscopy. PMID:24806101

  19. Surface extended x-ray absorption fine structure of low-Z absorbates using fluorescence detection

    SciTech Connect

    Stoehr, J.; Kollin, E.B.; Fischer, D.A.; Hastings, J.B.; Zaera, F.; Sette, F.

    1985-05-01

    Comparison of x-ray fluorescence yield (FY) and electron yield surface extended x-ray absorption fine structure spectra above the S K-edge for c(2 x 2) S on Ni(100) reveals an order of magnitude higher sensitivity of the FY technique. Using FY detection, thiophene (C/sub 4/H/sub 4/S) chemisorption on Ni(100) is studied with S coverages down to 0.08 monolayer. The molecule dissociates at temperatures as low as 100K by interaction with fourfold hollow Ni sites. Blocking of these sites by oxygen leaves the molecule intact.

  20. Single-molecule spectroscopy and femtosecond transient absorption studies on the excitation energy transfer process in ApcE(1-240) dimers.

    PubMed

    Long, Saran; Zhou, Meng; Tang, Kun; Zeng, Xiao-Li; Niu, Yingli; Guo, Qianjin; Zhao, Kai-Hong; Xia, Andong

    2015-05-28

    ApcE(1-240) dimers with one intrinsic phycocyanobilin (PCB) chromophore in each monomer that is truncated from the core-membrane linker (ApcE) of phycobilisomes (PBS) in Nostoc sp. PCC 7120 show a sharp and significantly red-shifted absorption. Two explanations either conformation-dependent Förster resonance energy transfer (FRET) or the strong exciton coupling limit have been proposed for red-shifted absorption. This is a classic example of the special pair in the photosynthetic light harvesting proteins, but the mechanism of this interaction is still a matter of intense debate. We report the studies using single-molecule and transient absorption spectra on the interaction in the special pair of ApcE dimers. Our results demonstrate the presence of conformation-dependent FRET between the two PCB chromophores in ApcE dimers. The broad distributions of fluorescence intensities, lifetimes and polarization difference from single-molecule measurements reveal the heterogeneity of local protein-pigment environments in ApcE dimers, where the same molecular structures but different protein environments are the main reason for the two PCB chromophores with different spectral properties. The excitation energy transfer rate between the donor and the acceptor about (110 ps)(-1) is determined from transient absorption measurements. The red-shifted absorption in ApcE dimers could result from more extending conformation, which shows another type of absorption redshift that does not depend on strong exciton coupling. The results here stress the importance of conformation-controlled spectral properties of the chemically identical chromophores, which could be a general feature to control energy/electron transfer, widely existing in the light harvesting complexes. PMID:25925197

  1. Imaging electronic trap states in perovskite thin films with combined fluorescence and femtosecond transient absorption microscopy

    DOE PAGES

    Xiao, Kai; Ma, Ying -Zhong; Simpson, Mary Jane; Doughty, Benjamin; Yang, Bin

    2016-04-22

    Charge carrier trapping degrades the performance of organometallic halide perovskite solar cells. To characterize the locations of electronic trap states in a heterogeneous photoactive layer, a spatially resolved approach is essential. Here, we report a comparative study on methylammonium lead tri-iodide perovskite thin films subject to different thermal annealing times using a combined photoluminescence (PL) and femtosecond transient absorption microscopy (TAM) approach to spatially map trap states. This approach coregisters the initially populated electronic excited states with the regions that recombine radiatively. Although the TAM images are relatively homogeneous for both samples, the corresponding PL images are highly structured. Themore » remarkable variation in the PL intensities as compared to transient absorption signal amplitude suggests spatially dependent PL quantum efficiency, indicative of trapping events. Furthermore, detailed analysis enables identification of two trapping regimes: a densely packed trapping region and a sparse trapping area that appear as unique spatial features in scaled PL maps.« less

  2. Fluorescence excitation and propagation through brain phantom gelatins: measurements and potential applications

    SciTech Connect

    Allison, Stephen W; Gillies, George

    2010-01-01

    We have investigated the utility of 0.6% agarose gels as surrogate materials for brain tissues in optical propagation studies for possible diagnostic and therapeutic applications. Centimeter-scale layers of the gel exhibited a Beer's law attenuation factor, , of 0.2 mm 1 for incident illumination via a pulsed LED (100 Hz) at 405 nm. This result was different by only about a factor of 3 from the effective penetration depth at similar wavelengths through in vitro samples of the gray (cortical) matter of human brain, as measured by others. Then, films of the thermographic phosphors La2O2S:Eu, Mg4FGeO6:Mn, YAG:Cr and variants of the latter were formed on aluminum substrates and the fluorescence of these samples was stimulated and observed through layers of the gel up to 4 cm thick. In all cases, the fluorescence was easily excited and distinguishable above the background. The results demonstrate that this gel might serve as an inexpensive and robust test bed for exploratory studies of neurological modalities involving propagation of optical signals within brain tissues.

  3. Effect of Solvation on Electron Detachment and Excitation Energies of a Green Fluorescent Protein Chromophore Variant.

    PubMed

    Bose, Samik; Chakrabarty, Suman; Ghosh, Debashree

    2016-05-19

    Hybrid quantum mechanics/molecular mechanics (QM/MM) is applied to the fluorinated green fluorescent protein (GFP) chromophore (DFHBDI) in its deprotonated form to understand the solvatochromic shifts in its vertical detachment energy (VDE) and vertical excitation energy (VEE). This variant of the GFP chromophore becomes fluorescent in an RNA environment and has a wide range of applications in biomedical and biochemical fields. From microsolvation studies, we benchmark (with respect to full QM) the accuracy of our QM/MM calculations with effective fragment potential (EFP) as the MM method of choice. We show that while the solvatochromic shift in the VEE is minimal (0.1 eV blue shift) and its polarization component is only 0.03 eV, the effect of the solvent on the VDE is quite large (3.85 eV). We also show by accurate calculations on the solvatochromic shift of the VDE that polarization accounts for ∼0.23 eV and therefore cannot be neglected. The effect of the counterions on the VDE of the deprotonated chromophore in solvation is studied in detail, and a charge-smearing scheme is suggested for charged chromophores. PMID:27116477

  4. Triggered Excited-State Intramolecular Proton Transfer Fluorescence for Selective Triplex DNA Recognition.

    PubMed

    Wang, Ying; Hu, Yuehua; Wu, Tao; Zhou, Xiaoshun; Shao, Yong

    2015-12-01

    The triplex DNA has received much interest due to its various applications in gene regulation, molecular switch, and sensor development. However, realizing a highly selective recognition using a fluorescence probe specific only for the triplex topology is still a great challenge. Herein, we found that relative to the structural analogues of natural robinetin, myricetin, quercetin, kaempferol, morin, rutin, baicalin, luteolin, naringenin, genistein, chrysin, galangin, isorhamnetin, and several synthetic flavonoids, fisetin (FIS) is the brightest emitter when targeting the triplex DNA in contrast to binding with ss-DNA, ds-DNA (with or without an abasic site), i-motif, and DNA/RNA G-quadruplexes. Only the triplex association triggers the FIS green fluorescence that is relaxed from the tautomer favorable for excited-state intramolecular proton transfer (ESIPT). FIS can stabilize the triplex structure and primarily interact with the two terminals of the triplex via a 2:1 binding mode. This work demonstrates the potential of FIS as a DNA structure-selective switch-on ESIPT probe when evolving the triplex-forming oligonucleotides and developing the novel triplex-based sensors and switches. PMID:26556582

  5. Measurement of radical-species concentrations and polycyclic aromatic hydrocarbons in flames by fluorescence and absorption using a tunable dye laser. Progress report, March 1, 1980-February 28, 1981

    SciTech Connect

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

    1981-03-01

    A theoretical and experimental investigation of OH saturated fluorescence is described. The goal of the research is to develop a saturated fluorescence technique which will yield accurate molecular number densities over a wide range of flame pressure, temperature, and composition. Experimentally, OH is excited by a ten nanosecond pulse from a Nd:YAG-pumped dye laser tuned to an isolated rotational transition in the (0,0) band of the A/sup 2/..sigma../sup +/-X/sup 2/ pi electronic system. The resulting fluorescence signal is resolved both spectrally and temporally. Total OH number densities are calculated by collecting fluorescence from the directly excited upper rotational level, and using the balanced cross-rate model to analyze the experimental data. Fluorescence measurements of OH number density agree to within a factor of three with the results of independent OH absorption measurements. Significantly, the ratio of the fluorescence signal to the number density measured by absorption is nearly the same in 30, 100 and 250 torr H/sub 2//O/sub 2//N/sub 2/ flat flames, demonstrating the insensitivity of the saturated fluorescence signal to the quenching environment of the radical. Collisional transfer in excited OH is studied by recording the time development of OH fluorescence spectrum. The experimental spectra are compared with the results of time-dependent computer modeling. By varying rotational transfer rates until the calculated and experimental spectra agree, rotational transfer cross sections can be calculated. The signal processing system was thoroughly checked by comparing the photomultiplier output to that of a fast photodiode, and by comparing single pulse Rayleigh scattering and fluorescence traces with sampling oscilloscope traces.

  6. Nanosecond two-photon excitation fluorescence imaging with a multi color fiber MOPA laser

    NASA Astrophysics Data System (ADS)

    Karpf, Sebastian; Eibl, Matthias; Huber, Robert

    2015-07-01

    A system is presented that uses a fiber based Master Oscillator Power Amplifier (MOPA) with nanosecond-range pulses for two-photon excitation fluorescence (TPEF) imaging. The robust laser in the extended near infrared is based on an actively modulated electro-optical modulator (EOM), enabling free synchronization of the pulses to any other light source or detection unit. Pulses with a freely programmable duration between 0.4 and 10 ns are generated and then amplified to up to kilowatts of peak power with ytterbium doped fiber amplifiers (YDFA). Since we achieve peak power and duty cycles comparable to standard femto- and picosecond setups, the TPEF signal levels are similar, but realized with a robust and inexpensive fiber-based setup. The delivery fiber is further used as an optional, electronically controllable Raman shifter to effectively shift the 1064 nm light to 1122 nm and to 1186 nm. This allows imaging of a manifold of fluorophores, like e.g. TexasRed, mCherry, mRaspberry and many more. We show TPEF imaging of the autofluorescence of plant leaves of moss and algae, acquired in epi-direction. This modular laser unit can be integrated into existing systems as either a fiber-based, alignment free excitation laser or an extension for multi-modal imaging.

  7. Ultrafast excited-state dynamics at interfaces: fluorescent DNA probes at the dodecane/water interface

    NASA Astrophysics Data System (ADS)

    Licari, Giuseppe; Vauthey, Eric

    2015-08-01

    Although the interfaces between two isotropic media are of primary importance in many areas of science and technology, their properties are only partially understood. Our strategy to obtain an insight into these properties is to investigate the ultrafast excited-state dynamics of environment-sensitive molecular probes at liquid interfaces using time-resolved surface second harmonic generation, and to compare it with the dynamics of the same molecules in bulk solutions. Additionally, this approach gives rich information on how the chemical reactivity may change when going from the bulk phase to the interface. This is illustrated by an investigation performed with a series of fluorescent DNA probes at the dodecane/water interface without and with the presence of DNA in the aqueous phase. Substantial differences in the conformation of these cyanine dyes (aggregated or not) and in the excited-state dynamics are observed when going from bulk solutions to the interface. Moreover, the presence of double-stranded DNA in the aqueous phase induces some chirality at the interface.

  8. Intracavity laser excitation of NCO fluorescence in an atmospheric pressure flame

    NASA Astrophysics Data System (ADS)

    Anderson, W. R.; Vanderhoff, J. A.; Kotlar, A. J.; Dewilde, M. A.; Beyer, R. A.

    1983-09-01

    Laser excited fluorescence of the NCO radical has been obtained using discrete prism selected lines of an argon ion laser pump source. To our knowledge, this is the first time NCO fluorescence has been obtained in a flame environment. NCO was formed in a slightly rich atmospheric pressure CH4/N2O flame. This flame was placed inside the extended cavity of the argon laser to take advantage of the much higher light intensity levels. All of the available laser lines pump vibrational hot bands of the NCO A doublet-sigma- plus to X doublet-pi system. The 4658 Angstom line appears to be the most useful for probing NCO densities. This line pumps from vl=1 in the X state to the lowest vibrational level of the A state. NCO is pumped to N prime = 31 by this line, probably via the Q2(31) transition although the R2(30) and P(32) transitions could not be ruled out in the present analysis. The 4658 Angstrom line was used to determine a relative NCO density profile through the reaction zone of a CH4/N2O flame. Profiles of C2, CN, and temperature were also obtained in this flame and are compared with the NCO profile. A lower limit of approximately 3x10 to the 14th power CM-3 was placed on the peak NCO density in the flame. Attempts to find NCO or CN fluorescence in a CH4/air flame failed, indicated probable differences in nitrogen chemistry for the two flames.

  9. Laser excitation spectrum and the long path length absorption spectrum of formyl cyanide, CHOCN

    NASA Astrophysics Data System (ADS)

    Judge, R. H.; Moule, D. C.; Biernacki, A.; Benkel, M.; Ross, J. M.; Rustenburg, J.

    1986-04-01

    Formyl cyanide has been prepared for the first time by the flash pyrolysis of methoxyacetonitrile. The compound was found to be unstable and had a half-life of about 8 min in the vapor phase in the laser experiments. The near-UV absorption spectrum was photographed under conditions of long path length (56 m) at modest dispersion (1.5 nm/mm between 368 and 390 nm, and 0.75 nm/mm between 368 and 350 nm). Excitation spectra were recorded over the 386- to 360-nm region with a N 2 pumped dye laser. The observed spectrum proved to have an open vibrational and rotational fine structure and was assigned to the n → π ∗Ã1A″ ← X˜1A' electronic transition. Vibrational assignments were made in terms of ν' 3 (CO), ν' 4 (CHO), ν' 5 (CC), ν' 6 (CCO), ν' 7 (CCN), ν' 8 (H wag), ν' 9 (CCN), ν″ 7 (CCN), and ν″ 9 (CCN). The vibrational frequencies in both states were found to correlate closely to those of the propynal molecule.

  10. Trapping and two-photon fluorescence excitation of microscopic objects using ultrafast single-fiber optical tweezers.

    PubMed

    Mishra, Yogeshwar N; Ingle, Ninad; Mohanty, Samarendra K

    2011-10-01

    Analysis of trapped microscopic objects using fluorescence and Raman spectroscopy is gaining considerable interest. We report on the development of single fiber ultrafast optical tweezers and its use in simultaneous two-photon fluorescence (TPF) excitation of trapped fluorescent microscopic objects. Using this method, trapping depth of a few centimeters was achieved inside a colloidal sample with TPF from the trapped particle being visible to the naked eye. Owing to the propagation distance of the Bessel-like beam emerging from the axicon-fiber tip, a relatively longer streak of fluorescence was observed along the microsphere length. The cone angle of the axicon was engineered so as to provide better trapping stability and high axial confinement of TPF. Trapping of the floating objects led to stable fluorescence emission intensity over a long period of time, suitable for spectroscopic measurements. Furthermore, the stability of the fiber optic trapping was confirmed by holding and maneuvering the fiber by hand so as to move the trapped fluorescent particle in three dimensions. Apart from miniaturization capability into lab-on-a-chip microfluidic devices, the proposed noninvasive microaxicon tipped optical fiber can be used in multifunctional mode for in-depth trapping, rotation, sorting, and ablation, as well as for two-photon fluorescence excitation of a motile sample.

  11. X-ray fluorescence and absorption analysis of krypton in irradiated nuclear fuel

    NASA Astrophysics Data System (ADS)

    Degueldre, Claude; Mieszczynski, Cyprian; Borca, Camelia; Grolimund, Daniel; Martin, Matthias; Bertsch, Johannes

    2014-10-01

    The analysis of krypton in irradiated uranium dioxide fuel has been successfully achieved by X-ray fluorescence and X-ray absorption. The present study focuses on the analytical challenge of sample and sub-sample production to perform the analysis with the restricted conditions dictated by the radioprotection regulations. It deals also with all potential interferences that could affect the quality of the measurement in fluorescence as well as in absorption mode. The impacts of all dissolved gases in the fuel matrix are accounted for the analytical result quantification. The krypton atomic environment is ruled by the presence of xenon. Other gases such as residual argon and traces of helium or hydrogen are negligible. The results are given in term of density for krypton (∼3 nm-3) and xenon (∼20 nm-3). The presence of dissolved, interstitial and nano-phases are discussed together with other analytical techniques that could be applied to gain information on fission gas behaviour in nuclear fuels.

  12. Detection limits of 405 nm and 633 nm excited PpIX fluorescence for brain tumor detection during stereotactic biopsy

    NASA Astrophysics Data System (ADS)

    Markwardt, Niklas; Götz, Marcus; Haj-Hosseini, Neda; Hollnburger, Bastian; Sroka, Ronald; Stepp, Herbert; Zelenkov, Petr; Rühm, Adrian

    2016-04-01

    5-aminolevulinic-acid-(5-ALA)-induced protoporphyrin IX (PpIX) fluorescence may be used to improve stereotactic brain tumor biopsies. In this study, the sensitivity of PpIX-based tumor detection has been investigated for two potential excitation wavelengths (405 nm, 633 nm). Using a 200 μm fiber in contact with semi-infinite optical phantoms containing ink and Lipovenös, PpIX detection limits of 4.0 nM and 200 nM (relating to 1 mW excitation power) were determined for 405 nm and 633 nm excitation, respectively. Hence, typical PpIX concentrations in glioblastomas of a few μM should be well detectable with both wavelengths. Additionally, blood layers of selected thicknesses were placed between fiber and phantom. Red excitation was shown to be considerably less affected by blood interference: A 50 μm blood layer, for instance, blocked the 405- nm-excited fluorescence completely, but reduced the 633-nm-excited signal by less than 50%. Ray tracing simulations demonstrated that - without blood layer - the sensitivity advantage of 405 nm rises for decreasing fluorescent volume from 50-fold to a maximum of 100-fold. However, at a tumor volume of 1 mm3, which is a typical biopsy sample size, the 633-nm-excited fluorescence signal is only reduced by about 10%. Further simulations revealed that with increasing fiber-tumor distance, the signal drops faster for 405 nm. This reduces the risk of detecting tumor tissue outside the needle's coverage, but diminishes the overlap between optically and mechanically sampled volumes. While 405 nm generally offers a higher sensitivity, 633 nm is more sensitive to distant tumors and considerably superior in case of blood-covered tumor tissue.

  13. A two photon absorption laser induced fluorescence diagnostic for fusion plasmasa)

    NASA Astrophysics Data System (ADS)

    Magee, R. M.; Galante, M. E.; McCarren, D.; Scime, E. E.; Boivin, R. L.; Brooks, N. H.; Groebner, R. J.; Hill, D. N.; Porter, G. D.

    2012-10-01

    The quality of plasma produced in a magnetic confinement fusion device is influenced to a large extent by the neutral gas surrounding the plasma. The plasma is fueled by the ionization of neutrals, and charge exchange interactions between edge neutrals and plasma ions are a sink of energy and momentum. Here we describe a diagnostic capable of measuring the spatial distribution of neutral gas in a magnetically confined fusion plasma. A high intensity (5 MW/cm2), narrow bandwidth (0.1 cm-1) laser is injected into a hydrogen plasma to excite the Lyman β transition via the simultaneous absorption of two 205 nm photons. The absorption rate, determined by measurement of subsequent Balmer α emission, is proportional to the number of particles with a given velocity. Calibration is performed in situ by filling the chamber to a known pressure of neutral krypton and exciting a transition close in wavelength to that used in hydrogen. We present details of the calibration procedure, including a technique for identifying saturation broadening, measurements of the neutral density profile in a hydrogen helicon plasma, and discuss the application of the diagnostic to plasmas in the DIII-D tokamak.

  14. A two photon absorption laser induced fluorescence diagnostic for fusion plasmas

    SciTech Connect

    Magee, R. M.; Galante, M. E.; McCarren, D.; Scime, E. E.; Boivin, R. L.; Brooks, N. H.; Groebner, R. J.; Hill, D. N.; Porter, G. D.

    2012-10-15

    The quality of plasma produced in a magnetic confinement fusion device is influenced to a large extent by the neutral gas surrounding the plasma. The plasma is fueled by the ionization of neutrals, and charge exchange interactions between edge neutrals and plasma ions are a sink of energy and momentum. Here we describe a diagnostic capable of measuring the spatial distribution of neutral gas in a magnetically confined fusion plasma. A high intensity (5 MW/cm{sup 2}), narrow bandwidth (0.1 cm{sup -1}) laser is injected into a hydrogen plasma to excite the Lyman {beta} transition via the simultaneous absorption of two 205 nm photons. The absorption rate, determined by measurement of subsequent Balmer {alpha} emission, is proportional to the number of particles with a given velocity. Calibration is performed in situ by filling the chamber to a known pressure of neutral krypton and exciting a transition close in wavelength to that used in hydrogen. We present details of the calibration procedure, including a technique for identifying saturation broadening, measurements of the neutral density profile in a hydrogen helicon plasma, and discuss the application of the diagnostic to plasmas in the DIII-D tokamak.

  15. Intramolecular charge transfer of 4-(dimethylamino)benzonitrile probed by time-resolved fluorescence and transient absorption: No evidence for two ICT states and a pisigma( *) reaction intermediate.

    PubMed

    Zachariasse, Klaas A; Druzhinin, Sergey I; Kovalenko, Sergey A; Senyushkina, Tamara

    2009-12-14

    For the double exponential fluorescence decays of the locally excited (LE) and intramolecular charge transfer (ICT) states of 4-(dimethylamino)benzonitrile (DMABN) in acetonitrile (MeCN) the same times tau(1) and tau(2) are observed. This means that the reversible LE<==>ICT reaction, starting from the initially excited LE state, can be adequately described by a two state mechanism. The most important factor responsible for the sometimes experimentally observed differences in the nanosecond decay time, with tau(1)(LE)fluorescence response functions with a time resolution of 0.5 ps/channel in 1200 channels reliable kinetic and thermodynamic data can be obtained. The arguments presented in the literature in favor of a pisigma( *) state with a bent CN group as an intermediate in the ICT reaction of DMABN are discussed. From the appearance of an excited state absorption (ESA) band in the spectral region between 700 and 800 nm in MeCN for N,N-dimethylanilines with CN, Br, F, CF(3), and C(=O)OC(2)H(2) p-substituents, it is concluded that this ESA band cannot be attributed to a pisigma( *) state, as only the C-C[Triple Bond]N group can undergo the required 120 degrees bending. PMID:20001042

  16. Two-photon absorption laser induced fluorescence measurement of atomic oxygen density in an atmospheric pressure air plasma jet

    NASA Astrophysics Data System (ADS)

    Conway, J.; Gogna, G. S.; Gaman, C.; Turner, M. M.; Daniels, S.

    2016-08-01

    Atomic oxygen number density [O] is measured in an air atmospheric pressure plasma jet (APPJ) using two-photon absorption laser induced fluorescence (TALIF). Gas flow is fixed at 8 slpm, the RF power coupled into the plasma jet varied between 5 W and 20 W, and the resulting changes in atomic oxygen density measured. Photolysis of molecular oxygen is employed to allow in situ calibration of the TALIF system. During calibration, O2 photo-dissociation and two-photon excitation of the resulting oxygen atoms are achieved within the same laser pulse. The atomic oxygen density produced by photolysis is time varying and spatially non-uniform which needs to be corrected for to calibrate the TALIF system for measurement of atomic oxygen density in plasma. Knowledge of the laser pulse intensity I 0(t), wavelength, and focal spot size allows correction factors to be determined using a rate equation model. Atomic oxygen is used for calibration and measurement, so the laser intensity can be increased outside the TALIF quadratic laser power dependence region without affecting the calibration reliability as the laser power dependence will still be the same for both. The atomic O density results obtained are not directly benchmarked against other known density measurement techniques. The results show that the plasma jet atomic oxygen content increases as the RF power coupled into the plasma increases.

  17. In vivo micro-vascular imaging and flow cytometry in zebrafish using two-photon excited endogenous fluorescence.

    PubMed

    Zeng, Yan; Yan, Bo; Sun, Qiqi; He, Sicong; Jiang, Jun; Wen, Zilong; Qu, Jianan Y

    2014-03-01

    Zebrafish has rapidly evolved as a powerful vertebrate model organism for studying human diseases. Here we first demonstrate a new label-free approach for in vivo imaging of microvasculature, based on the recent discovery and detailed characterization of the two-photon excited endogenous fluorescence in the blood plasma of zebrafish. In particular, three-dimensional reconstruction of the microvascular networks was achieved with the depth-resolved two-photon excitation fluorescence (TPEF) imaging. Secondly, the blood flow images, obtained by perpendicularly scanning the focal point across the blood vessel, provided accurate information for characterizing the hemodynamics of the circulatory system. The endogenous fluorescent signals of reduced nicotinamide adenine dinucleotide (NADH) enabled visualization of the circulating granulocytes (neutrophils) in the blood vessel. The development of acute sterile inflammation could be detected by the quantitative counting of circulating neutrophils. Finally, we found that by utilizing a short wavelength excitation at 650 nm, the commonly used fluorescent proteins, such as GFP and DsRed, could be efficiently excited together with the endogenous fluorophores to achieve four-color TPEF imaging of the vascular structures and blood cells. The results demonstrated that the multi-color imaging could potentially yield multiple view angles of important processes in living biological systems.

  18. Spectral fluorescence signature techniques and absorption measurements for continuous monitoring of biofuel-producing microalgae cultures

    NASA Astrophysics Data System (ADS)

    Martín de la Cruz, M. C.; Gonzalez Vilas, L.; Yarovenko, N.; Spyrakos, E.; Torres Palenzuela, J. M.

    2013-08-01

    Biofuel production from microalgae can be both sustainable and economically viable. Particularly in the case of algal growth in wastewater an extra benefit is the removal or biotransformation of pollutants from these types of waters. A continuous monitoring system of the microalgae status and the concentration of different wastewater contaminants could be of great help in the biomass production and the water characterisation. In this study we present a system where spectral fluorescence signature (SFS) techniques are used along with absorption measurements to monitor microalgae cultures in wastewater and other mediums. This system aims to optimise the microalgae production for biofuel applications or other uses and was developed and tested in prototype indoor photo-bioreactors at the University of Vigo. SFS techniques were applied using the fluorescence analyser INSTAND-SCREENER developed by Laser Diagnostic Instruments AS. INSTAND-SCREENER permits wavelength scanning in two modes, one in UV and another in VIS. In parallel, it permits the on-line monitoring and rapid analysis of both water quality and phytoplankton status without prior treatment of the sample. Considering that different contaminants and microalgae features (density, status etc.) have different spectral signatures of fluorescence and absorption properties, it is possible to characterise them developing classification libraries. Several algorithms were used for the classification. The implementation of this system in an outdoor raceway reactor in a Spanish wastewater treatment plant is also discussed. This study was part of the Project EnerBioAlgae (http://www.enerbioalgae.com/), which was funded by the Interreg SUDOE and led by the University of Vigo.

  19. Determination of mercury distribution inside spent compact fluorescent lamps by atomic absorption spectrometry.

    PubMed

    Rey-Raap, Natalia; Gallardo, Antonio

    2012-05-01

    In this study, spent compact fluorescent lamps were characterized to determine the distribution of mercury. The procedure used in this research allowed mercury to be extracted in the vapor phase, from the phosphor powder, and the glass matrix. Mercury concentration in the three phases was determined by the method known as cold vapor atomic absorption spectrometry. Median values obtained in the study showed that a compact fluorescent lamp contained 24.52±0.4ppb of mercury in the vapor phase, 204.16±8.9ppb of mercury in the phosphor powder, and 18.74±0.5ppb of mercury in the glass matrix. There are differences in mercury concentration between the lamps since the year of manufacture or the hours of operation affect both mercury content and its distribution. The 85.76% of the mercury introduced into a compact fluorescent lamp becomes a component of the phosphor powder, while more than 13.66% is diffused through the glass matrix. By washing and eliminating all phosphor powder attached to the glass surface it is possible to classified the glass as a non-hazardous waste.

  20. Ultraviolet high-excitation Fe II fluorescence lines excited by O VI, C IV, and H I resonance emission as seen in IUE spectra

    NASA Technical Reports Server (NTRS)

    Feibelman, Walter A.; Bruhweiler, Frederick C.; Johansson, Sveneric

    1991-01-01

    Archival high-dispersion spectra from the IUE are used in a search for Bowen emission lines of Fe II excited by the stronger transition of the O VI resonance doublet. The possibility of using these Fe II emission lines as a diagnostic of the strength of the far-ultraviolet emission of O VI at 1032 A is explored. It is found that the Fe II emission lines are quite common and strong in symbiotic stars, particularly those of the type known as 'symbiotic novae', as well as in normal novae. The lines are observed in central stars of some planetary nebulae of the O VI sequence besides a few central stars of type WR. High density, high excitation, and high temperature are suggested to be requirements for the excitation of the Fe II fluorescence lines. It is pointed out that while these lines were observed in PG 1159-035 and K1-16, they were not observed in AGNs.

  1. First-principles calculation of ground and excited-state absorption spectra of ruby and alexandrite considering lattice relaxation

    NASA Astrophysics Data System (ADS)

    Watanabe, Shinta; Sasaki, Tomomi; Taniguchi, Rie; Ishii, Takugo; Ogasawara, Kazuyoshi

    2009-02-01

    We performed first-principles calculations of multiplet structures and the corresponding ground-state absorption and excited-state absorption spectra for ruby (Cr3+:α-Al2O3) and alexandrite (Cr3+:BeAl2O4) which included lattice relaxation. The lattice relaxation was estimated using the first-principles total energy and molecular-dynamics method of the CASTEP code. The multiplet structure and absorption spectra were calculated using the configuration-interaction method based on density-functional calculations. For both ruby and alexandrite, the theoretical absorption spectra, which were already in reasonable agreement with experimental spectra, were further improved by consideration of lattice relaxation. In the case of ruby, the peak positions and peak intensities were improved through the use of models with relaxations of 11 or more atoms. For alexandrite, the polarization dependence of the U band was significantly improved, even by a model with a relaxation of only seven atoms.

  2. A reagentless enzymatic fluorescent biosensor for glucose based on upconverting glasses, as excitation source, and chemically modified glucose oxidase.

    PubMed

    Del Barrio, Melisa; Cases, Rafael; Cebolla, Vicente; Hirsch, Thomas; de Marcos, Susana; Wilhelm, Stefan; Galbán, Javier

    2016-11-01

    Upon near-infrared excitation Tm(3+)+Yb(3+) doped fluorohafnate glasses present upconversion properties and emit visible light. This property permits to use these glasses (UCG) as excitation sources for fluorescent optical biosensors. Taking this into account, in this work a fluorescent biosensor for glucose determination is designed and evaluated. The biosensor combines the UCG and the fluorescence of the enzyme glucose oxidase chemically modified with a fluorescein derivative (GOx-FS), whose intensity is modified during the enzymatic reaction with glucose. Optical parameters have been optimized and a mathematical model describing the behavior of the analytical signal is suggested. Working in FIA mode, the biosensor responds to glucose concentrations up to, at least, 15mM with a limit of detection of 1.9mM. The biosensor has a minimum lifetime of 9 days and has been applied to glucose determination in drinks. The applicability of the sensor was tested by glucose determination in two fruit juices.

  3. Separation and determination of amino acids by micellar electrokinetic chromatography coupling with novel multiphoton excited fluorescence detection.

    PubMed

    Chen, Sheng; Xu, Youzhi; Xu, Fei; Feng, Xiaojun; Du, Wei; Luo, Qingming; Liu, Bi-Feng

    2007-08-31

    In this article, it was demonstrated that separation and determination of 20 amino acids were accomplished by micellar electrokinetic chromatography (MEKC) coupling with novel multiphoton excited fluorescence (MPEF) detection method. Different from MPEF achieved by expensive fs laser, continuous wave (CW) diode laser of ultra-low cost was uniquely employed in our MPEF system. Amino acids were fluorescently labeled with fluorescein isothiocyanate (FITC), and were subjected to sodium dodecyl sulfate (SDS)-based MEKC separation and CW-based MPEF detection. The result was compared with that by single photon excited fluorescence (SPEF), which indicated that MPEF had the advantages of better mass detectability and higher separation selectivity over SPEF. Quantitative analysis was performed and revealed linear dynamic range of over 2 orders of magnitude, with mass detection limit down to ymole level. To evaluate the reliability, this method was successfully applied for analyzing a commercial nutrition supplement liquid.

  4. Laser-excited fluorescence of rare earth elements in fluorite: Initial observations with a laser Raman microprobe

    USGS Publications Warehouse

    Burruss, R.C.; Ging, T.G.; Eppinger, R.G.; Samson, a.M.

    1992-01-01

    Fluorescence emission spectra of three samples of fluorite containing 226-867 ppm total rare earth elements (REE) were excited by visible and ultraviolet wavelength lines of an argon ion laser and recorded with a Raman microprobe spectrometer system. Narrow emission lines ( 0.9 for Eu2+ and 0.99 for Er3+. Detection limits for three micrometer spots are about 0.01 ppm Eu2+ and 0.07 ppm Er3+. These limits are less than chondrite abundance for Eu and Er, demonstrating the potential microprobe analytical applications of laser-excited fluorescence of REE in fluorite. However, application of this technique to common rock-forming minerals may be hampered by competition between fluorescence emission and radiationless energy transfer processes involving lattice phonons. ?? 1992.

  5. Two-photon excitation and excited-state absorption cross sections for H2 E,F 1Sigma sub g (v=6) - Measurement and calculations

    NASA Astrophysics Data System (ADS)

    Buck, Jesse D.; Robie, Daniel C.; Hickman, A. P.; Bamford, Douglas J.; Bischel, William K.

    1989-04-01

    Observations of the E,F 1Sigma sub g (+) (v-prime=6)-to-B 1Sigma sub u (+) (v-double-prime=0) fluorescence transition at about 750 nm have been conducted to determine the absolute two-photon excitation cross section for the H2 E,F 1Sigma sub g (v-prime=6)-from-X 1Sigma sub g (v-double-prime=0) Q(1) transition at 193 nm. The 2.0 + or - 0.9 x 10 to the -36th cm exp 4 integrated two-photon excitation cross-section obtained is in good agreement with Huo and Jaffe's (1983) theoretical calculation result of 2.8 x 10 to the -36th cm exp 4.

  6. Two-photon excited fluorescence spectroscopy and imaging of melanin in vitro and in vivo

    NASA Astrophysics Data System (ADS)

    Krasieva, Tatiana B.; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L.; Tromberg, Bruce J.

    2012-03-01

    The ability to detect early melanoma non-invasively would improve clinical outcome and reduce mortality. Recent advances in two-photon excited fluorescence (TPEF) in vivo microscopy offer a powerful tool in early malignant melanoma diagnostics. The goal of this work was to develop a TPEF optical index for measuring relative concentrations of eumelanin and pheomelanin since ex vivo studies show that changes in this ratio have been associated with malignant transformation. We acquired TPEF emission spectra (λex=1000 nm) of melanin from several specimens, including human hair, malignant melanoma cell lines, and normal melanocytes and keratinocytes in different skin layers (epidermis, papillary dermis) in five healthy volunteers in vivo. We found that the pheomelanin emission peaks at around 620 nm and is blue-shifted from the eumelanin with broad maximum at 640-680nm. We defined "optical melanin index" (OMI) as a ratio of fluorescence signal intensities measured at 645 nm and 615nm. The measured OMI for a melanoma cell line MNT-1 was 1.6+/-0.2. The MNT-46 and MNT-62 lines (Mc1R gene knockdown) showed an anticipated change in melanins production ratio and had OMI of 0.55+/-0.05 and 0.17+/-0.02, respectively, which strongly correlated with HPLC data obtained for these lines. Average OMI measured for basal cells layers (melanocytes and keratinocytes) in normal human skin type I, II-III (not tanned and tanned) in vivo was 0.5, 1.05 and 1.16 respectively. We could not dependably detect the presence of pheomelanin in highly pigmented skin type V-VI. These data suggest that a non-invasive TPEF index could potentially be used for rapid melanin ratio characterization both in vitro and in vivo, including pigmented lesions.

  7. Picosecond excite-and-probe absorption measurement of the 4T2 state nonradiative lifetime in ruby

    NASA Technical Reports Server (NTRS)

    Gayen, S. K.; Wang, W. B.; Petricevic, V.; Dorsinville, R.; Alfano, R. R.

    1985-01-01

    In a picosecond excite-and-probe absorption measurement, a 527-nm picosecond pulse excites the 4T2 state of the Cr(3+) ion in ruby and a 3.4-micron picosecond probe pulse monitors the growth and decay of population in the 2E state as a function of pump-probe delay. From the growth of population in the metastable 2E state, an upper limit of 7 ps for the nonradiative lifetime of the 4T2 state is determined.

  8. Temperature-dependent excitonic photoluminescence excited by two-photon absorption in perovskite CsPbBr_3 quantum dots

    NASA Astrophysics Data System (ADS)

    Wei, Ke; Xu, Zhongjie; Chen, Runze; Zheng, Xin; Cheng, Xiangai; Jiang, Tian

    2016-08-01

    Recently lead halide nanocrystals (quantum dots) have been reported with potential for photovoltaic and optoelectronic applications due to their excellent luminescent properties. Herein excitonic photoluminescence (PL) excited by two-photon absorption in perovskite CsPbBr3 quantum dots (QDs) have been studied across a broad temperature range from 80K to 380K. Two-photon absorption has been investigated with absorption coefficient up to 0.085 cm/GW at room temperature. Moreover, the photoluminescence excited by two-photon absorption shows a linear blue-shift (0.25meV/K) below temperature of ~220K and turned steady with fluctuation below 1nm (4.4meV) for higher temperature up to 380K. These phenomena are distinctly different from general red-shift of semiconductor and can be explained by the competition between lattice expansion and electron-phonon couplling.Our results reveal the strong nonlinear absorption and temperature-independent chromaticity in a large temperature range from 220K to 380K in the CsPbX3 QDs, which will offer new opportunities in nonlinear photonics, light-harvesting and light-emitting devices.

  9. Fluorescence Excitation Models of Ammonia and Amidogen Radical (NH2) in Comets: Application to Comet C/2004 Q2 (Machholz)

    NASA Technical Reports Server (NTRS)

    Kawakita, Hideyo; Mumma, Michael J.

    2011-01-01

    Ammonia is a major reservoir of nitrogen atoms in cometary materials. However, detections of ammonia in comets are rare, with several achieved at radio wavelengths. A few more detections were obtained through near-infrared observations (around the 3 m wavelength region), but moderate relative velocity shifts are required to separate emission lines of cometary ammonia from telluric absorption lines in the 3 micron wavelength region. On the other hand, the amidogen radical (NH2 -- a photodissociation product of ammonia in the coma) also shows rovibrational emission lines in the 3 micron wavelength region. Thus, gas production rates for ammonia can be determined from the rovibrational emission lines of ammonia (directly) and amidogen radical (indirectly) simultaneously in the near-infrared. In this article, we present new fluorescence excitation models for cometary ammonia and amidogen radical in the near-infrared, and we apply these models to the near-infrared high-dispersion spectra of comet C/2004 Q2 (Machholz) to determine the mixing ratio of ammonia to water in the comet. Based on direct detection of NH3 lines, the mixing ratio of NH3/H2O is 0.46% +/- 0.03% in C/2004 Q2 (Machholz), in agreement with other results. The mixing ratio of ammonia determined from the NH2 observations (0.31% -- 0.79%) is consistent but has relatively larger error, owing to uncertainty in the photodissociation rates of ammonia. At the present level of accuracy, we confirm that NH3 could be the sole parent of NH2 in this comet.

  10. Monitor RNA synthesis in live cell nuclei by using two-photon excited fluorescence lifetime imaging microscopy

    NASA Astrophysics Data System (ADS)

    Peng, Xiao; Lin, Danying; Wang, Yan; Qi, Jing; Yan, Wei; Qu, Junle

    2015-03-01

    Probing of local molecular environment in cells is of significant value in creating a fundamental understanding of cellular processes and molecular profiles of diseases, as well as studying drug cell interactions. In order to investigate the dynamically changing in subcellular environment during RNA synthesis, we applied two-photon excited fluorescence lifetime imaging microscopy (FLIM) method to monitor the green fluorescent protein (GFP) fused nuclear protein ASF/SF2. The fluorescence lifetime of fluorophore is known to be in inverse correlation with a local refractive index, and thus fluorescence lifetimes of GFP fusions provide real-time information of the molecular environment of ASF/SF2- GFP. The FLIM results showed continuous and significant fluctuations of fluorescence lifetimes of the fluorescent protein fusions in live HeLa cells under physiological conditions. The fluctuations of fluorescence lifetime values indicated the variations of activities of RNA polymerases. Moreover, treatment with pharmacological drugs inhibiting RNA polymerase activities led to irreversible decreases of fluorescence lifetime values. In summary, our study of FLIM imaging of GFP fusion proteins has provided a sensitive and real-time method to investigate RNA synthesis in live cell nuclei.

  11. Excitations, optical absorption spectra, and optical excitonic gaps of heterofullerenes: I. C60, C59N+ and C48N12

    SciTech Connect

    Xie, R; Bryant, G W; Sun, G; C.Nicklaus, M; Heringer, D; Frauenheim, T; Manaa, M R; Smith, Jr., V H; Araki, Y; Ito, O

    2003-10-02

    Low-energy excitations and optical absorption spectrum of C{sub 60} are computed by using time-dependent (TD) Hartree-Fock (HF), TD-density functional theory (TD-DFT), TD-DFT-based tight-binding (TD-DFT-TB) and a semiempirical ZINDO method. A detailed comparison of experiment and theory for the excitation energies, optical gap and absorption spectrum of C{sub 60} is presented. It is found that electron correlations and collective effects of exciton pairs play important roles in assigning accurately the spectral features of C{sub 60} and the TD-DFT method with non-hybrid functionals or a local spin density approximation leads to more accurate excitation energies than with hybrid functionals. The level of agreement between theory and experiment for C{sub 60} justifies similar calculations of the excitations and optical absorption spectrum of a monomeric azafullerene cation C{sub 59}N{sup +} exhibits distinguishing spectral features different from C{sub 60}: (1) the first singlet is dipole-allowed and the optical gap is redshifted by 1.44 eV; (2) several weaker absorption maxima occur in the visible region; (3) the transient triplet-triplet absorption at 1.60 eV (775 nm) is much broader and the decay of the triplet state is much faster. The calculated spectra of C{sub 59}N{sup +} characterize and explain well our measured ultraviolet-visible (UV-vis) and transient absorption spectra of the carborane anion salt [C{sub 59}N][Ag(CB{sub 11}H{sub 6}Cl{sub 6}){sub 2}]. For the most stable isomer of C{sub 48}N{sub 12}, we predict that the first singlet is dipole-allowed, the optical gap is redshifted by 1.22 eV relative to that of C{sub 60}, and optical absorption maxima occur at 585, 528, 443, 363, 340, 314 and 303 nm. We point out that the characterization of the UV-vis and transient absorption spectra of C{sub 48}N{sub 12} isomers is helpful in distinguishing the isomer structures required for applications in molecular electronics. For C{sub 59}N{sup +} and C{sub 48}N

  12. Fluorescence excitation and emission spectroscopy on single MEH-PPV chains at low temperature.

    PubMed

    Feist, Florian A; Basché, Thomas

    2008-08-14

    Fluorescence emission and excitation spectra of single MEH-PPV polymer molecules dispersed in thin PMMA films have been recorded at 1.2 and 20 K. We observe single as well as multichromophore emission in single chain emission spectra, whereby the relative fractions depend on the two different molecular weights (50 and 350 kDa) studied. The molecular weight also affects the distribution of peak emission maxima, which is monomodal (bimodal) for the low (high) molecular weight. The appearance of an additional "red" subpopulation for the high molecular weight sample is attributed to interactions of multiple chromophores from a sufficiently flexible single chain. The comparison of emission spectra appearing in the "blue" as well as "red" subpopulations suggests that these intrachain interactions rather lead to ground-state aggregates than excimers. Independent of the molecular weight, large variations in spectral shape and apparent line width in the emission spectra have been observed. Occasionally, we find very narrow purely electronic zero-phonon lines both in emission and in excitation spectra, with line widths down to the instrumental resolution. In accordance with earlier literature data it is argued that linear electron-phonon coupling should be quite strong for MEH-PPV in PMMA, leading to only a small fraction of chromophores exhibiting zero-phonon lines. In addition, spectral diffusion, which manifests itself by several time-dependent line shifting and broadening phenomena, contributes to the substantial variations of spectral shapes. Excitation experiments with particularly stable chromophores provide an upper limit for the optical line width (approximately 0.1 cm(-1)), which at 1.2 K can actually approach the lifetime-limited homogeneous width. Raising the temperature to 20 K leads to line broadening and typically, to disappearance of zero-phonon lines. The failure to observe zero-phonon lines of chromophores supposedly serving as donors in intramolecular

  13. An instrument for fast acquisition of fluorescence decay curves at picosecond resolution designed for ``double kinetics'' experiments: Application to fluorescence resonance excitation energy transfer study of protein folding

    NASA Astrophysics Data System (ADS)

    Ishay, Eldad Ben; Hazan, Gershon; Rahamim, Gil; Amir, Dan; Haas, Elisha

    2012-08-01

    The information obtained by studying fluorescence decay of labeled biopolymers is a major resource for understanding the dynamics of their conformations and interactions. The lifetime of the excited states of probes attached to macromolecules is in the nanosecond time regime, and hence, a series of snapshot decay curves of such probes might - in principle - yield details of fast changes of ensembles of labeled molecules down to sub-microsecond time resolution. Hence, a major current challenge is the development of instruments for the low noise detection of fluorescence decay curves within the shortest possible time intervals. Here, we report the development of an instrument, picosecond double kinetics apparatus, that enables recording of multiple fluorescence decay curves with picosecond excitation pulses over wide spectral range during microsecond data collection for each curve. The design is based on recording and averaging multiphoton pulses of fluorescence decay using a fast 13 GHz oscilloscope during microsecond time intervals at selected time points over the course of a chemical reaction or conformational transition. We tested this instrument in a double kinetics experiment using reference probes (N-acetyl-tryptophanamide). Very low stochastic noise level was attained, and reliable multi-parameter analysis such as derivation of distance distributions from time resolved FRET (fluorescence resonance excitation energy transfer) measurements was achieved. The advantage of the pulse recording and averaging approach used here relative to double kinetics methods based on the established time correlated single photon counting method, is that in the pulse recording approach, averaging of substantially fewer kinetic experiments is sufficient for obtaining the data. This results in a major reduction in the consumption of labeled samples, which in many cases, enables the performance of important experiments that were not previously feasible.

  14. Optical diagnosis of gallbladder cancers via two-photon excited fluorescence imaging of unstained histological sections.

    PubMed

    Hong, Zhipeng; Chen, Youting; Chen, Jing; Chen, Hong; Xu, Yahao; Zhu, Xiaoqin; Zhuo, Shuangmu; Shi, Zheng; Chen, Jianxin

    2015-01-01

    Two-photon excited fluorescence (TPEF) microscopy, based on signal from cells, can provide detailed information on tissue architecture and cellular morphology in unstained histological sections to generate subcellular-resolution images from tissue directly. In this paper, we used TPEF microscopy to image microstructure of human normal gallbladder and three types of differentiated carcinomas in order to investigate the morphological changes of tissue structure, cell, cytoplasm, and nucleus without hematoxylin and eosin (H&E) staining. It displayed that TPEF microscopy can well image the stratified normal gallbladder tissue, including the mucosa, the muscularis, and the serosa. The typical cancer cell, characterized by cellular and nuclear pleomorphism, enlarged nuclei, and augmented nucleolus, can be identified in histological sections without H-E staining as well. The quantitative results showed that the areas of the nucleus and the nucleolus in three types of cancerous cells were all significantly greater than those in normal gallbladder columnar epithelial cells derived from TPEF microscopic images. The studies demonstrated that TPEF microscopy has the ability to characterize tissue structures and cell morphology of gallbladder cancers differentiated from a normal gallbladder in a manner similar to traditional histological analysis. As a novel tool, it has the potential for future retrospective studies of tumor staging and migration by utilizing histological section specimens without H-E staining.

  15. Flow cytometric analysis of cellular endogenous fluorescence simultaneously with emission from exogenous fluorochromes, light scatter and absorption

    SciTech Connect

    Thorell, B.

    1981-07-01

    A cytofluorimeter capable of simultaneous 4-parameter analysis (i.e., endogenous fluorescence (NAD(P)H), exogenous fluorescent dyes, absorption and small angle scatter) is described. Analyses have been performed on isolated rat liver cells, white blood cells, bone marrow, thymus cells and yeast under different metabolic states. This allows correlations between cell function and concentration of cell substances, size, refractive indices, etc. Kinetic measurements are also possible.

  16. Absorption, fluorescence, and acid-base equilibria of rhodamines in micellar media of sodium dodecyl sulfate.

    PubMed

    Obukhova, Elena N; Mchedlov-Petrossyan, Nikolay O; Vodolazkaya, Natalya A; Patsenker, Leonid D; Doroshenko, Andrey O; Marynin, Andriy I; Krasovitskii, Boris M

    2017-01-01

    Rhodamine dyes are widely used as molecular probes in different fields of science. The aim of this paper was to ascertain to what extent the structural peculiarities of the compounds influence their absorption, emission, and acid-base properties under unified conditions. The acid-base dissociation (HR(+)⇄R+H(+)) of a series of rhodamine dyes was studied in sodium n-dodecylsulfate micellar solutions. In this media, the form R exists as a zwitterion R(±). The indices of apparent ionization constants of fifteen rhodamine cations HR(+) with different substituents in the xanthene moiety vary within the range of pKa(app)=5.04 to 5.53. The distinct dependence of emission of rhodamines bound to micelles on pH of bulk water opens the possibility of using them as fluorescent interfacial acid-base indicators.

  17. Synchrotron x-ray fluorescence and extended x-ray absorption fine structure analysis

    SciTech Connect

    Chen, J.R.; Gordon, B.M.; Hanson, A.L.; Jones, K.W.; Kraner, H.W.; Chao, E.C.T.; Minkin, J.A.

    1984-01-01

    The advent of dedicated synchrotron radiation sources has led to a significant increase in activity in many areas of science dealing with the interaction of x-rays with matter. Synchrotron radiation provides intense, linearly polarized, naturally collimated, continuously tunable photon beams, which are used to determine not only the elemental composition of a complex, polyatomic, dilute material but also the chemical form of the elements with improved accuracy. Examples of the application of synchrotron radiation include experiments in synchrotron x-ray fluorescence (SXRF) analysis and extended x-ray absorption fine structure (EXAFS) analysis. New synchrotron radiation x-ray microprobes for elemental analysis in the parts per billion range are under construction at several laboratories. 76 references, 24 figures.

  18. High resolution laser induced fluorescence Doppler velocimetry utilizing saturated absorption spectroscopy

    SciTech Connect

    Aramaki, Mitsutoshi; Ogiwara, Kohei; Etoh, Shuzo; Yoshimura, Shinji; Tanaka, Masayoshi Y.

    2009-05-15

    A high resolution laser induced fluorescence (LIF) system has been developed to measure the flow velocity field of neutral particles in an electron-cyclotron-resonance argon plasma. The flow velocity has been determined by the Doppler shift of the LIF spectrum, which is proportional to the velocity distribution function. Very high accuracy in velocity determination has been achieved by installing a saturated absorption spectroscopy unit into the LIF system, where the absolute value and scale of laser wavelength are determined by using the Lamb dip and the fringes of a Fabry-Perot interferometer. The minimum detectable flow velocity of a newly developed LIF system is {+-}2 m/s, and this performance remains unchanged in a long-time experiment. From the radial measurements of LIF spectra of argon metastable atoms, it is found that there exists an inward flow of neutral particles associated with neutral depletion.

  19. Absorption, fluorescence, and acid-base equilibria of rhodamines in micellar media of sodium dodecyl sulfate.

    PubMed

    Obukhova, Elena N; Mchedlov-Petrossyan, Nikolay O; Vodolazkaya, Natalya A; Patsenker, Leonid D; Doroshenko, Andrey O; Marynin, Andriy I; Krasovitskii, Boris M

    2017-01-01

    Rhodamine dyes are widely used as molecular probes in different fields of science. The aim of this paper was to ascertain to what extent the structural peculiarities of the compounds influence their absorption, emission, and acid-base properties under unified conditions. The acid-base dissociation (HR(+)⇄R+H(+)) of a series of rhodamine dyes was studied in sodium n-dodecylsulfate micellar solutions. In this media, the form R exists as a zwitterion R(±). The indices of apparent ionization constants of fifteen rhodamine cations HR(+) with different substituents in the xanthene moiety vary within the range of pKa(app)=5.04 to 5.53. The distinct dependence of emission of rhodamines bound to micelles on pH of bulk water opens the possibility of using them as fluorescent interfacial acid-base indicators. PMID:27423469

  20. Sub-10 ns single-shot dynamic recording in holographic polymeric medium by nonlinear absorption using excited state absorption process

    NASA Astrophysics Data System (ADS)

    Kamada, Kenji; Satoh, Kazuyuki; Tanaka, Yoshito

    2016-09-01

    Optical recording by a single shot of an 8 ns laser pulse in a dye-doped holographic polymeric medium was demonstrated with a peak power as low as 71 W at a wavelength of 402 nm. Nonlinear absorption triggered by two-photon absorption and enhanced by the succeeding process allows high-speed recording corresponding to 125 Mbps (3.4 times the recording speed of a Blu-ray Disc). The preformed holographic grating in the medium enhanced the readout signal through diffraction, resulting in a signal-to-noise ratio of about 15 dB. The recording capacity was estimated at ca. 80 GB/side for 100 µm/side by recording 20 layers, but that of over TB/side class would be possible for optical optimization. Theoretically, it can be improved to 10 TB/side for 800 µm/side by recording 400 layers.

  1. Investigating line- versus point-laser excitation for three-dimensional fluorescence imaging and tomography employing a trimodal imaging system

    NASA Astrophysics Data System (ADS)

    Cao, Liji; Peter, Jörg

    2013-06-01

    The adoption of axially oriented line illumination patterns for fluorescence excitation in small animals for fluorescence surface imaging (FSI) and fluorescence optical tomography (FOT) is being investigated. A trimodal single-photon-emission-computed-tomography/computed-tomography/optical-tomography (SPECT-CT-OT) small animal imaging system is being modified for employment of point- and line-laser excitation sources. These sources can be arbitrarily positioned around the imaged object. The line source is set to illuminate the object along its entire axial direction. Comparative evaluation of point and line illumination patterns for FSI and FOT is provided involving phantom as well as mouse data. Given the trimodal setup, CT data are used to guide the optical approaches by providing boundary information. Furthermore, FOT results are also being compared to SPECT. Results show that line-laser illumination yields a larger axial field of view (FOV) in FSI mode, hence faster data acquisition, and practically acceptable FOT reconstruction throughout the whole animal. Also, superimposed SPECT and FOT data provide additional information on similarities as well as differences in the distribution and uptake of both probe types. Fused CT data enhance further the anatomical localization of the tracer distribution in vivo. The feasibility of line-laser excitation for three-dimensional fluorescence imaging and tomography is demonstrated for initiating further research, however, not with the intention to replace one by the other.

  2. The correction fluorescence inner filter effect using a single excitation and dual-emission fiber optic probe.

    PubMed

    Zeng, Li-Hua; Wang, Cong; Wang, Tan; Li, Dao-Liang

    2016-09-21

    A significant disadvantage of fluorometry is the inner filter effect when the fluorophore concentration is high. A new simple fiber-optic probe was made to measure the concentration of fluorescent solutions. The proposed probe consists of one excitation fiber and two emission fibers. One emission fiber is parallel to the excitation fiber, and the other has a tilted angle with the excitation fiber. A numerical model was used to optimize the tilted angle and distance of the three fibers. There was a linear relationship between the fluorophore concentration and the ratio of the fluorescence intensity of the two emission fibers. Using our homemade probe, we measured Eosin Y, fluorescein and quinine sulfate solutions. The linear range of Eosin Y solution was up to 500 μM, which was approximately 7 times the range measured with a single emission probe. The results of fluorescein and quinine sulfate solutions also showed that the fluorescence intensity ratio method could correct the inner filter effect. The experimental results also indicated that the probe is robust when the excitation light fluctuates. PMID:27334633

  3. Synthesis, crystal structure and DFT studies of a dual fluorescent ketamine: Structural changes in the ground and excited states

    NASA Astrophysics Data System (ADS)

    Latha, V.; Balakrishnan, C.; Neelakantan, M. A.

    2015-07-01

    A fluorescent probe 2Z,2‧Z-3,3‧-(4,4‧-methylenebis(4,1-phenylene) bis(azanediyl))bis (1,3-diphenylprop-2-en-1-one) (L) was synthesized and characterized by IR, 1H NMR, ESI-mass, UV-visible and fluorescence spectral techniques. The single crystal analysis illustrates the existence of L in ketamine form. The crystal structure is stabilized by intramolecular and intermolecular hydrogen bonding. The thermal stability of L was studied by TG analysis. The fluorescence spectrum of L shows dual emission, and is due to excited state intramolecular proton transfer (ESIPT) process. This is supported by the high Stokes shift value. Electronic structure calculations of L in the ground and excited state have been carried out using DFT and TD-DFT at B3LYP/6-31G (d,p) level, respectively. The vibrational spectrum was computed at this level and compared with experimental values. Major orbital contributions for the electronic transitions were assigned with the help of TD-DFT. The changes in the Mulliken charge, bond lengths and bond angles between the ground and excited states of the tautomers demonstrate that twisted intramolecular charge transfer (TICT) process occurs along with ESIPT in the excited state.

  4. Attosecond XUV absorption spectroscopy of doubly excited states in helium atoms dressed by a time-delayed femtosecond infrared laser

    NASA Astrophysics Data System (ADS)

    Yang, Z. Q.; Ye, D. F.; Ding, Thomas; Pfeifer, Thomas; Fu, L. B.

    2015-01-01

    In the present paper, we investigate the time-resolved transient absorption spectroscopy of doubly excited states of helium atoms by solving the time-dependent two-electron Schrödinger equation numerically based on a one-dimensional model. The helium atoms are subjected to an extreme ultraviolet (XUV) attosecond pulse and a time-delayed infrared (IR) few-cycle laser pulse. A superposition of doubly excited states populated by the XUV pulse is identified, which interferes with the direct ionization pathway leading to Fano resonance profiles in the photoabsorption spectrum. In the presence of an IR laser, however, the Fano line profiles are strongly modified: A shifting, splitting, and broadening of the original absorption lines is observed when the XUV attosecond pulse and infrared few-cycle laser pulse overlap in time, which is in good agreement with recent experimental results. At certain time delays, we observe symmetric Lorentz, inverted Fano profiles, and even negative absorption cross sections indicating that the XUV light can be amplified during the interaction with atoms. We further prove that the above pictures are general for different doubly excited states by suitably varying the frequency of the IR field to coherently couple the corresponding states.

  5. pH-Induced changes in electronic absorption and fluorescence spectra of phenazine derivatives

    NASA Astrophysics Data System (ADS)

    Ryazanova, O. A.; Voloshin, I. M.; Makitruk, V. L.; Zozulya, V. N.; Karachevtsev, V. A.

    2007-04-01

    The visible electronic absorption and fluorescence spectra as well as fluorescence polarization degrees of imidazo-[4,5-d]-phenazine (F1), 2-methylimidazo-[4,5-d]-phenazine (F2), 2-trifluoridemethylimidazo-[4,5-d]-phenazine (F3), 1,2,3-triazole-[4,5-d]-phenazine (F4) and their glycosides, imidazo-[4,5-d]-phenazine-N1-β- D-ribofuranoside (F1rib), 1,2,3-triazole-[4,5-d]-phenazine-N1-β- D-glucopyranoside (F4gl), were investigated in aqueous buffered solutions over the pH range of 0-12, where the spectral transformations were found to be reversible. The effects of protonation and deprotonation on spectral properties of these dyes were studied. We have determined the ranges of pH, where individual ionic species are predominant. In aqueous buffered solutions the fluorescence was found only for neutral species of F1, F1rib, F2, and F4gl dyes, whereas for the ionic forms of these dyes, as well as for F3 and F4 ones, the fluorescence has not been detected. The concentrational deprotonation p Ka values were evaluated from experimental data. It was shown that donor-acceptor properties of the substituent group in the second position of the pentagonal ring substantially affect the values of the deprotonation constants and the character of protonation for chromophore. The substitution of a hydrogen atom in the NH-group by the sugar residue blocks the formation of the anionic species, and results in enhancement of the dye emission intensity. The steep emission dependence for F1 and F1rib over pH range of 0-7 with intensities ratio of IpH 7/ IpH 1 = 60 allows us to propose them as possible indicator dyes in luminescence based pH sensors for investigation of processes accompanied by acidification, e.g. as gastric pH-sensors. A comparative analysis of the studied dyes has shown that F4gl is the most promising compound to be used as a fluorescent probe for investigation of molecular hybridization of nucleic acids.

  6. Electron-acceptor-dependent light absorption, excited-state relaxation, and charge generation in triphenylamine dye-sensitized solar cells.

    PubMed

    Li, Renzhi; Zhang, Min; Yan, Cancan; Yao, Zhaoyang; Zhang, Jing; Wang, Peng

    2015-01-01

    By choosing a simple triphenylamine electron donor, we herein compare the influence of electron acceptors benzothiadiazole benzoic acid (BTBA) and cyanoacrylic acid (CA), on energy levels, light absorption, and dynamics of excited-state evolution and electron injection. DFT and time-dependent DFT calculations disclosed remarkable intramolecular conformational changes for the excited states of these two donor-acceptor dyes. Photoinduced dihedral angle variation occurs to the triphenylamine unit in the CA dye and backbone planarization happens to conjugated aromatic blocks in the BTBA dye. Femtosecond spectroscopic measurements suggested the crucial role of having a long excited-state lifetime in maintaining a high electron-injection yield because a reduced driving force for a low energy-gap dye can result in slower electron-injection dynamics.

  7. Real-time monitoring of river water quality using in-line continuous acquisition of fluorescence excitation and emission matrices

    NASA Astrophysics Data System (ADS)

    Carstea, E.; Baker, A.; Johnson, R.; Reynolds, D. M.

    2009-12-01

    In-line fluorescence EEM monitoring has been performed over an eleven-day period for Bournbrook River, Birmingham, UK. River water was diverted to a portable laboratory via a continuous flow pump and filter system. Fluorescence excitation-emission matrices data was recorded every 3 minutes using a flow cell (1cm pathlength) coupled to a fiber optic probe. This real-time fluorescence EEM data (Excitation, 225-400 nm at 5 nm steps, emission, 280-500 nm at 2 nm steps) was collected 'in-line'and directly compared with the spectrophotometric properties and physical and chemical parameters of river water samples collected off-line at known time intervals. Over the monitoring period, minor pollution pulses from cross connections were detected and identified hourly along with a random diesel pollution event. This work addresses the practicalities of measuring and detecting fluorescence EEM in the field and discusses the potential of this technological approach for further understanding important hydrological and biogeochemical processes. Problems associated with fouling and system failure are also reported. Example of the data generated from the continuous fluorescence EEM monitoring.

  8. One-Dimensional Fluorescent Silicon Nanorods Featuring Ultrahigh Photostability, Favorable Biocompatibility, and Excitation Wavelength-Dependent Emission Spectra.

    PubMed

    Song, Bin; Zhong, Yiling; Wu, Sicong; Chu, Binbin; Su, Yuanyuan; He, Yao

    2016-04-13

    We herein report a kind of one-dimensional biocompatible fluorescent silicon nanorods (SiNRs) with tunable lengths ranging ∼100-250 nm, which can be facilely prepared through one-pot microwave synthesis. In addition to the strong fluorescence (quantum yield value: ∼15%) and negligible toxicity, the resultant SiNRs exhibit excitation wavelength-dependent photoluminescence whose maximum emission wavelength ranges from ∼450 to ∼600 nm under serial excitation wavelengths from 390 to 560 nm, providing feasibility for multicolor biological imaging. More significantly, the SiNRs are ultrahighly photostable, preserving strong and nearly unchanged fluorescence under 400 min high-power UV irradiation, which is in sharp contrast to severe fluorescence quenching of organic dyes (e.g., FITC) or II-VI quantum dots (QDs) (e.g., CdTe QDs and CdSe/ZnS QDs) within 15 or 160 min UV treatment under the same experiment conditions, respectively. Taking advantage of these attractive merits, we further exploit the SiNRs as a novel type of color converters for the construction of white light-emitting diodes (LED), which is the first proof-of-concept demonstration of LED device fabricated using the one-dimensional fluorescent silicon nanostructures. PMID:27010956

  9. Determination of mercury distribution inside spent compact fluorescent lamps by atomic absorption spectrometry

    SciTech Connect

    Rey-Raap, Natalia

    2012-05-15

    Highlights: Black-Right-Pointing-Pointer New treatments for CFL are required considering the aim of Directive 202/96/CE. Black-Right-Pointing-Pointer It is shown that most of the mercury introduced into a CFL is in the phosphor powder. Black-Right-Pointing-Pointer Experimental conditions for microwave-assisted sample digestion followed by AAS measurements are described. Black-Right-Pointing-Pointer By washing the glass it is possible to reduce the concentration below legal limits. - Abstract: In this study, spent compact fluorescent lamps were characterized to determine the distribution of mercury. The procedure used in this research allowed mercury to be extracted in the vapor phase, from the phosphor powder, and the glass matrix. Mercury concentration in the three phases was determined by the method known as cold vapor atomic absorption spectrometry. Median values obtained in the study showed that a compact fluorescent lamp contained 24.52 {+-} 0.4 ppb of mercury in the vapor phase, 204.16 {+-} 8.9 ppb of mercury in the phosphor powder, and 18.74 {+-} 0.5 ppb of mercury in the glass matrix. There are differences in mercury concentration between the lamps since the year of manufacture or the hours of operation affect both mercury content and its distribution. The 85.76% of the mercury introduced into a compact fluorescent lamp becomes a component of the phosphor powder, while more than 13.66% is diffused through the glass matrix. By washing and eliminating all phosphor powder attached to the glass surface it is possible to classified the glass as a non-hazardous waste.

  10. Study of nonlinear optical absorption properties of Sb2Se3 nanoparticles in the nanosecond and femtosecond excitation regime

    NASA Astrophysics Data System (ADS)

    Molli, Muralikrishna; Pradhan, Prabin; Dutta, Devarun; Jayaraman, Aditya; Bhat Kademane, Abhijit; Muthukumar, V. Sai; Kamisetti, Venkataramaniah; Philip, Reji

    2016-05-01

    In this work, we report for the first time, the nonlinear optical absorption properties of antimony selenide (Sb2Se3) nanoparticles synthesized through solvothermal route. X-ray diffraction results revealed the crystalline nature of the nanoparticles. Electron microscopy studies revealed that the nanoparticles are in the range of 10-40 nm. Elemental analysis was performed using EDAX. The nanosecond optical limiting effect was characterized by using fluence-dependent transmittance measurements with 15-ns laser pulses at 532 and 1064 nm excitation wavelengths. Mechanistically, effective two-photon (2PA) absorption and nonlinear scattering processes were the dominant nonlinear processes at both the wavelengths. At 800 nm excitation in the femtosecond regime (100 fs), the nonlinear optical absorption was found to be a three-photon (3PA) process. Both 2PA and 3PA processes were explained using the band structure and density of states of Sb2Se3 obtained using density functional theory. These nanoparticles exhibit strong intensity-dependent nonlinear optical absorption and hence could be considered to have optical power-limiting applications in the visible range.

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

    PubMed

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

    2012-11-01

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

  12. A new visibly-excited fluorescent component in latent fingerprint residue induced by gaseous electrical discharge.

    PubMed

    Davies, L M; Jones, N E; Brennan, J S; Bramble, S K

    2000-11-01

    A technique that exposes fingerprint residue to a gaseous electrical discharge in nitrogen followed by treatment with ammonium hydrogen carbonate vapors to produce fluorescence is investigated. Particular attention is made to fluorescence observed via laser illumination at 514 nm. Insight into the nature of the fluorescent components is achieved through the use of thin-layer chromatography (TLC) of fingerprint residue. Results reported indicate the fluorescence observed is from previously non-fluorescent fractions of the fingerprint residue, and TLC results point towards lipid derivatives as a possible source of the fluorescence. PMID:11110185

  13. The effects of pH and surfactants on the absorption and fluorescence properties of ochratoxin A and zearalenone.

    PubMed

    Li, Taihua; Kim, Bo Bae; Ha, Tae Hwan; Shin, Yong-Beom; Kim, Min-Gon

    2015-11-01

    The pH and surfactant dependencies of the absorption and fluorescence properties of ochratoxin A (OTA) and zearalenone (ZEN), the main mycotoxins found as contaminants in foods and feeds, were evaluated. Three surfactants with different ionic properties were investigated, namely sodium dodecyl sulfate (SDS, anionic), Tween 20 (nonionic) and hexadecyltrimethylammonium bromide (CTAB, cationic). The results show that the effects of pH on the absorption wavelength maxima and fluorescence efficiencies of the mycotoxins, which are a consequence of the presence of acidic phenol and/or carboxyl containing fluorophores, are dependent on the ionic nature of the added surfactants. Specifically, the fluorescence responses to pH changes of OTA and ZEN are similar in the presence or absence of Tween 20 and SDS. By contrast, the pH-dependent fluorescence properties of these mycotoxins are altered when CTAB is present in the solutions. Moreover, unlike OTA, ZEN in aqueous solution displays almost no fluorescence. However, fluorescence enhancement takes place when surfactants are present in aqueous solutions of this mycotoxin. The results of this study demonstrate that the different microenvironments, present in the organized micellar systems created by the individual surfactants, can be potentially employed to modulate the sensitivities and selectivities of the fluorescence detection of OTA or ZEN.

  14. Excitation-resolved wide-field fluorescence imaging of indocyanine green visualizes the microenvironment properties in vivo via solvatochromic shift (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Cho, Jaedu; Kim, Chang-Seok; Gulsen, Gultekin

    2016-03-01

    Near-infrared fluorescence imaging (NIRF) is a powerful wide-field optical imaging tool that has a potential to visualize molecular-specific exogenous fluorescence agents, such as FDA approved Indocyanine Green (ICG), in thick tissue. Indeed, ICG is sensitive to biochemical environment such that it can be used to detect micro- or macroscopic environmental changes in tissue by solvatochromic shift that is defined by the dependence of absorption and emission spectra with the solvent polarity. For example, dimethyl sulfoxide (DMSO) is a very powerful drug carrier that can penetrate biological barriers such as the skin, the membranes, and the blood-brain-barrier. In presence of DMSO, ICG in tissue shows the excitation blue shift. However, NIRF imaging of microenvironment dependent changes of ICG has been challenging for the following reasons. First, the Stoke's shift of ICG is too small to separate the excitation and emission spectra easily. Second, the solvatochromic shift of ICG is too small to be detected by conventional NIRF techniques. Last but not least, the multiple scattering in tissue degrades not only the spatial information but also the spectral contents by the red-shift. We developed a wavelength-swept laser-based NIRF system that can resolve the excitation shift of ICG in tissue such that DMSO can be indirectly visualized. We plan to conduct an in-vivo lymph-node drug-delivery study in a mouse model to show feasibility of the indirect imaging of the drug-carrier with the wavelength-swept-laser based NIRF system.

  15. Laser excited analytical atomic and ionic fluorescence in flames, furnaces and inductively coupled plasmas—I. General considerations

    NASA Astrophysics Data System (ADS)

    Omenetto, N.; Human, H. G. C.

    Several important parameters for the analytical use of laser excited fluorescence spectrometry in flames, graphite furnaces and inductively coupled plasmas are discussed in some detail. These parameters include the laser characteristics such as peak power, pulse duration, spectral bandwidth and repetition rate, the choice of the excitation line, the optical arrangement and the detection system, this last one centred on the widespread use of the boxcar averager. It is shown that, if the ultimate sensitivity is the goal to be achieved, then the choice must be the electrothermal atomization. However, even for flames and inductively coupled plasmas, excellent results are possible provided that: (i) the laser system allows complete spectral coverage in the ultraviolet: (ii) saturation of the fluorescence signal can be approached over a large sample volume; and (iii) the gated detection parameters and the laser repetition frequency are optimized with respect to each other so as to reach the maximum signal-to-noise ratio.

  16. Fluorescence excitation and imaging of single molecules near dielectric-coated and bare surfaces: a theoretical study.

    PubMed

    Axelrod, Daniel

    2012-08-01

    Microscopic fluorescent samples of interest to cell and molecular biology are commonly embedded in an aqueous medium near a solid surface that is coated with a thin film such as a lipid multilayer, collagen, acrylamide, or a cell wall. Both excitation and emission of fluorescent single molecules near film-coated surfaces are strongly affected by the proximity of the coated surface, the film thickness, its refractive index and the fluorophore's orientation. For total internal reflection excitation, multiple reflections in the film can lead to resonance peaks in the evanescent intensity versus incidence angle curve. For emission, multiple reflections arising from the fluorophore's near field emission can create a distinct intensity pattern in both the back focal plane and the image plane of a high aperture objective. This theoretical analysis discusses how these features can be used to report film thickness and refractive index, and fluorophore axial position and orientation. PMID:22612666

  17. Standoff detection of biological agents using laser induced fluorescence-a comparison of 294 nm and 355 nm excitation wavelengths.

    PubMed

    Farsund, Oystein; Rustad, Gunnar; Skogan, Gunnar

    2012-11-01

    Standoff detection measuring the fluorescence spectra of seven different biological agents excited by 294 nm as well as 355 nm wavelength laser pulses has been undertaken. The biological warfare agent simulants were released in a semi-closed aerosol chamber at 210 m standoff distance and excited by light at either of the two wavelengths using the same instrument. Significant differences in several of the agents' fluorescence response were seen at the two wavelengths. The anthrax simulants' fluorescence responses were almost an order of magnitude stronger at the shorter wavelength excitation. However, most importantly, the fluorescence spectra were significantly more dissimilar at 294 nm than at 355 nm excitation with ~7 nm spectral resolution. This indicates that classification of the substances should be possible with a lower error rate for standoff detection using 294 nm rather than 355 nm excitation wavelength, or even better, utilizing both. PMID:23162732

  18. Microwave-assisted synthesis of water-soluble, fluorescent gold nanoclusters capped with small organic molecules and a revealing fluorescence and X-ray absorption study

    NASA Astrophysics Data System (ADS)

    Helmbrecht, C.; Lützenkirchen-Hecht, D.; Frank, W.

    2015-03-01

    Colourless solutions of blue light-emitting, water-soluble gold nanoclusters (AuNC) were synthesized from gold colloids under microwave irradiation using small organic molecules as ligands. Stabilized by 1,3,5-triaza-7-phosphaadamantane (TPA) or l-glutamine (GLU), fluorescence quantum yields up to 5% were obtained. AuNC are considered to be very promising for biological labelling, optoelectronic devices and light-emitting materials but the structure-property relationships have still not been fully clarified. To expand the knowledge about the AuNC apart from their fluorescent properties they were studied by X-ray absorption spectroscopy elucidating the oxidation state of the nanoclusters' gold atoms. Based on curve fitting of the XANES spectra in comparison to several gold references, optically transparent fluorescent AuNC are predicted to be ligand-stabilized Au5+ species. Additionally, their near edge structure compared with analogous results of polynuclear clusters known from the literature discloses an increasing intensity of the feature close to the absorption edge with decreasing cluster size. As a result, a linear relationship between the cluster size and the X-ray absorption coefficient can be established for the first time.Colourless solutions of blue light-emitting, water-soluble gold nanoclusters (AuNC) were synthesized from gold colloids under microwave irradiation using small organic molecules as ligands. Stabilized by 1,3,5-triaza-7-phosphaadamantane (TPA) or l-glutamine (GLU), fluorescence quantum yields up to 5% were obtained. AuNC are considered to be very promising for biological labelling, optoelectronic devices and light-emitting materials but the structure-property relationships have still not been fully clarified. To expand the knowledge about the AuNC apart from their fluorescent properties they were studied by X-ray absorption spectroscopy elucidating the oxidation state of the nanoclusters' gold atoms. Based on curve fitting of the XANES

  19. Biocompatible photoresistant far-red emitting, fluorescent polymer probes, with near-infrared two-photon absorption, for living cell and zebrafish embryo imaging.

    PubMed

    Adjili, Salim; Favier, Arnaud; Fargier, Guillaume; Thomas, Audrey; Massin, Julien; Monier, Karine; Favard, Cyril; Vanbelle, Christophe; Bruneau, Sylvia; Peyriéras, Nadine; Andraud, Chantal; Muriaux, Delphine; Charreyre, Marie-Thérèse

    2015-04-01

    Exogenous probes with far-red or near-infrared (NIR) two-photon absorption and fluorescence emission are highly desirable for deep tissue imaging while limiting autofluorescence. However, molecular probes exhibiting such properties are often hydrophobic. As an attractive alternative, we synthesized water-soluble polymer probes carrying multiple far-red fluorophores and demonstrated here their potential for live cell and zebrafish embryo imaging. First, at concentrations up to 10 μm, these polymer probes were not cytotoxic. They could efficiently label living HeLa cells, T lymphocytes and neurons at an optimal concentration of 0.5 μm. Moreover, they exhibited a high resistance to photobleaching in usual microscopy conditions. In addition, these polymer probes could be successfully used for in toto labeling and in vivo two-photon microscopy imaging of developing zebrafish embryos, with remarkable properties in terms of biocompatibility, internalization, diffusion, stability and wavelength emission range. The near-infrared two-photon absorption peak at 910 nm is particularly interesting since it does not excite the zebrafish endogenous fluorescence and is likely to enable long-term time-lapse imaging with limited photodamage.

  20. Unraveling the exciton quenching mechanism of quantum dots on antimony-doped SnO₂ films by transient absorption and single dot fluorescence spectroscopy.

    PubMed

    Song, Nianhui; Zhu, Haiming; Liu, Zheng; Huang, Zhuangqun; Wu, David; Lian, Tianquan

    2013-02-26

    Integrating quantum dots (QDs) into modern optoelectronic devices requires an understanding of how a transparent conducting substrate affects the properties of QDs, especially their excited-state dynamics. Here, the exciton quenching dynamics of core/multishell (CdSe/CdS(3ML)ZnCdS(2ML)ZnS(2ML)) quantum dots deposited on glass, tin oxide (SnO₂), and antimony (Sb)-doped tin oxide (ATO) films are studied by transient absorption and single QD fluorescence spectroscopic methods. By comparing ensemble-averaged fluorescence decay and transient absorption kinetics, we show that, for QDs on SnO₂, the exciton is quenched by electron transfer from the QD to SnO₂. At the QD-ATO interface, much faster exciton quenching rates are observed and attributed to fast Auger recombination in charged QDs formed by Fermi level equilibration between the QD and n-doped ATO. Single QDs on SnO₂ and ATO show similar blinking dynamics with correlated fluctuations of emission intensities and lifetimes. Compared to QDs on SnO₂, QDs on ATO films show larger variation of average exciton quenching rates, which is attributed to a broad distribution of the number of charges and nature of charging sites on the QD surface.

  1. Projection imaging of photon beams using Čerenkov-excited fluorescence

    PubMed Central

    Glaser, Adam K.; Davis, Scott C.; Voigt, William H.A.; Zhang, Rongxiao; Pogue, Brian W.; Gladstone, David J.

    2013-01-01

    Full 3D beam profiling and quality assurance (QA) of therapeutic megavoltage linear accelerator (LINAC) x-ray photon beams is not routinely performed due to the slow point-by-point measurement nature of conventional scanning ionization chamber systems. In this study we explore a novel optical-based dose imaging approach using a standard commercial camera, water tank, and fluorescent dye, which when excited by the Čerenkov emission induced by the radiation beam, allows 2D projection imaging in a fast timeframe, potentially leading towards 3D tomographic beam profiling. Detailed analysis was done to optimize the imaging parameters in the experimental setup. The results demonstrate that the captured images are linear with delivered dose, independent of dose rate, and comparison of experimentally captured images to a reference dose distribution for a 4×4 cm 6 MV x-ray photon beam yielded results with improved accuracy over a previous study which used direct imaging and Monte Carlo calibration of the Čerenkov emission itself. The agreement with the reference dose distribution was within 1-2% in the lateral direction, and ± 3 % in the depth direction. The study was restricted to single 2D image projection, with the eventual goal of creating full 3D profiles after tomographic reconstruction from multiple projections. Given the increasingly complex advances in radiation therapy, and the increased emphasis on patient-specific treatment plans, further refinement of the technique could prove to be an important tool for fast and robust QA of x-ray photon LINAC beams. PMID:23318469

  2. Sizeable red-shift of absorption and fluorescence of subporphyrazine induced by peripheral push and pull substitution.

    PubMed

    Liang, Xu; Shimizu, Soji; Kobayashi, Nagao

    2014-11-18

    Peripheral substitution with electron-donating (push) and electron-withdrawing (pull) substituents caused a sizeable red-shift of the Q band absorption and fluorescence of subporphyrazine, and the red-shift was controlled by the push substituents. Control of the chromophore symmetry and inherent molecular chirality arising from the pattern of substitution were also investigated.

  3. Myofibrillogenesis in live neonatal cardiomyocytes observed with hybrid two-photon excitation fluorescence-second harmonic generation microscopy

    PubMed Central

    Liu, Honghai; Qin, Wan; Shao, Yonghong; Ma, Zhen; Ye, Tong; Borg, Tom; Gao, Bruce Z.

    2011-01-01

    We developed a hybrid two-photon excitation fluorescence-second harmonic generation (TPEF-SHG) imaging system with an on-stage incubator for long-term live-cell imaging. Using the imaging system, we observed the addition of new sarcomeres during myofibrillogenesis while a cardiomyocyte was spreading on the substrate. The results suggest that the TPEF-SHG imaging system with an on-stage incubator is an effective tool for investigation of dynamic myofibrillogenesis. PMID:22191929

  4. Laser-excited fluorescence and electron-spin resonance of Er3+ in polycrystalline AlCl3

    NASA Astrophysics Data System (ADS)

    Ceotto, G.; Pires, M. A.; Sanjurjo, J. A.; Rettori, C.; Barberis, G. E.

    1990-07-01

    The green fluorescence transitions among the levels corresponding to the 4S3/2 and 4I15/2 configurations of Er3+ diluted in AlCl3 have been measured using laser excitation. The data allow us to determine the crystalline-field splittings of these levels and, in turn, the spin-Hamiltonian parameters. The electron-paramagnetic-resonance spectrum observed at low temperatures is in good agreement with that expected from these parameters.

  5. The ultraviolet continuum and the fluorescent H2 lines in low-excitation Herbig-Haro objects

    NASA Technical Reports Server (NTRS)

    Boehm, K. H.; Scott, D. M.; Solf, J.

    1991-01-01

    Long-exposure IUE observations are used here to determine the spatial distribution of the fluorescent H2 line emission in the low-excitation objects HH 43 and HH 47. In HH 43, the spatial distribution of the fluorescent H2 line emission has a width equal to or smaller than the width of the point-spread function of IUE, while both the short-wavelength UV continuum and the optical forbidden S II lines show distributions which are wider by more than a factor of two. In HH 47, the results are qualitatively analogous, but the differences between the fluorescent HH 23 emission distribution on the one hand, and the UV continuum and optical forbidden S II distribution on the other, are smaller. These distributions indicate that the appropriate conditions for the formation of fluorscent H2 lines are fulfilled only in rather narrow regions for these low-excitation objects. A continuous energy distribution in HH 43 and HH 47 is found which differs only slightly from that in the high-excitation objects HH 1 and HH 2.

  6. Intercomparison of OH Radical Measurements by Long-Path Absorption and Laser Induced Fluorescence in the Atmosphere Simulation Chamber SAPHIR

    NASA Astrophysics Data System (ADS)

    Dorn, H.-P.; Brauers, T.; Greif, J.; Häseler, R.; Hofzumahaus, A.; Holland, F.; Rupp, L.

    2003-04-01

    A striking advantage of the SAPHIR chamber is the availability of two spectroscopic detection instruments for OH radicals: Laser-Induced Fluorescence Spectroscopy (LIF) and Long-Path Differential Optical Laser Absorption Spectroscopy (DOAS). Both instruments have already been compared in 1994 during the field measurement campaign POPCORN. They agreed well with a correlation coefficient of r=0.90 and a weighted linear fit with a slope of 1.09 +- 0.12. However, OH measurements in the simulation chamber differ significantly from measurements in ambient air. While DOAS measures OH as an integral value along the central longitudinal axis of SAPHIR, LIF samples the air locally and close (2 cm) to the floor of the chamber. Thus, the LIF measurements might be possibly affected by local concentration gradients caused by insufficient mixing of the chamber air or by deposition to the wall. On the other hand, if turbulent mixing of the chamber air is weak and high concentrations of ozone are used in experiments, the DOAS instrument might be subject to artificial formation of OH radicals in the air volume which is illuminated by the detection laser. This interference results from laser induced photolysis of ozone and the subsequent reaction of water vapor with the excited oxygen atoms formed. Thus it is of decisive importance to compare OH measurements from both instruments in order to investigate potential disturbing effects due to the specific sampling properties of both instruments within SAPHIR. We report on OH measurements accomplished simultaneously with both instruments using different trace gas compositions and experimental conditions.

  7. Probing Cu(I) in homogeneous catalysis using high-energy-resolution fluorescence-detected X-ray absorption spectroscopy.

    PubMed

    Walroth, Richard C; Uebler, Jacob W H; Lancaster, Kyle M

    2015-06-18

    Metal-to-ligand charge transfer excitations in Cu(I) X-ray absorption spectra are introduced as spectroscopic handles for the characterization of species in homogeneous catalytic reaction mixtures. Analysis is supported by correlation of a spectral library to calculations and to complementary spectroscopic parameters.

  8. Study of nonlinear optical absorption properties of V2O5 nanoparticles in the femtosecond excitation regime

    NASA Astrophysics Data System (ADS)

    Molli, Muralikrishna; Bhat Kademane, Abhijit; Pradhan, Prabin; Sai Muthukumar, V.

    2016-08-01

    In this work, we report for the first time, the nonlinear optical absorption properties of vanadium pentoxide (V2O5) nanoparticles in the femtosecond excitation regime. V2O5 nanoparticles were synthesized through solution combustion technique. The as-synthesized samples were further characterized using XRD, FESEM, EDAX, TEM and UV-visible spectroscopy. X-ray diffraction results revealed the crystalline nature of the nanoparticles. Electron microscopy studies showed the size of the nanoparticles to be ~200 nm. Open-aperture z-scan technique was employed to study the nonlinear optical absorption behavior of the synthesized samples using a 100-fs laser pulses at 800 nm from a regeneratively amplified Ti: sapphire laser. The mechanism of nonlinear absorption was found to be a three-photon absorption process which was explained using the density of states of V2O5 obtained using density functional theory. These nanoparticles exhibit strong intensity-dependent nonlinear optical absorption and hence could be considered for optical-power-limiting applications.

  9. A benzo-15-crown-5-modifying ratiometric-absorption and fluorescent OFF-ON chemosensor for Cu2 +

    NASA Astrophysics Data System (ADS)

    Chen, Yuting; Wang, Xinxin; Wang, Kaili; Zhang, Xiuling

    2016-05-01

    One new benzo-15-crown-5-modifying fluorene Schiff base (FBC), together with the Cdbnd N-linked fluorene-3,4-dimethoxybenzene (FBDMO) and fluorene-benzene (FB) references, has been designed and facilely synthesized. The binding of Cu2 + with nitrogen atom of Cdbnd N moiety in these three compounds can inhibit the photo-induced electronic transition process and induce the ratiometric-absorption and fluorescent OFF-ON response to Cu2 +. Whereas the employment of benzo-15-crown-5 moiety in FBC as additional binding platform for Cu2 + not only amplifies the fluorescent enhancement of FBCvia preventing the isomerization of Cdbnd N moiety, but also endows this compound high selectivity and rapid response towards Cu2 + over the references FB and FBDMO. These results render FBC highly sensitive ratiometric-absorption and fluorescent OFF-ON detecting potential for Cu2 + with the detection limit of 3.91 × 10- 6 M.

  10. Picosecond fluorescence of cryptomonad biliproteins. Effects of excitation intensity and the fluorescence decay times of phycocyanin 612, phycocyanin 645, and phycoerythrin 545.

    PubMed Central

    Guard-Friar, D; MacColl, R; Berns, D S; Wittmershaus, B; Knox, R S

    1985-01-01

    The fluorescence of purified biliproteins (phycocyanin 645, phycocyanin 612, and phycoerythrin 545) from three cryptomonads, Chroomonas species, Hemiselmis virescens, and Rhodomonas lens, and C-phycocyanin from Anacystis nidulans has been time resolved in the picosecond region with a streak camera system having less than or equal to 2-ps jitter. The fluorescence lifetimes of phycocyanins from Chroomonas species and Hemiselmis virescens are 1.5 +/- 0.2 ns and 2.3 +/- 0.2 ns, respectively, regardless of the fluence of the 30 ps, 532-nm excitation pulse. (Fluence [or photons/cm2] = f intensity [photons/cm2s]dt.). In contrast, that of C-phycocyanin is 2.3 +/- 0.2 ns when the excitation fluence is 8.2 X 10(11) photons/cm2 and decreases to a decay approximated by an exponential decay time of 0.65 +/- 0.1 ns at 7.2 X 10(16) photons/cm2. The cryptomonad phycoerythrin fluorescence decay lifetime is also dependent on intensity, having a decay time of 1.5 +/- 0.1 ns at low fluences and becoming clearly biphasic at higher fluences (greater than 10(15) photons/cm2). We interpret the shortening of decay times for C-phycocyanin and phycoerythrin 545 in terms of exciton annihilation, and have discussed the applicability of exciton annihilation theories to the high fluence effects. PMID:3926017

  11. Comparative analysis of the vibrational structure of the absorption spectra of acrolein in the excited ( S 1) electronic state

    NASA Astrophysics Data System (ADS)

    Koroleva, L. A.; Tyulin, V. I.; Matveev, V. K.; Pentin, Yu. A.

    2012-04-01

    The assignments of absorption bands of the vibrational structure of the UV spectrum are compared with the assignments of bands obtained by the CRDS method in a supersonic jet from the time of laser radiation damping for the trans isomer of acrolein in the excited ( S 1) electronic state. The ν00 trans = 25861 cm-1 values and fundamental frequencies, including torsional vibration frequency, obtained by the two methods were found to coincide in the excited electronic state ( S 1) for this isomer. The assignments of several absorption bands of the vibrational structure of the spectrum obtained by the CRDS method were changed. Changes in the assignment of (0-v') transition bands of the torsional vibration of the trans isomer in the Deslandres table from the ν00 trans trans origin allowed the table to be extended to high quantum numbers v'. The torsional vibration frequencies up to v' = 5 were found to be close to the frequencies found by analyzing the vibrational structure of the UV spectrum and calculated quantum-mechanically. The coincidence of the barrier to internal rotation (the cis-trans transition) in the one-dimensional model with that calculated quantum-mechanically using the two-dimensional model corresponds to a planar structure of the acrolein molecule in the excited ( S 1) electronic state.

  12. Calculation and interpretation of vibronic absorption and fluorescence spectra of the first electronic nπ* transitions of pyridine and pyrimidine

    NASA Astrophysics Data System (ADS)

    Ten, G. N.; Kadrov, D. M.; Berezin, M. K.; Baranov, V. I.

    2014-11-01

    We have calculated vibronic spectra of the first electronic nπ* transitions of pyridine and pyrimidine in the isolated state using the DFT method in the Franck-Condon approximation. Vibrational spectra for the ground and excited states have been calculated in the anharmonic approximation, which allowed us to refine the assignment of normal vibrations of pyridine and pyrimidine. We have done a complete interpretation of the vibrational structure of the absorption and fluorescence spectra of pyridine and pyrimidine. It has been shown that Fermi resonances between fundamental and combination vibrations and overtones 12 and 16 b + 4, 6 a and 2 × 16 b affect the formation of the vibrational structure of electronic spectra of pyrimidine. Good agreement between calculated and experimental spectra confirms the correctness of the models of the two molecules in their ground and excited states, which makes it possible to use the models in further investigations of various properties of these molecules in electronically excited states, e.g., tautomerism of pyrimidine bases of nucleic acids.

  13. Effect of planar dielectric interfaces on fluorescence emission and detection. Evanescent excitation with high-aperture collection.

    PubMed Central

    Burghardt, T P; Thompson, N L

    1984-01-01

    We consider the effect of planar dielectric interfaces (e.g., solid/liquid) on the fluorescence emission of nearby probes. First, we derive an integral expression for the electric field radiated by an oscillating electric dipole when it is close to a dielectric interface. The electric field depends on the refractive indices of the interface, the orientation of the dipole, the distance from the dipole to the interface, and the position of observation. We numerically calculate the electric field intensity for a dipole on an interface, as a function of observation position. These results are applicable to fluorescent molecules excited by the evanescent field of a totally internally reflected laser beam and thus very close to a solid/liquid interface. Next, we derive an integral expression for the electric field radiated when a second dielectric interface is also close to the fluorescent molecule. We numerically calculate this intensity as observed through the second interface. These results are useful when the fluorescence is collected by a high-aperture microscope objective. Finally, we define and calculate a "dichroic factor," which describes the efficiency of collection, in the two-interface system, of polarized fluorescence. The limit when the first interface is removed is applicable for any high-aperture collection of polarized or unpolarized fluorescence. The limit when the second interface is removed has application in the collection of fluorescence with any aperture from molecules close to a dielectric interface. The results of this paper are required for the interpretation of order parameter measurements on fluorescent probes in supported phospholipid monolayers (Thompson, N.L., H. M. McConnell, and T. P. Burghardt, 1984, Biophys. J., 46:739-747). PMID:6518253

  14. Electrochemical immobilization of Fluorescent labelled probe molecules on a FTO surface for affinity detection based on photo-excited current

    NASA Astrophysics Data System (ADS)

    Haruyama, Tetsuya; Wakabayashi, Ryo; Cho, Takeshi; Matsuyama, Sho-taro

    2011-10-01

    Photo-excited current can be generated at a molecular interface between a photo-excited molecules and a semi-conductive material in appropriate condition. The system has been recognized for promoting photo-energy devices such as an organic dye sensitized solar-cell. The photo-current generated reactions are totally dependent on the interfacial energy reactions, which are in a highly fluctuated interfacial environment. The authors investigated the photo-excited current reaction to develop a smart affinity detection method. However, in order to perform both an affinity reaction and a photo-excited current reaction at a molecular interface, ordered fabrications of the functional (affinity, photo-excitation, etc.) molecules layer on a semi-conductive surface is required. In the present research, we would like to present the fabrication and functional performance of photo-excited current-based affinity assay device and its application for detection of endocrine disrupting chemicals. On the FTO surface, fluorescent pigment labelled affinity peptide was immobilized through the EC tag (electrochemical-tag) method. The modified FTO produced a current when it was irradiated with diode laser light. However, the photo current decreased drastically when estrogen (ES) coexisted in the reaction solution. In this case, immobilized affinity probe molecules formed a complex with ES and estrogen receptor (ER). The result strongly suggests that the photo-excited current transduction between probe molecule-labelled cyanine pigment and the FTO surface was partly inhibited by a complex that formed at the affinity oligo-peptide region in a probe molecule on the FTO electrode. The bound bulky complex may act as an impediment to perform smooth transduction of photo-excited current in the molecular interface. The present system is new type of photo-reaction-based analysis. This system can be used to perform simple high-sensitive homogeneous assays.

  15. Atomic resolution mapping of the excited-state electronic structure of Cu2O with time-resolved x-ray absorption spectroscopy

    SciTech Connect

    Hillyard, P. W.; Kuchibhatla, S. V. N. T.; Glover, T. E.; Hertlein, M. P.; Huse, Nils; Nachimuthu, P.; Saraf, L. V.; Thevuthasan, S.; Gaffney, K. J.

    2010-05-02

    We have used time-resolved soft x-ray spectroscopy to investigate the electronic structure of optically excited cuprous oxide at the O K-edge and the Cu L3-edge. The 400 nm optical excitation shifts the Cu and O absorptions to lower energy, but does not change the integrated x-ray absorption significantly for either edge. The constant integrated x-ray absorption cross-section indicates that the conduction-band and valence-band edges have very similar Cu 3d and O 2p orbital contributions. The 2.1 eV optical band gap of Cu2O significantly exceeds the one eV shift in the Cu L3- and O K-edges absorption edges induced by optical excitation, demonstrating the importance of core-hole excitonic effects and valence electron screening in the x-ray absorption process.

  16. Atomic Resolution Mapping of the Excited-State Electronic Structure of Cu2O with Time-Resolved X-Ray Absorption Spectroscopy

    SciTech Connect

    Hillyard, Patrick B.; Kuchibhatla, Satyanarayana V N T; Glover, T. E.; Hertlein, M. P.; Huse, N.; Nachimuthu, Ponnusamy; Saraf, Laxmikant V.; Thevuthasan, Suntharampillai; Gaffney, Kelly J.

    2009-09-29

    We have used time-resolved soft x-ray spectroscopy to investigate the electronic structure of optically excited cuprous oxide at the O K-edge and the Cu L3-edge. The 400 nm optical excitation shifts the Cu and O absorptions to lower energy, but does not change the integrated x-ray absorption significantly for either edge. The constant integrated x-ray absorption cross-section indicates that that the conduction band and valence band edges have very similar Cu 3d and O 2p orbital contributions. The 2.1 eV optical band gap of Cu2O significantly exceeds the one eV shift in the Cu L3- and O K-edges absorption edges induced by optical excitation, demonstrating the importance of core-hole excitonic effects and valence electron screening in the x-ray absorption process.

  17. Fluorescence yields from photodissociative excitation of chloromethanes by vacuum ultraviolet radiation

    NASA Technical Reports Server (NTRS)

    Lee, L. C.; Suto, Masako

    1987-01-01

    The photoabsorption and fluorescence cross sections of chloromethanes were measured in the 105-220 nm region using synchrotron radiation as a light source. The fluorescence threshold for CCl4 is at 152 nm with a maximum yield of 3 percent at 113 nm. The fluorescence results from the CCl2(A-X) system. For CHCl3, the fluorescence threshold is at 155 nm with a maximum yield of 0.6 percent at 110 nm. For CH2Cl2, the threshold is at 137 nm with a maximum yield of 0.35 percent at 107 nm. The fluorescence yield of CH3Cl is very small with an upper limit of 0.02 percent. The photodissociation processes are discussed in accord with the fluorescence data observed. Vibrational structures in CHCl3 and CH3Cl2 are observed and classified into progressions.

  18. Theoretical study of Raman chirped adiabatic passage by X-ray absorption spectroscopy: Highly excited electronic states and rotational effects

    SciTech Connect

    Engin, Selma; Sisourat, Nicolas Selles, Patricia; Taïeb, Richard; Carniato, Stéphane

    2014-06-21

    Raman Chirped Adiabatic Passage (RCAP) is an efficient method to climb the vibrational ladder of molecules. It was shown on the example of fixed-in-space HCl molecule that selective vibrational excitation can thus be achieved by RCAP and that population transfer can be followed by X-ray Photoelectron spectroscopy [S. Engin, N. Sisourat, P. Selles, R. Taïeb, and S. Carniato, Chem. Phys. Lett. 535, 192–195 (2012)]. Here, in a more detailed analysis of the process, we investigate the effects of highly excited electronic states and of molecular rotation on the efficiency of RCAP. Furthermore, we propose an alternative spectroscopic way to monitor the transfer by means of X-ray absorption spectra.

  19. FTIR and fluorescence studies on the ground and excited state hydrogen-bonding interactions between 1-methylindole and water in water triethylamine mixtures

    NASA Astrophysics Data System (ADS)

    Muñoz, María A.; Carmona, Carmen; Balón, Manuel

    2007-05-01

    The ground and excited state π-hydrogen-bonding interactions between 1-methylindole, MI, and water have been investigated in water-triethylamine, water-TEA, mixtures. FTIR measurements performed on the OH stretching bands of the water-TEA clusters show that, upon MI addition, the typical bands of the water-TEA system at 3348 cm -1, 3440 cm -1, 3545 cm -1 and 3682 cm -1 diminish, whereas two new absorption bands at 3316 cm -1 and 3654 cm -1 grow up. These spectral changes have been rationalised assuming the formation of only one 1:1 water-MI complex, in which the dangling protons in the water-TEA clusters are hydrogen bonded to the π-cloud of the MI aromatic ring. Steady state and time resolved fluorescence measurements provide additional proofs on the ground state formation of a fluorescent OH ⋯ π hydrogen bonded complex. The relevance that the present and the previously reported results could have on the indole ring photophysics is discussed.

  20. Graphene quantum dots from graphite by liquid exfoliation showing excitation-independent emission, fluorescence upconversion and delayed fluorescence.

    PubMed

    Sarkar, Suprabhat; Gandla, Dayakar; Venkatesh, Yeduru; Bangal, Prakriti Ranjan; Ghosh, Sutapa; Yang, Yang; Misra, Sunil

    2016-08-21

    Facile synthesis of 2-10 nm-sized graphene quantum dots (GQDs) from graphite powder by organic solvent-assisted liquid exfoliation using a sonochemical method is reported in this study. Synthesized GQDs are well dispersed in organic solvents like ethyl acetoacetate (EAA), dimethyl formamide (DMF) and also in water. MALDI-TOF mass spectrometry reveals its selective mass fragmentation. Detailed characterizations by various techniques like X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), Raman spectroscopy and high resolution transmission electron microscopy (HRTEM) confirm the formation of disordered, functional GQDs. Density functional theory (DFT) calculation confirms HOMO-LUMO energy gap variation with changing size and functionalities. Photoluminescence (PL) properties of as-prepared GQDs were studied in detail. The ensemble studies of GQDs showed excellent photoluminescence properties comprising normal and upconverted fluorescence, delayed fluorescence and room-temperature phosphorescence. PL decay dynamics of GQDs has been explored using time-correlated single-photon technique (TCSPC) as well as femtosecond fluorescence upconversion technique. In vitro cytotoxicity study reveals its biocompatibility and high cell viability (>91%) even at high concentration (400 μg mL(-1)) of GQDs in Chinese Hamster Ovary (CHO) cells. PMID:27302411

  1. Fluorescence Excitation Spectra of Photo-Fragmented Nitrobenzene Using a Picosecond Laser: Potential Evidence for no Produced by Two Distinct Channels.

    NASA Astrophysics Data System (ADS)

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

    2013-06-01

    Upon absorption of a UV photon, nitrobenzene can dissociate into C_6H_5O and NO through two different mechanisms. Evidence for these mechanisms was obtained from velocity map imaging (VMI) studies and theoretical calculations. VMI experiments showed NO produced with two distinct rotational distributions, which the calculations explained as a fast and a slow channel for NO production. We have recorded high resolution fluorescence excitation spectra of the 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. This laser system allowed us to vary the delay between the photolysis and excitation pulses. At longer delays (>1 ns), we observed an increase in the population of NO, which may be evidence that at least two photolysis channels produce NO. We present the spectra we recorded at various photolysis/probe delays ranging from 0.025 to 1.5 ns. The spectral subtraction method we used to observe the production increase is introduced. Hause, M. L.; Herath, N.; Zhu, R.; Lin, M. C. and Suits, A. G. Nat Chem, Nature Publishing Group, 2011, 3, 932-937

  2. Study on the interaction of methylene blue with cyclodextrin derivatives by absorption and fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Zhang, Guomei; Shuang, Shaomin; Dong, Chuan; Pan, Jinghao

    2003-11-01

    The ability of β-cyclodextrin (β-CD), hydroxypropyl-β-cyclodextrin (HP-β-CD), and carboxymethyl-β-cyclodextrin (CM-β-CD) to break the aggregate of the methylene blue (MB) and to form 1:1 inclusion complexes has been studied by absorption and fluorescence spectroscopy. Experimental conditions including concentrations of various cyclodextrins (β-CD, HP-β-CD and CM-β-CD) and media acidity were investigated for the inclusion formation in detail. The formation constants are calculated by using steady-state fluorimetry, from which the inclusion capacity of different cyclodextrins (CDs) is compared. The results suggest that the charged β-cyclodextrin (CM-β-CD) is more suitable for inclusion of the cationic dye MB than the neutral β-cyclodextrins (β-CD, HP-β-CD) at pH>5. A mechanism is proposed which is consistent with the stronger binding of MB with CM-β-CD compared with the other CDs at pH>5.

  3. Americium characterization by X-ray fluorescence and absorption spectroscopy in plutonium uranium mixed oxide

    NASA Astrophysics Data System (ADS)

    Degueldre, Claude; Cozzo, Cedric; Martin, Matthias; Grolimund, Daniel; Mieszczynski, Cyprian

    2013-06-01

    Plutonium uranium mixed oxide (MOX) fuels are currently used in nuclear reactors. The actinides in these fuels need to be analyzed after irradiation for assessing their behaviour with regard to their environment and the coolant. In this work the study of the atomic structure and next-neighbour environment of Am in the (Pu,U)O2 lattice in an irradiated (60 MW d kg-1) MOX sample was performed employing micro-X-ray fluorescence (µ-XRF) and micro-X-ray absorption fine structure (µ-XAFS) spectroscopy. The chemical bonds, valences and stoichiometry of Am (˜0.66 wt%) are determined from the experimental data gained for the irradiated fuel material examined in its peripheral zone (rim) of the fuel. In the irradiated sample Am builds up as Am3+ species within an [AmO8]13- coordination environment (e.g. >90%) and no (<10%) Am(IV) or (V) can be detected in the rim zone. The occurrence of americium dioxide is avoided by the redox buffering activity of the uranium dioxide matrix.

  4. Immunochemical parameters of some commercial conjugates for the fluorescent treponemal antibody-absorption test.

    PubMed

    Hunter, E F; Smith, J F; Lewis, J S; McGrew, B E; Schmale, J D

    1972-06-01

    Fluorescein-labeled anti-human globulins were examined to determine the need for standardization of conjugates used in the fluorescent treponemal antibody-absorption (FTA-ABS) test. Twenty-one of 33 conjugates submitted by commercial manufacturers to the Reagents Control Activity, Venereal Disease Research Laboratory, for evaluation in the FTA-ABS test were available for study. Conjugates, after evaluation in FTA-ABS performance tests, were examined by immunoelectrophoresis, by titration against immunoglobulins G and M (IgG, IgM) with FTA-ABS techniques, and by the biuret protein and fluorescein diacetate methods for determining fluorescein to protein (F/P) ratios. The conjugates were predominately anti-IgG globulin with anti-light-chain activity. Differences were noted in the ability of some conjugates to detect IgM antibody. The F/P ratios of those conjugates that could be determined varied from 2.6 to 17.8 mug of fluorescein per mg of protein. The need to identify and standardize both the immunologic capabilities and the optimum F/P ratio for FTA-ABS test conjugates is presented.

  5. A multi-channel monolithic Ge detector system for fluorescence x-ray absorption spectroscopy

    SciTech Connect

    Bucher, J.J.; Allen, P.G.; Edelstein, N.M.; Shuh, D.K.; Madden, N.W.; Cork, C.; Luke, P.; Pehl, D.; Malone, D.

    1995-03-01

    Construction and performance of a monolithic quad-pixel Ge detector for fluorescence x-ray absorption spectroscopy (XAS) at synchrotron radiation sources are described. The detector semiconductor element has an active surface area of 4.0 cm{sup 2} which is electrically separated into four 1.0 cm{sup 2} pixels, with little interfacial dead volume. Spatial response of the array shows that cross-talk between adjacent pixels is < 10% for 5.9 keV photons that fall within 0.5 mm of the pixel boundaries. The detector electronics system uses pre-amplifiers built at LBNL with commercial Tennelec Model TC 244 amplifiers. Using an {sup 55}Fe test source (MnK{sub {alpha}}, 5.9 keV), energy resolution of better than 200 eV is achieved with a 4 {mu}sec peaking time. At 0.5 {mu}sec peaking time, pulse pileup results in a 75% throughput efficiency for an incoming count rate of 100 kHz. Initial XAS fluoresncece measurements at the beamline 4 wiggler end stations at SSRL show that the detector system has several advantages over commercial x-ray spectrometers for low-concentration counting.

  6. [Vermicomposting of different organic materials and three-dimensional excitation emission matrix fluorescence spectroscopic characterization of their dissolved organic matter].

    PubMed

    Yang, Wei; Wang, Dong-sheng; Liu, Man-qiang; Hu, Feng; Li, Hui-xin; Huang, Zhong-yang; Chang, Yi-jun; Jiao, Jia-guo

    2015-10-01

    In this experiment, different proportions of the cattle manure, tea-leaf, herb and mushroom residues, were used as food for earthworm (Eisenia fetida) to study the growth of the earth-worm. Then the characteristics and transformation of nutrient content and three-dimensional excitation emission matrix fluorescence (3DEEM) of dissolved organic matter (DOM) during vermistabilization were investigated by means of chemical and spectroscopic methods. The result showed that the mixture of different ratios of cattle manure with herb residue, and cattle manure with tea-leaf were conducive to the growth of earthworm, while the materials compounded with mushroom residue inhibited the growth of earthworm. With the increasing time of verimcomposting, the pH in vermicompost tended to be circumneutral and weakly acidic, and there were increases in electrical conductivity, and the contents of total nitrogen, total phosphorus, available nitrogen, and available phosphorus, while the total potassium and available potassium increased first and then decreased, and the organic matter content decreased. 3DEEM and fluorescence regional integration results indicated that, the fluorescence of protein-like fluorescence peaks declined significantly, while the intensity of humic-like fluorescence peak increased significantly in DOM. Vermicomposting process might change the compositions of DOM with elevated concentrations of humic acid and fulvic acid in the organics. In all, this study suggested the suitability of 3DEEM for monitoring the organics transformation and assessing the maturity in the vermicomposting. PMID:26995929

  7. Superior optical nonlinearity of an exceptional fluorescent stilbene dye

    SciTech Connect

    He, Tingchao; Sreejith, Sivaramapanicker; Zhao, Yanli; Gao, Yang; Grimsdale, Andrew C.; Lin, Xiaodong E-mail: hdsun@ntu.edu.sg; Sun, Handong E-mail: hdsun@ntu.edu.sg

    2015-03-16

    Strong multiphoton absorption and harmonic generation in organic fluorescent chromophores are, respectively, significant in many fields of research. However, most of fluorescent chromophores fall short of the full potential due to the absence of the combination of such different nonlinear upconversion behaviors. Here, we demonstrate that an exceptional fluorescent stilbene dye could exhibit efficient two- and three-photon absorption under the excitation of femtosecond pulses in solution phase. Benefiting from its biocompatibility and strong excited state absorption behavior, in vitro two-photon bioimaging and superior optical limiting have been exploited, respectively. Simultaneously, the chromophore could generate efficient three-photon excited fluorescence and third-harmonic generation (THG) when dispersed into PMMA film, circumventing the limitations of classical fluorescent chromophores. Such chromophore may find application in the production of coherent light sources of higher photon energy. Moreover, the combination of three-photon excited fluorescence and THG can be used in tandem to provide complementary information in biomedical studies.

  8. Characterization of CDOM from urban waters in Northern-Northeastern China using excitation-emission matrix fluorescence and parallel factor analysis.

    PubMed

    Zhao, Ying; Song, Kaishan; Li, Sijia; Ma, Jianhang; Wen, Zhidan

    2016-08-01

    Chromophoric dissolved organic matter (CDOM) plays an important role in aquatic systems, but high concentrations of organic materials are considered pollutants. The fluorescent component characteristics of CDOM in urban waters sampled from Northern and Northeastern China were examined by excitation-emission matrix fluorescence and parallel factor analysis (EEM-PARAFAC) to investigate the source and compositional changes of CDOM on both space and pollution levels. One humic-like (C1), one tryptophan-like component (C2), and one tyrosine-like component (C3) were identified by PARAFAC. Mean fluorescence intensities of the three CDOM components varied spatially and by pollution level in cities of Northern and Northeastern China during July-August, 2013 and 2014. Principal components analysis (PCA) was conducted to identify the relative distribution of all water samples. Cluster analysis (CA) was also used to categorize the samples into groups of similar pollution levels within a study area. Strong positive linear relationships were revealed between the CDOM absorption coefficients a(254) (R (2) = 0.89, p < 0.01); a(355) (R (2) = 0.94, p < 0.01); and the fluorescence intensity (F max) for the humic-like C1 component. A positive linear relationship (R (2) = 0.77) was also exhibited between dissolved organic carbon (DOC) and the F max for the humic-like C1 component, but a relatively weak correlation (R (2) = 0.56) was detected between DOC and the F max for the tryptophan-like component (C2). A strong positive correlation was observed between the F max for the tryptophan-like component (C2) and total nitrogen (TN) (R (2) = 0.78), but moderate correlations were observed with ammonium-N (NH4-N) (R (2) = 0.68), and chemical oxygen demand (CODMn) (R (2) = 0.52). Therefore, the fluorescence intensities of CDOM components can be applied to monitor water quality in real time compared to that of traditional approaches. These results demonstrate

  9. Laser-Induced Fluorescence in Gaseous [I[subscript]2] Excited with a Green Laser Pointer

    ERIC Educational Resources Information Center

    Tellinghuisen, Joel

    2007-01-01

    A green laser pointer could be used in a flashy demonstration of laser-induced fluorescence in the gas phase by directing the beam of the laser through a cell containing [I[subscript]2] at its room temperature vapor pressure. The experiment could be used to provide valuable insight into the requirements for laser-induced fluorescence (LIF) and the…

  10. A reagentless enzymatic fluorescent biosensor for glucose based on upconverting glasses, as excitation source, and chemically modified glucose oxidase.

    PubMed

    Del Barrio, Melisa; Cases, Rafael; Cebolla, Vicente; Hirsch, Thomas; de Marcos, Susana; Wilhelm, Stefan; Galbán, Javier

    2016-11-01

    Upon near-infrared excitation Tm(3+)+Yb(3+) doped fluorohafnate glasses present upconversion properties and emit visible light. This property permits to use these glasses (UCG) as excitation sources for fluorescent optical biosensors. Taking this into account, in this work a fluorescent biosensor for glucose determination is designed and evaluated. The biosensor combines the UCG and the fluorescence of the enzyme glucose oxidase chemically modified with a fluorescein derivative (GOx-FS), whose intensity is modified during the enzymatic reaction with glucose. Optical parameters have been optimized and a mathematical model describing the behavior of the analytical signal is suggested. Working in FIA mode, the biosensor responds to glucose concentrations up to, at least, 15mM with a limit of detection of 1.9mM. The biosensor has a minimum lifetime of 9 days and has been applied to glucose determination in drinks. The applicability of the sensor was tested by glucose determination in two fruit juices. PMID:27591654

  11. Determination of X-ray excitation spectra in micro X-ray fluorescence spectrometry with capillary optics

    NASA Astrophysics Data System (ADS)

    Perez, R. D.; Sosa, C.; Sbarato, V.; Leani, J.; Sánchez, H. J.

    2016-03-01

    The quantitative X-ray fluorescence microanalysis by fundamental parameters requires the knowledge of the energy distribution of the excitation beam. When this beam is produced by capillary optics, its high intensity and anisotropy complicate a direct determination. An alternative is an indirect determination based on measurement of induced X-ray fluorescence in a set of targets. In this work the X-ray excitation spectrum is determined by an iterative deconvolution process of the fundamental parameter expression for the X-ray fluorescence intensities. The method has the advantage that it does not require any assumption about the energy distribution of the X-ray source or the energy dependence of the lens transmission. Numerous XRF targets of pure elements with emission lines covering the full energy range of the X-ray source are employed. The only requirement on the targets is a high homogeneity in its composition. In fact, it does not impose any condition on the sample thickness. The accuracy observed in the validation process implemented with reference materials is similar to that reported with alternative approaches: 5% for main components, 10% for minor elements and 15% for trace elements.

  12. High-speed laser-induced fluorescence and spark plug absorption sensor diagnostics for mixing and combustion studies in engines

    SciTech Connect

    Cundy, Michael; Schucht, Torsten; Thiele, Olaf; Sick, Volker

    2009-02-01

    Simultaneous high-speed in-cylinder measurements of laser-induced fluorescence of biacetyl as a fuel tracer and mid-infrared broadband absorption of fuel and combustion products (water and carbon dioxide) using a spark plug probe are compared in an optical engine. The study addresses uncertainties and the applicability of absorption measurements at a location slightly offset to the spark plug when information about mixing at the spark plug is desired. Absorbance profiles reflect important engine operation events, such as valve opening and closing, mixing, combustion, and outgassing from crevices.

  13. Forensic analysis of laser printed ink by X-ray fluorescence and laser-excited plume fluorescence.

    PubMed

    Chu, Po-Chun; Cai, Bruno Yue; Tsoi, Yeuk Ki; Yuen, Ronald; Leung, Kelvin S Y; Cheung, Nai-Ho

    2013-05-01

    We demonstrated a minimally destructive two-tier approach for multielement forensic analysis of laser-printed ink. The printed document was first screened using a portable-X-ray fluorescence (XRF) probe. If the results were not conclusive, a laser microprobe was then deployed. The laser probe was based on a two-pulse scheme: the first laser pulse ablated a thin layer of the printed ink; the second laser pulse at 193 nm induced multianalytes in the desorbed ink to fluoresce. We analyzed four brands of black toners. The toners were printed on paper in the form of patches or letters or overprinted on another ink. The XRF probe could sort the four brands if the printed letters were larger than font 20. It could not tell the printing sequence in the case of overprints. The laser probe was more discriminatory; it could sort the toner brands and reveal the overprint sequence regardless of font size while the sampled area was not visibly different from neighboring areas even under the microscope. In terms of general analytical performance, the laser probe featured tens of micrometer lateral resolution and tens to hundreds of nm depth resolution and atto-mole mass detection limits. It could handle samples of arbitrary size and shape and was air compatible, and no sample pretreatment was necessary. It will prove useful whenever high-resolution and high sensitivity 3D elemental mapping is required.

  14. Rapid and quantitative imaging of excitation polarized fluorescence reveals ordered septin dynamics in live yeast.

    PubMed

    DeMay, Bradley S; Noda, Naoki; Gladfelter, Amy S; Oldenbourg, Rudolf

    2011-08-17

    We report an imaging method for fast, sensitive analysis of the orientation of fluorescent molecules by employing a liquid-crystal based universal polarizer in the optical path of a wide-field light microscope. We developed specific acquisition and processing algorithms for measuring the anisotropy and for correcting artifacts caused by fluorescence bleaching, background light, and differential transmission of optical components. We call this approach the Fluorescence LC-PolScope and we used it to analyze the architectural dynamics of septin-green fluorescent protein (septin-GFP) constructs in the neck region of budding yeast. We describe three different states of highly anisotropic septin arrays in which the prevailing orientation of GFP dipoles was either parallel or perpendicular to the mother-bud axis. The transitions between these ordered states were characterized by transient isotropic states. To analyze the patterns of polarized fluorescence, we modeled the alignment of septin-GFP constructs in different stages of septin ring formation. Based on our model, our experimental data are consistent with the formation of paired rather than single filaments and the axis of the α-helical septin terminus linked to a GFP molecule is likely oriented normal to the cell surface. The Fluorescence LC-PolScope combines the molecular specificity of fluorescence tagging with the structural specificity of polarized light analysis.

  15. Americium characterization by X-ray fluorescence and absorption spectroscopy in plutonium uranium mixed oxide

    SciTech Connect

    Degueldre, Claude Cozzo, Cedric; Martin, Matthias; Grolimund, Daniel; Mieszczynski, Cyprian

    2013-06-01

    Plutonium uranium mixed oxide (MOX) fuels are currently used in nuclear reactors. The actinides in these fuels need to be analyzed after irradiation for assessing their behaviour with regard to their environment and the coolant. In this work the study of the atomic structure and next-neighbour environment of Am in the (Pu,U)O₂ lattice in an irradiated (60 MW d kg⁻¹) MOX sample was performed employing micro-X-ray fluorescence (µ-XRF) and micro-X-ray absorption fine structure (µ-XAFS) spectroscopy. The chemical bonds, valences and stoichiometry of Am (~0.66 wt%) are determined from the experimental data gained for the irradiated fuel material examined in its peripheral zone (rim) of the fuel. In the irradiated sample Am builds up as Am³⁺ species within an [AmO₈]¹³⁻ coordination environment (e.g. >90%) and no (<10%) Am(IV) or (V) can be detected in the rim zone. The occurrence of americium dioxide is avoided by the redox buffering activity of the uranium dioxide matrix. - Graphical abstract: Americium LIII XAFS spectra recorded for the irradiated MOX sub-sample in the rim zone for a 300 μm×300 μm beam size area investigated over six scans of 4 h. The records remain constant during multi-scan. The analysis of the XAFS signal shows that Am is found as trivalent in the UO₂ matrix. This analytical work shall open the door of very challenging analysis (speciation of fission product and actinides) in irradiated nuclear fuels. - Highlights: • Americium was characterized by microX-ray absorption spectroscopy in irradiated MOX fuel. • The americium redox state as determined from XAS data of irradiated fuel material was Am(III). • In the sample, the Am³⁺ face an AmO₈¹³⁻coordination environment in the (Pu,U)O₂ matrix. • The americium dioxide is reduced by the uranium dioxide matrix.

  16. Transient absorption in water-micellar solutions of rhodamine 6G with flash lamp excitation

    SciTech Connect

    Levin, M.B.; Cherkasov, A.S.

    1986-06-01

    This paper studies the kinetics of transient losses in water-micellar solutions of rhodamine 6G by using flash lamp excitation. During the experiments, the laser radiation energy was measured, the time evolution of stimulated emission spectra was recorded; pulse shape was monitored by an oscillograph. The change of generation characteristics of water-micellar solutions of rhodamine 6G as a function of cyclooctatetraene concentration is shown.

  17. Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function

    PubMed Central

    Alonzo, Carlo Amadeo; Karaliota, Sevasti; Pouli, Dimitra; Liu, Zhiyi; Karalis, Katia P.; Georgakoudi, Irene

    2016-01-01

    Current methods for evaluating adipose tissue function are destructive or have low spatial resolution. These limit our ability to assess dynamic changes and heterogeneous responses that occur in healthy or diseased subjects, or during treatment. Here, we demonstrate that intrinsic two-photon excited fluorescence enables functional imaging of adipocyte metabolism with subcellular resolution. Steady-state and time-resolved fluorescence from intracellular metabolic co-factors and lipid droplets can distinguish the functional states of excised white, brown, and cold-induced beige fat. Similar optical changes are identified when white and brown fat are assessed in vivo. Therefore, these studies establish the potential of non-invasive, high resolution, endogenous contrast, two-photon imaging to identify distinct adipose tissue types, monitor their functional state, and characterize heterogeneity of induced responses. PMID:27491409

  18. Lung alveolar wall disruption in three-dimensional space identified using second-harmonic generation and multiphoton excitation fluorescence

    NASA Astrophysics Data System (ADS)

    Abraham, Thomas; Hogg, James

    2010-02-01

    Second harmonic generation and multiphoton excited fluorescence microscopy methods were used to examine structural remodeling of the extracellular matrix in human lung alveolar walls undergoing emphysematous destruction. Fresh lung samples removed from a patient undergoing lung transplantation for very severe chronic obstructive pulmonary disease were compared to similar samples from an unused donor lung that served as a control. The generated spatially resolved 3D images show the spatial distribution of collagen, elastin and other endogenously fluorescent tissue components such as macrophages. In the case of control lung tissue, we found well ordered alveolar walls with composite type structure made up of collagen matrix and relatively fine elastic fibers. In contrast, lung tissue undergoing emphysematous destruction was highly disorganized with increased alveolar wall thickness compared to control lung tissue.

  19. Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function

    NASA Astrophysics Data System (ADS)

    Alonzo, Carlo Amadeo; Karaliota, Sevasti; Pouli, Dimitra; Liu, Zhiyi; Karalis, Katia P.; Georgakoudi, Irene

    2016-08-01

    Current methods for evaluating adipose tissue function are destructive or have low spatial resolution. These limit our ability to assess dynamic changes and heterogeneous responses that occur in healthy or diseased subjects, or during treatment. Here, we demonstrate that intrinsic two-photon excited fluorescence enables functional imaging of adipocyte metabolism with subcellular resolution. Steady-state and time-resolved fluorescence from intracellular metabolic co-factors and lipid droplets can distinguish the functional states of excised white, brown, and cold-induced beige fat. Similar optical changes are identified when white and brown fat are assessed in vivo. Therefore, these studies establish the potential of non-invasive, high resolution, endogenous contrast, two-photon imaging to identify distinct adipose tissue types, monitor their functional state, and characterize heterogeneity of induced responses.

  20. Efficiently tuning the absorption and fluorescence spectroscopy of the novel branched p-nitro-stilbene derivatives with chemical strategy.

    PubMed

    Gao, Fang; Yang, Liufeng; Yang, Long; Li, Hongru; Zhang, Shengtao

    2010-01-01

    Suitable chemical strategy is a useful approach on the tuning color and photoluminescence of organic dyes. This paper presented tuning novel branched p-nitro-stilbene derivatives efficiently with a new chemical strategy through variation of chemical bridged bond. Linking bonds played significant effects on the absorption and fluorescence spectroscopy of the branched p-nitro-stilbene derivatives. A change from "D-pi-A" to "A-pi-A" chemical structural characteristics occurred for the branched p-nitro-stilbene derivatives as ester bond was attached. This led to not only large hypsochromic shift of the maximal absorption wavelength of the branched p-nitro-stilbene derivatives, but considerable reduction of the fluorescence intensity. While in contrast, the branched p-nitro-stilbene derivatives with ether bond exhibited longer wavelength absorption and much stronger fluorescence emission in modest polar solvent. The cyclic voltammograms of these branched p-nitro-stilbene derivatives were determined. Different electrochemistry processes were observed for the branched p-nitro-stilbene derivatives with various linking bonds. The energies of frontier orbital of the branched p-nitro-stilbene derivatives were estimated from their corresponding redox potentials. Molecular geometry optimization of the branched p-nitro-stilbene derivatives was performed, and the electron density distribution of frontier orbital was analyzed. Thermal stabilities of these branched nitro-stilbene derivatives were investigated via the analysis of the differential scanning calorimetry (DSC) and thermograving (TGA) curves. This paper presented strong evidences that the absorption and fluorescence spectroscopy of the branched stilbene derivatives could be mediated efficiently by chemical strategy. PMID:19953312

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

    NASA Astrophysics Data System (ADS)

    Galante, Matthew

    2013-10-01

    Neutral particles play a critical role in nearly all plasmas, from the pedestal region of a tokamak fusion plasma to industrial plasma processing systems. In fusion plasmas, neutrals at the edge serve as both a source of particles and also a sink of momentum and energy. Control of the edge plasma density in tokamaks is critical for the transition to H-mode plasmas and the role of neutrals in modifying the plasma rotation in the edge is an area of active research. However, few methods exist to make localized, direct neutral density measurements. We have developed a new diagnostic based on two-photon absorption laser induced fluorescence (TALIF). We use a high intensity (5 MW/cm2), narrow bandwidth (0.1 cm-1) laser to probe the ground state of neutral hydrogen, deuterium and krypton with spatial resolution better than 0.2 cm, a time resolution of 10 ns, and a measurement cadence of 20 Hz. In this talk I will describe proof-of-principle measurements in a helicon plasma source that demonstrate the TALIF diagnostic is capable of measuring neutral densities spanning four orders of magnitude; comparable to the edge neutral gradients predicted in the tokamak pedestal. The measurements are performed in hydrogen and deuterium plasmas and absolute calibration is accomplished through TALIF measurements in neutral krypton. The optical configuration employed is confocal, i.e., both light injection and collection are accomplished through a single optical port in the vacuum vessel. The wavelength resolution of the diagnostic is sufficient to separate hydrogen and deuterium spectra and I will present measurements from mixed hydrogen and deuterium plasmas that demonstrate isotopic abundance measurements are feasible with the TALIF system. Time and spatially resolved measurements also allow us to explore the effects of wall recycling and pulse repetition rates on the neutral density profile in the plasma source. Work supported in part by US DOE under DE-FC02-04ER54698.

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

    NASA Astrophysics Data System (ADS)

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

    2014-05-01

    We have developed a new diagnostic based on two-photon absorption laser induced fluorescence (TALIF). We use a high intensity (5 MW/cm2), narrow bandwidth (0.1 cm-1) laser to probe the ground state of neutral hydrogen, deuterium and krypton with spatial resolution better than 0.2 cm, a time resolution of 10 ns, and a measurement cadence of 20 Hz. Here, we describe proof-of-principle measurements in a helicon plasma source that demonstrate the TALIF diagnostic is capable of measuring neutral densities spanning four orders of magnitude; comparable to the edge neutral gradients predicted in the DIII-D tokamak pedestal. The measurements are performed in hydrogen and deuterium plasmas and absolute calibration is accomplished through TALIF measurements in neutral krypton. The optical configuration employed is confocal, i.e., both light injection and collection are accomplished with a single lens through a single optical port in the vacuum vessel. The wavelength resolution of the diagnostic is sufficient to separate hydrogen and deuterium spectra and we present measurements from mixed hydrogen and deuterium plasmas that demonstrate isotopic abundance measurements are feasible. Time resolved measurements also allow us to explore the evolution of the neutral hydrogen density and temperature and effects of wall recycling. We find that the atomic neutral density grows rapidly at the initiation of the discharge, reaching the steady-state value within 1 ms. Additionally, we find that neutral hydrogen atoms are born with 0.08 eV temperatures, not 2 eV as is typically assumed.

  3. Two photon absorption laser induced fluorescence measurements of neutral density in a helicon plasma

    SciTech Connect

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

    2014-05-15

    We have developed a new diagnostic based on two-photon absorption laser induced fluorescence (TALIF). We use a high intensity (5 MW/cm{sup 2}), narrow bandwidth (0.1 cm{sup −1}) laser to probe the ground state of neutral hydrogen, deuterium and krypton with spatial resolution better than 0.2 cm, a time resolution of 10 ns, and a measurement cadence of 20 Hz. Here, we describe proof-of-principle measurements in a helicon plasma source that demonstrate the TALIF diagnostic is capable of measuring neutral densities spanning four orders of magnitude; comparable to the edge neutral gradients predicted in the DIII-D tokamak pedestal. The measurements are performed in hydrogen and deuterium plasmas and absolute calibration is accomplished through TALIF measurements in neutral krypton. The optical configuration employed is confocal, i.e., both light injection and collection are accomplished with a single lens through a single optical port in the vacuum vessel. The wavelength resolution of the diagnostic is sufficient to separate hydrogen and deuterium spectra and we present measurements from mixed hydrogen and deuterium plasmas that demonstrate isotopic abundance measurements are feasible. Time resolved measurements also allow us to explore the evolution of the neutral hydrogen density and temperature and effects of wall recycling. We find that the atomic neutral density grows rapidly at the initiation of the discharge, reaching the steady-state value within 1 ms. Additionally, we find that neutral hydrogen atoms are born with 0.08 eV temperatures, not 2 eV as is typically assumed.

  4. Ultrashort Two-Photon-Absorption Laser-Induced Fluorescence in Nanosecond-Duration, Repetitively Pulsed Discharges

    NASA Astrophysics Data System (ADS)

    Schmidt, Jacob Brian

    Absolute number densities of atomic species produced by nanosecond duration, repetitively pulsed electric discharges are measured by two-photon absorption laser-induced fluorescence (TALIF). Relatively high plasma discharge pulse energies (=1 mJ/pulse) are used to generate atomic hydrogen, oxygen, and nitrogen in a variety of discharge conditions and geometries. Unique to this work is the development of femtosecond-laser-based TALIF (fs-TALIF). Fs-TALIF offers a number of advantages compared to more conventional ns-pulse-duration laser systems, including better accuracy of direct quenching measurements in challenging environments, significantly reduced photolytic interference including photo-dissociation and photo-ionization, higher signal and increased laser-pulse bandwidth, the ability to collect two-dimensional images of atomic species number densities with far greater spatial resolution compared with more conventional diagnostics, and much higher laser repetition rates allowing for more efficient and accurate measurements of atomic species number densities. In order to fully characterize the fs-TALIF diagnostic and compare it with conventional ns-TALIF, low pressure (100 Torr) ns-duration pulsed discharges are operated in mixtures of H2, O2, and N2 with different buffer gases including argon, helium, and nitrogen. These discharge conditions are used to demonstrate the capability for two-dimensional imaging measurements. The images produced are the first of their kind and offer quantitative insight into spatially and temporally resolved kinetics and transport in ns-pulsed discharge plasmas. The two-dimensional images make possible comparison with high-fidelity plasma kinetics models of the presented data. The comparison with the quasi-one-dimensional kinetic model show good spatial and temporal agreement. The same diagnostics are used at atmospheric pressure, when atomic oxygen fs-TALIF is performed in an atmospheric-pressure plasma jet (APPJ). Here, the

  5. Excited-state molecular structures captured by X-ray transient absorption spectroscopy: a decade and beyond.

    PubMed

    Chen, Lin X; Zhang, Xiaoyi; Lockard, Jenny V; Stickrath, Andrew B; Attenkofer, Klaus; Jennings, Guy; Liu, Di-Jia

    2010-03-01

    Transient molecular structures along chemical reaction pathways are important for predicting molecular reactivity, understanding reaction mechanisms, as well as controlling reaction pathways. During the past decade, X-ray transient absorption spectroscopy (XTA, or LITR-XAS, laser-initiated X-ray absorption spectroscopy), analogous to the commonly used optical transient absorption spectroscopy, has been developed. XTA uses a laser pulse to trigger a fundamental chemical process, and an X-ray pulse(s) to probe transient structures as a function of the time delay between the pump and probe pulses. Using X-ray pulses with high photon flux from synchrotron sources, transient electronic and molecular structures of metal complexes have been studied in disordered media from homogeneous solutions to heterogeneous solution-solid interfaces. Several examples from the studies at the Advanced Photon Source in Argonne National Laboratory are summarized, including excited-state metalloporphyrins, metal-to-ligand charge transfer (MLCT) states of transition metal complexes, and charge transfer states of metal complexes at the interface with semiconductor nanoparticles. Recent developments of the method are briefly described followed by a future prospective of XTA. It is envisioned that concurrent developments in X-ray free-electron lasers and synchrotron X-ray facilities as well as other table-top laser-driven femtosecond X-ray sources will make many breakthroughs and realise dreams of visualizing molecular movies and snapshots, which ultimately enable chemical reaction pathways to be controlled. PMID:20164647

  6. Excited-state molecular structures captured by x-ray transient absorption spectroscopy : a decade and beyond.

    SciTech Connect

    Chen, L. X.; Zhang, X.; Lockard, J. V.; Stickrath, A. B.; Attenkofer, K.; Jennings, G.; Liu, D.-J.; Northwestern Univ.

    2010-03-02

    Transient molecular structures along chemical reaction pathways are important for predicting molecular reactivity, understanding reaction mechanisms, as well as controlling reaction pathways. During the past decade, X-ray transient absorption spectroscopy (XTA, or LITR-XAS, laser-initiated X-ray absorption spectroscopy), analogous to the commonly used optical transient absorption spectroscopy, has been developed. XTA uses a laser pulse to trigger a fundamental chemical process, and an X-ray pulse(s) to probe transient structures as a function of the time delay between the pump and probe pulses. Using X-ray pulses with high photon flux from synchrotron sources, transient electronic and molecular structures of metal complexes have been studied in disordered media from homogeneous solutions to heterogeneous solution-solid interfaces. Several examples from the studies at the Advanced Photon Source in Argonne National Laboratory are summarized, including excited-state metalloporphyrins, metal-to-ligand charge transfer (MLCT) states of transition metal complexes, and charge transfer states of metal complexes at the interface with semiconductor nanoparticles. Recent developments of the method are briefly described followed by a future prospective of XTA. It is envisioned that concurrent developments in X-ray free-electron lasers and synchrotron X-ray facilities as well as other table-top laser-driven femtosecond X-ray sources will make many breakthroughs and realise dreams of visualizing molecular movies and snapshots, which ultimately enable chemical reaction pathways to be controlled.

  7. Study on the interaction between fluoroquinolones and erythrosine by absorption, fluorescence and resonance Rayleigh scattering spectra and their application

    NASA Astrophysics Data System (ADS)

    Wang, Jian; Liu, Zhongfang; Liu, Jiangtao; Liu, Shaopu; Shen, Wei

    2008-03-01

    In pH 4.4-4.5 Britton-Robinson (BR) buffer solution, fluoroquinolone antibiotics (FLQs) including ciprofloxacin (CIP), norfloxacin (NOR), levofloxacin (LEV) and lomefloxacin (LOM) could react with erythrosine (Ery) to form 1:1 ion-association complexes, which not only resulted in the changes of the absorption spectra and the quenching of fluorescence, but also resulted in the great enhancement of resonance Rayleigh scattering (RRS). These offered some indications of the determination of fluoroquinolone antibiotics by spectrophotometric, fluorescence and resonance Rayleigh scattering methods. The detection limits for fluoroquinolone antibiotics were in the range of 0.097-0.265 μg/mL for absorption methods, 0.022-0.100 μg/mL for fluorophotometry and 0.014-0.027 μg/mL for RRS method, respectively. Among them, the RRS method had the highest sensitivity. In this work, the spectral characteristics of the absorption, fluorescence and RRS, the optimum conditions of the reactions and the properties of the analytical chemistry were investigated. The methods have been successfully applied to determination of some fluoroquinolone antibiotics in human urine samples and tablets. Taking CIP-Ery system as an example, the charge distribution, the enthalpy of formation and the mean polarizability were calculated by density function theory (DFT) method. In addition, the reasons for the enhancement of scattering spectra were discussed.

  8. Nonlinear optical absorption and fluorescence of phosphine-substituted bithiophenes in the violet-blue spectral region

    NASA Astrophysics Data System (ADS)

    Wang, Jianwei; Zhao, Qun; Lawson, Christopher M.; Gray, Gary M.

    2011-06-01

    The nonlinear optical absorptions of two 5,5‧-bis(diphenylphosphino)-2,2‧-bithiophene derivatives, Ph2(X)P(C4H2S)2P(X)Ph2 (X = O, 1; S, 2), have been investigated by direct transmission measurement with both picosecond and nanosecond laser pulses from 420 nm to 480 nm. Saturated dichloromethane solutions of 1 and 2 exhibit strong nonlinear optical absorptions in this violet-blue spectral region with that of 2 being stronger at all wavelengths. In the picosecond regime, at 420 nm, the transmittance rapidly falls to 50% when the incident fluence is 0.22 J/cm2 for 1 and 0.11 J/cm2 for 2. Two-photon absorption appears to be the primary mechanism for this nonlinear absorption. The two-photon absorption coefficients β for 1 (2.1 cm/GW) and 2 (4.4 cm/GM) were obtained by fitting the measurement of transmittance as the function of incident beam intensity at 420 nm. These β values are comparable with some of the best results obtained for organic materials in the green, red and infrared spectral region. Both compounds also show fluorescence with an emission peak at 390 nm for 1 and 400 nm for 2. The fluorescence of 1 is considerably stronger than is that of 2. The combination of the wide band gap and strong fluorescence emission of 1 makes it a promising candidate as a host material for blue organic light emitting diodes.

  9. Log spiral of revolution highly oriented pyrolytic graphite monochromator for fluorescence x-ray absorption edge fine structure

    SciTech Connect

    Pease, D. M.; Daniel, M.; Budnick, J. I.; Rhodes, T.; Hammes, M.; Potrepka, D. M.; Sills, K.; Nelson, C.; Heald, S. M.; Brewe, D. I.

    2000-09-01

    We have constructed an x-ray monochromator based on a log spiral of revolution covered with highly oriented pyrolytic graphite. Such a monochromator is used for obtaining x-ray absorption edge fine structure by the fluorescence method, and is particularly useful for measuring the fine structure of dilute element A in a concentrated matrix of element B, where B is to the left of A in the Periodic Table. Using the log spiral monochromator, we measure good Cr x-ray fine structure in an alloy of 1% Cr in a V matrix, whereas the corresponding spectrum is severely distorted by the V background if nonmonochromatized fluorescence is used. We also obtain excellent rejection of Mn fluorescence relative to Cr fluorescence in a Cr{sub 80}Mn{sub 20} alloy, and can tune the monochromator such that the entire Mn step height is significantly smaller than the Cr x-ray absorption edge fine structure oscillations for this system. (c) 2000 American Institute of Physics.

  10. Time-resolved spectroscopic fluorescence imaging, transient absorption and vibrational spectroscopy of intact and photo-inhibited photosynthetic tissue.

    PubMed

    Lukins, Philip B; Rehman, Shakil; Stevens, Gregory B; George, Doaa

    2005-01-01

    Fluorescence, absorption and vibrational spectroscopic techniques were used to study spinach at the photosystem II (PS II), chloroplast and cellular levels and to determine the effects and mechanisms of ultraviolet-B (UV-B) photoinhibition of these structures. Two-photon fluorescence spectroscopic imaging of intact chloroplasts shows significant spatial variations in the component fluorescence spectra in the range 640-740 nm, indicating that the type and distribution of chlorophylls vary markedly with position in the chloroplast. The chlorophyll distributions and excitonic behaviour in chloroplasts and whole plant tissue were studied using picosecond time-gated fluorescence imaging, which also showed UV-induced kinetic changes that clearly indicate that UV-B induces both structural and excitonic uncoupling of chlorophylls within the light-harvesting complexes. Transient absorption measurements and low-frequency infrared and Raman spectroscopy show that the predominant sites of UV-B damage in PS II are at the oxygen-evolving centre (OEC) itself, as well as at specific locations near the OEC-binding sites.

  11. Zero-field splitting of the ground and excited triplet states of 2-naphthylphenylcarbene studied by hole-burning on triplet-triplet fluorescence excitation spectra

    NASA Astrophysics Data System (ADS)

    Kozankiewicz, B.; Bernard, J.; Migirdicyan, E.; Orrit, M.; Platz, M. S.

    1995-11-01

    Spectral holes were burned in the two main lines of the triplet-triplet 0-0 fluorescence excitation spectrum of 2-napththylphenylcarbene in n-hexane at 1.8 K. For the line assigned to pseudo-E/trans conformer, the central hole at the frequency of burning light has several narrow components separated by 0.3 ± 0.05 and 1.0 ± 0.1 GHz and a satellite doublet line on the low energy side, 14.5 ± 0.5 and 15.5 ± 0.5 GHz away. For the line assigned to the psuedo-Z/cis conformer, the central hole is accompanied by holes burned on the low energy side, 0.8 ± 0.1, 5.2 ± 0.1, 11.9 ± 0.1 and 16.2 ± 0.1 GHz away. The hole-burning pattern is explained by a model taking into account the zero-field splitting (ZFS) of the ground T 0 and excited T 1 triplet states as well as the selectivity of excitation relaxation by the intersystem crossing channel. Analysis provides information about the ZFS parameters of the T 1 state. For the pseudo-E/trans 2-NPC they are: D0 = 0.47 ± 0.02 cm -1, E0 = 0.017 ± 0.003 cm -1, D1 = 0.038 ± 0.003 cm -1 and E1 = 0.005 ± 0.001 cm -1.

  12. A time-resolved fluorescence study of electronic excitation energy transport in concentrated dye solutions

    NASA Astrophysics Data System (ADS)

    Scully, A. D.; Matsumoto, A.; Hirayama, S.

    1991-11-01

    Singlet-state radiative and nonradiative energy transport among randomly distributed donor and acceptor molecules in solutions of ethylene glycol has been investigated by using time-resolved fluorescence spectroscopy. Radiative energy transfer in moderately concentrated solutions of rhodamine 6G contained in a 1 cm pathlength cuvette results in nonexponential fluorescence decay curves which can be described in terms of the relative fluorescence emission yield of the ith transfer process, φ i, the average number of transfer steps, < n >, and the apparent fluorescence lifetime, <τ >. The fluorescence decay profiles measured from thin films (⩽ 20 μm) of solutions of rhodamine 6G donors in the presence of malachite green acceptors are also nonexponential and at low donor concentrations (1.0 × 10 -4 M) the decay curves can be described by using Förster's dipole-dipole model for nonradiative energy transfer, where the value for the critical transfer distance, R0, is calculated to be (5.9±0.1) nm. At higher donor concentrations (3.0 × 10 -3 M) the Förster model is inappropriate. However, the model proposed by Loring, Anderson and Fayer (LAF), which allows for the effects of nonradiative energy transfer among donors, provides excellent fits to the experimentally determined fluorescence decay curves for this donor concentration and results in a value of R0 for nonradiative energy transfer between rhodamine 6G chromophores of (5.8±0.1) nm. The LAF model also provides a satisfactory description of the kinetics of the quenching by rhodamine 6G dimers of the fluorescence from thin films of highly concentrated solutions of rhodamine 6G. The value of R0 for nonradiative energy transfer from monomer to dimer and the equilibrium constant for rhodamine 6G dimerization are calculated to be (3.2±0.1) nm and 19 M -1, respectively.

  13. Analysis of dilute aqueous multifluorophoric mixtures using excitation-emission matrix fluorescence (EEMF) and total synchronous fluorescence (TSF) spectroscopy: a comparative evaluation.

    PubMed

    Kumar, Keshav; Mishra, Ashok Kumar

    2013-12-15

    Excitation-emission matrix fluorescence (EEMF) and total synchronous fluorescence (TSF) spectroscopy are two conceptually different fluorescence techniques that have been used to map the fluorescence responses of the fluorophores present in a multifluorophoric mixture. EEMF was introduced four decades back and most of the fluorimeters have the suitable computer program which allows the acquisition EEMF spectra. Recently introduced TSF spectroscopy has been shown to possess good application potential in analytical fluorimetry and has started attracting the attention of analytical chemists. TSF data structure, however, is intrinsically different from EEMF data structure and a better understanding of TSF data structure is crucial to utilising its application potential. In the present work, a comprehensive comparative study between EEMF and TSF spectroscopic data set was performed by taking aqueous mixtures containing low concentrations of benzo[a]pyrene, chrysene, and pyrene as test case. The EEMF and TSF data structures were clearly explained by taking pyrene as an example. The effects of Rayleigh and Raman scattering on the quality of EEMF and TSF data sets were studied. EEMF and TSF data sets of dilute aqueous mixtures of benzo[a]pyrene, chrysene, and pyrene were subjected to three chemometric techniques PARAFAC, N-PLS, and MCR-ALS analysis. TSF data set in particular was found to be highly attuned to MCR-ALS analysis. Obtained results of chemometric analyses on EEMF and TSF data sets show that TSF data of dilute aqueous mixtures provides more accurate spectral and concentration information than EEMF data sets. Therefore, TSF spectroscopy could be considered as an alternate to the EEMF for the analyses of dilute multifluorophoric mixtures.

  14. Interactions of cyanine dyes with nucleic acids. XXIV. Aggregation of monomethine cyanine dyes in presence of DNA and its manifestation in absorption and fluorescence spectra

    NASA Astrophysics Data System (ADS)

    Ogul'chansky, T. Yu; Losytskyy, M. Yu; Kovalska, V. B.; Yashchuk, V. M.; Yarmoluk, S. M.

    2001-06-01

    Absorption, fluorescence emission and excitation spectra of benzothiazole cyanine dyes — thiazole orange (TO) and 7-methyl-6-(3-methyl-2,3-dihydro-1,3-benzothiazol-2-ylidenmethyl) [1,3] dioxolo [4',5':4,5] benzo [d] [1,3] thiazolium methylmethosulfate (Cyan 13) — were investigated over a wide concentration range. The dyes form aggregates with a 'sandwich'-like structure in water solution. At low dye to DNA concentrations ratios, Cyan 13 and TO monomers appear to interact with the DNA. On increasing the dye to DNA concentrations ratio, free dye molecules aggregate with the DNA-bound ones. The spectra of the free dye aggregates and the aggregates formed on the DNA, are characterized by an anomalously large (more than 100 nm) Stokes shift. This suggests, that the π-electron systems of the aggregates undergo substantial changes in excited state, compared to those of the monomers. The formation of aggregates consisting of the free and DNA-bound dye molecules can be explained using the half-intercalation model of the interaction of the cyanine dye monomers with the DNA.

  15. Absorption and fluorescence properties of the eastern Bering Sea in the summer with special reference to the influence of a Cold Pool

    NASA Astrophysics Data System (ADS)

    D'Sa, E. J.; Goes, J. I.; Gomes, H.; Mouw, C.

    2013-12-01

    The absorption and fluorescence properties of chromophoric dissolved organic matter (CDOM) are reported for the inner shelf, slope waters and outer shelf regions of the eastern Bering Sea during the summer of 2008, when a warm, thermally stratified surface mixed layer lay over a Cold Pool (< 2 °C) that occupied the entire middle shelf. CDOM absorption at 355 nm (ag355) and its spectral slope (S) in conjunction with excitation emission matrix (EEM) fluorescence and parallel factor analysis (PARAFAC) revealed large variability in the characteristics of CDOM in different regions of the Bering Sea. PARAFAC analysis aided in the identification of three humic-like (components 1, 2 and 5) and two protein-like (a tyrosine-like component 3, and a tryptophan-like component 4) components. In the extensive shelf region, average absorption coefficients at 355 nm (ag355 m-1) and DOC concentrations (μM) were highest in the inner shelf (0.342 ± 0.11 m-1, 92.67 ± 14.60 μM) and lower in the middle (0.226 ± 0.05 m-1, 78.38 ± 10.64 μM) and outer (0.176 ± 0.05 m-1, 80.73 ± 18.11 μM) shelves, respectively. Mean spectral slopes S were elevated in the middle shelf (24.38 ± 2.25 μm-1) especially in the surface waters (26.87 ± 2.39 μm-1) indicating high rates of photodegradation in the highly stratified surface mixed layer, which intensified northwards in the northern middle shelf likely contributing to greater light penetration and to phytoplankton blooms at deeper depths. The fluorescent humic-like components 1, 2, and 5 were most elevated in the inner shelf most likely from riverine inputs. Measurements at depth in slope waters (> 250 m) revealed low values of ag355 (0.155 ± 0.03 m-1) and S (15.45 ± 1.78 μm-1) indicative of microbial degradation of CDOM in deep waters. DOC concentrations, however were not significantly different suggesting CDOM sources and sinks to be uncoupled from DOC. Along the productive "green belt" in the outer shelf/slope region, absorption and

  16. Absorption and fluorescence properties of chromophoric dissolved organic matter of the eastern Bering Sea in the summer with special reference to the influence of a cold pool

    NASA Astrophysics Data System (ADS)

    D'Sa, E. J.; Goes, J. I.; Gomes, H.; Mouw, C.

    2014-06-01

    The absorption and fluorescence properties of chromophoric dissolved organic matter (CDOM) are reported for the inner shelf, slope waters and outer shelf regions of the eastern Bering Sea during the summer of 2008, when a warm, thermally stratified surface mixed layer lay over a cold pool (< 2 °C) that occupied the entire middle shelf. CDOM absorption at 355 nm (ag355) and its spectral slope (S) in conjunction with excitation-emission matrix (EEM) fluorescence and parallel factor analysis (PARAFAC) revealed large variability in the characteristics of CDOM in different regions of the Bering Sea. PARAFAC analysis aided in the identification of three humic-like (components one, two and five) and two protein-like (a tyrosine-like component three, and a tryptophan-like component four) components. In the extensive shelf region, average absorption coefficients at 355 nm (ag355, m-1) and DOC concentrations (μM) were highest in the inner shelf (0.342 ± 0.11 m-1, 92.67 ± 14.60 μM) and lower in the middle (0.226 ± 0.05 m-1, 78.38 ± 10.64 μM) and outer (0.185 ± 0.05 m-1, 79.24 ± 18.01 μM) shelves, respectively. DOC concentrations, however were not significantly different, suggesting CDOM sources and sinks to be uncoupled from DOC. Mean spectral slopes S were elevated in the middle shelf (24.38 ± 2.25 μm-1) especially in the surface waters (26.87 ± 2.39 μm-1) indicating high rates of photodegradation in the highly stratified surface mixed layer, which intensified northwards in the northern middle shelf likely contributing to greater light penetration and to phytoplankton blooms at deeper depths. The fluorescent humic-like components one, two, and five were most elevated in the inner shelf most likely from riverine inputs. Along the productive "green belt" in the outer shelf/slope region, absorption and fluorescence properties indicated the presence of fresh and degraded autochthonous DOM. Near the Unimak Pass region of the Aleutian Islands, low DOC and ag355

  17. Effects of annealing treatment and gamma irradiation on the absorption and fluorescence spectra of Cr:GSGG laser crystal

    NASA Astrophysics Data System (ADS)

    Sun, D. L.; Luo, J. Q.; Xiao, J. Z.; Zhang, Q. L.; Jiang, H. H.; Yin, S. T.; Wang, Y. F.; Ge, X. W.

    2008-09-01

    The influence of annealing treatments and gamma-ray irradiation on the absorption and fluorescence spectra of Cr:GSGG crystals grown by the Czochralski method has been investigated. Two absorption bands located near 686 nm and 1050 nm were weakened markedly after the crystal was re-annealed in H2 atmosphere, which is due to the Cr4+ ions being de-oxidized into Cr3+ ions. The other two weak additional absorption bands induced by gamma-ray irradiation appearing near 310 nm and 480 nm are ascribed to the Fe2+ ions and F-type color centers, respectively. In particular, the gamma-ray irradiation with a dose of 100 Mrad has an effect of improving slightly the luminescence properties of Cr:GSGG crystals. The improvement mechanism is analyzed and discussed.

  18. Decay kinetics and quantum yields of fluorescence in photosystem I from Synechococcus elongatus with P700 in the reduced and oxidized state: are the kinetics of excited state decay trap-limited or transfer-limited?

    PubMed Central

    Byrdin, M; Rimke, I; Schlodder, E; Stehlik, D; Roelofs, T A

    2000-01-01

    Transfer and trapping of excitation energy in photosystem I (PS I) trimers isolated from Synechococcus elongatus have been studied by an approach combining fluorescence induction experiments with picosecond time-resolved fluorescence measurements, both at room temperature (RT) and at low temperature (5 K). Special attention was paid to the influence of the oxidation state of the primary electron donor P700. A fluorescence induction effect has been observed, showing a approximately 12% increase in fluorescence quantum yield upon P700 oxidation at RT, whereas at temperatures below 160 K oxidation of P700 leads to a decrease in fluorescence quantum yield ( approximately 50% at 5 K). The fluorescence quantum yield for open PS I (with P700 reduced) at 5 K is increased by approximately 20-fold and that for closed PS I (with P700 oxidized) is increased by approximately 10-fold, as compared to RT. Picosecond fluorescence decay kinetics at RT reveal a difference in lifetime of the main decay component: 34 +/- 1 ps for open PS I and 37 +/- 1 ps for closed PS I. At 5 K the fluorescence yield is mainly associated with long-lived components (lifetimes of 401 ps and 1.5 ns in closed PS I and of 377 ps, 1.3 ns, and 4.1 ns in samples containing approximately 50% open and 50% closed PS I). The spectra associated with energy transfer and the steady-state emission spectra suggest that the excitation energy is not completely thermally equilibrated over the core-antenna-RC complex before being trapped. Structure-based modeling indicates that the so-called red antenna pigments (A708 and A720, i.e., those with absorption maxima at 708 nm and 720 nm, respectively) play a decisive role in the observed fluorescence kinetics. The A720 are preferentially located at the periphery of the PS I core-antenna-RC complex; the A708 must essentially connect the A720 to the reaction center. The excited-state decay kinetics turn out to be neither purely trap limited nor purely transfer (to the trap

  19. A laser-excited synchronous fluorescence spectrometer for benzo(a)pyrene

    SciTech Connect

    Huebedr, D.M.; Iruela Del Olmo, M.; Stevenson, C.L.; Vo-Dinh, T.

    1995-09-01

    A laser-based synchronous scanning fluorimeter was developed for detecting polycyclic aromatic hydrocarbons (PAHs). Although extensive sample cleanup and separation are usually required, fluorescence has long been used for determination of PAHs. Amount of sample pretreatment can often be reduced when synchronous fluorescence (SF) is used, and the sensitivity of SF can be increased by the high power and narrow profile of a laser source. A small, portable, pulsed dye laser was used in this study. The instrument and its application to the determination of the carcinogen benzo(a)pyrene are described.

  20. Development of an electrothermal atomization laser-excited atomic fluorescence spectrometry procedure for direct measurements of arsenic in diluted serum.

    PubMed

    Swart, D J; Simeonsson, J B

    1999-11-01

    A procedure for the direct determination of arsenic in diluted serum by electrothermal atomization laser-excited atomic fluorescence spectrometry (ETA-LEAFS) is reported. Laser radiation needed to excite As at 193.696 and 197.197 nm is generated as the second anti-Stokes stimulated Raman scattering output of a frequency-doubled dye laser operating near 230.5 and 235.5 nm, respectively. Two different LEAFS schemes have been utilized and provide limits of detection of 200-300 fg for As in aqueous standards. When measurements of serum samples diluted 1:10 with deionized water are performed, a stable background signal is observed that can be accounted for by taking measurements with the laser tuned off-wavelength. No As is detected in any of the bovine or human serum samples analyzed. Measurements of 100 pg/mL standard additions of As to a diluted bovine serum sample utilizing either inorganic or organic As species demonstrate a linear relationship of the fluorescence signal to As spike concentration, but exhibit a sensitivity of approximately half that observed in pure aqueous standards. The limit of detection for As in 1:10 diluted serum samples is 65 pg/mL or 650 fg absolute mass, which corresponds to 0.65 ng/mL As in undiluted serum. To our knowledge, the ETA-LEAFS procedure is currently the only one capable of directly measuring As in diluted serum at these levels.