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Sample records for absorption fluorescence excitation

  1. Excitation energy dependence of excited states dynamics in all- trans-carotenes determined by femtosecond absorption and fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Kosumi, Daisuke; Yanagi, Kazuhiro; Nishio, Tomohiro; Hashimoto, Hideki; Yoshizawa, Masayuki

    2005-06-01

    Ultrafast relaxation kinetics in β-carotene and lycopene has been investigated by femtosecond absorption and fluorescence spectroscopies using tunable excitation pulses. The transient signals induced by the photoexcitation with larger excess energy have broader bands and longer lifetimes both in the 11Bu+and21Ag- excited states. The excess vibrational energy remains longer than several picoseconds and slows the relaxation kinetics in carotenoids.

  2. 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.

  3. Donor-acceptor substituted phenylethynyltriphenylenes – excited state intramolecular charge transfer, solvatochromic absorption and fluorescence emission

    PubMed Central

    Nandy, Ritesh

    2010-01-01

    Summary Several 2-(phenylethynyl)triphenylene derivatives bearing electron donor and acceptor substituents on the phenyl rings have been synthesized. The absorption and fluorescence emission properties of these molecules have been studied in solvents of different polarity. For a given derivative, solvent polarity had minimal effect on the absorption maxima. However, for a given solvent the absorption maxima red shifted with increasing conjugation of the substituent. The fluorescence emission of these derivatives was very sensitive to solvent polarity. In the presence of strongly electron withdrawing (–CN) and strongly electron donating (–NMe2) substituents large Stokes shifts (up to 130 nm, 7828 cm−1) were observed in DMSO. In the presence of carbonyl substituents (–COMe and –COPh), the largest Stokes shift (140 nm, 8163 cm−1) was observed in ethanol. Linear correlation was observed for the Stokes shifts in a Lippert–Mataga plot. Linear correlation of Stokes shift was also observed with E T(30) scale for protic and aprotic solvents but with different slopes. These results indicate that the fluorescence emission arises from excited state intramolecular charge transfer in these molecules where the triphenylene chromophore acts either as a donor or as an acceptor depending upon the nature of the substituent on the phenyl ring. HOMO–LUMO energy gaps have been estimated from the electrochemical and spectral data for these derivatives. The HOMO and LUMO surfaces were obtained from DFT calculations. PMID:21085512

  4. 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.

  5. Absolute excited-state absorption cross section and fluorescence quantum efficiency of Cr/sup 3 +/: gadolinium scandium gallium garnet

    SciTech Connect

    Seelert, W.; Strauss, E.

    1987-10-01

    Excited-state properties of the laser material Cr/sup 3 +/:Gd/sub 3/Sc/sub 2/(GaO/sub 4/)/sub 3/ were determined by a photocaloric technique. The excited-state absorption cross section at 650 nm is (3.6 +- 0.6)10/sup -20/ cm/sup 2/, and the fluorescence quantum efficiency at ambient temperature is (91 +- 1)%.

  6. 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

  7. Excited state dipole moments of N, N-dimethylaniline from thermochromic effect on electronic absorption and fluorescence spectra

    NASA Astrophysics Data System (ADS)

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

    2006-01-01

    The effect of temperature on absorption and fluorescence spectra of N, N-dimethylaniline (DMA) in ethyl acetate has been studied for temperature ranging from 293 to 388 K. The permittivity ɛ and refractive index n of the solvent decrease with temperature increase and the absorption and fluorescence bands are blue shifted (so-called "thermochromic shift"). Based on this phenomenon, the dipole moment μe in the excited singlet state and the Onsager interaction radius a for DMA were determined using the Bilot and Kawski theory [L. Bilot, A. Kawski, Z. Naturforsch. 17a (1962) 621; 18a (1963) 10, 256]. For the known dipole moment in the ground state μg = 1.61 D and for α/ a3 = 0.54 ( α is the polarizability of the solute) the average value of μe = 3.55 D and a = 3.1 Å were determined. The obtained values for DMA are compared with the experimental values determined by other authors.

  8. 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.

  9. Asymmetry in ground and excited states in styryls and methoxystyryls detected by NMR (13C), absorption, fluorescence and fluorescence excitation spectroscopy

    NASA Astrophysics Data System (ADS)

    Stanova, A. V.; Ryabitsky, A. B.; Yashchuk, V. M.; Kachkovsky, O. D.; Gerasov, A. O.; Prostota, Ya. O.; Kropachev, O. V.

    2011-03-01

    Combined quantum-chemical and spectral study of electron structure features of styryls and their oxyanalogues containing benzothiazolium, benzooxazolium, indoleninium, pyridium, quinolinium residues has been fulfilled. It showed that asymmetry degree of molecular geometry and charge distribution in the chromophore of styryls and methoxystyryls considerably differ in the ground and excited states. It was established that two the lowest transitions in styryls are splitting and involve both donor levels, similarly to symmetrical cyanines. If compare with methoxystyryls the long-wave high intensive absorption band is shifted bathochromically due to considerable interaction between the donor quasi-local chromophores. In contrary, because of the low position of a lone electron pair of oxygen in methoxystyryls, only one donor quasi-local chromophore is effective, hence such unsymmetrical dyes absorb appreciably higher.

  10. 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

    2016-09-08

    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.

  11. Anharmonic Franck-Condon simulation of the absorption and fluorescence spectra for the low-lying S1 and S2 excited states of pyridine.

    PubMed

    Wang, Huan; Zhu, Chaoyuan; Yu, Jian-Guo; Lin, Sheng Hsien

    2009-12-31

    Anharmonic effects of the absorption and fluorescence spectra of pyridine molecule are studied and analyzed for the two-low lying singlet excited states S(1)((1)B(1)) and S(2)((1)B(2)). The complete active space self-consistent field (CASSCF) method is utilized to compute equilibrium geometries and all 27 vibrational normal-mode frequencies for the ground state and the two excited states. The present calculations show that the frequency differences between the ground and two excited states are small for the ten totally symmetric vibrational modes so that the displaced oscillator approximation can be used for spectrum simulations. The Franck-Condon factors within harmonic approximation basically grasp the main features of molecular spectra, but simulated 0-0 transition energy position and spectrum band shapes are not satisfactorily good for S(1)((1)B(1)) absorption and fluorescence spectra in comparison with experiment observation. As the first-order anharmonic correction added to Franck-Condon factors, both spectrum positions and band shapes can be simultaneously improved for both absorption and fluorescence spectra. It is concluded that the present anharmonic correction produces a significant dynamic shifts for spectrum positions and improves spectrum band shapes as well. The detailed structures of absorption spectrum of S(2)((1)B(2)) state observed from experiment can be also reproduced with anharmonic Franck-Condon simulation, and these were not shown in the harmonic Franck-Condon simulation with either distorted or Duschinsky effects in the literature.

  12. 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.

  13. Multiphoton excitation of fluorescent DNA base analogs.

    PubMed

    Katilius, Evaldas; Woodbury, Neal W

    2006-01-01

    Multiphoton excitation was used to investigate properties of the fluorescent DNA base analogs, 2-aminopurine (2AP) and 6-methylisoxanthopterin (6MI). 2-aminopurine, a fluorescent analog of adenine, was excited by three-photon absorption. Fluorescence correlation measurements were attempted to evaluate the feasibility of using three-photon excitation of 2AP for DNA-protein interaction studies. However, high excitation power and long integration times needed to acquire high signal-to-noise fluorescence correlation curves render three-photon excitation FCS of 2AP not very useful for studying DNA base dynamics. The fluorescence properties of 6-methylisoxanthopterin, a guanine analog, were investigated using two-photon excitation. The two-photon absorption cross-section of 6MI was estimated to be about 2.5 x 10(-50) cm(4)s (2.5 GM units) at 700 nm. The two-photon excitation spectrum was measured in the spectral region from 700 to 780 nm; in this region the shape of the two-photon excitation spectrum is very similar to the shape of single-photon excitation spectrum in the near-UV spectral region. Two-photon excitation of 6MI is suitable for fluorescence correlation measurements. Such measurements can be used to study DNA base dynamics and DNA-protein interactions over a broad range of time scales.

  14. Estimation of ground- and excited-state dipole moments of Nile Red dye from solvatochromic effect on absorption and fluorescence spectra

    NASA Astrophysics Data System (ADS)

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

    2008-10-01

    The effect of various nonpolar and polar solvents on the location of absorption and fluorescence of Nile Red at room temperature was investigated. Based on the batochromic shift of electronic spectra of Nile Red and Bilot-Kawski theory the following values of ground- and excited-state dipole moments μg = 8.2 ± 1.0 D and μe = 10.0 ± 1.0 D were found. The reasons of high discrepancy between the literature results were discussed.

  15. [Two-photon excitation fluorescence spectrum of the light-harvesting complex LH2 from Chromatium minutissimum within 650-745 nm range is determined by two-photon absorption of bacteriochlorophyll rather than of carotenoids].

    PubMed

    Krikunova, M A; Leupold, D; Rini, M; Voigt, B; Moskalenko, A A; Toropygina, O A; Razzhivin, A P

    2002-01-01

    Two-photon fluorescence excitation spectra of the peripheral light-harvesting complex LH2 from the purple photosynthetic bacterium Chromatium minutissimum were examined within the expected spectral range of the optically forbidden S1 singlet state of carotenoids. LH2 preparations isolated from wild-type and carotenoid-depleted cells were used. 100-fs laser pulses in the range of 1300-1490 nm with an energy of 7-9 mW (corresponding to one-photon absorption between 650 and 745 nm) were used for two-photon fluorescence excitation. It was shown that two-photon fluorescence excitation spectra of LH2 complex from wild and carotenoid-depleted cells are very similar to each other and to the two-photon fluorescence excitation spectrum of bacteriochlorophyll a in acetone. It was concluded that direct two-photon excitation of bacteriochlorophyll a determines the fluorescence of both samples within the 650-745 nm spectral range.

  16. Quantitative treatment of the solvent effects on the electronic absorption and fluorescence spectra of acridines and phenazines. The ground and first excited singlet-state dipole moments

    NASA Astrophysics Data System (ADS)

    Aaron, Jean Jacques; Maafi, Mounir; Párkányi, Cyril; Boniface, Christian

    1995-04-01

    Electronic absorption and fluorescence excitation and emission spectra of four acridines (acridine, Acridine Yellow, 9-aminoacridine and proflavine) and three phenazines (phenazine, neutral Red and safranine) are determined at room temperature (298 K) in several solvents of various polarities (dioxane, chloroform, ethyl ether, ethyl acetate, 1-butanol, 2-propanol, ethanol, methanol, dimethylformamide, acetonitrile and dimethyl sulfoxide). The effect of the solvent upon the spectral characteristics of the above compounds, is studied. In combination with the ground-state dipole moments of these compounds, the spectral data are used to evaluate their first excited singlet-state dipole moments by means of the solvatochromic shift method (Bakhshiev's and Kawski-Chamma-Viallet's correlations). The theoretical ground and excited singlet-state dipole moments for acridines and phenazines are also calculated as a vector sum of the π-component (obtained by the PPP method) and the σ-component (obtained from σ-bond moments). For most acridines and phenazines under study, the experimental excited singlet-state dipole moments are found to be higher than their ground state counterpart. The application of the Kamlet-Abboud-Taft solvatochromic parameters to the solvent effect on spectral properties of acridine and phenazine derivatives is discussed.

  17. Solvent effect on the absorption and fluorescence spectra of 7-acetoxy-6-(2,3-dibromopropyl)-4,8-dimethylcoumarin: determination of ground and excited state dipole moments.

    PubMed

    Gülseven Sıdır, Yadigar; Sıdır, Isa

    2013-02-01

    The ground state (μ(g)) and excited state (μ(e)) dipole moments of 7-acetoxy-6-(2,3-dibromopropyl)-4,8-dimethylcoumarin (abbreviated as 7ADDC) are estimated from solvatochromic shifts of absorption and fluorescence spectra as a function of the dielectric constant (ε) and refractive index (n). While the ground state dipole moment is determined by using Bilot-Kawski method, the excited state dipole moment is calculated by using Bilot-Kawski, Lippert-Mataga, Bakhshiev, Kawski-Chamma-Viallet and Reichardt correlation methods. Excited state dipole moment is observed as larger than the ground state dipole moment due to substantial π-electron density redistribution. The ground state and excited state dipole moments are observed as parallel to each other with angle of 0°. Solute-solvent interactions are analyzed by means of linear solvation free energy relationships (LSER) using dielectric constant function (f(ε)), refractive index function (f(n)) and Kamlet-Taft parameters (α and β). Atomic charges, electron densities and molecular orbitals are calculated in vacuum and with solvent effect by using both DFT and TDDFT methods. Solvent accessible surface, molecular electrostatic potential (MEP) and electrostatic potential (ESP) are visualized as a result of DFT calculations.

  18. Quantitative treatment of the effect of solvent on the electronic absorption and fluorescence spectra of substituted coumarins: Evaluation of the first excited singlet-state dipole moments.

    PubMed

    Aaron, J J; Buna, M; Parkanyi, C; Antonious, M S; Tine, A; Cisse, L

    1995-12-01

    The electronic absorption and fluorescence spectra of coumarin and 11 substituted coumarins were measured in several solvents (dioxane, ethyl ether, ethyl acetate, ethanol, dimethylformamide, acetonitrile, and dimethyl sulfoxide). Ground-state dipole moments were determined in dioxane at 298 K. The results were used to obtain the first excited singlet-state dipole moments of the coumarins under study by the solvatochromic shift method (Bakhshiev, Kawski-Chamma-Viallet, McRae, and Suppan correlations). Also, the ground- and the first excited singlet-state dipole moments were calculated using a combination of the PPP method (π-contribution) and the vector sum of the σ-bond and group moments (σ-contribution). In general, the first excited singlet-state dipole moments of the coumarins are noticeably higher than the corresponding ground-state values, indicating a substantial redistribution of theπ-electron densities resulting in a more polar excited state. There is a reasonably good agreement between the calculated and the experimental dipole moments.

  19. Solvent effect on the absorption and fluorescence spectra of 7-acetoxy-6-(2,3-dibromopropyl)-4,8-dimethylcoumarin: Determination of ground and excited state dipole moments

    NASA Astrophysics Data System (ADS)

    Gülseven Sıdır, Yadigar; Sıdır, İsa

    2013-02-01

    The ground state (μg) and excited state (μe) dipole moments of 7-acetoxy-6-(2,3-dibromopropyl)-4,8-dimethylcoumarin (abbreviated as 7ADDC) are estimated from solvatochromic shifts of absorption and fluorescence spectra as a function of the dielectric constant (ɛ) and refractive index (n). While the ground state dipole moment is determined by using Bilot-Kawski method, the excited state dipole moment is calculated by using Bilot-Kawski, Lippert-Mataga, Bakhshiev, Kawski-Chamma-Viallet and Reichardt correlation methods. Excited state dipole moment is observed as larger than the ground state dipole moment due to substantial π-electron density redistribution. The ground state and excited state dipole moments are observed as parallel to each other with angle of 0°. Solute-solvent interactions are analyzed by means of linear solvation free energy relationships (LSER) using dielectric constant function (f(ɛ)), refractive index function (f(n)) and Kamlet-Taft parameters (α and β). Atomic charges, electron densities and molecular orbitals are calculated in vacuum and with solvent effect by using both DFT and TDDFT methods. Solvent accessible surface, molecular electrostatic potential (MEP) and electrostatic potential (ESP) are visualized as a result of DFT calculations.

  20. 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.

  1. 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.

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

    PubMed

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

    2016-02-05

    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 (μ(e)). It is observed that dipole moment value of excited state (μ(e)) 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.

  3. Solvent effects on the absorption and fluorescence spectra of some laser dyes: estimation of ground and excited-state dipole moments.

    PubMed

    Thipperudrappa, J; Biradar, D S; Manohara, S R; Hanagodimath, S M; Inamadar, S R; Manekutla, R J

    2008-03-01

    The absorption and fluorescence spectra of three extensively used laser dyes namely 1,1,4,4-tetraphenyl-1,3-butadiene (TPB), 2-(4'-t-butylphenyl)-5-(4''-biphenylyl)-1-oxa-3,4-diazole (BPBD), 1,4-bis[2-(2-methylphenyl)ethenyl]-benzene (Bis-MSB) have been recorded at room temperature (300K) in solvents of different polarities. The effects of the solvents upon the spectral properties are discussed. The ground-state dipole moments (mu(g)) were determined experimentally by Guggenheim and Higasi method separately and were compared with theoretical values obtained using quantum chemical method. The ground-state dipole moments obtained by using Guggenheim method were then used in the estimation of excited-state dipole moments (mu(e)) by using Lippert's, Bakhshiev's and Kawski-Chamma-Viallet's equations. In all the above three equations the variation of the Stokes shift with the solvent dielectric constant and refractive index was made use of. It was observed that dipole moments of excited state were higher than those of the ground state for all the dyes.

  4. Solvent effects on the absorption and fluorescence spectra of some laser dyes: Estimation of ground and excited-state dipole moments

    NASA Astrophysics Data System (ADS)

    Thipperudrappa, J.; Biradar, D. S.; Manohara, S. R.; Hanagodimath, S. M.; Inamadar, S. R.; Manekutla, R. J.

    2008-03-01

    The absorption and fluorescence spectra of three extensively used laser dyes namely 1,1,4,4-tetraphenyl-1,3-butadiene (TPB), 2-(4'- t-butylphenyl)-5-(4″-biphenylyl)-1-oxa-3,4-diazole (BPBD), 1,4-bis[2-(2-methylphenyl)ethenyl]-benzene (Bis-MSB) have been recorded at room temperature (300 K) in solvents of different polarities. The effects of the solvents upon the spectral properties are discussed. The ground-state dipole moments ( μg) were determined experimentally by Guggenheim and Higasi method separately and were compared with theoretical values obtained using quantum chemical method. The ground-state dipole moments obtained by using Guggenheim method were then used in the estimation of excited-state dipole moments ( μe) by using Lippert's, Bakhshiev's and Kawski-Chamma-Viallet's equations. In all the above three equations the variation of the Stokes shift with the solvent dielectric constant and refractive index was made use of. It was observed that dipole moments of excited state were higher than those of the ground state for all the dyes.

  5. Two-photon excitation fluorescence microscopy.

    PubMed

    So, P T; Dong, C Y; Masters, B R; Berland, K M

    2000-01-01

    Two-photon fluorescence microscopy is one of the most important recent inventions in biological imaging. This technology enables noninvasive study of biological specimens in three dimensions with submicrometer resolution. Two-photon excitation of fluorophores results from the simultaneous absorption of two photons. This excitation process has a number of unique advantages, such as reduced specimen photodamage and enhanced penetration depth. It also produces higher-contrast images and is a novel method to trigger localized photochemical reactions. Two-photon microscopy continues to find an increasing number of applications in biology and medicine.

  6. On the Estimation of Excited-State Dipole Moments from Solvatochromic Shifts of Absorption and Fluorescence Spectra

    NASA Astrophysics Data System (ADS)

    Kawski, A.

    2002-05-01

    The effect of the solvent polarity parameters f (Ɛ, n) and ᵩ(Ɛ, n) appearing in solvatochromic theories, and the effect of the molecular shape of the Onsager cavity (sphere, ellipsoid of revolution) on the determined electric dipole moments μe in the singlet excited state are studied. It is found that the shape of the solute does not exhibit a significant effect on the determined values of μe, but only on the solvent parameters f (Ɛ, n) and ᵩ(Ɛ, n) as well as on the Onsager radius a. Passing from a sphere to an ellipsoid leads to such a change in the scale that induces a proportional change in the slope coefficients m1 and m2. Also the effect of α/a3 (a and a are the mean isotropic polarizability of the solute and the Onsager cavity radius in a homogeneous dielectric, respectively) on the determined values of m e has been studied, and it is found that the assumption α/a3 = 1/2 is valid in many cases.

  7. Intramolecular charge transfer with the planarized 4-cyanofluorazene and its flexible counterpart 4-cyano-N-phenylpyrrole. Picosecond fluorescence decays and femtosecond excited-state absorption.

    PubMed

    Druzhinin, Sergey I; Kovalenko, Sergey A; Senyushkina, Tamara A; Demeter, Attila; Machinek, Reinhard; Noltemeyer, Mathias; Zachariasse, Klaas A

    2008-09-11

    The fluorescence spectrum of the rigidified 4-cyanofluorazene (FPP4C) in n-hexane consists of a dual emission from a locally excited (LE) and an intramolecular charge-transfer (ICT) state, with an ICT/LE fluorescence quantum yield ratio of Phi'(ICT)/Phi(LE) = 3.3 at 25 degrees C. With the flexible 4-cyano- N-phenylpyrrole (PP4C) in n-hexane, such an ICT reaction also takes place, with Phi'(ICT)/Phi(LE) = 1.5, indicating that for this reaction, a perpendicular twist of the pyrrole and benzonitrile moieties is not required. The ICT emission band of FPP4C and PP4C in n-hexane has vibrational structure, but a structureless band is observed in all other solvents more polar than the alkanes. The enthalpy difference Delta H of the LE --> ICT reaction in n-hexane, -11 kJ/mol for FPP4C and -7 kJ/mol for PP4C, is determined by analyzing the temperature dependence of Phi'(ICT)/Phi(LE). Using these data, the energy E(FC,ICT) of the Franck-Condon ground state populated by the ICT emission is calculated, 41 (FPP4C) and 40 kJ/mol (PP4C). These large values for E(FC,ICT) lead to the conclusion that with FPP4C and PP4C, direct ICT excitation, bypassing LE, does not take place. FPP4C has an ICT dipole moment of 15 D, similar to that of PP4C (16 D). Picosecond fluorescence decays allow the determination of the ICT lifetime, from which the radiative rate constant k'(f)(ICT) is derived, with comparable values for FPP4C and PP4C. This shows that an argument for a twisted ICT state of PP4C cannot come from k'(f)(ICT). After correction for the solvent refractive index and the energy of the emission maximum nu(max)(ICT), it appears that k'(f)(ICT) is solvent-polarity-independent. Femtosecond transient absorption with FPP4C and PP4C in n-hexane reveals that the ICT state is already nearly fully present at 100 fs after excitation, in rapid equilibrium with LE. In MeCN, the ICT state of FPP4C and PP4C is likewise largely developed at this delay time, and the reaction is limited by dielectric

  8. Two-photon absorption properties of fluorescent proteins

    PubMed Central

    Drobizhev, Mikhail; Makarov, Nikolay S.; Tillo, Shane E.; Hughes, Thomas E.; Rebane, Aleksander

    2016-01-01

    Two-photon excitation of fluorescent proteins is an attractive approach for imaging living systems. Today researchers are eager to know which proteins are the brightest, and what the best excitation wavelengths are. Here we review the two-photon absorption properties of a wide variety of fluorescent proteins, including new far-red variants, to produce a comprehensive guide to choosing the right FP and excitation wavelength for two-photon applications. PMID:21527931

  9. Control of excitation in the fluorescence microscope.

    PubMed

    Lea, D J; Ward, D J

    1979-01-01

    In fluorescence microscopy image brightness and contrast and the rate of fading depend upon the intensity of illumination of the specimen. An iris diaphragm or neutral density filters may be used to reduce fluorescence excitation. Also the excitation bandwidth may be varied by using a broad band exciter filter with a set of interchangeable yellow glass filters at the lamphouse.

  10. 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.

  11. Three-photon excitation in fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Hell, Stefan W.; Bahlmann, Karsten; Schrader, Martin; Soini, Aleksi; Malak, Henryk; Gryczynski, Ignacy; Lakowicz, Joseph R.

    1996-01-01

    We show experiments proving the feasibility of scanning fluorescence microscopy by three-photon excitation. Three-photon excitation fluorescence axial images are shown of polystyrene beads stained with the fluorophore 2,5- bis(4-biphenyl)oxazole (BBO). Three-photon excitation is performed at an excitation wavelength of 900 nm and with pulses of 130 fs duration provided by a mode-locked titanium-sapphire laser. Fluorescence is collected between 350 and 450 nm. The fluorescence image signal features a third-order dependence on the excitation power, also providing intrinsic 3-D imaging. The resolution of a three-photon excitation microscope is increased over that of a comparable two-photon excitation microscope.

  12. Multiphoton excited fluorescence spectroscopy of biomolecular systems

    NASA Astrophysics Data System (ADS)

    Birch, David J. S.

    2001-09-01

    Recent work on the emerging application of multiphoton excitation to fluorescence studies of biomolecular dynamics and structure is reviewed. The fundamental principles and experimental techniques of multiphoton excitation are outlined, fluorescence lifetimes, anisotropy and spectra in membranes, proteins, hydrocarbons, skin, tissue and metabolites are featured, and future opportunities are highlighted.

  13. Application of excitation-emission fluorescence matrices and UV/Vis absorption to monitoring the photocatalytic degradation of commercial humic acid.

    PubMed

    Valencia, Sergio; Marín, Juan M; Restrepo, Gloria; Frimmel, Fritz H

    2013-01-01

    This study reports the use of excitation-emission matrix (EEM) fluorescence and UV/Vis spectroscopy to monitor the changes in the composition and reactivity of Aldrich humic acids (Aldrich HA) as a model compound for natural organic matter (NOM) during photocatalytic degradation. Degussa P-25 titanium dioxide (TiO(2)) and a solar UV-light simulator (a batch reactor) were used. The photocatalysis shifted the fluorescence maxima of EEMs of Aldrich HA toward shorter wavelengths, which implied that the photocatalytic degradation of commercial Aldrich HA caused the breakdown of high molecular weight components and the formation of lower molecular weight fractions. In addition, the fluorescence intensity of fulvic- and humic-like Aldrich HA presented a strong correlation with dissolved organic carbon (DOC), specific UV absorbance (SUVA) parameters, trihalomethane formation potential (THMFP), and organically bound halogens absorbable on activated carbon formation potential (AOXFP). Fluorescence spectroscopy was shown to be a powerful tool for monitoring of the photocatalytic degradation of HA.

  14. Two-photon fluorescence excitation spectroscopy of biological molecules

    NASA Astrophysics Data System (ADS)

    Meshalkin, Yuri P.; Alfimov, E. E.; Groshev, D. E.; Makukha, V. K.

    1996-06-01

    The UV fluorescence spectra of aromatic amino-acids and some proteins at two photon excitation by second harmonic of Nd:YAG laser are received. Two-photon absorption cross sections of tryptophan, tyrosine, phenylalanine and proteins: bovine serum albumin, lysozyme, trypsin, (alpha) - chymotrypsinogen and pepsin at wavelength 532 nm were measured by means of the two-quantum standard method.

  15. Intravital Fluorescence Excitation in Whole-Animal Optical Imaging

    PubMed Central

    Bixler, Joel N.; Kong, Ying; Cirillo, Jeffrey D.; Maitland, Kristen C.

    2016-01-01

    Whole-animal fluorescence imaging with recombinant or fluorescently-tagged pathogens or cells enables real-time analysis of disease progression and treatment response in live animals. Tissue absorption limits penetration of fluorescence excitation light, particularly in the visible wavelength range, resulting in reduced sensitivity to deep targets. Here, we demonstrate the use of an optical fiber bundle to deliver light into the mouse lung to excite fluorescent bacteria, circumventing tissue absorption of excitation light in whole-animal imaging. We present the use of this technology to improve detection of recombinant reporter strains of tdTomato-expressing Mycobacterium bovis BCG (Bacillus Calmette Guerin) bacteria in the mouse lung. A microendoscope was integrated into a whole-animal fluorescence imager to enable intravital excitation in the mouse lung with whole-animal detection. Using this technique, the threshold of detection was measured as 103 colony forming units (CFU) during pulmonary infection. In comparison, the threshold of detection for whole-animal fluorescence imaging using standard epi-illumination was greater than 106 CFU. PMID:26901051

  16. Femtosecond two-photon-excited fluorescence of melanin

    NASA Astrophysics Data System (ADS)

    Teuchner, Klaus; Mueller, Susanne; Freyer, Wolfgang; Leupold, Dieter; Altmeyer, Peter; Stuecker, Markus; Hoffmann, Klaus

    2003-02-01

    Spectral and time-resolved fluorescence studies of different eumelanins (natural, synthetic, enzymatic) in solution have been carried out by two-photon excitation at 800 nm, using 80 fs pulses with photon flux densities <= 1027 cm-2.s-1. Whereas all samples show monotonously decreasing absorption between near UV and near IR, their fluorescence behavior indicates strong heterogeneity. With respect to the also measured one-photon excited fluorescence (OPF) of melanin at 400 nm, the overall spectral shape of the two-photon excited fluorescence (TPF) is red-shifted. Both OPF and TPF exhibit three-exponential decay with a shortest component # 200 ps. As is also shown, the fluorescence properties of melanin are dependent on the micro-environment. This allows the hypothesis, that the process of malignant transformation in skin tissue could be reflected in the fluorescence, provided the melanin in skin is selectively excited. The latter is realized by the described stepwise absorption of two 800 nm photons. In this way, indeed characteristic differences between the TPF spectra of healthy tissue, nevus cell nevi and malignant melanoma have been found.

  17. Intravital Fluorescence Excitation in Whole-Animal Optical Imaging.

    PubMed

    Nooshabadi, Fatemeh; Yang, Hee-Jeong; Bixler, Joel N; Kong, Ying; Cirillo, Jeffrey D; Maitland, Kristen C

    2016-01-01

    Whole-animal fluorescence imaging with recombinant or fluorescently-tagged pathogens or cells enables real-time analysis of disease progression and treatment response in live animals. Tissue absorption limits penetration of fluorescence excitation light, particularly in the visible wavelength range, resulting in reduced sensitivity to deep targets. Here, we demonstrate the use of an optical fiber bundle to deliver light into the mouse lung to excite fluorescent bacteria, circumventing tissue absorption of excitation light in whole-animal imaging. We present the use of this technology to improve detection of recombinant reporter strains of tdTomato-expressing Mycobacterium bovis BCG (Bacillus Calmette Guerin) bacteria in the mouse lung. A microendoscope was integrated into a whole-animal fluorescence imager to enable intravital excitation in the mouse lung with whole-animal detection. Using this technique, the threshold of detection was measured as 103 colony forming units (CFU) during pulmonary infection. In comparison, the threshold of detection for whole-animal fluorescence imaging using standard epi-illumination was greater than 106 CFU.

  18. Application of excitation and emission matrix fluorescence (EEM) and UV-vis absorption to monitor the characteristics of Alizarin Red S (ARS) during electro-Fenton degradation process.

    PubMed

    Lai, Bo; Zhou, Yuexi; Wang, Juling; Yang, Zhishan; Chen, Zhiqiang

    2013-11-01

    Oxidative degradation of Alizarin Red S (ARS) in aqueous solutions by using electro-Fenton was studied. At first, effect of operating parameters such as current density, aeration rate and initial pH on the degradation of ARS were studied by using UV-vis spectrum, respectively. Then, under the optimal operating conditions (current density: 10.0mAcm(-2), aeration rate: 1000mLmin(-1), initial pH: 2.8), the identification of degradation products of ARS was carried out by using GC-MS and HPLC, meanwhile its degradation pathway was proposed according to the intermediates. Considering the location, intensity and intensity ratio of fluorescence center peak of the ARS in aqueous solution, a convenient and quick monitoring method by using excitation-emission matrix fluorescence spectrum technology was developed to monitor the degradation degree of ARS through electro-Fenton process. Furthermore, it is suggested that the developed method would be promising for the quick analysis and evaluation of the degradation degree of the pollutants with π-conjugated system.

  19. Photosynthetic light-harvesting complexes: fluorescent and absorption spectroscopy under two-photon (1200-1500 nm) and one-photon (600-750 nm) excitation by laser femtosecond pulses

    NASA Astrophysics Data System (ADS)

    Stepanenko, Il'ya A.; Kompanets, Viktor O.; Chekalin, Sergey V.; Makhneva, Zoya K.; Moskalenko, Andrey A.; Razjivin, Andrei P.

    2010-09-01

    The pathways of excitation energy transfer (EET) via pigments of the light-harvesting antenna are still in discussion. The bacteriochlorophyll fluorescence of peripheral light-harvesting complexes (LH2) from purple bacteria can be observed upon two-photon excitation (TPE) within 1200-1500 nm spectral range (a broad band near 1300 nm). Earlier the occurrence of this band was taken as an evidence for the participation of "dark" carotenoid S1 state in EET processes (see [Walla et al., Proc. Nat. Acad. Sci. U.S.A. 97, 10808-10813 (2000)] and references in it). However we showed that TPE spectrum of LH2 fluorescence within 1200-1500 nm is not associated with carotenoids [Stepanenko et al., J. Phys. Chem. B. 113(34), 11720-11723 (2009)]. Here we present TPE spectra of fluorescence for chromatophores and lightharvesting complexes LH2 and LH1 from wild-type cells and from carotenoid-depleted or carotenoidless mutant cells of several purple bacteria. The broad band within 1300-1400 nm was found for all preparations. Absorption pump-probe femtosecond spectroscopy applied to LH2 complex from Rb. sphaeroides revealed the similar spectral and kinetic patterns for TPE at 1350 nm and one-photon excitation at 675 nm. Analysis of pigment composition of this complex by high-pressure liquid chromatography showed that even under mild isolation conditions some bacteriochlorophyll molecules were oxidized to 3-acetyl-chlorophyll molecules having the long-wavelength absorption peak in the 650-700 nm range. It is proposed that these 3-acetyl-chlorophyll molecules are responsible for the broad band in TPE spectra within the 1200-1500 nm region.

  20. Photosynthetic light-harvesting complexes: fluorescent and absorption spectroscopy under two-photon (1200-1500 nm) and one-photon (600-750 nm) excitation by laser femtosecond pulses

    NASA Astrophysics Data System (ADS)

    Stepanenko, Il'ya A.; Kompanets, Viktor O.; Chekalin, Sergey V.; Makhneva, Zoya K.; Moskalenko, Andrey A.; Razjivin, Andrei P.

    2011-02-01

    The pathways of excitation energy transfer (EET) via pigments of the light-harvesting antenna are still in discussion. The bacteriochlorophyll fluorescence of peripheral light-harvesting complexes (LH2) from purple bacteria can be observed upon two-photon excitation (TPE) within 1200-1500 nm spectral range (a broad band near 1300 nm). Earlier the occurrence of this band was taken as an evidence for the participation of "dark" carotenoid S1 state in EET processes (see [Walla et al., Proc. Nat. Acad. Sci. U.S.A. 97, 10808-10813 (2000)] and references in it). However we showed that TPE spectrum of LH2 fluorescence within 1200-1500 nm is not associated with carotenoids [Stepanenko et al., J. Phys. Chem. B. 113(34), 11720-11723 (2009)]. Here we present TPE spectra of fluorescence for chromatophores and lightharvesting complexes LH2 and LH1 from wild-type cells and from carotenoid-depleted or carotenoidless mutant cells of several purple bacteria. The broad band within 1300-1400 nm was found for all preparations. Absorption pump-probe femtosecond spectroscopy applied to LH2 complex from Rb. sphaeroides revealed the similar spectral and kinetic patterns for TPE at 1350 nm and one-photon excitation at 675 nm. Analysis of pigment composition of this complex by high-pressure liquid chromatography showed that even under mild isolation conditions some bacteriochlorophyll molecules were oxidized to 3-acetyl-chlorophyll molecules having the long-wavelength absorption peak in the 650-700 nm range. It is proposed that these 3-acetyl-chlorophyll molecules are responsible for the broad band in TPE spectra within the 1200-1500 nm region.

  1. Determination of subnanogram per cubic meter concentrations of metals in the air of a trace metal clean room by impaction graphite furnace atomic absorption and laser excited atomic fluorescence spectrometry.

    PubMed

    Liang, Z W; Wei, G T; Irwin, R L; Walton, A P; Michel, R G; Sneddon, J

    1990-07-15

    Air, drawn by vacuum through a jet, was impacted against the inside surface of an atomic absorption graphite electrothermal atomizer (ETA). The amounts of the particles thus collected were determined at the ng m-3 level by graphite furnace atomic absorption or at the pg m-3 level by laser excited atomic fluorescence. The overall reproducibility of two sets of measurements, made 7 months apart, was 23%, with no significant difference between the two sets of data, based on Student's "t" test at the 95% confidence level. Short-term reproducibility varied from 13% to 34% depending upon the air concentration of the metal. The method shows promise for monitoring long-term effectiveness of the filtering systems in trace metal clean rooms. It was not possible to test for accuracy, due to the low concentrations involved, but accuracy was expected to be within a factor of 2 or 3 of the actual value, based on theoretical aspects of impaction.

  2. 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.

  3. 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.

  4. A quantitative study of the effect of solvent on the electronic absorption and fluorescence spectra of substituted phenothiazines: evaluation of their ground and excited singlet-state dipole moments

    NASA Astrophysics Data System (ADS)

    Párkányi, C.; Boniface, C.; Aaron, J. J.; Maafi, M.

    1993-11-01

    Electronic absorption and fluorescence excitation and emission spectra of five phenothiazines (phenothiazine, promethazine, thionine, methylene blue and Azure A) were determined at room temperature (293 K) in several solvents of various polarities (cyclohexane, dioxane, ethyl ether, chloroform, ethyl acetate, 1-butanol, 2-propanol, ethanol, methanol, acetonitrile, dimethylformamide and dimethyl sulfoxide). The effect of the solvents upon the spectral characteristics was studied. In combination with the ground state dipole moments of these phenothiazines, the spectral data were used to evaluate their first excited singlet-state dipole moments by means of the solvatochromic shift method (Bakhshiev's and Kawski—Chamma—Viallet correlations). The theoretical ground and excited singlet-state dipole moments for phenothiazines were calculated as a vector sum of the π component (obtained by the Pariser—Parr—Pople method) and the σ component (obtained from σ-bond moments). A reasonable agreement was found with the experimental values. For most phenothiazines under study, excited singlet-state dipole moments were found to be significantly higher than their ground-state counterparts. The application of the Kamlet—Abboud—Taft solvatochromic parameters to the solvent effect on spectral properties of phenothiazines is discussed.

  5. Fluorescence lifetime excitation cytometry by kinetic dithering.

    PubMed

    Li, Wenyan; Vacca, Giacomo; Castillo, Maryann; Houston, Kevin D; Houston, Jessica P

    2014-07-01

    Flow cytometers are powerful high-throughput devices that capture spectroscopic information from individual particles or cells. These instruments provide a means of multi-parametric analyses for various cellular biomarkers or labeled organelles and cellular proteins. However, the spectral overlap of fluorophores limits the number of fluorophores that can be used simultaneously during experimentation. Time-resolved parameters enable the quantification of fluorescence decay kinetics, thus circumventing common issues associated with intensity-based measurements. This contribution introduces fluorescence lifetime excitation cytometry by kinetic dithering (FLECKD) as a method to capture multiple fluorescence lifetimes using a hybrid time-domain approach. The FLECKD approach excites fluorophores by delivering short pulses of light to cells or particles by rapid dithering and facilitates measurement of complex fluorescence decay kinetics by flow cytometry. Our simulations demonstrated a resolvable fluorescence lifetime value as low as 1.8 ns (±0.3 ns) with less than 20% absolute error. Using the FLECKD instrument, we measured the shortest average fluorescence lifetime value of 2.4 ns and found the system measurement error to be ±0.3 ns (SEM), from hundreds of monodisperse and chemically stable fluorescent microspheres. Additionally, we demonstrate the ability to detect two distinct excited state lifetimes from fluorophores in single cells using FLECKD. This approach presents a new ability to resolve multiple fluorescence lifetimes while retaining the fluidic throughput of a cytometry system. The ability to discriminate more than one average fluorescence lifetime expands the current capabilities of high-throughput and intensity-based cytometry assays as the need to tag one single cell with multiple fluorophores is now widespread.

  6. Solvent effect on absorption and fluorescence spectra of three biologically active carboxamides (C1, C2 and C3). Estimation of ground and excited state dipole moment from solvatochromic method using solvent polarity parameters.

    PubMed

    Patil, N R; Melavanki, R M; Kapatkar, S B; Ayachit, N H; Saravanan, J

    2011-05-01

    The absorption and fluorescence spectra of three Carboxamides namely (E)-2-(4-Chlorobenzylideneamino)-N-(2-chlorophenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxamide (C(1)), (E)-N-(3-Chlorophenyl)-2-(3, 4-dimethoxybenzylideneamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxamide (C(2)) and (E)-N-(3-Chlorophenyl)-2-(3,4,5-trimethoxybenzylideneamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxamide (C(3)) have been recorded at room temperature in solvents of different polarities using dielectric constant (ε) and refractive index (n). Experimental ground (μ(g)) and excited (μ(e)) state dipole moments are estimated by means of solvatochromic shift method and also the excited dipole moments are estimated in combination with ground state dipole moments. It was estimated that dipole moments of the excited state were higher than those of the ground state of all three molecules. Further, the changes in dipole moment (Δμ) were calculated both from solvatochromic shift method and on the basis of microscopic empirical solvent polarity parameter (E(N)(T)) and the values are compared.

  7. 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.

  8. Fluorescence Spectroscopy with Surface Plasmon Excitation

    NASA Astrophysics Data System (ADS)

    Neumann, T.; Kreiter, M.; Knoll, W.

    In recent years, much effort has been directed towards the development of optical biosensors. While direct sensors are capable of monitoring the presence of an analyte without the use of labelling groups, the class of indirect sensors exploits the signal enhancement caused by bound marker molecules. Surface plasmon spectroscopy (SPS) as a direct detection method [1] is known to lack sensitivity for monitoring of low molecular mass analytes. In order to enhance the sensitivity and to improve the detection limit the technique was combined with fluorescence detection schemes in surface plasmon fluorescence spectroscopy (SPFS), as described recently [2]. Here, we briefly review the theory of plasmon excitation and the experimental realization of SPFS.

  9. Transient absorption of vibrationally excited water

    NASA Astrophysics Data System (ADS)

    Bakker, H. J.; Nienhuys, H.-K.; Gallot, G.; Lascoux, N.; Gale, G. M.; Leicknam, J.-C.; Bratos, S.

    2002-02-01

    We study the spectral response of the transition between the first and the second excited state of the O-H stretch vibration of HDO dissolved in liquid D2O with two-color femtosecond mid-infrared spectroscopy. The spectral response of this transition differs strongly from the fundamental absorption spectrum of the O-H stretch vibration. In addition, excitation of the O-H stretch vibration is observed to lead to a change of the hydrogen-bond dynamics of liquid water. We show that both these observations can be described with a refined quantum-mechanical version of the Lippincott-Schroeder model for hydrogen-bonded OH⋯O systems.

  10. Review of applications of fluorescence excitation spectroscopy to dermatology.

    PubMed

    Franco, W; Gutierrez-Herrera, E; Kollias, N; Doukas, A

    2016-03-01

    Endogenous molecules that exhibit fluorescence hold the potential to serve as reporters of tissue structure, activity and physiology. Fluorescence excitation spectroscopy is one means to measure and express tissue's innate fluorescence. This review focuses on the application of endogenous fluorescence ultraviolet excitation spectroscopy to dermatology.

  11. Fluorescence excitation-emission matrix (EEM) spectroscopy and cavity ring-down (CRD) absorption spectroscopy of oil-contaminated jet fuel using fiber-optic probes.

    PubMed

    Omrani, Hengameh; Barnes, Jack A; Dudelzak, Alexander E; Loock, Hans-Peter; Waechter, Helen

    2012-06-21

    Excitation emission matrix (EEM) and cavity ring-down (CRD) spectral signatures have been used to detect and quantitatively assess contamination of jet fuels with aero-turbine lubricating oil. The EEM spectrometer has been fiber-coupled to permit in situ measurements of jet turbine oil contamination of jet fuel. Parallel Factor (PARAFAC) analysis as well as Principal Component Analysis and Regression (PCA/PCR) were used to quantify oil contamination in a range from the limit of detection (10 ppm) to 1000 ppm. Fiber-loop cavity ring-down spectroscopy using a pulsed 355 nm laser was used to quantify the oil contamination in the range of 400 ppm to 100,000 ppm. Both methods in combination therefore permit the detection of oil contamination with a linear dynamic range of about 10,000.

  12. Non-radiative excitation fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Riachy, Lina; Vézy, Cyrille; Jaffiol, Rodolphe

    2016-03-01

    Non-radiative Excitation Fluorescence Microscopy (NEFM) constitutes a new way to observe biological samples beyond the diffraction limit. Non-radiative excitation of the samples is achieved by coating the substrate with donor species, such as quantum dots (QDs). Thus the dyes are not excited directly by the laser source, as in common fluorescence microscopy, but through a non-radiative energy transfer. To prevent dewetting of the donor film, we have recently implemented a silanization process to covalently bond the QDs on the substrate. An homogeneous monolayer of QDs was then deposited on only one side of the coverslips. Atomic force microscopy was then used to characterize the QD layer. We highlight the potential of our method through the study of Giant Unilamellar Vesicles (GUVs) labeled with DiD as acceptor, in interaction with surface functionalized with poly-L-lysine. In the presence of GUVs, we observed a quenching of QDs emission, together with an emission of DiD located in the membrane, which clearly indicated that non-radiative energy transfer from QDs to DiD occurs.

  13. 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.

  14. 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.

  15. Two-Photon-Excited Fluorescence-Encoded Infrared Spectroscopy.

    PubMed

    Mastron, Joseph N; Tokmakoff, Andrei

    2016-11-23

    We report on a method for performing ultrafast infrared (IR) vibrational spectroscopy using fluorescence detection. Vibrational dynamics on the ground electronic state driven by femtosecond mid-infrared pulses are detected by changes in fluorescence amplitude resulting from modulation of a two-photon visible transition by nuclear motion. We examine a series of coumarin dyes and study the signals as a function of solvent and excitation pulse parameters. The measured signal characterizes the relaxation of vibrational populations and coherences but yields different information than conventional IR transient absorption measurements. These differences result from the manner in which the ground-state dynamics are projected by the two-photon detection step. Extensions of this method can be adapted for a variety of increased-sensitivity IR measurements.

  16. 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

    2013-06-04

    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.

  17. New insights in the interpretation of tryptophan fluorescence : origin of the fluorescence lifetime and characterization of a new fluorescence parameter in proteins: the emission to excitation ratio.

    PubMed

    Albani, J R

    2007-07-01

    Origin of tryptophan fluorescence is still up to these days a quiz which is not completely solved. Fluorescence emission properties of tryptophan within proteins are in general considered as the result of fluorophore interaction within its environment. For example, a low fluorescence quantum yield is supposed to be the consequence of an important fluorophore-environment interaction. However, are we sure that the fluorophore has been excited upon light absorption? What if fluorophore excitation did not occur as the result of internal conformation specific to the fluorophore environment? Are we sure that all absorbed energy is used for the excitation process? Fluorescence lifetimes of Trp residues are considered to originate from rotamers or conformers resulting from the rotation of the indole ring within the peptide bonds. However, how can we explain the fact that in most of the proteins, the two lifetimes 0.5 and 3 ns, attributed to the conformers, are also observed for free tryptophan in solution? The present work, performed on free tryptophan and tyrosine in solution and on different proteins, shows that absorption and excitation spectra overlap but their intensities at the different excitation wavelengths are not necessarily equal. Also, we found that fluorescence emission intensities recorded at different excitation wavelengths depend on the intensities at these excitation wavelengths and not on the optical densities. Thus, excitation is not equal to absorption. In our interpretation of the data, we consider that absorbed photons are not necessary used only for the excitation, part of them are used to reorganize fluorophore molecules in a new state (excited structure) and another part is used for the excitation process. A new parameter that characterizes the ratio of the number of emitted photons over the real number of photons used to excite the fluorophore can be defined. We call this parameter, the emission to excitation ratio. Since our results were

  18. Absorption and fluorescent spectral studies of imidazophenazine derivatives

    NASA Astrophysics Data System (ADS)

    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 3.7 D, respectively. The changes in dipole moments upon the optical excitation for all derivatives estimated using Lippert equation were found to be Δ μ=9 D. The energies of the electronic S 1←S 0 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.

  19. Electric-field-induced changes in absorption and fluorescence of the green fluorescent protein chromophore in a PMMA film.

    PubMed

    Nakabayashi, Takakazu; Hino, Kazuyuki; Ohta, Yuka; Ito, Sayuri; Nakano, Hirofumi; Ohta, Nobuhiro

    2011-07-07

    External electric field effects on absorption, fluorescence, and fluorescence decay of p-HBDI that is a model compound of the chromophore of GFP have been examined in a poly(methyl methacrylate) film. The electroabsorption spectrum is similar in shape to the first derivative of the absorption spectrum, which results from the difference in molecular polarizability between the ground state and the Franck-Condon excited state. The electrophotoluminescence spectrum is dominated by the corresponding fluorescence spectrum, indicating the enhancement of the fluorescence intensity in the presence of external electric fields. The direct measurements of the electric field effect on the fluorescence decay profile suggest that the field-induced deceleration of the nonradiative process contributes to the increase in the fluorescence intensity in the presence of electric fields.

  20. Ultra-Fast Excited State Dynamics in Green Fluorescent Protein: Multiple States and Proton Transfer

    NASA Astrophysics Data System (ADS)

    Chattoraj, Mita; King, Brett A.; Bublitz, Gerold U.; Boxer, Steven G.

    1996-08-01

    The green fluorescent protein (GFP) of the jellyfish Aequorea Victoria has attracted widespread interest since the discovery that its chromophore is generated by the autocatalytic, posttranslational cyclization and oxidation of a hexapeptide unit. This permits fusion of the DNA sequence of GFP with that of any protein whose expression or transport can then be readily monitored by sensitive fluorescence methods without the need to add exogenous fluorescent dyes. The excited state dynamics of GFP were studied following photo-excitation of each of its two strong absorption bands in the visible using fluorescence upconversion spectroscopy (about 100 fs time resolution). It is shown that excitation of the higher energy feature leads very rapidly to a form of the lower energy species, and that the excited state interconversion rate can be markedly slowed by replacing exchangeable protons with deuterons. This observation and others lead to a model in which the two visible absorption bands correspond to GFP in two ground-state conformations. These conformations can be slowly interconverted in the ground state, but the process is much faster in the excited state. The observed isotope effect suggests that the initial excited state process involves a proton transfer reaction that is followed by additional structural changes. These observations may help to rationalize and motivate mutations that alter the absorption properties and improve the photo stability of GFP.

  1. 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.

  2. 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-...

  3. 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.

  4. Absorption and fluorescence properties of aryl substituted porphyrins in different media

    NASA Astrophysics Data System (ADS)

    Bozkurt, Serap Seyhan; Merdivan, Melek; Ayata, Sevda

    2010-02-01

    Absorption and fluorescence properties of aryl substituted porphyrins, 5,10,15,20-tetra-4-oxy(aceticacid)phenylporphyrin (TAPP), 5,10,15,20-tetra-(4-phenoxyphenyl) porphyrin (TPPP), 5,10,15,20-tetra-(3-bromo-4-hydroxyphenyl) porphyrin (TBHPP), and 5,10,15,20-tetra-p-chloromethylphenyl porphyrin (CMPP) were investigated. The UV/vis absorption, fluorescence and excited spectra as the fluorescence quantum yields and fluorescence lifetimes for the compounds were measured in organic solvents (chloroform (CHCl 3), tetrahydrofuran (THF)) and immobilized media (PVC film, sol-gel matrix). The fluorescence quantum yields of TAPP and TPPP were higher than the others. The fluorescence lifetimes of all studied porphyrin derivates were found to be fifty percent lower and their fluorescence intensities were increased fifty percent more in both of immobilized mediums, as compared to organic solvents.

  5. 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.

  6. 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.

  7. Hyperspectral imaging fluorescence excitation scanning for colon cancer detection

    NASA Astrophysics Data System (ADS)

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

    2016-10-01

    Optical spectroscopy and hyperspectral imaging have shown the potential to discriminate between cancerous and noncancerous tissue with high sensitivity and specificity. However, to date, these techniques have not been effectively translated to real-time endoscope platforms. Hyperspectral imaging of the fluorescence excitation spectrum represents 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 goal of this study was to evaluate the initial feasibility of using fluorescence excitation-scanning hyperspectral imaging for measuring changes in fluorescence excitation spectrum concurrent with colonic adenocarcinoma using a small pre-pilot-scale sample size. Ex vivo analysis was performed using resected pairs of colorectal adenocarcinoma and normal mucosa. Adenocarcinoma was confirmed by histologic evaluation of hematoxylin and eosin (H&E) permanent sections. Specimens were imaged using a custom hyperspectral imaging fluorescence excitation-scanning microscope system. Results demonstrated consistent spectral differences between normal and cancerous tissues over the fluorescence excitation range of 390 to 450 nm that could be the basis for wavelength-dependent detection of colorectal cancers. Hence, excitation-scanning hyperspectral imaging may offer an alternative approach for discriminating adenocarcinoma from surrounding normal colonic mucosa, but further studies will be required to evaluate the accuracy of this approach using a larger patient cohort.

  8. Two-color excited-state absorption imaging of melanins

    NASA Astrophysics Data System (ADS)

    Fu, Dan; Ye, Tong; Matthews, Thomas E.; Yurtsever, Gunay; Hong, Lian; Simon, John D.; Warren, Warren S.

    2007-02-01

    We have demonstrated a new method for imaging melanin with two-color excited state absorption microscopy. If one of two synchronized mode-locked pulse trains at different colors is intensity modulated, the modulation transfers to the other pulse train when nonlinear absorption takes place in the medium. We can easily measure 10 -6 absorption changes caused by either instantaneous two-photon absorption or relatively long lived excited state absorption with a RF lock-in amplifier. Eumelanin and pheomelanin exhibit similar excited state dynamics. However, their difference in excited state absorption and ground state absorption leads to change in the phase of the transient absorption signal. Scanning microscopic imaging is performed with B16 cells, melanoma tissue to demonstrate the 3D high resolution imaging capability. Different melanosome samples are also imaged to illustrate the differences between eumelanin and pheomelanin signals. These differences could enable us to image their respective distribution in tissue samples and provide us with valuable information in diagnosing malignant transformation of melanocytes.

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

    PubMed

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

    2005-03-03

    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.

  10. 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.

  11. Two-photon excited UV fluorescence for protein crystal detection

    SciTech Connect

    Madden, Jeremy T.; DeWalt, Emma L.; Simpson, Garth J.

    2011-10-01

    Complementary measurements using SONICC and TPE-UVF allow the sensitive and selective detection of protein crystals. Two-photon excited ultraviolet fluorescence (TPE-UVF) microscopy is explored for sensitive protein-crystal detection as a complement to second-order nonlinear optical imaging of chiral crystals (SONICC). Like conventional ultraviolet fluorescence (UVF), TPE-UVF generates image contrast based on the intrinsic fluorescence of aromatic residues, generally producing higher fluorescence emission within crystals than the mother liquor by nature of the higher local protein concentration. However, TPE-UVF has several advantages over conventional UVF, including (i) insensitivity to optical scattering, allowing imaging in turbid matrices, (ii) direct compatibility with conventional optical plates and windows by using visible light for excitation, (iii) elimination of potentially damaging out-of-plane UV excitation, (iv) improved signal to noise through background reduction from out-of-plane excitation and (v) relatively simple integration into instrumentation developed for SONICC.

  12. 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.

  13. Transient absorption of vibrationally excited ice Ih

    NASA Astrophysics Data System (ADS)

    Dokter, Adriaan M.; Bakker, Huib J.

    2008-01-01

    The ultrafast dynamics of HDO :D2O ice Ih at 180K is studied by midinfrared ultrafast pump-probe spectroscopy. The vibrational relaxation of HDO :D2O ice is observed to proceed via an intermediate state, which has a blueshifted absorption spectrum. Polarization resolved measurements reveal that the intermediate state is part of the intramolecular relaxation pathway of the HDO molecule. In addition, slow dynamics on a time scale of the order of 10-100ps is observed, related to thermally induced collective reorganizations of the ice lattice. The transient absorption line shape is analyzed within a Lippincott-Schroeder model for the OH-stretch potential. This analysis identifies the main mechanism behind the strong spectral broadening of the vOH=1→2 transition.

  14. Synchronous fluorescence and excitation emission characteristics of transformer oil ageing.

    PubMed

    Deepa, Subbiah; Sarathi, R; Mishra, Ashok K

    2006-11-15

    This paper describes the evaluation of synchronous fluorescence spectroscopy (SFS) and excitation emission matrix fluorescence (EEMF) spectroscopy as means of monitoring transformer oil degradation. When accelerated thermal ageing method is used, the onset of degradation of transformer oil on 17th day and transformer oil with polypropylene and cellulosic paper on 23rd and 27th days is sensitively reflected in the SFS and EEMF fluorescence spectral characteristics.

  15. 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.

  16. Moving in on the Action: An Experimental Comparison of Fluorescence Excitation and Photodissociation Action Spectroscopy.

    PubMed

    Wellman, Sydney M J; Jockusch, Rebecca A

    2015-06-18

    Photodissociation action spectroscopy is often used as a proxy for measuring gas-phase absorption spectra of ions in a mass spectrometer. Although the potential discrepancy between linear optical and photodissociation spectra is generally acknowledged, direct experimental comparisons are lacking. In this work, we use a quadrupole ion trap that has been modified to enable both photodissociation and laser-induced fluorescence to assess how closely the visible photodissociation action spectrum of a fluorescent dye reflects its fluorescence excitation spectrum. Our results show the photodissociation action spectrum of gaseous rhodamine 110 is both substantially narrower and slightly red-shifted (∼120 cm(-1)) compared to its fluorescence excitation spectrum. Power dependence measurements reveal that the photodissociation of rhodamine 110 requires, on average, the absorption of three photons whereas fluorescence is a single-photon process. These differing power dependences are the key to interpreting the differences in the measured spectra. The experimental results provide much-needed quantification and insight into the differences between action spectra and linear optical spectra, and emphasize the utility of fluorescence excitation spectra to provide a more reliable benchmark for comparison with theory.

  17. Excited-state dynamics of bacteriorhodopsin probed by broadband femtosecond fluorescence spectroscopy.

    PubMed

    Schmidt, B; Sobotta, C; Heinz, B; Laimgruber, S; Braun, M; Gilch, P

    2005-01-07

    The impact of varying excitation densities (approximately 0.3 to approximately 40 photons per molecule) on the ultrafast fluorescence dynamics of bacteriorhodopsin has been studied in a wide spectral range (630-900 nm). For low excitation densities, the fluorescence dynamics can be approximated biexponentially with time constants of <0.15 and approximately 0.45 ps. The spectrum associated with the fastest time constant peaks at 650 nm, while the 0.45 ps component is most prominent at 750 nm. Superimposed on these kinetics is a shift of the fluorescence maximum with time (dynamic Stokes shift). Higher excitation densities alter the time constants and their amplitudes. These changes are assigned to multi-photon absorptions.

  18. Time-resolved Hyperspectral Fluorescence Spectroscopy using Frequency Modulated Excitation

    SciTech Connect

    ,; Neill, M

    2012-07-01

    An intensity-modulated excitation light source is used together with a micro channel plate intensified CCD (ICCD) detector gated at a slightly different frequency to generate a beat frequency from a fluorescent sample. The addition of a spectrograph produces a hyperspectral time-resolved data product where the resulting beat frequency is detected with a low frame rate camera. Measuring the beat frequency of the spectrum as a function of time allows separation of the excited fluorescence from ambient constant light sources. The excitation and detector repetition rates are varied over a range of discrete frequencies, and the phase shift of the beat wave maps out the emission decay rate(s).

  19. 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.

  20. Fluorescence yields from photodissociative excitation of HCOOH, HCOOCH3, and CH3COOH in the vacuum-ultraviolet region

    NASA Technical Reports Server (NTRS)

    Suto, Masako; Wang, Xiuyan; Lee, L. C.

    1988-01-01

    The photoexcitation processes of HCOOH, HCOOCH3, and CH3COOH were studied in the vacuum-ultraviolet region by using synchroton radiation and a pulsed discharge lamp as light sources. The absorption and fluorescence cross sections of these molecules were measured in the 106-250-nm region. Fluorescences were detected from photoexcitation of HCOOH and HCOOCH3, but not from CH3COOH. Fluorescence produced at 123.9 nm was dispersed and identified as the excited OH and HCOO radicals. Fluorescence quantum yields of HCOOH and HCOOCH3 increase with decreasing excitation wavelengths with maxima of 5 and 0.3 percent at 106 nm, respectively.

  1. Comparison of absorption, fluorescence, and polarization spectroscopy of atomic rubidium

    NASA Astrophysics Data System (ADS)

    Ashman, Seth; Stifler, Cayla; Romero, Joaquin

    2015-05-01

    An ongoing spectroscopic investigation of atomic rubidium utilizes a two-photon, single-laser excitation process. Transitions accessible with our tunable laser include 5P1 / 2F' <-- 5S1 / 2 F and 5P3 / 2F' <-- 5S1 / 2 F . The laser is split into a pump and probe beam to allow for Doppler-free measurements of transitions between hyperfine levels. The pump and probe beams are overlapped in a counter-propagating geometry and the laser frequency scans over a transition. Absorption, fluorescence and polarization spectroscopy techniques are applied to this basic experimental setup. The temperature of the vapor cell and the power of the pump and probe beams have been varied to explore line broadening effects and signal-to-noise of each technique. This humble setup will hopefully grow into a more robust experimental arrangement in which double resonance, two-laser excitations are used to explore hyperfine state changing collisions between rubidium atoms and noble gas atoms. Rb-noble gas collisions can transfer population between hyperfine levels, such as 5P3 / 2 (F' = 3) <-- Collision 5P3 / 2 (F ' = 2) , and the probe beam couples 7S1 / 2 (F'' = 2) <-- 5P3 / 2 (F' = 3) . Polarization spectroscopy signal depends on the rate of population transfer due to the collision as well as maintaining the orientation created by the pump laser. Fluorescence spectroscopy relies only on transfer of population due to the collision. Comparison of these techniques yields information regarding the change of the magnetic sublevels, mF, during hyperfine state changing collisions.

  2. Selective two-photon fluorescence suppression by ultrafast pulse-pair excitation: control by selective one-color stimulated emission.

    PubMed

    Kumar De, Arijit; Roy, Debjit; Goswami, Debabrata

    2011-10-01

    Controlling two-photon molecular fluorescence leading to selective fluorophore excitation has been a long sought after goal in fluorescence microscopy. In this letter, we thoroughly explore selective fluorescence suppression through simultaneous two-photon absorption by two different fluorophores followed by selective one-photon stimulated emission for one particular fluorophore. We achieve this by precisely controlling the time delay between two identical ultrafast near infrared laser pulses.

  3. Fluorescence imaging of the propagation of excitability in gastrointestinal muscles

    NASA Astrophysics Data System (ADS)

    Publicover, Nelson G.; Smith, Terence K.; Stevens, Randel J.

    1999-07-01

    Fluorescence imaging is a useful tool to study the sequence an dynamics of the spread of excitability in biological tissues. Gastrointestinal muscles are particularly amenable to imaging using standard video rates because the frequency of events i slow and propagation velocities are slow. Calcium-sensitive fluorescent indicators such as fluo-3 provide effective markers of excitability because optically they exhibit high quantum yields and calcium plays important biological roles including regulating intracellular signaling and muscle contraction. Video sequences of gastrointestinal tissues demonstrate the existence of multiple preferred locations to indicate excitability. The spatial and temporal resolution of microscope-based imagin system allows pacing sites to be identified within single muscle bundles. Anisotropic conduction velocities result in spatially complex patterns of excitability where the range of propagation appears to be limited by 'collisions' with neighboring excitable events or recently activated regions. Although standard video rates are generally not sufficient to monitor more rapid excitable event such as nerve action potentials, fluorescence imaging can be used to investigate excitability mechanisms in tissues such as smooth muscles where event frequencies and propagation velocities are low.

  4. Substituent Effects on the Absorption and Fluorescence Properties of Anthracene.

    PubMed

    Abou-Hatab, Salsabil; Spata, Vincent A; Matsika, Spiridoula

    2017-02-16

    Substitution can be used to efficiently tune the photophysical properties of chromophores. In this study, we examine the effect of substituents on the absorption and fluorescence properties of anthracene. The effects of mono-, di-, and tetrasubstitution of electron-donating and -withdrawing functional groups were explored. In addition, the influence of a donor-acceptor substituent pair and the position of substitution were investigated. Eleven functional groups were varied on positions 1, 2, and 9 of anthracene, and on position 6 of 2-methoxyanthracene and 2-carboxyanthracene. Moreover, the donor-acceptor pair NH2/CO2H was added on different positions of anthracene for additional studies of doubly substituted anthracenes. Finally, we looked into quadruple substitutions on positions 1,4,5,8 and 2,3,6,7. Vertical excitation energies and oscillator strengths were computed using density functional theory with the hybrid CAM-B3LYP functional and 6-311G(d) basis set. Correlations between the excitation energies or oscillator strengths of the low-lying bright La state and the Hammett sigma parameter, σp(+), of the substituents were examined. The energy is red-shifted for all cases of substitution. Oscillator strengths increase when substituents are placed along the direction of the transition dipole moment of the bright La excited state. Substitution of long chain conjugated groups significantly increases the oscillator strength in comparison to the cases for other substituents. In addition, the results of quadruply substituted geometries reveal symmetric substitution at the 1,4,5,8 positions significantly increases the oscillator strength and can lower the band gap compared to that of the unsubstituted anthracene molecule by up to 0.5 eV.

  5. Excited-state intramolecular proton transfer in 3-hyroxyflavone isolated in solid argon: fluorescence and fluorescence-excitation spectra and tautomer fluorescence rise time

    SciTech Connect

    Dick, B.; Ernsting, N.P.

    1987-07-30

    The fluorescence properties of 3-hydroxyflavone isolated in solid argon at 15 K have been investigated. Upon electronic excitation the molecules undergo rapid intramolecular proton transfer. No fluorescence from the excited state of the normal form of the molecule could be detected. Perturbations due to hydrogen-bonding impurities which produce serious experimental problems in hydrocarbon glasses are largely suppressed in argon matrices. The rise of the green fluorescence of the tautomer was studied with excitation pulses of 230-fs duration and streak camera detection. An apparent tautomer fluorescence rise time of 2.7 ps was obtained by deconvolution. A comparative measurement of the dye coumarine 6 yielded an apparent fluorescence rise time of 2.5 ps, which can be entirely attributed to the group velocity dispersion of the streak camera optics. This indicates a rate constant for excited-state intramolecular proton transfer in 3-hydroxyflavone of greater than 10/sup 12/ s/sup -1/.

  6. 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.

  7. Excitation dynamics in Phycoerythrin 545: modeling of steady-state spectra and transient absorption with modified Redfield theory.

    PubMed

    Novoderezhkin, Vladimir I; Doust, Alexander B; Curutchet, Carles; Scholes, Gregory D; van Grondelle, Rienk

    2010-07-21

    We model the spectra and excitation dynamics in the phycobiliprotein antenna complex PE545 isolated from the unicellular photosynthetic cryptophyte algae Rhodomonas CS24. The excitonic couplings between the eight bilins are calculated using the CIS/6-31G method. The site energies are extracted from a simultaneous fit of the absorption, circular dichroism, fluorescence, and excitation anisotropy spectra together with the transient absorption kinetics using the modified Redfield approach. Quantitative fit of the data enables us to assign the eight exciton components of the spectra and build up the energy transfer picture including pathways and timescales of energy relaxation, thus allowing a visualization of excitation dynamics within the complex.

  8. Fluorescence following excited-state protonation of riboflavin at N(5).

    PubMed

    Quick, Martin; Weigel, Alexander; Ernsting, Nikolaus P

    2013-05-09

    Excited-state protonation of riboflavin in the oxidized form is studied in water. In the -1 < pH < 2 range, neutral and N(1)-protonated riboflavin coexist in the electronic ground state. Transient absorption shows that the protonated form converts to the ground state in <40 fs after optical excitation. Broadband fluorescence upconversion is therefore used to monitor solvation and protonation of the neutral species in the excited singlet state exclusively. A weak fluorescence band around 660 nm is assigned to the product of protonation at N(5). Its radiative rate and quantum yield relative to neutral riboflavin are estimated. Protonation rates agree with proton diffusion times for H(+) concentrations below 5 M but increase at higher acidities, where the average proton distance is below the diameter of the riboflavin molecule.

  9. A search for molecular hydrogen fluorescence near 100 km. [excitation by solar extreme UV radiation

    NASA Technical Reports Server (NTRS)

    Feldman, P. D.; Takacs, P. Z.

    1975-01-01

    The fluorescence of H2 in the Lyman band system, excited by solar extreme ultraviolet radiation, provides a means for the optical detection of H2 in the upper atmosphere. In particular, the Ly beta line of hydrogen is nearly degenerate with the (6,0) P1 transition, and absorption in this line produces fluorescence in the v-prime = 6 progression, principally at 1265, 1366, 1462 and 1608 A. Absorption by O2 rapidly attenuates the Ly beta from an overhead sun below 100 km and also significantly attenuates the fluorescent radiation. Far-ultraviolet dayglow spectra from 1130 to 1510 A obtained from an Aerobee rocket experiment on 11 December 1972 give an upper limit for any H2 emission which is a factor of 5 higher than expected according to recent hydrogen models.

  10. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2012-02-01

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

  12. Cyanines as new fluorescent probes for DNA detection and two-photon excited bioimaging.

    PubMed

    Feng, Xin Jiang; Wu, Po Lam; Bolze, Frédéric; Leung, Heidi W C; Li, King Fai; Mak, Nai Ki; Kwong, Daniel W J; Nicoud, Jean-François; Cheah, Kok Wai; Wong, Man Shing

    2010-05-21

    A series of cyanine fluorophores based on fused aromatics as an electron donor for DNA sensing and two-photon bioimaging were synthesized, among which the carbazole-based biscyanine exhibits high sensitivity and efficiency as a fluorescent light-up probe for dsDNA, which shows selective binding toward the AT-rich regions. The synergetic effect of the bischromophoric skeleton gives a several-fold enhancement in a two-photon absorption cross-section as well as a 25- to 100-fold enhancement in two-photon excited fluorescence upon dsDNA binding.

  13. Optical sensor instrumentation using absorption- and fluorescence-based capillary waveguide optrodes

    NASA Astrophysics Data System (ADS)

    Weigl, Bernhard H.; Draxler, Sonja; Kieslinger, Dietmar; Lehmann, H.; Trettnak, Wolfgang; Wolfbeis, Otto S.; Lippitsch, Max E.

    1995-09-01

    An analytical instrument comprising absorption- and fluorescence-based capillary waveguide optrodes (CWOs) is described. Glass capillaries with a chemically sensitive coating on the inner surface are used for optical chemical sensing in gaseous and liquid samples. In case of absorption-based CWOs, light from a LED is coupled into and out of the capillary under a defined angle via a rigid waveguide and an immersion coupler. The coated glass capillary forms an inhomogeneous waveguide, in which the light is guided in both the glass and the coating. The portion of the light which is absorbed in the chemically sensitive coating is proportional to a chemcial concentration or activity. This principle is demonstrated with a pCO2-sensitive inner coating. Typical relative light intensity signal changes with this type of optical interrogation are 98%, with an active capillary length of 10 mm. For fluorescence- based CWOs, the excitation light from an LED is coupled diffusely into the glass capillary and the optical sensor layer. A major portion of the excited fluorescence light is then collected within the coated capillary, and guided to the photodiode, which is located on the distal end of the capillary waveguide. Hereby, the excitation light is separated very efficiently from the fluorescent light. As an example, a CWO for pO2 is described. By applying this optical geometry, it was possible to utilize fluorescence decay time of the sensor layer as the transducer signal even when using solid state components (LEDs and photodiodes).

  14. Absorption and fluorescence spectra of poly(p-phenylenevinylene) (PPV) oligomers: an ab initio simulation.

    PubMed

    Cardozo, Thiago M; Aquino, Adélia J A; Barbatti, Mario; Borges, Itamar; Lischka, Hans

    2015-03-05

    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.

  15. 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

  16. Excited-state absorption measurements of Tm3+-doped crystals

    NASA Astrophysics Data System (ADS)

    Szela, J. W.; Mackenzie, J. I.

    2012-06-01

    High resolution, absolute excited-state absorption (ESA) spectra, at room temperature, from the long-lived 3F4 energy level of several crystals doped with trivalent thulium (Tm3+) ions have been measured employing high-brightness narrowband (FWHM <30 nm) light emitting diodes (LEDs) as a probe wavelength. The aim of this investigation was to determine the strength of ESA channels at wavelengths addressable by commercially available semiconductor laser diodes operating around 630-680 nm. The favourable lifetime of the 3F4 manifold and negligible ground-state absorption (GSA) for the red-wavelength second-step excitation, ensures a direct and efficient route for a dual-wavelength pumping scheme of the thulium ion, which will enable blue-green laser emission from its 1G4 upper-laser level.

  17. Excited State Absorption Measurements In Some Scintillator Dye Solutions

    NASA Astrophysics Data System (ADS)

    Dharamsi, A., N.; Jong, Shawpin; Hassam, A. B.

    1986-11-01

    Time-resolved excited state triplet-triplet absorption spectra were measured for solutions of 2,5 diphenyloxazole (PPO) and 2,1 napthyl, 5 phenyloxazole (aNPO) in several solvents. Concentration quenching effects due to excimer formation in nonaromatic solvents were observed. A numerical analysis of the experimental results yielded the rate constants for intersystem crossing, triplet quenching by 02, triplet self quenching and the formation of excimers.

  18. Excitation wavelength dependent fluorescence of graphene oxide controlled by strain.

    PubMed

    Cushing, Scott K; Ding, Weiqiang; Chen, Gang; Wang, Chao; Yang, Feng; Huang, Fuqiang; Wu, Nianqiang

    2017-02-09

    Unlike conventional fluorophores, the fluorescence emission of graphene oxide (GO) sheets can shift hundreds of nanometers as the excitation wavelength increases. The excitation wavelength dependent fluorescence is referred to as a giant red-edge effect and originates in a local reorganization potential slowing down the solvation dynamics of the excited state to the same time scale as the fluorescence lifetime. The present work has discovered that out-of-plane strain in the graphene oxide sheet leads to the intra-layer interaction necessary to slow down the solvation time scale. The oxygen percentage, dopant percentage, disorder, and strain are correlated with the presence and extent of the red-edge effect in oxygen, boron, nitrogen, and fluorine doped graphene oxide. Of these commonly cited possibilities, only out-of-plane strain is directly correlated to the red-edge effect. Furthermore, it is shown that the extent of the red-edge effect, or how far the emission wavelength can shift with increasing excitation wavelength, can be tuned by the electronegativity of the dopant. The present work interprets why the giant red-edge effect is present in some GO sheets but not in other GO sheets.

  19. Nonlinear absorption properties and excited state dynamics of ferrocene.

    PubMed

    Scuppa, Stefano; Orian, Laura; Dini, Danilo; Santi, Saverio; Meneghetti, Moreno

    2009-08-20

    We report on the first observation of reverse saturable absorption by ferrocene (Fc) in toluene using nanosecond pulses at 532 nm. Pump and probe experiments in the visible spectral region show the existence of an excited triplet state with an intersystem crossing quantum yield S1 --> T1 of 0.085 and a molar extinction coefficient epsilon(Fc)(T) of 5650 L mol(-1) cm(-1) at 700 nm. The full understanding of the nonlinear optical behavior of Fc cannot be obtained, however, with a model that includes only the one-photon absorption from T1, but it is mandatory to consider also a simultaneous two-photon absorption from an excited singlet state of Fc (two-photon absorption cross section: 2.4 x 10(-41) cm4 s ph(-1) mol(-1)). The optical spectrum of the ground and triplet state of Fc are calculated within a TD-DFT approach considering several functionals (PBE, BLYP, LDA, OPBE) for the optimization of molecular geometry.

  20. 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.

  1. 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

  2. Two-color two-photon excited fluorescence of indole: determination of wavelength-dependent molecular parameters.

    PubMed

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

    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 (1)La and (1)Lb 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 (1)Lb-(1)La inversion occurred under the conditions of our experiment. The two-photon absorption predominantly populated the (1)La state at all excitation wavelengths but in the 287-289 nm area which contained an absorption hump of the (1)Lb 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. 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.

  9. Selective suppression of two-photon fluorescence in laser scanning microscopy by ultrafast pulse-train excitation.

    PubMed

    De, Arijit Kumar; Roy, Debjit; Goswami, Debabrata

    2010-01-01

    Selective excitation of a particular fluorophore in the presence of others demands clever design of the optical field interacting with the molecules. We describe the use of 20- to 50-GHz pulse-train excitation leading to two-photon absorption, followed by successive one-photon stimulated emission as a potential technique in the context of controlling two-photon molecular fluorescence, with applications in microscopy.

  10. One- and two-photon excited fluorescence lifetimes and anisotropy decays of green fluorescent proteins.

    PubMed Central

    Volkmer, A; Subramaniam, V; Birch, D J; Jovin, T M

    2000-01-01

    We have used one- (OPE) and two-photon (TPE) excitation with time-correlated single-photon counting techniques to determine time-resolved fluorescence intensity and anisotropy decays of the wild-type Green Fluorescent Protein (GFP) and two red-shifted mutants, S65T-GFP and RSGFP. WT-GFP and S65T-GFP exhibited a predominant approximately 3 ns monoexponential fluorescence decay, whereas for RSGFP the main lifetimes were approximately 1.1 ns (main component) and approximately 3.3 ns. The anisotropy decay of WT-GFP and S65T-GFP was also monoexponential (global rotational correlation time of 16 +/- 1 ns). The approximately 1.1 ns lifetime of RSGFP was associated with a faster rotational depolarization, evaluated as an additional approximately 13 ns component. This feature we attribute tentatively to a greater rotational freedom of the anionic chromophore. With OPE, the initial anisotropy was close to the theoretical limit of 0.4; with TPE it was higher, approaching the TPE theoretical limit of 0.57 for the colinear case. The measured power dependence of the fluorescence signals provided direct evidence for TPE. The general independence of fluorescence decay times, rotation correlation times, and steady-state emission spectra on the excitation mode indicates that the fluorescence originated from the same distinct excited singlet states (A*, I*, B*). However, we observed a relative enhancement of blue fluorescence peaked at approximately 440 nm for TPE compared to OPE, indicating different relative excitation efficiencies. We infer that the two lifetimes of RSGFP represent the deactivation of two substates of the deprotonated intermediate (I*), distinguished by their origin (i.e., from A* or B*) and by nonradiative decay rates reflecting different internal environments of the excited-state chromophore. PMID:10692343

  11. Dipole moment of benzonitrile in its excited S 1 state from thermochromic shifts of fluorescence spectra

    NASA Astrophysics Data System (ADS)

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

    2006-02-01

    The effect of temperature T ranging from 293 to 393 K on absorption and fluorescence spectra of benzonitrile (BN) in ethyl acetate is studied. The absorption spectra of BN remain unchanged with increasing T. The analysis of fluorescence band shift by the Bilot and Kawski theory [L. Bilot, A. Kawski, Z. Naturforsch. 17a (1962) 621], for the known dipole moment value in the ground state μg = 4.18 D and α/ a3 = 0.5 ( α is the polarizability and a is the Onsager interaction radius of the solute), yield the average value of excited state dipole moment μe = 4.42 D. This value is in satisfactory agreement with 4.45, 4.51 and 4.57 D obtained previously from the Stark shift of rotational lines.

  12. Solar excited fluorescence of vegetation canopies: a unique look at photosynthesis

    NASA Astrophysics Data System (ADS)

    Stoll, M.; Moya, I.; Moreno, J.; Laurila, T.; Buschmann, C.; Court, A.; Smorenburg, K.; Visser, H.; Bazalgette, G.; Sioris, C.

    Although used for decades in the laboratory, it is only recently that the remote sensing biosphere community started being concerned in fluorescence and considering its role in advanced modeling of vegetation processes and vegetation function screening. Reasons are twofold : i) recent research works on photosynthesis that open the way to interpretation of chlorophyll fluorescence variations ; ii) developments towardhighly accurate radiative transfer models that need incorporating all radiative processes within the cover. This led to the FLEX- Fluorescence Explorer - mission idea submitted to ESA late 1998. It is well known that fluorescence is the radiative way of de-excitation of the photosynthetic apparatus for energy not consumed in photosynthesis, and that fluorescence is a very weak signal. An oversimplified, and often incorrect, view would be that anything that restricts the rate of electron transfer has increased fluorescence in counterpart. This is not so because of adaptation mechanisms of plant to environmental conditions, water and nutrients availability, etc. Nevertheless, link to photosynthetic capacity and CO2 absorption and plant function are fundamental characteristics of fluorescence. For solar excited passive stationary fluorescence, the only one relevant for remote sensing, the blue-green emission is strongly reduced due to prior absorption of UV radiation; thus, interest has been focussed on the chlorophyll red emission. To evaluate the potential and scientific usage of fluorescence for far-range remote sensing, there are steps and issues that must be carefully and thoroughly addressed, e.g.: i) interpretation of fluorescence variations against illumination and environmental conditions; ii) up-scaling from leaf to canopy level; iii) measurability of fluorescence signal. Investigations have already been carried out or are being undertaken: 1- FLEX preliminary instrument feasibility study: major drawbacks reside in the necessity to observing in

  13. 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)…

  14. Influence of transannular interaction over absorption and fluorescent properties of [2.2] paracyclophane and its phenyl derivatives

    NASA Astrophysics Data System (ADS)

    Nurmukhametov, R. N.; Shapovalov, A. V.; Antonov, D. Yu.

    2016-12-01

    A significant bathochromic shift of the fluorescent and long-wavelength absorption bands of [2.2] paracyclophane comparing to corresponding bands of alkyl-benzenes is due to a strong transannular interaction, resulting in formation of a principally new excited state of lower energy. It is concluded that the fluorescent levels for alkylbenzene excimers and for the macrocycle are of the same nature. Analysis of [2.2] paracyclophane mono- and diphenylderivatives spectra shows that their intensive absorption bands (230-310 nm) are originated from electron transitions of biphenyl groups and weak long wavelength absorption (310-340 nm) and fluorescent bands are governed by the same electron transitions between ground and excimer-like excited states as in the case of non-substituted macrocycle.

  15. Fluorescence and absorption spectroscopy of the weakly fluorescent chlorophyll a in cytochrome b6f of Synechocystis PCC6803.

    PubMed Central

    Peterman, E J; Wenk, S O; Pullerits, T; Pâlsson, L O; van Grondelle, R; Dekker, J P; Rögner, M; van Amerongen, H

    1998-01-01

    A spectroscopic characterization of the chlorophyll a (Chl) molecule in the monomeric cytochrome b6f complex (Cytb6f) isolated from the cyanobacterium Synechocystis PCC6803 is presented. The fluorescence lifetime and quantum yield have been determined, and it is shown that Chl in Cytb6f has an excited-state lifetime that is 20 times smaller than that of Chl in methanol. This shortening of the Chl excited state lifetime is not caused by an increased rate of intersystem crossing. Most probably it is due to quenching by a nearby amino acid. It is suggested that this quenching is a mechanism for preventing the formation of Chl triplets, which can lead to the formation of harmful singlet oxygen. Using site-selected fluorescence spectroscopy, detailed information on vibrational frequencies in both the ground and Qy excited states has been obtained. The vibrational frequencies indicate that the Chl molecule has one axial ligand bound to its central magnesium and accepts a hydrogen bond to its 13(1)-keto carbonyl. The results show that the Chl binds to a well-defined pocket of the protein and experiences several close contacts with nearby amino acids. From the site-selected fluorescence spectra, it is further concluded that the electron-phonon coupling is moderately strong. Simulations of both the site-selected fluorescence spectra and the temperature dependence of absorption and fluorescence spectra are presented. These simulations indicate that the Huang-Rhys factor characterizing the electron-phonon coupling strength is between 0.6 and 0.9. The width of the Gaussian inhomogeneous distribution function is 210 +/- 10 cm-1. PMID:9649396

  16. Excited state absorption spectra and intersystem crossing kinetics in diazanaphthalenes

    NASA Astrophysics Data System (ADS)

    Scott, Gary W.; Talley, Larry D.; Anderson, Robert W.

    1980-05-01

    Picosecond time-resolved, excited state absorption spectra in the visible following excitation at 355 nm are discussed for room temperature solutions of four diazanaphthalenes (DN)—quinoxaline (1,4-DN), quinazoline (1,3-DN), cinnoline (1,2-DN), and phthalazine (2,3-DN). Kinetics of singlet state decay are obtained by monitoring the decay of Sn←S1 bands. The intersystem crossing rate constant (kisc) is found to vary as kisc(1,4-DN)≳kisc(1,3-DN)≳kisc(1,2-DN). The kisc in phthalazine could not be determined from the weak, visible Sn←S1 absorption. Assuming rapid singlet vibrational relaxation and only minor effects due to energy gap variations, these experimental results agree with statistical limit predictions for the relative nonradiative rate. Calculations of the spin-orbit coupling matrix element βel= , using INDO wave functions, give the ordering βel(1,4-DN)≳βel(2,3-DN)≳βel(1,3-DN) ≳βel(1,2-DN).

  17. 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.

  18. Structural basis for dual excitation and photoisomerization of the Aequorea victoria green fluorescent protein.

    PubMed

    Brejc, K; Sixma, T K; Kitts, P A; Kain, S R; Tsien, R Y; Ormö, M; Remington, S J

    1997-03-18

    The 2.1-A resolution crystal structure of wild-type green fluorescent protein and comparison of it with the recently determined structure of the Ser-65 --> Thr (S65T) mutant explains the dual wavelength absorption and photoisomerization properties of the wild-type protein. The two absorption maxima are caused by a change in the ionization state of the chromophore. The equilibrium between these states appears to be governed by a hydrogen bond network that permits proton transfer between the chromophore and neighboring side chains. The predominant neutral form of the fluorophore maximally absorbs at 395 nm. It is maintained by the carboxylate of Glu-222 through electrostatic repulsion and hydrogen bonding via a bound water molecule and Ser-205. The ionized form of the fluorophore, absorbing at 475 nm, is present in a minor fraction of the native protein. Glu-222 donates its charge to the fluorophore by proton abstraction through a hydrogen bond network, involving Ser-205 and bound water. Further stabilization of the ionized state of the fluorophore occurs through a rearrangement of the side chains of Thr-203 and His-148. UV irradiation shifts the ratio of the two absorption maxima by pumping a proton relay from the neutral chromophore's excited state to Glu-222. Loss of the Ser-205-Glu-222 hydrogen bond and isomerization of neutral Glu-222 explains the slow return to the equilibrium dark-adapted state of the chromophore. In the S65T structure, steric hindrance by the extra methyl group stabilizes a hydrogen bonding network, which prevents ionization of Glu-222. Therefore the fluorophore is permanently ionized, causing only a 489-nm excitation peak. This new understanding of proton redistribution in green fluorescent protein should enable engineering of environmentally sensitive fluorescent indicators and UV-triggered fluorescent markers of protein diffusion and trafficking in living cells.

  19. Structural Basis for Dual Excitation and Photoisomerization of the Aequorea victoria Green Fluorescent Protein

    NASA Astrophysics Data System (ADS)

    Brejc, Katjusa; Sixma, Titia K.; Kitts, Paul A.; Kain, Steven R.; Tsien, Roger Y.; Ormo, Mats; Remington, S. James

    1997-03-01

    The 2.1- angstrom resolution crystal structure of wild-type green fluorescent protein and comparison of it with the recently determined structure of the Ser-65 --> Thr (S65T) mutant explains the dual wavelength absorption and photoisomerization properties of the wild-type protein. The two absorption maxima are caused by a change in the ionization state of the chromophore. The equilibrium between these states appears to be governed by a hydrogen bond network that permits proton transfer between the chromophore and neighboring side chains. The predominant neutral form of the fluorophore maximally absorbs at 395 nm. It is maintained by the carboxylate of Glu-222 through electrostatic repulsion and hydrogen bonding via a bound water molecule and Ser-205. The ionized form of the fluorophore, absorbing at 475 nm, is present in a minor fraction of the native protein. Glu-222 donates its charge to the fluorophore by proton abstraction through a hydrogen bond network, involving Ser-205 and bound water. Further stabilization of the ionized state of the fluorophore occurs through a rearrangement of the side chains of Thr-203 and His-148. UV irradiation shifts the ratio of the two absorption maxima by pumping a proton relay from the neutral chromophore's excited state to Glu-222. Loss of the Ser-205-Glu-222 hydrogen bond and isomerization of neutral Glu-222 explains the slow return to the equilibrium dark-adapted state of the chromophore. In the S65T structure, steric hindrance by the extra methyl group stabilizes a hydrogen bonding network, which prevents ionization of Glu-222. Therefore the fluorophore is permanently ionized, causing only a 489-nm excitation peak. This new understanding of proton redistribution in green fluorescent protein should enable engineering of environmentally sensitive fluorescent indicators and UV-triggered fluorescent markers of protein diffusion and trafficking in living cells.

  20. UV radiation at the fluorescence excitation maxima produces significant changes in the fluorescence of skin

    NASA Astrophysics Data System (ADS)

    Kollias, Nikiforos; Tian, W. D.; Zonios, George I.; Brancaleon, Lorenzo; Gillies, Robert

    1999-06-01

    Fluorescence excitation spectra of skin have been determined to be stable and reproducible. Three major bands dominate the wavelength range 280-400 nm. The major epidermal band due to tryptophan moieties appears at 295+/-5 nm and the major dermal bands due to collagen cross links appear at 335+/-5 nm and at 370+/-5 nm. The tryptophan fluorescence intensity has been found to increase with exposure to UV radiation; the UVB wavelengths are more effective than the UVA wavelengths. The PDCCL fluorescence intensity has been found to decrease dramatically with exposure to UVA in a wavelength specific way. The maximum of the action spectrum for this process is centered at the maximum of the excitation spectrum. The fluorescence of the skin recovers within 24 hours following exposure to UVA from single exposures. Multiple exposures produce permanent changes, in a follow-up of 8 weeks. the changes in the tryptophan fluorescence are probably due to changes in the molecular environment brought about by changes in the electrolyte balance in the epidermis following exposure. The changes in the dermis following UVA exposure appear to be associated with change in the collagen cross links, either through their association with other dermal species leading to quenching of the fluorescence or by the formation of stronger cross links with a smaller quantum efficiency. As these changes are immediate both alterations may provide the means for in vivo UV dosimetry.

  1. 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.

  2. Absorption into fluorescence. A method to sense biologically relevant gas molecules

    NASA Astrophysics Data System (ADS)

    Strianese, Maria; Varriale, Antonio; Staiano, Maria; Pellecchia, Claudio; D'Auria, Sabato

    2011-01-01

    In this work we present an innovative optical sensing methodology based on the use of biomolecules as molecular gating nano-systems. Here, as an example, we report on the detection ofanalytes related to climate change. In particular, we focused our attention on the detection ofnitric oxide (NO) and oxygen (O2). Our methodology builds on the possibility of modulating the excitation intensity of a fluorescent probe used as a transducer and a sensor molecule whose absorption is strongly affected by the binding of an analyte of interest used as a filter. The two simple conditions that have to be fulfilled for the method to work are: (a) the absorption spectrum of the sensor placed inside the cuvette, and acting as the recognition element for the analyte of interest, should strongly change upon the binding of the analyte and (b) the fluorescence dye transducer should exhibit an excitation band which overlaps with one or more absorption bands of the sensor. The absorption band of the sensor affected by the binding of the specific analyte should overlap with the excitation band of the transducer. The high sensitivity of fluorescence detection combined with the use of proteins as highly selective sensors makes this method a powerful basis for the development of a new generation of analytical assays. Proof-of-principle results showing that cytochrome c peroxidase (CcP) for NO detection and myoglobin (Mb) for O2 detection can be successfully used by exploiting our new methodology are reported. The proposed technology can be easily expanded to the determination of different target analytes.

  3. Absorption into fluorescence. A method to sense biologically relevant gas molecules.

    PubMed

    Strianese, Maria; Varriale, Antonio; Staiano, Maria; Pellecchia, Claudio; D'Auria, Sabato

    2011-01-01

    In this work we present an innovative optical sensing methodology based on the use of biomolecules as molecular gating nano-systems. Here, as an example, we report on the detection of analytes related to climate change. In particular, we focused our attention on the detection of nitric oxide (NO) and oxygen (O2). Our methodology builds on the possibility of modulating the excitation intensity of a fluorescent probe used as a transducer and a sensor molecule whose absorption is strongly affected by the binding of an analyte of interest used as a filter. The two simple conditions that have to be fulfilled for the method to work are: (a) the absorption spectrum of the sensor placed inside the cuvette, and acting as the recognition element for the analyte of interest, should strongly change upon the binding of the analyte and (b) the fluorescence dye transducer should exhibit an excitation band which overlaps with one or more absorption bands of the sensor. The absorption band of the sensor affected by the binding of the specific analyte should overlap with the excitation band of the transducer. The high sensitivity of fluorescence detection combined with the use of proteins as highly selective sensors makes this method a powerful basis for the development of a new generation of analytical assays. Proof-of-principle results showing that cytochrome c peroxidase (CcP) for NO detection and myoglobin (Mb) for O2 detection can be successfully used by exploiting our new methodology are reported. The proposed technology can be easily expanded to the determination of different target analytes.

  4. Two-step excitation and blue fluorescence under continuous-wave pumping in Nd:YLF

    NASA Technical Reports Server (NTRS)

    Fan, T. Y.; Byer, Robert L.

    1986-01-01

    Near-UV and blue fluorescence from the 4D3/2 and 4D5/2 manifolds in Nd:YLF has been observed at room temperature under CW pumping by a rhodamine 590 dye laser. Excitation to these manifolds is attributed to two-step excitation involving excited-state absorption from the 4F3/2 metastable level. A similar phenomenon has also been observed in Nd:YAG and Nd:glass. The effective excited-state absorption cross section is measured to be (2 + or - 1) x 10 to the -20th sq cm at 587.4 nm in the pi polarization, and the peak effective stimulated emission cross section is measured to be 5 x 10 to the -20th sq cm at 411.7 nm, also in the pi polarization. Estimated laser threshold at 411.7 nm for two-step pumping at 587.4 nm is 70 mW.

  5. A fluorescent benzothiazole probe with efficient two-photon absorption

    NASA Astrophysics Data System (ADS)

    Echevarria, Lorenzo; Moreno, Iván; Camacho, José; Salazar, Mary Carmen; Hernández, Antonio

    2012-11-01

    In this work, we report the two-photon absorption of 2-[4-(dimethylamino)phenyl]-1,3-benzothiazole-6-carbonitrile (DBC) in DMSO solution pumping at 779 nm with a 10 ns pulse laser-Nd:YAG system. The obtained two-photon absorption cross-section in DBC (407 ± 18 GM) is considerably high. Because DBC is a novel compound and have high values of fluorescence quantum yield, this result is expected to have an impact in biomolecules detection, diagnosis and treatment of cancer. Similar structures have previously been reported to show remarkable antitumour effects.

  6. 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.

  7. On the saturation of Tb phosphors under cathode-ray excitation. I. Excited-state absorption in Tb-activated phosphor powders

    NASA Astrophysics Data System (ADS)

    Nieuwesteeg, K. J. B. M.; Raue, R.; Busselt, W.

    1990-12-01

    Optical transitions between the 5D4 excited state of the Tb3+ (4f)8 configuration and the crystal-field split components of the (4f)7(5d) configuration are observed using high-resolution laser excitation of Tb-doped powder samples at room temperature. Excited-state absorption spectra of Tb3+ in YAG, YAGaG, Y2SiO5, and LaOBr are presented. Superimposed on the broadband excitation spectrum of these transitions we find the relatively narrow 4f→4f lines. We present a theoretical model for interpreting the 4f→5d transitions, which predicts a strict proportionality between the (4f)8 5D4→(4f)7(5d) and the (4f)8 7F6→(4f)7(5d) transitions. This model is used to estimate the optical-absorption cross section for the former transitions. Although the terminating 4f levels lie at the same energy as the 5d bands, the 4f→4f transitions can be interpreted on the basis of the Judd-Ofelt theory. We derived a method for calibration of the Tb3+ 4f→5d fluorescence-excitation spectra from powder samples. For YAG:Tb, the resulting optical cross sections were confirmed by direct-absorption measurements on a Tb3+ -doped epitaxial YAG layer. To our knowledge, this is the first time that the probabilities for excited-state absorption to highly excited 5d and 4f states in Tb3+ have been obtained theoretically and experimentally from Tb-doped powder materials.

  8. Solvent effects on the steady-state absorption and fluorescence spectra of uracil, thymine and 5-fluorouracil.

    PubMed

    Gustavsson, Thomas; Sarkar, Nilmoni; Bányász, Akos; Markovitsi, Dimitra; Improta, Roberto

    2007-01-01

    We report a comparison of the steady-state absorption and fluorescence spectra of three representative uracil derivatives (uracil, thymine and 5-fluorouracil) in alcoholic solutions. The present results are compared with those from our previous experimental and computational studies of the same compounds in water and acetonitrile. The effects of solvent polarity and hydrogen bonding on the spectra are discussed in the light of theoretical predictions. This comparative analysis provides a more complete picture of the solvent effects on the absorption and fluorescence properties of pyrimidine nucleobases, with special emphasis on the mechanism of the excited state deactivation.

  9. Solvent effects on the absorption and fluorescence spectra of rhaponticin: Experimental and theoretical studies

    NASA Astrophysics Data System (ADS)

    Sun, Yang; Liang, Xuhua; Zhao, Yingyong; Fan, Jun

    2013-02-01

    Rhaponticin (RH) possesses a variety of pharmacological activities including potent antitumor, antitumor-promoting, antithrombotic, antioxidant and vasorelaxant effects. The fundamental photophysics of RH is not well understood. In this work, solvent effect on the photoluminescence behavior of RH was studied by fluorescence and absorption spectra. The bathchromic shift was observed in absorption and fluorescence spectra with the increase of solvents polarity, which implied that transition involved was π → π*. A quantitative estimation of the contribution from different solvatochromic parameters, like normalized transition energy value (ETN), was made using the linear stokes shift (Δν) relationship based on the Lippert-Suppan equation. The ground state and excited state dipole moments were calculated by quantum-mechanical second-order perturbation method as a function of the dielectric constant (ɛ) and refractive index (n). The result was found to be 2.23 and 3.67 D in ground state and excited state respectively. The density functional theory (DFT) was used to obtain the most stable structure, electronic excitation energy, dipole moments and charge distribution. The analysis revealed that the RH exhibited strong photoinduced intramolecular charge transfer (ICT), and the intermolecular hydrogen bonding ability of the solvent was the most important parameter to characterize the photophysics behavior of RH. The hydrogen bonding effect occurred at the localized electron-acceptor oxygen at the glycoside bond. The experimental and theoretical results would help us better understand the photophysical properties of RH.

  10. 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.

  11. Dynamics of the higher lying excited states of cyanine dyes. An ultrafast fluorescence study.

    PubMed

    Guarin, Cesar A; Villabona-Monsalve, Juan P; López-Arteaga, Rafael; Peon, Jorge

    2013-06-20

    The electronic relaxation dynamics of the second singlet excited states of several cyanine dyes was studied through the femtosecond fluorescence up-conversion technique. Our interest in these molecules comes from the potential applications of systems with upper excited singlet states with a long lifetime, which can include electron and energy transfer from the higher lying singlets after one- or two-photon absorption. We studied three series of cyanines with 4-quinolyl, 2-quinolyl, or benzothiazolyl type end groups, each with varying sp(2) carbon conjugation lengths in the methinic bridge. The dynamics after electronic excitation to singlet states above the fluorescent state vary significantly as a function of cyanine structure and conjugation length. In particular, for the 4-quinolyl series the cyanine with an intermediate conjugation length (three methinic carbons) has the slowest S2 decays with lifetimes of 5.4 ps in ethanol and 6.6 ps in ethylene glycol. On the other hand, we observed that the 2-quinolyl family has S2 decay times in the subpicosecond range independent of the conjugation length between the end groups. The slowest internal conversion was observed for the benzothiazolyl type cyanine with five methinic carbons, with an S2 lifetime of 17.3 ps in ethanol. For the planar cyanines of this study we observed for the first time a clear systematic trend in the S2 decay times which closely follow the energy gap law. It was also demonstrated that a slow S2 decay is as well observed upon excitation through degenerate two-photon absorption with near-IR pulses. The present study isolates the most important variables for the design of cyanines with long S2 lifetimes.

  12. UV absorption and fluorescence properties of gas-phase p-difluorobenzene

    NASA Astrophysics Data System (ADS)

    Benzler, Thorsten; Dreier, Thomas; Schulz, Christof

    2017-01-01

    1,4-Difluorobenzene ( p-DFB) is a promising aromatic tracer for determining concentration, temperature, and O2 partial pressure in mixing gas flows based on laser-induced fluorescence (LIF). Signal quantification requires the knowledge of absorption and fluorescence properties as a function of environmental conditions. We report absorption and fluorescence spectra as well as fluorescence lifetimes of p-DFB in the temperature, pressure, and oxygen partial pressure range that is relevant for many applications including internal combustion engines. The UV absorption cross section, investigated between 296 and 675 K, has a peak value close to 266 nm and decreases with temperature, while still exceeding other single-ring aromatics. Time-resolved fluorescence spectra were recorded after picosecond laser excitation at 266 nm as a function of temperature (296-1180 K), pressure (1-10 bar), and O2 partial pressure (0-210 mbar) using a streak camera (temporal resolution 50 ps) coupled to a spectrometer. The fluorescence spectra red-shift ( 2 nm/100 K) and broaden (increase in full width at half maximum by 58% in the investigated temperature range) with temperature. In N2 as bath gas (1 bar), the fluorescence lifetime τ eff decreases with temperature by a factor of about 20 (from 7 ns at 298 K down to 0.32 ns at 1180 K), while at 8 bar the shortest lifetime at 975 K is 0.4 ns. A noticeable pressure dependence (i.e., reduced τ eff) is only visible at 675 K and above. Quenching of p-DFB LIF by O2 (for partial pressures up to 210 mbar) shortens the fluorescence lifetime significantly at room temperature (by a factor of 8), but much less at higher temperatures (by a factor of 1.8 at 970 K). For fixed O2 partial pressures (52 mbar and above), τ eff shows a plateau region with temperature which shifts toward higher temperatures at the higher O2 partial pressures. O2 quenching is less prominent for p-DFB compared to other aromatic compounds investigated so far. The temperature

  13. Effect of temperature on the {sup 127}I{sub 2} fluorescence excited by He-Ne laser radiation (0.63 {mu}m)

    SciTech Connect

    Kireev, S.V.; Shnyrev, S.L.

    1994-10-01

    Effect of temperature on the fluorescence of molecular iodine-127 vapors excited by the 0.63 {mu}m radiation of a {sup 3}He-{sup 22}Ne laser was studied. It was shown that heating iodine vapors can increase the sensitivity of the iodine-127 detection by the laser-fluorescent method by approximately an order of magnitude. The main contribution to the fluorescence signal is made by the absorption line 6-3 P(33). 20 refs., 4 figs.

  14. [Intensity loss of two-photon excitation fluorescence microscopy images of mouse oocyte chromosomes].

    PubMed

    Zhao, Feng-Ying; Wu, Hong-Xin; Chen, Die-Yan; Ma, Wan-Yun

    2014-07-01

    As an optical microscope with high resolution, two-photon excitation (TPE) fluorescence microscope is widely used in noninvasive 3D optical imaging of biological samples. Compared with confocal laser scanning microscope, TPE fluorescence microscope provides a deeper detecting depth. In spite of that, the image quality of sample always declines as the detecting depth increases when a noninvasive 3D optical imaging of thicker samples is performed. Mouse oocytes with a large diameter, which play an important role in clinical and biological fields, have obvious absorption and scattering effects. In the present paper, we performed compensation for two-photon fluorescence images of mouse oocyte chromosomes. Using volume as a parameter, the attenuation degree of these chromosomes was also studied. The result of our data suggested that there exists a severe axial intensity loss in two-photon microscopic images of mouse oocytes due to the absorption and scattering effects. It is necessary to make compensation for these images of mouse oocyte chromosomes obtained from two-photon microscopic system. It will be specially needed in studying the quantitative three-dimensional information of mouse oocytes.

  15. Electronically excited rubidium atom in helium clusters and films. II. Second excited state and absorption spectrum.

    PubMed

    Leino, Markku; Viel, Alexandra; Zillich, Robert E

    2011-01-14

    Following our work on the study of helium droplets and film doped with one electronically excited rubidium atom Rb(∗) ((2)P) [M. Leino, A. Viel, and R. E. Zillich, J. Chem. Phys. 129, 184308 (2008)], we focus in this paper on the second excited state. We present theoretical studies of such droplets and films using quantum Monte Carlo approaches. Diffusion and path integral Monte Carlo algorithms combined with a diatomics-in-molecule scheme to model the nonpair additive potential energy surface are used to investigate the energetics and the structure of Rb(∗)He(n) clusters. Helium films as a model for the limit of large clusters are also considered. As in our work on the first electronic excited state, our present calculations find stable Rb(∗)He(n) clusters. The structures obtained are however different with a He-Rb(∗)-He exciplex core to which more helium atoms are weakly attached, preferentially on one end of the core exciplex. The electronic absorption spectrum is also presented for increasing cluster sizes as well as for the film.

  16. 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.

  17. 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.

  18. Microfluorometric Detection of Catecholamines with Multiphoton-Excited Fluorescence

    NASA Astrophysics Data System (ADS)

    Balaji, J.; Reddy, Chandra S.; Kaushalya, S. K.; Maiti, Sudipta

    2004-04-01

    We demonstrate sensitive spatially resolved detection of physiological chromophores that emit in the ultraviolet (less than 330 nm). An atypical laser source (a visible wavelength femtosecond optical parametric oscillator), and an unconventional collection geometry (a lensless detector that detects the forward-emitted fluorescence) enable this detection. We report the excitation spectra of the catecholamines dopamine and norepinephrine, together with near-UV emitters serotonin and tryptophan, in the range of 550-595 nm. We estimate the molecular two-photon action cross section of dopamine, norepinephrine, and serotonin to be 1.2 mGM (1 GM, or Goppert Mayor, is equal to 10^-58 m^4 s^-1 photon^-1), 2 mGM, and 43 mGM, respectively, at 560 nm. The sensitivity achieved by this method holds promise for the microscopic imaging of vesicular catecholamines in live cells.

  19. The electronic excited states of green fluorescent protein chromophore models

    NASA Astrophysics Data System (ADS)

    Olsen, Seth Carlton

    We explore the properties of quantum chemical approximations to the excited states of model chromophores of the green fluorescent protein of A. victoria. We calculate several low-lying states by several methods of quantum chemical calculation, including state-averaged complete active space SCF (CASSCF) methods, time dependent density functional theory (TDDFT), equation-of motion coupled cluster (EOM-CCSD) and multireference perturbation theory (MRPT). Amongst the low-lying states we identify the optically bright pipi* state of the molecules and examine its properties. We demonstrate that the state is dominated by a single configuration function. We calculate zero-time approximations to the resonance Raman spectrum of GFP chromophore models, and assign published spectra based upon these.

  20. 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

  1. Polarized fluorescence and absorption of macroscopically aligned Light Harvesting Complex II.

    PubMed

    van Amerongen, H; Kwa, S L; van Bolhuis, B M; van Grondelle, R

    1994-08-01

    Polarized absorption and fluorescence measurements have been performed at 77 K on isotropic and anisotropic preparations of trimeric Light Harvesting Complex II (LHC-II) from spinach. The results enable a decomposition of the absorption spectrum into components parallel and perpendicular to the trimeric plane. For the first time, it is shown quantitatively that the strong absorption band around 676 nm is polarized essentially parallel to the plane of the trimer, i.e., the average angle between the corresponding transition dipole moments and this plane is at most 12 degrees. The different absorption bands for LHC-II should not be considered as corresponding to individual pigments but to collective excitations of different pigments. Nevertheless, the average angle between the Qy transition dipole moments of all chlorophyll a pigments in LHC-II and the trimeric plane could be determined and was found to be 17.5 degrees +/- 2.5 degrees. For the chlorophyll b pigments, this angle is significantly larger (close to 35 degrees). At 77 K, most of the fluorescence stems from a weak band above 676 nm and the corresponding transition dipole moments are oriented further out of plane than the dipole moments corresponding to the 676-nm band. The results are shown to be of crucial significance for understanding the relation between the LHC-II structure and its spectroscopy.

  2. Photoconversion of DAPI following UV or violet excitation can cause DAPI to fluoresce with blue or cyan excitation.

    PubMed

    Piterburg, M; Panet, H; Weiss, A

    2012-04-01

    4'-6-Diamidino-2-phenylindole is a fluorescent dye commonly used to visualize deoxyribonucleic acid or cell nuclei in fixed cell preparations, and is often used together with fluorescein or green fluorescent protein, which can be excited without exciting 4'-6-Diamidino-2-phenylindole. It is assumed that when using typical fluorescein or green fluorescent protein filter cubes, 4'-6-Diamidino-2-phenylindole will not be observed. In this paper, we show that following observation of 4'-6-Diamidino-2-phenylindole using UV or violet excitation, it may become sensitive to the blue/cyan excitation used in fluorescein/green fluorescent protein filter cubes. This has serious implications for the use of 4'-6-Diamidino-2-phenylindole together with widely used green fluorophores in double labelling experiments.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. Excited singlet-state absorption in laser dyes at the XeCl wavelength

    NASA Astrophysics Data System (ADS)

    Taylor, R. S.; Mihailov, S.

    1985-10-01

    The transmission properties of the laser dyes BBQ, PBD, BPBD, α-NPO, p-Quarterphenyl and PPO have been measured using a XeCl (308 nm) excimer laser. A model for the dye saturation which incorporates excited-state absorption was used to estimate the lifetime and the absorption cross section of the first excited singlet-state for each dye.

  8. Two-photon excitation of nitric oxide fluorescence as a temperature indicator in unsteady gas-dynamic processes

    NASA Technical Reports Server (NTRS)

    Mckenzie, R. L.; Gross, K. P.

    1980-01-01

    A laser induced fluorescence technique, suitable for measuring fluctuating temperatures in cold turbulent flows containing very low concentrations of nitric oxide is described. Temperatures below 300 K may be resolved with signal to noise ratios greater than 50 to 1 using high peak power, tunable dye lasers. The method relies on the two photon excitation of selected ro-vibronic transitions. The analysis includes the effects of fluorescence quenching and shows the technique to be effective at all densities below ambient. Signal to noise ratio estimates are based on a preliminary measurement of the two photon absorptivity for a selected rotational transition in the NO gamma (0,0) band.

  9. Ground state bromine atom density measurements by two-photon absorption laser-induced fluorescence

    NASA Astrophysics Data System (ADS)

    Sirse, N.; Foucher, M.; Chabert, P.; Booth, J.-P.

    2014-12-01

    Ground state bromine atom detection by two-photon absorption laser-induced fluorescence (TALIF) is demonstrated. The (4p5) {^2Po3/2} bromine atoms are excited by two-photon absorption at 252.594 nm to the (5p) {^4So3/2} state and detected by 635.25 nm fluorescence to the (5s) 4P5/2 state. The atoms are generated in a radio-frequency inductively-coupled plasma in pure HBr. The excitation laser also causes some photodissociation of HBr molecules, but this can be minimized by not focussing the laser beam, still giving adequate signal levels. We determined the natural lifetime of the emitting (5p) {^4So3/2} state, τf^Br*=30.9 +/- 1.4 ns and the rate constant for quenching of this state by collision with HBr molecules, k_HBrQ = 1.02 +/- 0.07× 10-15 m3 s-1 .

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

    PubMed

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

    2009-08-20

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

  11. Absorption and fluorescence of PRODAN in phospholipid bilayers: a combined quantum mechanics and classical molecular dynamics study.

    PubMed

    Cwiklik, Lukasz; Aquino, Adelia J A; Vazdar, Mario; Jurkiewicz, Piotr; Pittner, Jiří; Hof, Martin; Lischka, Hans

    2011-10-20

    Absorption and fluorescence spectra of PRODAN (6-propionyl-2-dimethylaminonaphthalene) were studied by means of the time-dependent density functional theory and the algebraic diagrammatic construction method. The influence of environment, a phosphatidylcholine lipid bilayer and water, was taken into account employing a combination of quantum chemical calculations with empirical force-field molecular dynamics simulations. Additionally, experimental absorption and emission spectra of PRODAN were measured in cyclohexane, water, and lipid vesicles. Both planar and twisted configurations of the first excited state of PRODAN were taken into account. The twisted structure is stabilized in both water and a lipid bilayer, and should be considered as an emitting state in polar environments. Orientation of the excited dye in the lipid bilayer significantly depends on configuration. In the bilayer, the fluorescence spectrum can be regarded as a combination of emission from both planar and twisted structures.

  12. Analysis of the excited-state absorption spectral bandshape of oligofluorenes

    NASA Astrophysics Data System (ADS)

    Hayes, Sophia C.; Silva, Carlos

    2010-06-01

    We present ultrafast transient absorption spectra of two oligofluorene derivatives in dilute solution. These spectra display a photoinduced absorption band with clear vibronic structure, which we analyze rigorously using a time-dependent formalism of absorption to extract the principal excited-state vibrational normal-mode frequencies that couple to the electronic transition, the configurational displacement of the higher-lying excited state, and the reorganization energies. We can model the excited-state absorption spectrum using two totally symmetric vibrational modes with frequencies 450 (dimer) or 400 cm-1 (trimer), and 1666 cm-1. The reorganization energy of the ground-state absorption is rather insensitive to the oligomer length at 230 meV. However, that of the excited-state absorption evolves from 58 to 166 meV between the oligofluorene dimer and trimer. Based on previous theoretical work [A. Shukla et al., Phys. Rev. B 67, 245203 (2003)], we assign the absorption spectra to a transition from the 1Bu excited state to a higher-lying mAg state, and find that the energy of the excited-state transition with respect to the ground-state transition energy is in excellent agreement with the theoretical predictions for both oligomers studied here. These results and analysis permit profound understanding of the nature of excited-state absorption in π-conjugated polymers, which are the subject of general interest as organic semiconductors in the solid state.

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

    PubMed

    Early, Kevin T; Nesbitt, David J

    2015-12-09

    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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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.

  1. Fluorescence Excitation Function Produced Through Photoexcitation of the Rydberg States b, cn, and o3 of N2 in the 80-100 nm Region

    NASA Astrophysics Data System (ADS)

    Wu, R. C.; Judge, D. L.; Singh, T. S.; Mu, X. L.; Nee, J. B.; Chiang, S. Y.; Fung, H. S.

    2006-12-01

    Fluorescence excitation functions produced through photoexcitation of N2 using synchrotron radiation in the spectral region between 80 and 100 nm have been studied. Two broadband detectors were employed to simultaneously monitor fluorescence in the 115-320 nm and 300-700 nm region, respectively. The peaks in the VUV Fluorescence excitation functions are found to correspond to excitation of absorption transitions from the ground electronic state to the Rydberg states of b, cn (with n= 4-9), and o3 of N2. The relative fluorescence production cross sections for the observed peaks are determined. No fluorescence has been produced through excitation of the most dominating absorption features of the b-X transition except the (1,0), (5,0), and (6,0) bands, in excellent agreement with the recent lifetime measurements [1] and theoretical calculations [2]. In addition, fluorescence peaks correlate with the long vibrational progression of the c4 (v = 0-7) has also been observed. Weak fluorescence excitation peaks are also found to correlate with the excitation of the (1,0), (2,0), and (3,0) bands of the o3 VX transition. The present results provide important information for further unraveling of complicated and intriguing interactions among the excited electronic states of N2. Furthermore, solar photon excitation of N2 leading to the production of c4(0) may provide useful data required for evaluation and analysis of dayglow models of the c4(0) issue of the Earth's atmosphere [3]. Detailed results will be presented. This research is based on work supported by NSF grant ATM-0096761. [1] Sprengers, J. P., W. Ubachs, and K. G. H. Baldwin, J. Chem. Phys., 122, 144301 (2005). [2] Lewis, B. R., S. T. Gibson, W. Zhang, H. Lefebvre-Brion, and J.-M. Robbe, J. Chem. Phys., 122, 144302 (2005). [3] e.g., Meier, R. R., Space Sci. Rev., 58, 1 (1991).

  2. Absorption and Fluorescence Lineshape Theory for Polynomial Potentials.

    PubMed

    Anda, André; De Vico, Luca; Hansen, Thorsten; Abramavičius, Darius

    2016-12-13

    The modeling of vibrations in optical spectra relies heavily on the simplifications brought about by using harmonic oscillators. However, realistic molecular systems can deviate substantially from this description. We develop two methods which show that the extension to arbitrarily shaped potential energy surfaces is not only straightforward, but also efficient. These methods are applied to an electronic two-level system with potential energy surfaces of polynomial form and used to study anharmonic features such as the zero-phonon line shape and mirror-symmetry breaking between absorption and fluorescence spectra. The first method, which constructs vibrational wave functions as linear combinations of the harmonic oscillator wave functions, is shown to be extremely robust and can handle large anharmonicities. The second method uses the cumulant expansion, which is readily solved, even at high orders, thanks to an ideally suited matrix theorem.

  3. Determination of biological toxins using capillary electrokinetic chromatography with multiphoton-excited fluorescence.

    PubMed

    Wei, J; Okerberg, E; Dunlap, J; Ly, C; Shear, J B

    2000-03-15

    We report a highly sensitive and rapid strategy for characterizing biological toxins based on capillary electrokinetic chromatography with multiphoton-excited fluorescence. In this approach, aflatoxins B1, B2, and G1 and the cholera toxin A-subunit are fractionated in approximately 80 s in a narrow-bore electrophoretic channel using the negatively charged pseudostationary phase, carboxymethyl-beta-cyclodextrin. The aflatoxins--highly mutagenic multiple-ringed heterocycles produced by Aspergillus fungi--are excited at the capillary outlet through the simultaneous absorption of two to three 750-nm photons to yield characteristic blue fluorescence; cholera toxin A-subunit, the catalytic domain of the bacterial protein toxin from Vibrio cholera, is excited through an unidentified multiphoton pathway that apparently includes photochemical transformation of an aromatic residue in the polypeptide. The anionic carboxymethyl-beta-cyclodextrin, used to chromatographically resolve the uncharged aflatoxins, enhances emission from these compounds without contributing substantially to the background. Detection limits for these toxins separated in 2.1-micron-i.d. capillaries range from 4.4 zmol (approximately 2700 molecules) for aflatoxin B2 to 3.4 amol for the cholera toxin A-subunit. Larger (16-micron-i.d.) separation capillaries provide concentration detection limits for aflatoxins in the 0.2-0.4 nM range, severalfold lower than achieved in 2.1-micron capillaries. These results represent an improvement of > 10(4) in mass detectability compared to previously published capillary separations of aflatoxins and demonstrate new possibilities for the analysis of proteins and peptides.

  4. Three-color femtosecond source for simultaneous excitation of three fluorescent proteins in two-photon fluorescence microscopy.

    PubMed

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

    2012-09-01

    We demonstrate a fiber-based, three-color femtosecond source for simultaneous imaging of three fluorescent proteins (FPs) using two-photon fluorescence microscopy (2PM). The three excitation wavelengths at 775 nm, 864 nm and 950 nm, are obtained through second harmonic generation (SHG) of the 1550-nm pump laser and the 1728-nm and 1900-nm solitons generated through soliton self-frequency shift (SSFS) in a large-mode-area (LMA) fiber. These energetic pulses are well matched to the two-photon excitation peaks of red, cyan and yellow fluorescent proteins (TagRFPs, TagCFPs, and TagYFPs) for efficient excitation. We demonstrate simultaneous 2PM of human melanoma cells expressing a "rainbow" combination of these three fluorescent proteins.

  5. Noninvasive fluorescence excitation spectroscopy during application of 5-aminolevulinic acid in vivo.

    PubMed

    Juzenas, Petras; Juzeniene, Asta; Kaalhus, Olav; Iani, Vladimir; Moan, Johan

    2002-10-01

    The fluorescence of PpIX induced by topical application of 5-aminolevulinic acid (ALA) in normal mouse skin was studied noninvasively by means of a fibre optic probe. The fluorescence excitation spectrum of PpIX exhibits five distinct peaks at around 408. 510, 543, 583 and 633 nm under fluorescence monitoring at the second emission peak of PpIX (705 nm). The transmission of the excitation light is wavelength dependent: the long wavelength light (>600 nm) penetrates deeper into the tissues by a factor of 6 compared with the short wavelength light (<590 nm). Thus, the fluorescence excitation spectrum of PpIX measured on the surface of the skin can be used to estimate the depth of the penetration of topically applied ALA. The fluorescence excitation spectra calculated for the depth 1.1 mm obtained the best fit with the experimentally measured spectra after topical application of ALA.

  6. 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.

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

    PubMed

    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.

  8. 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

  9. Laser-excited fluorescence for measuring atmospheric pollution

    NASA Technical Reports Server (NTRS)

    Menzies, R. T.

    1975-01-01

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

  10. 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.

  11. 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.

  12. 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.

  13. Theoretical studies on the vibrationally-resolved absorption and fluorescence spectra of H-Pyrene+ and H-Coronene+

    NASA Astrophysics Data System (ADS)

    Li, JunFeng; Tian, GuanJun; Luo, Yi; Cao, ZeXing

    2015-11-01

    H-Pyrene+ and H-Coronene+ are important carrier candidates for the diffuse interstellar band. In order to understand the observed absorption and fluorescence emission spectra of H-Pyrene+ and H-Coronene+, time-dependent density functional theory (TD-DFT) method and Franck-Condon approximation have been employed to simulate the corresponding vibrationally-resolved optical spectra. For H-Pyrene+, the calculated absorption, emission and 0-0 band energies are in good agreement with the experimental values. The strong absorption and emission vibrational peaks near the 0-0 band match well with the experiment peaks. A noticeable deviation for several weak peaks far away from the origin band is observed, as a result of the vibronic coupling with other excited states. For H-Coronene+, the predicted vibrationally resolved electronic absorption and emission spectra resemble very well their experimental counterparts spectra, allowing to fully assign the observed vibronic peaks.

  14. Excited state two photon absorption of a charge transfer radical dimer in the near infrared.

    PubMed

    Schiccheri, Nicola; Meneghetti, Moreno

    2005-06-02

    Nonlinear transmission measurements of a solution of radical dimers of tetramethyl-tetrathiafulvalene, (TMTTF+)2, recorded with 9 ns laser pulses at 1064 nm are reported and interpreted on the basis of a multiphoton absorption process. One finds that the process can be interpreted with a sequence of three photon absorption, the first being a one photon absorption related to the intermolecular charge transfer process characteristic of the dimers and the second a two photon absorption from the excited state created with the first process. A model calculation allows one to obtain the value of the two photon absorption cross section which is found to be several orders of magnitude larger than those usually found for two photon absorbing systems excited from the ground state. These results show the importance of an excited-state population for obtaining large nonlinear optical responses.

  15. Absorption and fluorescence spectroscopic characterization of BLUF domain of AppA from Rhodobacter sphaeroides

    NASA Astrophysics Data System (ADS)

    Zirak, P.; Penzkofer, A.; Schiereis, T.; Hegemann, P.; Jung, A.; Schlichting, I.

    2005-08-01

    The BLUF domain of the transcriptional anti-repressor protein AppA from the non-sulfur anoxyphototrophic purple bacterium Rhodobacter sphaeroides was characterized by absorption and emission spectroscopy. The BLUF domain constructs AppA 148 (consisting of amino-acid residues 1-148) and AppA 126 (amino-acid residues 1-126) are investigated. The cofactor of the investigated domains is found to consist of a mixture of the flavins riboflavin, FMN, and FAD. The dark-adapted domains exist in two different active receptor conformations (receptor states) with different sub-nanosecond fluorescence lifetimes (BLUF r,f and BLUF r,sl) and a small non-interacting conformation (BLUF nc). The active receptor conformations are transformed to putative signalling states (BLUF s,f and BLUF s,sl) of low fluorescence efficiency and picosecond fluorescence lifetime by blue-light excitation (light-adapted domains). In the dark at room temperature both signalling states recover back to the initial receptor states with a time constant of about 17 min. A quantum yield of signalling state formation of about 25% was determined by intensity dependent transmission measurements. A photo-cycle scheme is presented including photo-induced charge transfer complex formation, charge recombination, and protein binding pocket reorganisation.

  16. 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

  17. Invited review article: Imaging techniques for harmonic and multiphoton absorption fluorescence microscopy.

    PubMed

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

    2009-08-01

    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.

  18. Extending single molecule fluorescence observation time by amplitude-modulated excitation

    NASA Astrophysics Data System (ADS)

    Kisley, Lydia; Chang, Wei-Shun; Cooper, David; Mansur, Andrea P.; Landes, Christy F.

    2013-09-01

    We present a hardware-based method that can improve single molecule fluorophore observation time by up to 1500% and super-localization by 47% for the experimental conditions used. The excitation was modulated using an acousto-optic modulator (AOM) synchronized to the data acquisition and inherent data conversion time of the detector. The observation time and precision in super-localization of four commonly used fluorophores were compared under modulated and traditional continuous excitation, including direct total internal reflectance excitation of Alexa 555 and Cy3, non-radiative Förster resonance energy transfer (FRET) excited Cy5, and direct epi-fluorescence wide field excitation of Rhodamine 6G. The proposed amplitude-modulated excitation does not perturb the chemical makeup of the system or sacrifice signal and is compatible with multiple types of fluorophores. Amplitude-modulated excitation has practical applications for any fluorescent study utilizing an instrumental setup with time-delayed detectors.

  19. Excited-state absorption of Tm3 + -doped single crystals at photon-avalanche wavelengths

    NASA Astrophysics Data System (ADS)

    Garnier, N.; Moncorgé, R.; Manaa, H.; Descroix, E.; Laporte, P.; Guyot, Y.

    1996-04-01

    Excited-state absorption (ESA) spectra calibrated in units of cross sections are reported in the case of Tm3+-doped YAG, YAP, and Y2O3 single crystals around wavelengths at which photon-avalanche absorptions were observed recently. The peak ESA cross sections are compared with those obtained theoretically by using the Judd-Ofelt approach.

  20. Excited-state absorption in bacteriochlorophyll a-protein from the green photosynthetic bacterium Prosthecochloris aestuarii: Reinterpretation of the absorption difference spectrum

    SciTech Connect

    Amerongen, H. van; Struve, W.S. )

    1991-10-31

    Excited-state absorption arising from transitions between singly and doubly excited exciton components in strongly coupled photosynthetic antennae profoundly influences the absorption difference spectra observed in pump-probe spectroscopy. Model calculations of the absorption difference spectrum in the BChl a-protein complex from P. aestuarii are compared with the experimental spectrum.

  1. Reduced lifetimes are directly correlated with excitation irradiance in metal-enhanced fluorescence (MEF).

    PubMed

    Karolin, Jan O; Geddes, Chris D

    2012-11-01

    We describe a fundamental observation in Metal-Enhanced Fluorescence (MEF), which has become a leading technology in the life sciences today, namely, how the lifetime of fluorophores near-to metallic plasmon-supporting silver islands/nanoparticles, modulates as a function of excitation power irradiance. This finding is in stark contrast to that observed in classical far-field fluorescence spectroscopy, where excitation power does not influence fluorophore radiative decay/lifetime.

  2. 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.

  3. Two-Photon Excited Fluorescence from Biological Aerosol Particles

    DTIC Science & Technology

    2010-09-29

    multi-photon fluorescence from coumarin in ethanol droplets and tryptophan in nominally 50 m water droplets. Mode-locked fiber lasers are...43 2.0E-43 coumarin 450 in ethanol 3 1.1E-45 3.5E-44 Table. 1. The measured 2-photon fluorescence cross-section for a number of biological

  4. 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,"…

  5. 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

    2006-09-06

    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.

  6. 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.

  7. 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.

  8. 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.

  9. Capillary Electrophoresis and Fluorescence Excitation-Emission Matrix Characterization of Soil Mobile and Calcium Humates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Capillary electrophoresis (CE) and Excitation-emission matrix (EEM) fluorescence spectroscopy have been used in natural organic matter (NOM) studies. The mutual relevance of data collected from each of the two methods provides novel insight into the correlation of complex NOM fluorescence spectra to...

  10. Excitation-and-collection geometry insensitive fluorescence imaging of tissue-simulating turbid media.

    PubMed

    Qu, J Y; Huang, Z; Hua, J

    2000-07-01

    We present an imaging technique for the correction of geometrical effects in fluorescence measurement of optically thick, turbid media such as human tissue. Specifically, we use the cross-polarization method to reject specular reflection and enhance the diffusive backscattering of polarized fluorescence excitation light from the turbid media. We correct the nonuniformity of the image field caused by the excitation-and-collection geometry of a fluorescence imaging system by normalizing the fluorescence image to the cross-polarized reflection image. The ratio image provides a map of relative fluorescence yield, defined as the ratio of emerging fluorescence power to incident excitation, over the surface of an imaged homogeneous turbid medium when fluorescence excitation-and-collection geometries vary in a wide range. We investigate the mechanism of ratio imaging by using Monte Carlo modeling. Our findings show that this technique could have a potential use in the detection of early cancer, which usually starts from a superficial layer of tissue, based on the contrast in the tissue fluorescence of an early lesion and of the surrounding normal tissue.

  11. 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.

  12. Fluorescent resonant excitation energy transfer in linear polyenes

    NASA Astrophysics Data System (ADS)

    Das, Mousumi; Ramasesha, S.

    2010-03-01

    We have studied the dynamics of excitation transfer between two conjugated polyene molecules whose intermolecular separation is comparable to the molecular dimensions. We have employed a correlated electron model that includes both the charge-charge, charge-bond, and bond-bond intermolecular electron repulsion integrals. We have shown that the excitation transfer rate varies as inverse square of donor-acceptor separation R-2 rather than as R-6, suggested by the Förster type of dipolar approximation. Our time-evolution study also shows that the orientational dependence on excitation transfer at a fixed short donor-acceptor separation cannot be explained by Förster type of dipolar approximation beyond a certain orientational angle of rotation of an acceptor polyene with respect to the donor polyene. The actual excitation transfer rate beyond a certain orientational angle is faster than the Förster type of dipolar approximation rate. We have also studied the excitation transfer process in a pair of push-pull polyenes for different push-pull strengths. We have seen that, depending on the push-pull strength, excitation transfer could occur to other dipole coupled states. Our study also allows for the excitation energy transfer to optically dark states which are excluded by Förster theory since the one-photon transition intensity to these states (from the ground state) is zero.

  13. Quenching of nitrogen dioxide fluorescence excited by a He-Cd laser

    SciTech Connect

    Kireev, S.V.; Shnyrev, S.L.

    1994-06-01

    The effect of some buffer gases on nitrogen dioxide fluorescence excited by a 0.44-{mu}m He-Cd laser is investigated. The rate constants of fluorescence self-quenching by buffer gases are measured. The cross sections of collision quenching of fluorescence are determined. The obtained results can be used for nitrogen dioxide monitoring in the atmosphere. 12 refs., 3 figs., 1 tab.

  14. 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.

  15. 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

  16. Vibrational Spectrum of an Excited State and Huang-Rhys Factors by Coherent Wave Packets in Time-Resolved Fluorescence Spectroscopy.

    PubMed

    Lee, Gyeongjin; Kim, Junwoo; Kim, So Young; Kim, Dong Eon; Joo, Taiha

    2017-01-10

    Coherent nuclear wave packet motions in an electronic excited state of a molecule are measured directly by time-resolved spontaneous fluorescence spectroscopy with an unprecedented time resolution by using two-photon absorption excitation and fluorescence upconversion by noncollinear sum frequency generation. With an estimated time resolution of approximately 25 fs, wave packet motions of vibrational modes up to 1600 cm(-1) are recorded for coumarin 153 in ethanol. Two-color transient absorption at 13 fs time resolution are measured to confirm the result. Vibrational displacements between the ground and excited states and Huang-Rhys factors (HRFs) are calculated by quantum mechanical methods and are compared with the experimental results. HRFs calculated by density functional theory (DFT) and time-dependent DFT reproduce the experiment adequately. This fluorescence-based method provides a unique and direct way to obtain the vibrational spectrum of a molecule in an electronic excited state and the HRFs, as well as the dynamics of excited states, and it might provide information on the structure of an excited state through the HRFs.

  17. Excited-state symmetry breaking of linear quadrupolar chromophores: A transient absorption study

    NASA Astrophysics Data System (ADS)

    Dozova, Nadia; Ventelon, Lionel; Clermont, Guillaume; Blanchard-Desce, Mireille; Plaza, Pascal

    2016-11-01

    The photophysical properties of two highly symmetrical quadrupolar chromophores were studied by both steady-state and transient absorption spectroscopy. Their excited-state behavior is dominated by the solvent-induced Stokes shift of the stimulated-emission band. The origin of this shift is attributed to symmetry breaking that confers a non-vanishing dipole moment to the excited state of both compounds. This dipole moment is large and constant in DMSO, whereas symmetry breaking appears significantly slower and leading to smaller excited-state dipole in toluene. Time-dependant increase of the excited-state dipole moment induced by weak solvation is proposed to explain the results in toluene.

  18. Statistical analysis of excitation-emission matrices for laser-induced fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Maslov, N. A.; Papaeva, E. O.

    2016-07-01

    An algorithm for statistical processing of the set of multicomponent excitation-emission matrices for laser-induced fluorescence spectroscopy is proposed that is based on principal component analysis. It is shown for the first time that the fluorescence emission and excitation spectra of unknown fluorophores in optically thin samples can be calculated. Using the proposed algorithm, it is possible to pass from principal components with alternating signs to positive quantities corresponding to the spectra of real substances. The method is applied to a mixture of three fluorescent dyes, and it is demonstrated that the obtained spectra of principal components well reproduce the spectra of initial dyes.

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

    PubMed

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

    2014-05-05

    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.

  20. 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.

  1. Fluorescence enhancement and reflection of the excitation light observed with a multilayered substrate.

    PubMed

    Akimoto, Takuo; Yasuda, Mitsuru

    2010-01-01

    Fluorescence enhancement from a multilayered substrate fabricated with Ag and Al(2)O(3) was investigated using fluorescein, rhodamine B, Cy3, and Cy5 as fluorophores. The change in the fluorescence enhancement with Al(2)O(3) had two peaks and one valley in the range from 0 to 300 nm of Al(2)O(3) thickness, and such peaks and valley were found to appear periodically. Moreover, the reflection of the excitation light from the multilayered substrate was investigated. The reflection of the excitation light periodically changed depending on the Al(2)O(3) thickness as well, and the maximum reflection was observed near the Al(2)O(3) thickness of the peak fluorescence enhancement. It was found that the periodic changes of the fluorescence enhancement and the reflection of the excitation light could be explained, for the most part, with the integral multiples of the lambda/4 derived by a simple interference theory.

  2. 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.

  3. Interpretation of unusual absorption bandwidths and resonance Raman intensities in excited state mixed valence.

    PubMed

    Lockard, Jenny V; Valverde, Guadalupe; Neuhauser, Daniel; Zink, Jeffrey I; Luo, Yun; Weaver, Michael N; Nelsen, Stephen F

    2006-01-12

    Excited state mixed valence (ESMV) occurs in molecules in which the ground state has a symmetrical charge distribution but the excited state possesses two or more interchangeably equivalent sites that have different formal oxidation states. Although mixed valence excited states are relatively common in both organic and inorganic molecules, their properties have only recently been explored, primarily because their spectroscopic features are usually overlapped or obscured by other transitions in the molecule. The mixed valence excited state absorption bands of 2,3-di-p-anisyl-2,3-diazabicyclo[2.2.2]octane radical cation are well-separated from others in the absorption spectrum and are particularly well-suited for detailed analysis using the ESMV model. Excited state coupling splits the absorption band into two components. The lower energy component is broader and more intense than the higher energy component. The absorption bandwidths are caused by progressions in totally symmetric modes, and the difference in bandwidths is caused by the coordinate dependence of the excited state coupling. The Raman intensities obtained in resonance with the high and low energy components differ significantly from those expected based on the oscillator strengths of the bands. This unexpected observation is a result of the excited state coupling and is explained by both the averaging of the transition dipole moment orientation over all angles for the two types of spectroscopies and the coordinate-dependent coupling. The absorption spectrum is fit using a coupled two-state model in which both symmetric and asymmetric coordinates are included. The physical meaning of the observed resonance Raman intensity trends is discussed along with the origin of the coordinate-dependent coupling. The well-separated mixed valence excited state spectroscopic components enable detailed electronic and resonance Raman data to be obtained from which the model can be more fully developed and tested.

  4. Nonlinear optical properties of tetrapyrazinoporphyrazinato indium chloride complexes due to excited-state absorption processes.

    PubMed

    Dini, Danilo; Hanack, Michael; Meneghetti, Moreno

    2005-07-07

    The multiphoton absorption properties of the axially substituted tetrapyrazinotetraazaporphyrinato complex Pyz(4)TAPInCl (1) are reported and interpreted. In particular, the nonlinear optical transmission of the complex and the excited states involved in the nonlinear absorption have been determined at the frequency of the second harmonic generation of a Nd:YAG laser in the nanosecond time regime. Pyz(4)TAPInCl has an excited-state absorption cross section larger than its ground state in the 460-540 nm spectral region, and it shows an optical limiting (OL) behavior at 532 nm, which derives from a sequential two-photon absorption with a larger absorption cross section of the excited triplet state with respect to the ground state. It results that the absorption cross section of 1 in the excited triplet state is 7.8 x 10(-18) cm(2) vs 0.9 x 10(-18) cm(2) of the ground state at the wavelength of OL analysis.

  5. 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.

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

    PubMed

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

    2016-10-14

    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.

  7. 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.

  8. Biocompatible glutathione capped gold clusters as one- and two-photon excitation fluorescence contrast agents for live cells imaging.

    PubMed

    Polavarapu, Lakshminarayana; Manna, Manoj; Xu, Qing-Hua

    2011-02-01

    The one- and two-photon excitation emission properties of water soluble glutathione monolayer protected gold clusters were investigated. Strong two-photon emission was observed from the gold clusters. The two-photon absorption cross section of these gold clusters in water was deduced from the z-scan measurement to be 189 740 GM, which is much higher compared to organic fluorescent dyes and quantum dots. These gold clusters also showed high photo-stability. The MTT assay showed that these gold clusters have low toxicity even at high concentrations. We have successfully demonstrated their applications for both one and two-photon excitation live cell imaging. The exceptional properties of these gold clusters make them a promising alternative for one- and two-photon bio-imaging and other nonlinear optical applications.

  9. 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.

  10. 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.

  11. Simultaneous acquisition of absorption and fluorescence spectra of strong absorbers utilizing an evanescent supercontinuum.

    PubMed

    Kiefer, Johannes

    2016-12-15

    The determination of the absorption and emission spectra of strongly absorbing molecules is challenging, and the data can be biased by self-absorption of the fluorescence signal. To overcome this problem, a total internal reflection approach is proposed. The strongly absorbing sample is placed in an evanescent field of the radiation of a supercontinuum source. The collimated reflected light encodes the absorption spectrum, and the isotropic fluorescence emission is collected in a direction perpendicular to the surface at the same time. This ensures that the emitted light has a minimum possibility of self-absorption inside the sample.

  12. 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.

  13. Sensitivity of laser-induced upconversion fluorescence dynamics to exciting wavelength in Er3+-doped YAG

    NASA Astrophysics Data System (ADS)

    Du, S.; Jiang, L.; Zhang, W.; Gong, W.; Fu, Z.; Dai, Z.

    2011-06-01

    A novel sensitivity of laser-induced upconversion (UC) fluorescence dynamics to the exciting wavelength in Er3+:YAG crystal has been observed. The sensitivities to exciting wavelength variations are only 0.02 and 0.06 nm for 2 G 9/2 and 2 P 3/2 UC luminescence, respectively. The observation of this sensitivity reveals that when a certain level is populated by more than one UC mechanisms, a precise determination of suitable exciting wavelength is crucial because the competition between different UC mechanisms has a sensitive variation with exciting wavelength in near-resonant range.

  14. Biocompatible glutathione capped gold clusters as one- and two-photon excitation fluorescence contrast agents for live cells imaging

    NASA Astrophysics Data System (ADS)

    Polavarapu, Lakshminarayana; Manna, Manoj; Xu, Qing-Hua

    2011-02-01

    The one- and two-photon excitation emission properties of water soluble glutathione monolayer protected gold clusters were investigated. Strong two-photon emission was observed from the gold clusters. The two-photon absorption cross section of these gold clusters in water was deduced from the z-scan measurement to be 189 740 GM, which is much higher compared to organic fluorescent dyes and quantum dots. These gold clusters also showed high photo-stability. The MTT assay showed that these gold clusters have low toxicity even at high concentrations. We have successfully demonstrated their applications for both one and two-photon excitation live cell imaging. The exceptional properties of these gold clusters make them a promising alternative for one- and two-photon bio-imaging and other nonlinear optical applications.The one- and two-photon excitation emission properties of water soluble glutathione monolayer protected gold clusters were investigated. Strong two-photon emission was observed from the gold clusters. The two-photon absorption cross section of these gold clusters in water was deduced from the z-scan measurement to be 189 740 GM, which is much higher compared to organic fluorescent dyes and quantum dots. These gold clusters also showed high photo-stability. The MTT assay showed that these gold clusters have low toxicity even at high concentrations. We have successfully demonstrated their applications for both one and two-photon excitation live cell imaging. The exceptional properties of these gold clusters make them a promising alternative for one- and two-photon bio-imaging and other nonlinear optical applications. Electronic supplementary information (ESI) available: Control experiments; blinking studies of gold clusters. See DOI: 10.1039/c0nr00458h

  15. 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.

  16. Excited state absorption in glasses activated with rare earth ions: Experiment and modeling

    NASA Astrophysics Data System (ADS)

    Piatkowski, Dawid; Mackowski, Sebastian

    2012-10-01

    We present semiempirical approach based on the Judd-Ofelt theory and apply it for modeling the spectral properties of fluoride glasses activated with the rare earth (RE) ions. This method provide a powerful tool for simulating both ground state absorption (GSA) and excited state absorption (ESA) spectra of RE ions, e.g. Nd3+, Ho3+, Er3+ and Tm3+ in the ZBLAN glass matrix. The results of theoretical calculations correspond to the experimentally measured data. We also demonstrate that the spectra obtained using the presented approach are applicable in the analysis of up-conversion excitation schemes in these optoelectronically relevant materials.

  17. Dye-doped sol-gel materials for two-photon absorption induced fluorescence

    NASA Astrophysics Data System (ADS)

    Canva, Michael; Roger, Gisèle; Cassagne, Florence; Lévy, Yves; Brun, Alain; Chaput, Frédéric; Boilot, Jean-Pierre; Rapaport, Alexandra; Heerdt, Céline; Bass, Michael

    2002-01-01

    Two-photon absorption (TPA) and subsequent fluorescence properties of laser dyes are retained when doped into solid state sol-gel materials. These properties were demonstrated to be applicable in true 3D displays.

  18. 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.

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

    PubMed

    Moreno, J; Dobryakov, A L; Ioffe, I N; Granovsky, A A; Hecht, S; Kovalenko, S A

    2015-07-14

    1-photon (382 nm) and 2-photon (752 nm) excitations to the S1 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 S1 → Sn due to resonant absorption of a third pump photon. Subsequent Sn → S1 internal conversion (with τ1 = 1 ps) prepares a very hot S1 state which cools down with τ2 = 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(2) = 32 ⋅ 10(-50) cm(4) s at 752 nm are evaluated from the bleach signal.

  20. 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.

  1. 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.

  2. Cine: Line excitation by infrared fluorescence in cometary atmospheres

    NASA Astrophysics Data System (ADS)

    de Val-Borro, Miguel; Cordiner, Martin A.; Milam, Stefanie N.; Charnley, Steven B.

    2017-03-01

    CINE is a Python module for calculating infrared pumping efficiencies that can be applied to the most common molecules found in cometary comae such as water, hydrogen cyanide or methanol. Excitation by solar radiation of vibrational bands followed by radiative decay to the ground vibrational state is one of the main mechanisms for molecular excitation in comets. This code calculates the effective pumping rates for rotational levels in the ground vibrational state scaled by the heliocentric distance of the comet. Line transitions are queried from the latest version of the HITRAN spectroscopic repository using the astroquery affiliated package of astropy. Molecular data are obtained from the LAMDA database. These coefficients are useful for modeling rotational emission lines observed in cometary spectra at sub-millimeter wavelengths. Combined with computational methods to solve the radiative transfer equations based, e.g., on the Monte Carlo algorithm, this model can retrieve production rates and rotational temperatures from the observed emission spectrum.

  3. Plasmon-Modulated Excitation-Dependent Fluorescence from Activated CTAB Molecules Strongly Coupled to Gold Nanoparticles.

    PubMed

    Ding, Si-Jing; Nan, Fan; Liu, Xiao-Li; Hao, Zhong-Hua; Zhou, Li; Zeng, Jie; Xu, Hong-Xing; Zhang, Wei; Wang, Qu-Quan

    2017-03-07

    Excitation-dependent fluorophores (EDFs) have been attracted increasing attention owing to their high tunability of emissions and prospective applications ranging from multicolor patterning to bio-imaging. Here, we report tunable fluorescence with quenching dip induced by strong coupling of exciton and plasmon in the hybrid nanostructure of CTAB* EDFs and gold nanoparticles (AuNPs). The quenching dip in the fluorescence spectrum is tuned by adjusting excitation wavelength as well as plasmon resonance and concentration of AuNPs. The observed excitation-dependent emission spectra with quenching dip are theoretically reproduced and revealed to be induced by resonant energy transfer from multilevel EDFs with wider width channels to plasmonic AuNPs. These findings provide a new approach to prepare EDF molecules and a strategy to modulate fluorescence spectrum via exciton-to-plasmon energy transfer.

  4. Plasmon-Modulated Excitation-Dependent Fluorescence from Activated CTAB Molecules Strongly Coupled to Gold Nanoparticles

    NASA Astrophysics Data System (ADS)

    Ding, Si-Jing; Nan, Fan; Liu, Xiao-Li; Hao, Zhong-Hua; Zhou, Li; Zeng, Jie; Xu, Hong-Xing; Zhang, Wei; Wang, Qu-Quan

    2017-03-01

    Excitation-dependent fluorophores (EDFs) have been attracted increasing attention owing to their high tunability of emissions and prospective applications ranging from multicolor patterning to bio-imaging. Here, we report tunable fluorescence with quenching dip induced by strong coupling of exciton and plasmon in the hybrid nanostructure of CTAB* EDFs and gold nanoparticles (AuNPs). The quenching dip in the fluorescence spectrum is tuned by adjusting excitation wavelength as well as plasmon resonance and concentration of AuNPs. The observed excitation-dependent emission spectra with quenching dip are theoretically reproduced and revealed to be induced by resonant energy transfer from multilevel EDFs with wider width channels to plasmonic AuNPs. These findings provide a new approach to prepare EDF molecules and a strategy to modulate fluorescence spectrum via exciton-to-plasmon energy transfer.

  5. Plasmon-Modulated Excitation-Dependent Fluorescence from Activated CTAB Molecules Strongly Coupled to Gold Nanoparticles

    PubMed Central

    Ding, Si-Jing; Nan, Fan; Liu, Xiao-Li; Hao, Zhong-Hua; Zhou, Li; Zeng, Jie; Xu, Hong-Xing; Zhang, Wei; Wang, Qu-Quan

    2017-01-01

    Excitation-dependent fluorophores (EDFs) have been attracted increasing attention owing to their high tunability of emissions and prospective applications ranging from multicolor patterning to bio-imaging. Here, we report tunable fluorescence with quenching dip induced by strong coupling of exciton and plasmon in the hybrid nanostructure of CTAB* EDFs and gold nanoparticles (AuNPs). The quenching dip in the fluorescence spectrum is tuned by adjusting excitation wavelength as well as plasmon resonance and concentration of AuNPs. The observed excitation-dependent emission spectra with quenching dip are theoretically reproduced and revealed to be induced by resonant energy transfer from multilevel EDFs with wider width channels to plasmonic AuNPs. These findings provide a new approach to prepare EDF molecules and a strategy to modulate fluorescence spectrum via exciton-to-plasmon energy transfer. PMID:28266619

  6. CF2 and CFCl fluorescence from VUV excitation of C2F3Cl

    NASA Technical Reports Server (NTRS)

    Nee, J. B.; Wang, Xiuyan; Suto, Masako; Lee, Long C.

    1986-01-01

    The photoexcitation process of C2F3Cl molecule was investigated in the 106 to 230 nm region using synchrotron radiation as a light source. Photoabsorption and fluorescence cross sections were measured and used to determine the fluorescence quantum yield. Fluorescence yield starts to appear at 170 nm and increases to about 2% at 155 nm. The fluorescence spectra were dispersed to identify the emitting species. At the excitation wavelength of 155 nm, the emission system is CFCl (Hermitian conjugate of A - Hermitian conjugate of X), and at 123.9 nm, both the CF2 (Hermitian conjugate of A - Hermitian conjugate of X) and CFCl (Hermitian conjugate of A - Hermitian conjugate of X) systems are observed. The dissociation processes that produced these excited species are discussed.

  7. Two-photon excited fluorescence microendoscopic imaging using a GRIN lens

    NASA Astrophysics Data System (ADS)

    Yan, Wei; Peng, Xiao; Lin, Danying; Wang, Qi; Gao, Jian; Zhou, Jie; Ye, Tong; Qu, Junle; Niu, Hanben

    2015-03-01

    With the rapid development of life sciences, there is an increasing demand for intravital fluorescence imaging of small animals. However, large dimensions and limited working distances of objective lenses in traditional fluorescence microscopes have limited the imaging applications mostly to superficial tissues. To overcome this disadvantage, researchers have developed the graded-index (GRIN) probes with small diameters for imaging internal organs of small animals in a minimally invasive fashion. Here, we present the development of a fluorescence endoscopic imaging system based on a GRIN lens using two-photon excitation. Experimental results showed that this system could perform dynamic fluorescence microendoscopic imaging and monitor the blood flow in anesthetized living mice using two-photon excitation.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. Two Photon Absorption Laser Induced Fluorescence for Neutral Hydrogen Profile Measurements

    SciTech Connect

    Scime, Earl E.

    2016-09-23

    The magnitude and spatial dependence of neutral density in magnetic confinement fusion experiments is a key physical parameter, particularly in the plasma edge. Modeling codes require precise measurements of the neutral density to calculate charge-exchange power losses and drag forces on rotating plasmas. However, direct measurements of the neutral density are problematic. In this work, we proposed to construct a laser-based diagnostic capable of providing spatially resolved measurements of the neutral density in the edge of plasma in the DIII-D tokamak. The diagnostic concept is based on two-photon absorption laser induced fluorescence (TALIF). By injecting two beams of 205 nm light (co or counter propagating), ground state hydrogen (or deuterium or tritium) can be excited from the n = 1 level to the n = 3 level at the location where the two beams intersect. Individually, the beams experience no absorption, and therefore have no difficulty penetrating even dense plasmas. After excitation, a fraction of the hydrogen atoms decay from the n = 3 level to the n = 2 level and emit photons at 656 nm (the Hα line). Calculations based on the results of previous TALIF experiments in magnetic fusion devices indicated that a laser pulse energy of approximately 3 mJ delivered in 5 ns would provide sufficient signal-to-noise for detection of the fluorescence. In collaboration with the DIII-D engineering staff and experts in plasma edge diagnostics for DIII-D from Oak Ridge National Laboratory (ORNL), WVU researchers designed a TALIF system capable of providing spatially resolved measurements of neutral deuterium densities in the DIII-D edge plasma. The laser systems were specified, purchased, and assembled at WVU. The TALIF system was tested on a low-power hydrogen discharge at WVU and the plan was to move the instrument to DIII-D for installation in collaboration with ORNL researchers. After budget cuts at DIII-D, the DIII-D facility declined to support

  13. 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.

  14. Attosecond Transient Absorption Spectroscopy of doubly-excited states in helium

    NASA Astrophysics Data System (ADS)

    Argenti, Luca; Ott, Christian; Pfeifer, Thomas; Martín, Fernando

    2014-04-01

    Theoretical calculations of the XUV attosecond transient absorption spectrum (ATAS) of helium in the doubly-excited state region reproduce recent high-precision measurements, reveal novel means of controlling the dynamics of transiently-bound electronic wavepackets in intense laser fields, and indicates a possible extension of 2D-spectroscopies to the XUV range.

  15. A comparison of the fluorescence dynamics of single molecules of a green fluorescent protein: one- versus two-photon excitation.

    PubMed

    Cotlet, Mircea; Goodwin, Peter M; Waldo, Geoffrey S; Werner, James H

    2006-01-16

    We report on the dynamics of fluorescence from individual molecules of a mutant of the wild-type green fluorescent protein (GFP) from Aequorea victoria, super folder GFP (SFGFP). SFGFP is a novel and robust variant designed for in vivo high-throughput screening of protein expression levels. It shows increased thermal stability and is able to retain its fluorescence when fused to poorly folding proteins. We use a recently developed single-molecule technique which combines fluorescence-fluctuation spectroscopy and time-correlated single photon counting in order to characterize the photophysical properties of SFGFP under one- (OPE) and two- (TPE) photon excitation conditions. We use Rhodamine 110 as a model chromophore to validate the methodology and to explain the single-molecule results of SFGFP. Under OPE, single SFGFP molecules undergo fluorescence flickering on the time scale of micros and tens of micros due to triplet formation and ground-state protonation-deprotonation, respectively, as demonstrated by excitation intensity- and pH-dependent experiments. OPE single-molecule fluorescence lifetimes indicate heterogeneity in the population of SFGFP, indicating the presence of the deprotonated I and B forms of the SFGFP chromophore. TPE of single SFGFP molecules results in the photoconversion of the chromophore. TPE of single SFGFP molecules show fluorescence flickering on the time scale of micros due to triplet formation. A flicker connected with protonation-deprotonation of the SFGFP chromophore is detected only at low pH. Our results show that SFGFP is a promising fusion reporter for intracellular applications using OPE and TPE microscopy.

  16. Large excited state two photon absorptions in the near infrared region of surprisingly stable radical cations of (ferrocenyl)indenes.

    PubMed

    Orian, Laura; Scuppa, Stefano; Santi, Saverio; Meneghetti, Moreno

    2013-08-21

    Multiphoton absorptions are important non-linear optical processes which allow us to explore excited states with low energy photons giving rise to new possibilities for photoinduced processes. Among these processes, multiphoton absorptions from excited states are particularly interesting because of the large susceptibilities characteristic of excited states. Here we explore the nonlinear transmission measurements recorded with 9 ns laser pulses at 1064 nm of the radical cations of (2-ferrocenyl)indene and of (2-ferrocenyl)-hexamethylindene, two interesting very stable molecules. The non-linear transmission data can be interpreted with a multiphoton sequence of three photon absorptions, the first being a one photon absorption related to the intramolecular charge transfer and the second a two photon absorption from the excited state created with the first process. The two photon absorption cross section is found to be several orders of magnitude larger than those usually found for two photon absorbing systems excited from the ground state.

  17. Ultrafast relaxation dynamics of 5,10,15,20-meso-tetrakis pentafluorophenyl porphyrin studied by fluorescence up-conversion and transient absorption spectroscopy.

    PubMed

    Kumar, P Hemant; Venkatesh, Yeduru; Siva, Doddi; Ramakrishna, B; Bangal, Prakriti Ranjan

    2015-02-26

    The ultrafast photophysical characterization of 5,10,15,20-meso-tetrakis pentafluorophenyl porphyrin (H2F20TPP) in 4:1 dichloromethane (DCM) and tetrahydrofuran (THF) solution has been done in the femtosecond-picosecond time domain, by combining fluorescence up-conversion and femtosecond transient absorption spectroscopy. Fluorescence up-conversion studies on H2F20TPP were done demonstrating fluorescence dynamics over the whole spectral range from 440 to 650 nm when excited at 405 nm, 360.5 cm(-1) excess vibrational energy of Soret band (411 nm). Single-exponential decay with ∼160 ± 50 fs lifetime of Soret fluorescence (also called S2 fluorescence or B band fluorescence) at around 440 nm was observed. On going from 440 nm, S2 fluorescence to S1 fluorescence, (Q-band) around 640 nm (wavelength of 0-0 transition in the stationary spectrum), single-exponential fluorescence time profile turns into a multiexponential time profile and it could be resolved critically into five-exponential components. An ultrafast rise component with ∼160 ± 50 fs followed by two decay components: a very fast decay component with 200 ± 50 fs time constant and another relatively slower 1.8 ± 0.5 ps decay component. Next, a very prominent rise component with 105 ± 30 ps lifetime followed by long-lived 10 ns decay component. The initial rise of S1 (Q-band) fluorescence around 640 nm agreed with the decay time of S2 (Soret or B band) fluorescence indicates that internal conversion (IC) from relaxed S2 to vibrationally excited S1 occurs in the ∼160 fs time scale and subsequent very fast decay with 200 fs time constant, which is assigned to be intramolecular vibrational dephasing or redistribution. The 1.8 ps decay component of S1 fluorescence is attributed to be "hot" fluorescence from vibrationally excited S1 state, and it reveals the vibrational relaxation time induced by elastic or quasi-elastic collision with solvent molecules. The 105 ps rise component is the creation time of the

  18. 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%.

  19. Absorption and Luminescence Studies of Some Highly Fluorescent Derivatives of Vitamin B1; Solvent and pH Effects

    NASA Astrophysics Data System (ADS)

    Marciniak, B.; Koput, J.; Kozubek, H.

    1990-08-01

    The influence of solvent on the UV-visible absorption and luminescence spectra of some highly fluorescent vitamin B1 derivatives, the products of the reaction of N-methylated vitamin B1 with cytidine (I), adenosine (II) and 2-amino-4-methylpyridine (III) is studied. Spectroscopic manifestations of protonation of I and II are also investigated using a semiempirical INDO/S CI method. Singlet and triplet energy levels of the free ion and several protonated species are calculated, and transition energies and oscillator strengths are compared with the experimental spectra. Calculated charge densities on heteroatoms in the ground and excited singlet and triplet states are correlated with changes of the experimental pKa values with excitation. The results for I and II are compared with those for the trimethylated pyrichrominium ion (III) previously studied

  20. 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.

  1. Role of non-Condon vibronic coupling and conformation change on two-photon absorption spectra of green fluorescent protein

    NASA Astrophysics Data System (ADS)

    Ai, Yuejie; Tian, Guangjun; Luo, Yi

    2013-07-01

    Two-photon absorption spectra of green fluorescent proteins (GFPs) often show a blue-shift band compared to their conventional one-photon absorption spectra, which is an intriguing feature that has not been well understood. We present here a systematic study on one- and two-photon spectra of GFP chromophore by means of the density functional response theory and complete active space self-consistent field (CASSCF) methods. It shows that the popular density functional fails to provide correct vibrational progression for the spectra. The non-Condon vibronic coupling, through the localised intrinsic vibrational modes of the chromophore, is responsible for the blue-shift in the TPA spectra. The cis to trans isomerisation can be identified in high-resolution TPA spectra. Our calculations demonstrate that the high level ab initio multiconfigurational CASSCF method, rather than the conventional density functional theory is required for investigating the essential excited-state properties of the GFP chromophore.

  2. Absorption and fluorescence spectroscopic characterisation of the circadian blue-light photoreceptor cryptochrome from Drosophila melanogaster (dCry)

    NASA Astrophysics Data System (ADS)

    Shirdel, J.; Zirak, P.; Penzkofer, A.; Breitkreuz, H.; Wolf, E.

    2008-09-01

    The absorption and fluorescence behaviour of the circadian blue-light photoreceptor cryptochrome from Drosophila melanogaster (dCry) in a pH 8 aqueous buffer solution is studied. The flavin adenine dinucleotide (FAD) cofactor of dCry is identified to be present in its oxidized form (FAD ox), and the 5,10-methenyltetrahydrofolate (MTHF) cofactor is found to be hydrolyzed and oxidized to 10-formyldihydrofolate (10-FDHF). The absorption and the fluorescence behaviour of dCry is investigated in the dark-adapted (receptor) state, the light-adapted (signalling) state, and under long-time violet light exposure. Photo-excitation of FAD ox in dCry causes a reductive electron transfer to the formation of anionic FAD semiquinone (FAD rad - ), and photo-excitation of the generated FAD rad - causes an oxidative electron transfer to the back formation of FAD ox. In light adapted dCry a photo-induced equilibrium between FAD ox and FAD rad - exists. The photo-cycle dynamics of signalling state formation and recovery is discussed. Quantum yields of photo-induced signalling state formation of about 0.2 and of photo-induced back-conversion of about 0.2 are determined. A recovery of FAD rad - to FAD ox in the dark with a time constant of 1.6 min at room temperature is found.

  3. Saturated two-photon excitation fluorescence microscopy with core-ring illumination.

    PubMed

    Oketani, Ryosuke; Doi, Atsushi; Smith, Nicholas I; Nawa, Yasunori; Kawata, Satoshi; Fujita, Katsumasa

    2017-02-01

    We demonstrated resolution improvement in two-photon excitation microscopy by combining saturated excitation (SAX) of fluorescence and pupil manipulation. We theoretically estimated the resolution improvement and the sidelobe effect in the point spread function with various pupil designs and found that the combination of SAX and core-ring illumination can effectively enhance the spatial resolution in 3D and suppress sidelobe artifacts. The experimental demonstration shows that the proposed technique is effective for observation with a depth of 100 μm in a tissue phantom and can be applied to 3D observations of tissue samples with higher spatial resolution than conventional two-photon excitation microscopy.

  4. Adaptive Control of Two-Photon Excitation of Green Fluorescent Protein with Shaped Femtosecond Pulses

    NASA Astrophysics Data System (ADS)

    Kawano, Hiroyuki; Nabekawa, Yasuo; Suda, Akira; Oishi, Yu; Mizuno, Hideaki; Miyawaki, Atsushi; Midorikawa, Katsumi

    For many years, it has been believed that a Fourier-transform-limited (FTL) laser pulse is the most effective light source for the generation of nonlinear phenomena, since the FTL pulse has the shortest pulse duration, that is, the highest intensity, that can be limited by the spectral width due to the principle of uncertainty. Recently, many reports have been published on the adaptive control of nonlinear phenomena with shaped femtosecond excitation laser pulses [1, 2]. Their reports have shown that the modification of the spectral and temporal phases of excitation pulses can increase or decrease the probabilities and efficiencies of such nonlinear phenomena. This method has been widely applied to studies on the active control of molecular motions or chemical reactions [3,4]. Considering further novel biological applications, we focus on the two-photon excited fluorescence (TPEF) of the green fluorescent protein (GFP) from the jellyfish Aequorea victoria. GFP is spontaneously fluorescent and is relatively nontoxic compared with other organic dyes used as optical markers. Therefore, it has been widely used as a "tag" material for the fluorescence observation of living cells [5]. Two-photon excitation microscopy (TPEM) is a powerful tool for biological real-time observation due to its various advantages, such as a clear contrast, good S/N ratio, and high spatial resolution [7]. From a practical point of view, however, there is a serious problem with TPEM, which is the photobleaching of a dye. The intensity of a fluorescence signal decreases significantly during observation. One of the reasons for this is that the chromophore structure is degraded by intense excitation laser pulses that are required for efficient two-photon excitation. In this study, therefore, we attempted to determine the optimal phase for maximizing the fluorescence efficiency of a GFP variant with excitation laser pulses of minimal intensity. We considered that GFP can be an ideal dye for the

  5. A reversibly photoswitchable GFP-like protein with fluorescence excitation decoupled from switching.

    PubMed

    Brakemann, Tanja; Stiel, Andre C; Weber, Gert; Andresen, Martin; Testa, Ilaria; Grotjohann, Tim; Leutenegger, Marcel; Plessmann, Uwe; Urlaub, Henning; Eggeling, Christian; Wahl, Markus C; Hell, Stefan W; Jakobs, Stefan

    2011-09-11

    Photoswitchable fluorescent proteins have enabled new approaches for imaging cells, but their utility has been limited either because they cannot be switched repeatedly or because the wavelengths for switching and fluorescence imaging are strictly coupled. We report a bright, monomeric, reversibly photoswitchable variant of GFP, Dreiklang, whose fluorescence excitation spectrum is decoupled from that for optical switching. Reversible on-and-off switching in living cells is accomplished at illumination wavelengths of ∼365 nm and ∼405 nm, respectively, whereas fluorescence is elicited at ∼515 nm. Mass spectrometry and high-resolution crystallographic analysis of the same protein crystal in the photoswitched on- and off-states demonstrate that switching is based on a reversible hydration/dehydration reaction that modifies the chromophore. The switching properties of Dreiklang enable far-field fluorescence nanoscopy in living mammalian cells using both a coordinate-targeted and a stochastic single molecule switching approach.

  6. Excited state dynamics and nonlinear absorption of a pyrazinoporphyrazine macrocycle carrying externally appended pyridine rings.

    PubMed

    Villano, Massimo; Amendola, Vincenzo; Sandonà, Giancarlo; Donzello, Maria Pia; Ercolani, Claudio; Meneghetti, Moreno

    2006-12-07

    The multiphoton absorption properties of the tetrakis-2,3-[5,6-di(2-pyridyl)pyrazino]porphyrazinato(monoacquo)-Mg(II) complex [Py(8)TPyzPzMg(H(2)O)] (1) are reported and interpreted. The nonlinear optical behavior of 1 and the characterization of the excited states important for the nonlinear absorption process were studied at the pump frequency of the second harmonic generation of a Nd:YAG laser in the nanosecond time regime. It was found that complex 1 shows a very good optical limiting performance at 532 nm, which derives from two processes: (a) a reverse-saturable absorption process, which involves a triplet excited state at low intensities, and (b) a two-photon absorption process at higher intensities, which is due to the formation of the radical monoanion of 1, [Py(8)TPyzPzMg(H2O)](.-), during the photoreduction of the triplet state. The participation of a monoanion in determining the overall nonlinear absorption behavior of 1 is found, for the first time, for a tetrapyrrolic system. One can deduce that the involvement of the monoanion derives from the electron-withdrawing effect of the dipyridinopyrazino fragments externally attached to the porphyrazine core which make the reduced form of 1 easily accessible. These results suggest a modification of tetrapyrrolic systems with new nonlinear absorption properties.

  7. 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, ...

  8. Comparison of laser excited fluorescence and photoacoustic limits of detection for static and flow cells

    SciTech Connect

    Voigtman, E.; Jurgensen, A.; Winefordner, J.D.

    1981-10-01

    The fluorescence and photoacoustic characteristics of a windowless flow cell intended for liquid chromatographic applications are compared with respective characteristics of a static cuvette cell. In addition, a photoionization mode of operation for the flow cell is exhibited which utilizes the ionization products of two-photon excitation of polynuclear aromatic hydrocarbons in n-alkanes to effect a sensitive detection of those PAHs.

  9. Synthesis, spectral behaviour and photophysics of donor-acceptor kind of chalcones: Excited state intramolecular charge transfer and fluorescence quenching studies

    NASA Astrophysics Data System (ADS)

    Pannipara, Mehboobali; Asiri, Abdullah M.; Alamry, Khalid A.; Arshad, Muhammad N.; El-Daly, Samy A.

    2015-02-01

    The spectral and photophysical properties of two chalcones containing electron donating and accepting groups with intramolecular charge transfer characteristics were synthesized and characterized by 1H NMR, 13C NMR and X-ray crystallography. Both compounds show very strong solvent polarity dependent changes in their photophysical characteristics, namely, remarkable red shift in the emission spectra with increasing solvent polarity, large change in Stokes shift, significant reduction in the fluorescence quantum yield; indicating that the fluorescence states of these compounds are of intramolecular charge transfer (ICT) character. The solvent effect on the photophysical parameters such as singlet absorption, molar absorptivity, oscillator strength, dipole moment, fluorescence spectra, and fluorescence quantum yield of both compounds have been investigated comprehensively. For both dyes, Lippert-Mataga and Reichardt's correlations were used to estimate the difference between the excited and ground state dipole moments (Δμ). The interactions of dyes with colloidal silver nanoparticles (Ag NPs) were also studied in ethanol using steady state fluorescence quenching measurements. The fluorescence quenching data reveal that dynamic quenching and energy transfer play a major role in the fluorescence quenching of dyes by Ag NPs.

  10. 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.

  11. B1-based specific energy absorption rate determination for nonquadrature radiofrequency excitation.

    PubMed

    Katscher, Ulrich; Findeklee, Christian; Voigt, Tobias

    2012-12-01

    The current gold standard to estimate local and global specific energy absorption rate for MRI involves numerically modeling the patient and the transmit radiofrequency coil. Recently, a patient-individual method was presented, which estimated specific energy absorption rate from individually measured B(1) maps. This method, however, was restricted to quadrature volume coils due to difficulties distinguishing phase contributions from radiofrequency transmission and reception. In this study, a method separating these two phase contributions by comparing the electric conductivity reconstructed from different transmit channels of a parallel radiofrequency transmission system is presented. This enables specific energy absorption rate estimation not only for quadrature excitation but also for the nonquadrature excitation of the single elements of the transmit array. Though the contributions of the different phases are known, unknown magnetic field components and tissue boundary artifacts limit the technique. Nevertheless, the high agreement between simulated and experimental results found in this study is promising. B(1)-based specific energy absorption rate determination might become possible for arbitrary radiofrequency excitation on a patient-individual basis.

  12. 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.

  13. Lifetime of fluorescence from light-harvesting chlorophyll a/b proteins: excitation intensity dependence

    SciTech Connect

    Nordlund, T.M.; Knox, W.H.

    1981-10-01

    The fluorescence from a purified, aggregate form of the light-harvesting chlorophyll a/b protein has a lifetime of 1.2 +/- 0.5 ns at low excitation intensity, but the lifetime decreases significantly when the intensity of the 20-ps, 5300nm excitation pulse is increased above about 10/sup 16/ photons/cm/sup 2/. A solubilized, monomeric form of the protein, on the other hand, has a fluorescence lifetime of 3.1 +/- 0.3 ns independent of excitation intensity from 10/sup 14/-10/sup 18/ photons/cm/sup 2//pulse. We interpret the lifetime shortening in the aggregates and the lack of shortening in monomers in terms of exciton annihilation, facilitated in the aggregate by the larger population of interacting chlorophylls.

  14. Fluorescence anisotropy excitation by polarization-shaped laser pulses after transmission through a kagome fiber

    NASA Astrophysics Data System (ADS)

    Otto, J.; Patas, A.; Althoff, J.; Lindinger, A.

    2016-08-01

    We report improved fluorescence contrast between dyes by two-photon excitation with polarization-shaped laser pulses after transmission through a kagome fiber utilizing the anisotropy of the dye molecules. Particularly phase- and polarization-tailored pulse shapes are employed for two-photon excited fluorescence of dyes in a liquid environment at the distal end of the kagome fiber. The distortions due to the optical fiber properties are precompensated in order to receive predefined polarization-shaped laser pulses after the kagome fiber. This enables to optimally excite one dye in one polarization direction and simultaneously the other dye in the other polarization direction. The presented method has a high potential for endoscopic applications due to the unique properties of kagome fibers for guiding ultrashort laser pulses.

  15. 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.

  16. 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.

  17. 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

  18. Rapid prototyping of electrochromatography chips for improved two-photon excited fluorescence detection.

    PubMed

    Hackl, Claudia; Beyreiss, Reinhild; Geissler, David; Jezierski, Stefan; Belder, Detlev

    2014-04-15

    In the present study, we introduce two-photon excitation at 532 nm for label-free fluorescence detection in chip electrochromatography. Two-photon excitation at 532 nm offers a promising alternative to one-photon excitation at 266 nm, as it enables the use of economic chip materials instead of fused silica. In order to demonstrate these benefits, one-photon and two-photon induced fluorescence detection are compared in different chip layouts and materials with respect to the achievable sensitivity in the detection of polycyclic aromatic hydrocarbons (PAHs). Customized chromatography chips with cover or bottom slides of different material and thickness are produced by means of a rapid prototyping method based on liquid-phase lithography. The design of thin bottom chips (180 μm) enables the use of high-performance immersion objectives with low working distances, which allows one to exploit the full potential of two-photon excitation for a sensitive detection. The developed method is applied for label-free analysis of PAHs separated on a polymer monolith inside polymer glass sandwich chips made from fused silica or soda-lime glass. The obtained limits of detection range from 40 nM to 1.95 μM, with similar sensitivities in fused silica thin bottom chips for one-photon and two-photon excitation. In deep-UV non- or less-transparent devices two-photon excitation is mandatory for label-free detection of aromatics with high sensitivity.

  19. Light absorption and excitation energy transfer calculations in primitive photosynthetic bacteria

    NASA Astrophysics Data System (ADS)

    Komatsu, Yu; Kayanuma, Megumi; Shoji, Mitsuo; Yabana, Kazuhiro; Shiraishi, Kenji; Umemura, Masayuki

    2015-06-01

    In photosynthetic organisms, light energy is converted into chemical energy through the light absorption and excitation energy transfer (EET) processes. These processes start in light-harvesting complexes, which contain special photosynthetic pigments. The exploration of unique mechanisms in light-harvesting complexes is directly related to studies, such as artificial photosynthesis or biosignatures in astrobiology. We examined, through ab initio calculations, the light absorption and EET processes using cluster models of light-harvesting complexes in purple bacteria (LH2). We evaluated absorption spectra and energy transfer rates using the LH2 monomer and dimer models to reproduce experimental results. After the calibration tests, a LH2 aggregation model, composed of 7 or 19 LH2s aligned in triangle lattice, was examined. We found that the light absorption is red shifted and the energy transfer becomes faster as the system size increases. We also found that EET is accelerated by exchanging the central pigments to lower energy excited pigments. As an astrobiological application, we calculated light absorptions efficiencies of the LH2 in different photoenvironments.

  20. 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.

  1. Excitation-wavelength Dependent Fluorescence of Ethyl 5-(4-aminophenyl)-3-amino-2,4-dicyanobenzoate

    PubMed Central

    Heldt, Janina R.

    2010-01-01

    The excitation wavelength dependence of the steady-state and time-resolved emission spectra of ethyl 5-(4-aminophenyl)-3-amino-2,4-dicyanobenzoate (EAADCy) in tetrahydrofuran (THF) at room temperature has been examined. It is found that the ratio of the fluorescence intensity of the long-wavelength and short-wavelength fluorescence bands strongly depends on the excitation wavelength, whereas the wavelengths of the fluorescence excitation and fluorescence bands maxima are independent on the observation/excitation wavelengths. The dynamic Stokes shift of fluorophore in locally excited (LE) and intramolecular charge transfer (ICT) states has been studied with a time resolution about 30 ps. The difference between Stokes shift in the LE and ICT states was attributed to the solvent response to the large photoinduced dipole moment of EAADCy in the fluorescent charge transfer state. On this base we can state that, the relaxation of the polar solvent molecules around the fluorophore was observed. PMID:20798980

  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.

  3. Multi-excitation near infrared (NIR) spectral fluorescence imaging using organic fluorophores

    NASA Astrophysics Data System (ADS)

    Kobayashi, Hisataka; Koyama, Yoshinori; Barrett, Tristan; Hama, Yukihiro; Choyke, Peter L.

    2008-02-01

    The ability to obtain multi-color fluorescent imaging in vivo simultaneously using multi-targeted imaging probes could be of potential benefit from both a research and a clinical perspective. However, the simultaneous acquisition of more than 2 separate organic fluorophores usually requires more than one excitation source, since a single excitation source may not optimally excite all the fluorophores. In this study, we employed a multi-excitation approach in order to acquire optimized images with multiple near infrared (NIR) organic fluorophores at the same time. Using 3 sets of excitation filters (595+/-20nm, 640+/-25nm, 688+/-17nm) to acquire 3 distinct spectra and spectral unmixing software (CRi, Woburn, MA), it was possible to resolve the emission spectra of each of the NIR fluorophores using commercial software (Nuance, CRi, Woburn, MA) To demonstrate the utility of this approach 2 mouse models were investigated; In one model, mice bearing four implanted malignancies were injected with a cocktail of 3 fluorescently labeled monoclonal antibodies, each with its own distinct NIR fluorophore. In the second model five different lymph node drainage basins were imaged with 5-color dendrimer-based lymphatic imaging agents tagged with 5 different NIR fluorophores. We successfully detected each of the targeted tumors in the first model and all of the lymph nodes by their distinct color in the second model; neither of which would have been possible using the single excitation method. In conclusion, multi-excitation NIR spectral fluorescence imaging is feasible in a reasonable time frame and opens the possibility for in vivo immunohistochemical imaging (IHCi).

  4. 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.

  5. Excitation-dependent visible fluorescence in decameric nanoparticles with monoacylglycerol cluster chromophores

    PubMed Central

    Lee, Kwang-Ming; Cheng, Wan-Yin; Chen, Cheng-Yu; Shyue, Jing-Jong; Nieh, Chih-Chun; Chou, Chen-Fu; Lee, Jia-Rong; Lee, Ya-Yun; Cheng, Chih-Yang; Chang, Sarah Y.; Yang, Thomas C.; Cheng, Mei-Ching; Lin, Bi-Yun

    2013-01-01

    Organic fluorescent nanoparticles, excitation-dependent photoluminescence, hydrogen-bonded clusters and lysobisphosphatidic acid are four interesting individual topics in materials and biological sciences. They have attracted much attention not only because of their unique properties and important applications, but also because the nature of their intriguing phenomena remained unclear. Here we report a new type of organic fluorescent nanoparticles with intense blue and excitation-dependent visible fluorescence in the range of 410–620 nm. The nanoparticles are composed of ten bis(monoacylglycerol)bisphenol-A molecules and the self-assembly occurs only in elevated concentrations of 2-monoacylglycerol via radical-catalysed 3,2-acyl migration from 3-monoacylglycerol in neat conditions. The excitation-dependent fluorescence behaviour is caused by chromophores composed of hydrogen-bonded monoacylglycerol clusters, which are linked by an extensive hydrogen-bonding network between the ester carbonyl groups and the protons of the alcohols with collective proton motion and HO···C=O (n→π*) interactions. PMID:23443572

  6. Non-typical fluorescence studies of excited and ground state proton and hydrogen transfer

    NASA Astrophysics Data System (ADS)

    Gil, Michał; Kijak, Michał; Piwoński, Hubert; Herbich, Jerzy; Waluk, Jacek

    2017-03-01

    Fluorescence studies of tautomerization have been carried out for various systems that exhibit single and double proton or hydrogen translocation in various environments, such as liquid and solid condensed phases, ultracold supersonic jets, and finally, polymer matrices with single emitters. We focus on less explored areas of application of fluorescence for tautomerization studies, using porphycene, a porphyrin isomer, as an example. Fluorescence anisotropy techniques allow investigations of self-exchange reactions, where the reactant and product are formally identical. Excitation with polarized light makes it possible to monitor tautomerization in single molecules and to detect their three-dimensional orientation. Analysis of fluorescence from single vibronic levels of jet-isolated porphycene not only demonstrates coherent tunneling of two internal protons, but also indicates that the process is vibrational mode-specific. Next, we present bifunctional proton donor-acceptor systems, molecules that are able, depending on the environment, to undergo excited state single intramolecular or double intermolecular proton transfer. For molecules that have donor and acceptor groups located in separate moieties linked by a single bond, excited state tautomerization can be coupled to mutual twisting of the two subunits.

  7. Strongly Enhanced Laser Absorption and Electron Acceleration via Resonant Excitation of Surface Plasma Waves

    NASA Astrophysics Data System (ADS)

    Raynaud, M.; Riconda, C.; Adam, J. C.; Heron, A.

    2010-02-01

    The possibility of creating enhanced fast electron bunches via the excitation of surface plasma waves (SPW) in laser overdense plasma interaction has been investigated by mean of relativistic one dimension motion of a test electron in the field of the surface plasma wave study and with two-dimensional (2D) Particle-In-Cell (PIC) numerical simulations. Strong electron acceleration together with a dramatic increase, up to 70%, of light absorption by the plasma is observed.

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

    NASA Astrophysics Data System (ADS)

    Li, Tanping; Kumar, Revati

    2015-11-01

    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.

  9. 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.

  10. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2013-11-01

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

  12. 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.

  13. 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

  14. Detection of pretreated fingerprint fluorescence using an LED-based excitation system.

    PubMed

    Takatsu, Masahisa; Shimoda, Osamu; Onishi, Kyoko; Onishi, Akira; Oguri, Naoki

    2008-07-01

    Optimization of a light emitting diode (LED)-based excitation system for the detection of pretreated fingerprint fluorescence is described. Fluorescent ridges can usually be excited by irradiation with forensic light sources such as xenon arc lamps or quartz-halogen lamps with high-power output and suitable filters. However, they are too expensive for many crime laboratories in smaller organizations. We concentrated on LEDs which have advantages over conventional light sources in that they are simpler and of lower cost, but the power output and quality of each individual LED unit is not sufficient for the detection of weak fluorescent ridges. To resolve this subject, blue and green LED arrays composed of ninety LED units were adopted and suitable low pass filters for them were designed. An experimental system, consisting of blue and green LED arrays with the suitable low pass filters for illumination, high pass filters for viewing, a digital camera and a computer, was tested. The fluorescent images of cyanoacrylate ester fumed/rhodamine 6G stained fingerprint on white polyethylene sheet and weak fluorescent ridges of ninhydrin/indium chloride treated fingerprint on white paper were successfully detected and photographed. It was shown that the improvement of LED beam in intensity and quality can compensate the disadvantages, resulting in well-contrasted images.

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

    NASA Astrophysics Data System (ADS)

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

    2017-04-01

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

  16. 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

  17. 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

    2015-03-06

    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.

  18. 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.

  19. Switching of the fluorescence emission of single molecules between the locally excited and charge transfer states

    NASA Astrophysics Data System (ADS)

    Angeles Izquierdo, M.; Bell, Toby D. M.; Habuchi, Satoshi; Fron, Eduard; Pilot, Roberto; Vosch, Tom; De Feyter, Steven; Verhoeven, Jan; Jacob, Josemon; Müllen, Klaus; Hofkens, Johan; De Schryver, Frans C.

    2005-01-01

    A novel perylene imide and oligo-pentaphenyl bisfluorene containing molecule is shown to undergo electron transfer to form an emissive charge transfer state in di-benzyl ether and THF. At the single molecule level in a PMMA film, fluorescence spectra characteristic of both emissive states (locally excited and charge transfer) are observed with 44% of the molecules studied showing switching between the two states. These results demonstrate that charge transfer fluorescence from single molecules can be used to report on the properties and dynamics of a molecule's immediate surroundings or nano-environment.

  20. Quantitation of DNA and RNA with Absorption and Fluorescence Spectroscopy.

    PubMed

    Gallagher, Sean R

    2017-02-02

    Quantitation of nucleic acids is a fundamental tool in molecular biology that requires accuracy, reliability, and the use of increasingly smaller sample volumes. This unit describes the traditional absorbance measurement at 260 nm and three more sensitive fluorescence techniques employing Hoechst 33258, ethidium bromide, and PicoGreen. The range of the assays covers 25 pg/ml to 50 µg/ml. Absorbance at 260 nm has an effective range from 1 to 50 µg/ml; Hoechst 33258 from 0.01 to 15 µg/ml; ethidium bromide from 0.1 to 10 µg/ml; and PicoGreen from 25 to 1000 pg/ml. © 2017 by John Wiley & Sons, Inc.

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

    PubMed

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

    2010-03-01

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

  2. Electronic excited states responsible for dimer formation upon UV absorption directly by thymine strands: joint experimental and theoretical study.

    PubMed

    Banyasz, Akos; Douki, Thierry; Improta, Roberto; Gustavsson, Thomas; Onidas, Delphine; Vayá, Ignacio; Perron, Marion; Markovitsi, Dimitra

    2012-09-12

    The study addresses interconnected issues related to two major types of cycloadditions between adjacent thymines in DNA leading to cyclobutane dimers (T<>Ts) and (6-4) adducts. Experimental results are obtained for the single strand (dT)(20) by steady-state and time-resolved optical spectroscopy, as well as by HPLC coupled to mass spectrometry. Calculations are carried out for the dinucleoside monophosphate in water using the TD-M052X method and including the polarizable continuum model; the reliability of TD-M052X is checked against CASPT2 calculations regarding the behavior of two stacked thymines in the gas phase. It is shown that irradiation at the main absorption band leads to cyclobutane dimers (T<>Ts) and (6-4) adducts via different electronic excited states. T<>Ts are formed via (1)ππ* excitons; [2 + 2] dimerization proceeds along a barrierless path, in line with the constant quantum yield (0.05) with the irradiation wavelength, the contribution of the (3)ππ* state to this reaction being less than 10%. The formation of oxetane, the reaction intermediate leading to (6-4) adducts, occurs via charge transfer excited states involving two stacked thymines, whose fingerprint is detected in the fluorescence spectra; it involves an energy barrier explaining the important decrease in the quantum yield of (6-4) adducts with the irradiation wavelength.

  3. Absorption and fluorescence characteristics of photo-activated adenylate cyclase nano-clusters from the amoeboflagellate Naegleria gruberi NEG-M strain

    NASA Astrophysics Data System (ADS)

    Penzkofer, A.; Stierl, M.; Hegemann, P.; Kateriya, S.

    2012-01-01

    The spectroscopic characteristics of BLUF (BLUF = sensor of blue light using flavin) domain containing soluble adenylate cyclase (nPAC = Naegleria photo-activated cyclase) samples from the amoeboflagellate Naegleria gruberi NEG-M strain is studied at room temperature. The absorption and fluorescence spectroscopic development in the dark was investigated over two weeks. Attenuation coefficient spectra, fluorescence quantum distributions, fluorescence quantum yields, and fluorescence excitation distributions were measured. Thawing of frozen nPAC samples gave solutions with varying protein nano-cluster size and varying flavin, tyrosine, tryptophan, and protein color-center emission. Protein color-center emission was observed in the wavelength range of 360-900 nm with narrow emission bands of small Stokes shift and broad emission bands of large Stokes shift. The emission spectra evolved in time with protein nano-cluster aging.

  4. Self-absorption Effects on Alpha-Induced Atmospheric Nitrogen Fluorescence Yield

    SciTech Connect

    Bachelor, Paula P.; Jordan, David V.; Harper, Warren W.; Cannon, Bret D.; Finn, Erin C.

    2009-12-01

    Nitrogen fluorescence induced by alpha, beta and gamma radiation can be used to detect the presence of radioactive contamination in the environment. Successful measurement of fluorescence yield involves a number of factors, including: known fluorescence signal rate during the measurement; the effective alpha spectrum of the radioactive sources used in the measurement; optical attenuation length of the fluorescence signal in air during the measurement; the absolute throughput of the instrumentation; calibration of the instrumentation; and radiation transport modeling of the "effective" array exposure rate given the spectrum of the alpha particles. Field testing of optical instrumentation was conducted to measure the nitrogen fluorescence yield from the alpha radiation generated from americium-241 (241Am) decay. The 241Am test sources were prepared by direct evaporation of ~1 mCi in nitric acid solution, and some solids were visible on the surface of the sources. A laboratory study was conducted with lower activities of 241Am to determine whether the presence of solids on the surface of the sources prepared both by direct evaporation and by electrodeposition onto stainless steel disks produced sufficient self-absorption to cause a decrease in expected fluorescence. Alpha spectroscopy was used to determine the apparent activity of the sources versus the known activity deposited on the surface. Results from the self-absorption laboratory studies were used to correct the activity values in the model and calculate the nitrogen fluorescence generated by the 241Am during the field experiments.

  5. Fluorescence imaging of single molecules and photosynthetic membranes with two-photon excitation

    SciTech Connect

    Sanchez, E.J.; Novotny, L.; Xie, X.S.

    1997-12-31

    We report the imaging of single-molecule fluorescence induced by two-photon excitation in ambient conditions. Using an inverted fluorescence microscope, we obtained the two-photon images of different single fluorophores (Rhodamine B, Sulforhodamine 101, Coumarin 535 on poly-methyl methacrylate films) and biological membrane fragments by Faster scanning the sample with respect to a diffraction limited focus of a mode-locked Ti: sapphire laser beam. The signal to background ratio was as high as 50:1 and the full width at half maximum (250nm) of a single-molecule peak was significantly shorter than that for one photon excitation. With its high sensitivity and simplicity, the two-photon experiment offers a valuable approach for spectroscopic studies on individual immobilized molecules.

  6. Plastic lab-on-a-chip for fluorescence excitation with integrated organic semiconductor lasers.

    PubMed

    Vannahme, Christoph; Klinkhammer, Sönke; Lemmer, Uli; Mappes, Timo

    2011-04-25

    Laser light excitation of fluorescent markers offers highly sensitive and specific analysis for bio-medical or chemical analysis. To profit from these advantages for applications in the field or at the point-of-care, a plastic lab-on-a-chip with integrated organic semiconductor lasers is presented here. First order distributed feedback lasers based on the organic semiconductor tris(8-hydroxyquinoline) aluminum (Alq3) doped with the laser dye 4-dicyanomethylene-2-methyl-6-(p-dimethylaminostyril)-4H-pyrane (DCM), deep ultraviolet induced waveguides, and a nanostructured microfluidic channel are integrated into a poly(methyl methacrylate) (PMMA) substrate. A simple and parallel fabrication process is used comprising thermal imprint, DUV exposure, evaporation of the laser material, and sealing by thermal bonding. The excitation of two fluorescent marker model systems including labeled antibodies with light emitted by integrated lasers is demonstrated.

  7. Optical tuning of the fluorescence spectrum of a π-conjugated polymer through excitation power.

    PubMed

    Deichmann, Vitor Angelo Fonseca; Yakutkin, Vladimir; Baluschev, Stanislav; Akcelrud, Leni

    2011-05-26

    The photophysical properties of a π-conjugated polymer containing 2,2'-bipyridyl alternated with 2,5-dihexyloxyphenylene units (PBPyDHP) are investigated experimentally in terms of the conditions used (solvent, concentration, presence or absence of molecular oxygen, and optical excitation power). The experimental results have suggested that the fluorescence from PBPyDHP can be tuned by proper selection of the experimental conditions showing only one or two emission peaks: 445 nm (blue) and 555 nm (green). The observed effects were interpreted in terms of the twisted intramolecular charge transfer (TICT) theory. This is the first experimental report showing the interconversion of an usual fluorescence, called locally emission state (LE), to a TICT state in second scale time by varying the excitation power; that is, even though the torsion of only one fluorophore occurs in a nano or picosecond scale, the global change (the interconversion for all fluorophores) has occurred in the second time scale.

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

    PubMed Central

    Zhao, Ming; Li, Yu; Peng, Leilei

    2014-01-01

    We present a novel excitation-emission multiplexed fluorescence lifetime microscopy (FLIM) method that surpasses current FLIM techniques in multiplexing capability. The method employs Fourier multiplexing to simultaneously acquire confocal fluorescence lifetime images of multiple excitation wavelength and emission color combinations at 44,000 pixels/sec. The system is built with low-cost CW laser sources and standard PMTs with versatile spectral configuration, which can be implemented as an add-on to commercial confocal microscopes. The Fourier lifetime confocal method allows fast multiplexed FLIM imaging, which makes it possible to monitor multiple biological processes in live cells. The low cost and compatibility with commercial systems could also make multiplexed FLIM more accessible to biological research community. PMID:24921725

  9. Excited-State Dynamics of Melamine and Its Lysine Derivative Investigated by Femtosecond Transient Absorption Spectroscopy.

    PubMed

    Zhang, Yuyuan; Beckstead, Ashley A; Hu, Yuesong; Piao, Xijun; Bong, Dennis; Kohler, Bern

    2016-11-30

    Melamine may have been an important prebiotic information carrier, but its excited-state dynamics, which determine its stability under UV radiation, have never been characterized. The ability of melamine to withstand the strong UV radiation present on the surface of the early Earth is likely to have affected its abundance in the primordial soup. Here, we studied the excited-state dynamics of melamine (a proto-nucleobase) and its lysine derivative (a proto-nucleoside) using the transient absorption technique with a UV pump, and UV and infrared probe pulses. For melamine, the excited-state population decays by internal conversion with a lifetime of 13 ps without coupling significantly to any photochemical channels. The excited-state lifetime of the lysine derivative is slightly longer (18 ps), but the dominant deactivation pathway is otherwise the same as for melamine. In both cases, the vast majority of excited molecules return to the electronic ground state on the aforementioned time scales, but a minor population is trapped in a long-lived triplet state.

  10. Absorption Spectrum of the Green Fluorescent Protein Chromophore Anion In Vacuo

    NASA Astrophysics Data System (ADS)

    Nielsen, S. B.; Lapierre, A.; Andersen, J. U.; Pedersen, U. V.; Tomita, S.; Andersen, L. H.

    2001-11-01

    A sensitive photoabsorption technique for studies of gas-phase biomolecules has been used at the ELISA electrostatic heavy-ion storage ring. We show that the anion form of the chromophore of the green fluorescent protein in vacuo has an absorption maximum at 479 nm, which coincides with one of the two absorption peaks of the protein. Its absorption characteristics are therefore ascribed to intrinsic chemical properties of the chromophore. Evidently, the special β-can structure of the protein provides shielding of the chromophore from the surroundings without significantly changing the electronic structure of the chromophore through interactions with amino acid side chains.

  11. Analysis of Spectral Features of Seawaterbiooptical Components Fluorescence from the Excitation-emission Matrix

    NASA Astrophysics Data System (ADS)

    Salyuk, P. A.; Nagorny, I. G.

    The paper presents the method for processing of excitation-emission matrix of sea water and the allocation of the spectral characteristics of different types of colored dissolved organic matter (CDOM) and phytoplankton cells in seawater. The method consists of identification of regularly observed fluorescence peaks of CDOM in marine waters of different type and definition of the spectral ranges, where the predominant influence of these peaks are observed.

  12. 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.

  13. Influence of different environments on the excited-state proton transfer and dual fluorescence of fisetin

    NASA Astrophysics Data System (ADS)

    Guharay, Jayanti; Dennison, S. Moses; Sengupta, Pradeep K.

    1999-05-01

    The influence of different protic and aprotic solvent environments on the excited-state intramolecular proton transfer (ESIPT) leading to a dual fluorescence behaviour of a biologically important, naturally occurring, polyhydroxyflavone, fisetin (3,3',4',7-tetrahydroxyflavone), has been investigated. The normal fluorescence band, in particular, is extremely sensitive to solvent polarity with νmax shifting from 24 510 cm -1 in dioxane ( ET(30)=36.0) to 20 790 cm -1 in methanol ( ET(30)=55.5). This is rationalized in terms of solvent dipolar relaxation process, which also accounts for the red edge excitation shifts (REES) observed in viscous environments such as glycerol at low temperatures. Significant solvent dependence of the tautomer fluorescence properties ( νmax, yield and decay kinetics) reveals the influence of external hydrogen bonding perturbation on the internal hydrogen bond of the molecule. These excited-state relaxation phenomena and their relevant parameters have been used to probe the microenvironment of fisetin in a membrane mimetic system, namely AOT reverse micelles in n-heptane at different water/surfactant molar ratio ( w0).

  14. Fluorescent molecular probes based on excited state prototropism in lipid bilayer membrane

    NASA Astrophysics Data System (ADS)

    Mohapatra, Monalisa; Mishra, Ashok K.

    2012-03-01

    Excited state prototropism (ESPT) is observed in molecules having one or more ionizable protons, whose proton transfer efficiency is different in ground and excited states. The interaction of various ESPT molecules like naphthols and intramolecular ESPT (ESIPT) molecules like hydroxyflavones etc. with different microheterogeneous media have been studied in detail and excited state prototropism as a probe concept has been gaining ground. The fluorescence of different prototropic forms of such molecules, on partitioning to an organized medium like lipid bilayer membrane, often show sensitive response to the local environment with respect to the local structure, physical properties and dynamics. Our recent work using 1-naphthol as an ESPT fluorescent molecular probe has shown that the incorporation of monomeric bile salt molecules into lipid bilayer membranes composed from dipalmitoylphosphatidylcholine (DPPC, a lung surfactant) and dimyristoylphosphatidylcholine (DMPC), in solid gel and liquid crystalline phases, induce appreciable wetting of the bilayer up to the hydrocarbon core region, even at very low (<= 1 mM) concentrations of the bile salts. The incorporation and location of fisetin, an ESIPT molecule having antioxidant properties, in lipid bilayer membrane has been sensitively monitored from its intrinsic fluorescence behaviour.

  15. A visible-light-excited fluorescence method for imaging protein crystals without added dyes.

    PubMed

    Lukk, Tiit; Gillilan, Richard E; Szebenyi, Doletha M E; Zipfel, Warren R

    2016-02-01

    Fluorescence microscopy methods have seen an increase in popularity in recent years for detecting protein crystals in screening trays. The fluorescence-based crystal detection methods have thus far relied on intrinsic UV-inducible tryptophan fluorescence, nonlinear optics or fluorescence in the visible light range dependent on crystals soaked with fluorescent dyes. In this paper data are presented on a novel visible-light-inducible autofluorescence arising from protein crystals as a result of general stabilization of conjugated double-bond systems and increased charge delocalization due to crystal packing. The visible-light-inducible autofluorescence serves as a complementary method to bright-field microscopy in beamline applications where accurate crystal centering about the rotation axis is essential. Owing to temperature-dependent chromophore stabilization, protein crystals exhibit tenfold higher fluorescence intensity at cryogenic temperatures, making the method ideal for experiments where crystals are cooled to 100 K with a cryostream. In addition to the non-damaging excitation wavelength and low laser power required for imaging, the method can also serve a useful role for differentiating protein crystals from salt crystals in screening trays.

  16. A visible-light-excited fluorescence method for imaging protein crystals without added dyes

    PubMed Central

    Lukk, Tiit; Gillilan, Richard E.; Szebenyi, Doletha M. E.; Zipfel, Warren R.

    2016-01-01

    Fluorescence microscopy methods have seen an increase in popularity in recent years for detecting protein crystals in screening trays. The fluorescence-based crystal detection methods have thus far relied on intrinsic UV-inducible tryptophan fluorescence, nonlinear optics or fluorescence in the visible light range dependent on crystals soaked with fluorescent dyes. In this paper data are presented on a novel visible-light-inducible autofluorescence arising from protein crystals as a result of general stabilization of conjugated double-bond systems and increased charge delocalization due to crystal packing. The visible-light-inducible autofluorescence serves as a complementary method to bright-field microscopy in beamline applications where accurate crystal centering about the rotation axis is essential. Owing to temperature-dependent chromophore stabilization, protein crystals exhibit tenfold higher fluorescence intensity at cryogenic temperatures, making the method ideal for experiments where crystals are cooled to 100 K with a cryostream. In addition to the non-damaging excitation wavelength and low laser power required for imaging, the method can also serve a useful role for differentiating protein crystals from salt crystals in screening trays. PMID:26937240

  17. [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.

  18. 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.

  19. How to Collect National Institute of Standards and Technology (NIST) Traceable Fluorescence Excitation and Emission Spectra.

    PubMed

    Gilmore, Adam Matthew

    2014-01-01

    Contemporary spectrofluorimeters comprise exciting light sources, excitation and emission monochromators, and detectors that without correction yield data not conforming to an ideal spectral response. The correction of the spectral properties of the exciting and emission light paths first requires calibration of the wavelength and spectral accuracy. The exciting beam path can be corrected up to the sample position using a spectrally corrected reference detection system. The corrected reference response accounts for both the spectral intensity and drift of the exciting light source relative to emission and/or transmission detector responses. The emission detection path must also be corrected for the combined spectral bias of the sample compartment optics, emission monochromator, and detector. There are several crucial issues associated with both excitation and emission correction including the requirement to account for spectral band-pass and resolution, optical band-pass or neutral density filters, and the position and direction of polarizing elements in the light paths. In addition, secondary correction factors are described including (1) subtraction of the solvent's fluorescence background, (2) removal of Rayleigh and Raman scattering lines, as well as (3) correcting for sample concentration-dependent inner-filter effects. The importance of the National Institute of Standards and Technology (NIST) traceable calibration and correction protocols is explained in light of valid intra- and interlaboratory studies and effective spectral qualitative and quantitative analyses including multivariate spectral modeling.

  20. Photoswitchable Fluorescent Proteins: Ten Years of Colorful Chemistry and Exciting Applications

    PubMed Central

    Zhou, Xin X.; Lin, Michael Z.

    2013-01-01

    Reversibly photoswitchable fluorescent proteins (RSFPs) are fluorescent proteins whose fluorescence, upon excitation at a certain wavelength, can be switched on or off by light in a reversible manner. In the last ten years, many new RSFPs have been developed and novel applications in cell imaging discovered that rely on their photoswitching properties. This review will describe research on the mechanisms of reversible photoswitching and recent applications using RSFPs. While cis-trans isomerization of the chromophore is believed to be the general mechanism for most RSFPs, structural studies reveal diversity in the details of photoswitching mechanisms, including different effects of protonation, chromophore planarity, and pocket flexibility. Applications of RSFPs include new types of live-cell superresolution imaging, tracking of protein movements and interactions, information storage, and optical control of protein activity. PMID:23876529

  1. Second- and third-harmonic generation and multiphoton excitation fluorescence microscopy for simultaneous imaging of cardiomyocytes

    NASA Astrophysics Data System (ADS)

    Barzda, Virginijus; Greenhalgh, Catherine; Aus der Au, Juerg; Squier, Jeffrey A.; Elmore, Steven; van Beek, Johannes H.

    2004-06-01

    Simultaneous detection of second harmonic generation (SHG), third harmonic generation (THG) and multiphoton excitation fluorescence with ultrafast laser pulses from a Nd:Glass laser was used to image isolated adult rat cardiomyocytes. The simultaneous detection enabled visualization of different organelles of cardiomyocytes, based on the different contrast mechanisms. It was found that SHG signal depicted characteristic patterns of sarcomeres in a myofilament lattice. The regular pattern of the THG signal, which was anticorrelated with the SHG signal, suggested that the third harmonic is generated within mitochondria. By labeling the cardiomyocytes with the mitochondrial dye tetramethylrhodamine methyl ester (TMRM), comparisons could be made between the TMRM fluorescence, THG, and SHG images. The TMRM fluorescence had significant correlation with THG signal confirming that part of the THG signal originates from mitochondria.

  2. 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.

  3. Population inversion in fluorescing fragments of super-excited molecules inside an air filament

    NASA Astrophysics Data System (ADS)

    See-Leang, Chin; Huai-Liang, Xu

    2015-01-01

    An original idea is reviewed. When a molecule is pumped into a super-excited state, one of its decay channels is neutral dissociation. One or more of the neutral fragments will fluoresce. Hence, if a lower state of such fluorescing fragments was populated through other channels but with a lower probability, population inversion of the fluorescing fragments would be naturally realized. This idea seems to be validated, so far, by comparing published work on three hydrocarbon molecules, CH4, C2H2, C2H4, and water vapor, H2O. After super-excitation in a femtosecond laser filament in air mixed with these molecules, the fluorescence from the CH or OH fragments exhibits population inversion, i.e., amplified spontaneous emission was observed in the backscattering direction of the filament. Project supported by the Canada Research Chairs, the Natural Science and Engineering Research Council of Canada (NSERC), the FRQNT, the Canada Foundation for Innovation (CFI), the National Basic Research Program of China (Grant No. 2014CB921300), the National Natural Science Foundation of China (Grant No. 61235003), the Research Fund for the Doctoral Program of Higher Education of China, and the Scientific Research Foundation for Returned Scholars, Ministry of Education of China.

  4. Pressure broadening of atomic oxygen two-photon absorption laser induced fluorescence

    NASA Astrophysics Data System (ADS)

    Marinov, Daniil; Drag, Cyril; Blondel, Christophe; Guaitella, Olivier; Golda, Judith; Klarenaar, Bart; Engeln, Richard; Schulz-von der Gathen, Volker; Booth, Jean-Paul

    2016-12-01

    Atomic oxygen, considered to be a determining reactant in plasma applications at ambient pressure, is routinely detected by two-photon absorption laser induced fluorescence (TALIF). Here, pressure broadening of the (2p 4 3 P 2  →  3p 3 P J=0,1,2) two-photon transition in oxygen atoms was investigated using a high-resolution TALIF technique in normal and Doppler-free configurations. The pressure broadening coefficients determined were {γ{{\\text{O}2}}}   =  0.40  ±  0.08  cm-1/bar for oxygen molecules and {γ\\text{He}}   =  0.46  ±  0.03 cm-1/bar for helium atoms. These correspond to pressure broadening rate constants k\\text{PB}{{\\text{O}2}}   =  9 · 10-9 cm3 s-1 and k\\text{PB}\\text{He}   =  4 · 10-9 cm3 s-1, respectively. The well-known quenching rate constants of O(3p 3 P J ) by O2 and He are at least one order of magnitude smaller, which signifies that non-quenching collisions constitute the main line-broadening mechanism. In addition to providing new insights into collisional processes of oxygen atoms in electronically excited 3p 3 P J state, reported pressure broadening parameters are important for quantification of oxygen TALIF line profiles when both collisional and Doppler broadening mechanisms are important. Thus, the Doppler component (and hence the temperature of oxygen atoms) can be accurately determined from high resolution TALIF measurements in a broad range of conditions.

  5. Two-photon fluorescence excitation spectroscopy by pulse shaping ultrabroad-bandwidth femtosecond laser pulses

    SciTech Connect

    Xu Bingwei; Coello, Yves; Lozovoy, Vadim V.; Dantus, Marcos

    2010-11-10

    A fast and automated approach to measuring two-photon fluorescence excitation (TPE) spectra of fluorophores with high resolution ({approx}2 nm ) by pulse shaping ultrabroad-bandwidth femtosecond laser pulses is demonstrated. Selective excitation in the range of 675-990 nm was achieved by imposing a series of specially designed phase and amplitude masks on the excitation pulses using a pulse shaper. The method eliminates the need for laser tuning and is, thus, suitable for non-laser-expert use. The TPE spectrum of Fluorescein was compared with independent measurements and the spectra of the pH-sensitive dye 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) in acidic and basic environments were measured for the first time using this approach.

  6. 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.

  7. [Rapid identification of potato cultivars using NIR-excited fluorescence and Raman spectroscopy].

    PubMed

    Dai, Fen; Bergholt, Mads Sylvest; Benjamin, Arnold Julian Vinoj; Hong, Tian-Sheng; Zhiwei, Huang

    2014-03-01

    Potato is one of the most important food in the world. Rapid and noninvasive identification of potato cultivars plays a important role in the better use of varieties. In this study, The identification ability of optical spectroscopy techniques, including near-infrared (NIR) Raman spectroscopy and NIR fluorescence spectroscopy, for invasive detection of potato cultivars was evaluated. A rapid NIR Raman spectroscopy system was applied to measure the composite Raman and NIR fluorescence spectroscopy of 3 different species of potatoes (98 samples in total) under 785 nm laser light excitation. Then pure Raman and NIR fluorescence spectroscopy were abstracted from the composite spectroscopy, respectively. At last, the partial least squares-discriminant analysis (PLS-DA) was utilized to analyze and classify Raman spectra of 3 different types of potatoes. All the samples were divided into two sets at random: the calibration set (74samples) and prediction set (24 samples), the model was validated using a leave-one-out, cross-validation method. The results showed that both the NIR-excited fluorescence spectra and pure Raman spectra could be used to identify three cultivars of potatoes. The fluorescence spectrum could distinguish the Favorita variety well (sensitivity: 1, specificity: 0.86 and accuracy: 0.92), but the result for Diamant (sensitivity: 0.75, specificity: 0.75 and accuracy: 0. 75) and Granola (sensitivity: 0.16, specificity: 0.89 and accuracy: 0.71) cultivars identification were a bit poorer. We demonstrated that Raman spectroscopy uncovered the main biochemical compositions contained in potato species, and provided a better classification sensitivity, specificity and accuracy (sensitivity: 1, specificity: 1 and accuracy: 1 for all 3 potato cultivars identification) among the three types of potatoes as compared to fluorescence spectroscopy.

  8. Large-scale correlated study of excited state absorptions in naphthalene and anthracene.

    PubMed

    Sony, Priya; Shukla, Alok

    2009-07-07

    In this paper, we report theoretical calculations of the photoinduced absorption (PA) spectrum of naphthalene and anthracene, with the aim of understanding those excited states, which are invisible in the linear optical absorption. The excited state absorption spectra are computed from the 1B(2u)(+) and the 1B(3u)(+) states and a detailed analysis of the many-body character of the states contributing to various peaks in the spectra is presented. The calculations are performed using the Pariser-Parr-Pople (PPP) Hamiltonian, along with the full configuration interaction technique. The role of Coulomb parameters used in the PPP Hamiltonian is examined by considering standard Ohno parameters, as well as a screened set of parameters. The results of our calculations are extensively compared with the experimental data where available and very good agreement has been obtained. Moreover, our calculations predict the presence of high intensity features which, to the best of our knowledge, have not been explored earlier. We also present concrete predictions on the polarization properties of the PA spectrum, which can be verified in experiments performed on oriented samples.

  9. Excitation spectroscopy in multispectral optical fluorescence tomography: methodology, feasibility and computer simulation studies

    NASA Astrophysics Data System (ADS)

    Chaudhari, Abhijit J.; Ahn, Sangtae; Levenson, Richard; Badawi, Ramsey D.; Cherry, Simon R.; Leahy, Richard M.

    2009-08-01

    Molecular probes used for in vivo optical fluorescence tomography (OFT) studies in small animals are typically chosen such that their emission spectra lie in the 680-850 nm wavelength range. This is because tissue attenuation in this spectral band is relatively low, allowing optical photons even from deep sites in tissue to reach the animal surface and consequently be detected by a CCD camera. The wavelength dependence of tissue optical properties within the 680-850 nm band can be exploited for emitted light by measuring fluorescent data via multispectral approaches and incorporating the spectral dependence of these optical properties into the OFT inverse problem—that of reconstructing underlying 3D fluorescent probe distributions from optical data collected on the animal surface. However, in the aforementioned spectral band, due to only small variations in the tissue optical properties, multispectral emission data, though superior for image reconstruction compared to achromatic data, tend to be somewhat redundant. A different spectral approach for OFT is to capitalize on the larger variations in the optical properties of tissue for excitation photons than for the emission photons by using excitation at multiple wavelengths as a means of decoding source depth in tissue. The full potential of spectral approaches in OFT can be realized by a synergistic combination of these two approaches, that is, exciting the underlying fluorescent probe at multiple wavelengths and measuring emission data multispectrally. In this paper, we describe a method that incorporates both excitation and emission spectral information into the OFT inverse problem. We describe a linear algebraic formulation of the multiple wavelength illumination-multispectral detection forward model for OFT and compare it to models that use only excitation at multiple wavelengths or those that use only multispectral detection techniques. This study is carried out in a realistic inhomogeneous mouse atlas

  10. 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).

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

    PubMed

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

    2015-01-01

    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).

  12. A new near-infrared absorption and fluorescent probe based on bombesin for molecular imaging

    NASA Astrophysics Data System (ADS)

    Kujala, Naresh; Zhai, Huifang; Smith, Charles; Prasanphanich, Adam; Sieckman, Gary; Hoffman, Timothy; Volkert, Wynn; Ma, Lixin; Yu, Ping

    2009-02-01

    We have developed a series of new dye bombesin conjugates for site-specific absorption and fluorescence imaging of human prostate and breast cancers. Bombesin (BBN), an amphibian analog to the endogenous ligand, binds to the gastrin releasing peptide (GRP) receptors with high specificity and affinity. Previously, we developed an Alexa Fluor 680-GGG-BBN peptide conjugate which demonstrated high binding affinity and specificity for breast cancer cells in the in vitro and in vivo tests (Ref: Ma et al., Molecular Imaging, vol. 6, no. 3, 2007: 171-180). This probe can not be used as an absorption probe in near-infrared imaging because its absorption peak is in the visible wavelength range. In addition, site specific longer wavelength fluorescent probe is desired for in vivo molecular imaging because long wavelength photons penetrate deeper into tissue. The new absorption and fluorescent probe we developed is based on the last eight-residues of BBN, -Q-W-A-V-G-H-L-M-(NH2), and labeled with AlexaFluor750 through a chemical linker, beta-alanine. The new probe, Alexa Fluor 750-BetaAla-BBN(7-14)NH2, exhibits optimal pharmacokinetics for specific targeting and optical imaging of the GRP receptor over-expressing cancer cells. Absorption spectrum has been measured and showed absorption peaks at 690nm, 720nm and 735nm. Fluorescent band is located at 755nm. In vitro and in vivo investigations have demonstrated the effectiveness of the new conjugates to specifically target human cancer cells overexpressing GRP receptors and tumor xenografts in severely compromised immunodeficient mouse model.

  13. Excited state absorption and energy transfer in Ho3+-doped indium fluoride glass

    NASA Astrophysics Data System (ADS)

    Gomes, Laercio; Fortin, Vincent; Bernier, Martin; Maes, Frédéric; Vallée, Réal; Poulain, Samuel; Poulain, Marcel; Jackson, Stuart D.

    2017-04-01

    This investigation examines in detail the rates of energy transfer relevant to the 5I5 → 5I6 transition (at 3930 nm) in Ho3+-doped InF3 glass as a function of the Ho3+ concentration. The decay times, branching ratios and rate parameters for energy transfer were measured in this investigation for Ho3+ (x)-doped InF3 glass with x = 2, 4 and 10 mol.% and they were used as the input parameters for a rate equation analysis. Excited state absorption (ESA) initiating from the lower laser level is included in the study. Numerical simulation of CW laser emission at 3.9 μm was performed using two pump wavelengths, one for upper laser level excitation (i.e., 5I8 → 5I5 = λP1) and the other for lower laser level de-excitation (i.e., 5I6 → 5S2 = λP2). The pump wavelength λP2 = 962 nm was chosen based on the measurements of ESA and the application of the McCumber method. Critically, the estimated ESA cross section at λP2 = 962 nm (σESA = 7.1 × 10-21 cm2) is approximately sixteen times larger than ground state (5I8) absorption cross section (σGSA = 4.3 × 10-22 cm2) and ESA does not overlap with any ground state absorption process. Our calculations suggest that even for high Ho3+ concentration in which cross relaxation has been shown in a previous study to quench the 5I5 level, ESA is nevertheless strong enough to allow a sufficient population inversion required for practical CW emission.

  14. Excitation relaxation dynamics and energy transfer in pigment-protein complexes of a dinoflagellate, revealed by ultrafast fluorescence spectroscopy.

    PubMed

    Tanaka, Kazunori; Iida, Satoko; Takaichi, Shinichi; Mimuro, Mamoru; Murakami, Akio; Akimoto, Seiji

    2016-12-01

    Photosynthetic light-harvesting complexes, found in aquatic photosynthetic organisms, contain a variety of carotenoids and chlorophylls. Most of the photosynthetic dinoflagellates possess two types of light-harvesting antenna complexes: peridinin (Peri)-chlorophyll (Chl) a/c-protein, as an intrinsic thylakoid membrane complex protein (iPCP), and water-soluble Peri-Chl a-protein, as an extrinsic membrane protein (sPCP) on the inner surface of the thylakoid. Peri is a unique carotenoid that has eight C=C bonds and one C=O bond, which results in a characteristic absorption band in the green wavelength region. In the present study, excitation relaxation dynamics of Peri in solution and excitation energy transfer processes of sPCP and the thylakoid membranes, prepared from the photosynthetic dinoflagellate, Symbiodinium sp., are investigated by ultrafast time-resolved fluorescence spectroscopy. We found that Peri-to-Chl a energy transfer occurs via the Peri S1 state with a time constant of 1.5 ps or 400 fs in sPCP or iPCP, respectively, and that Chl c-to-Chl a energy transfer occurs in the time regions of 350-400 fs and 1.8-2.6 ps.

  15. Local conformations and excited state dynamics of porphyrins and nucleic acids by 2-dimensional fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Widom, Julia R.

    Biological systems present many challenges to researchers attempting to study them using spectroscopy. Low specificity, low sensitivity, and broad and overlapping lineshapes limit the amount of information that can be obtained in experiments. Two-dimensional fluorescence spectroscopy (2D FS) is a highly sensitive and information-rich spectroscopic technique that was developed to study the conformations and excited state dynamics of systems exhibiting exciton coupling. In this dissertation, I describe a variety of extensions of 2D FS that further increase its utility for the study of biological systems. I describe experiments on a dimer of zinc tetraphenylporphyrin embedded in a membrane, in which the signals from two conformational subpopulations were separated in order to study the thermodynamics of their interconversion. I present proof-of-principle experiments on nucleic acids that utilize fluorescence resonance energy transfer to separate signals from different subpopulations. I also describe experiments in which 2D FS was performed using ultraviolet excitation to determine the conformation of a dinucleotide of a fluorescent analogue of the nucleic acid base adenine. I discuss experiments on porphyrin dimers in which 2D FS was used as a probe of excited state dynamics. Finally, I present model calculations for a proposed variation of 2D FS in which entangled photons would be used as the excitation source. These calculations suggest that this approach has the potential to yield significantly narrower spectral lineshapes than conventional 2D FS. These experiments and calculations yield new insight into the systems investigated and establish a `toolbox' of variations of 2D FS that can be used to gain as much information as possible from experiments on challenging systems such as protein-DNA complexes.

  16. 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.

  17. 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...

  18. 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/.

  19. Increasing efficiency of two-photon excited fluorescence and second harmonic generation using ultrashort pulses

    NASA Astrophysics Data System (ADS)

    Tang, Shuo; Krasieva, Tatiana B.; Chen, Zhongping; Tempea, Gabriel; Tromberg, Bruce J.

    2006-02-01

    Multiphoton microscopy (MPM) has become an important tool for high-resolution and non-invasive imaging in biological tissues. However, the efficiencies of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) are relatively low because of their nonlinear nature. Therefore, it is critical to optimize laser parameters for most efficient excitation of MPM. Reducing the pulse duration can increase the peak intensity of excitation and thus potentially increase the excitation efficiency. In this paper, a multiphoton microscopy system using a 12 fs Ti:Sapphire laser is reported. With adjustable dispersion pre-compensation, the pulse duration at the sample location can be varied from 400 fs to sub-20 fs. The efficiencies of TPEF and SHG are studied for the various pulse durations, respectively. Both TPEF and SHG are found to increase proportionally to the inverse of the pulse duration for the entire tested range. To transmit most of the SHG and TPEF signals, the spectral transmission widow of the detection optics needs to be carefully considered. Limitation from phase-matching in SHG generation is not significant because the effective interaction length for SHG is less than 10 μm at the focal depth of the objectives. These results are important in improving MPM excitation efficiency using ultrashort pulses. MPM images from human artery wall are also demonstrated.

  20. Excited states and absorption spectra of β-diketonate complexes of boron difluoride with aromatic substituents

    NASA Astrophysics Data System (ADS)

    Vovna, V. I.; Kazachek, M. V.; L'vov, I. B.

    2012-04-01

    In the approximation of the time-dependent electron density functional theory, we have studied using the quantum-chemical method the nature of excited states of boron difluoride acetylacetonate F2BAA and its substituted derivatives that contain aromatic groups with one or two benzene cycles in the β-position. Optimization of the geometry of complexes show coplanar positions of cycles for all compounds, except for that with the substituent C6H3(CH3)2. Based on the calculated transition energies and oscillator strengths, we have simulated the absorption spectra in the prevacuum range. The calculated absorption spectra have been compared with the experimental spectra in the gas phase or in solutions. We show that, in the absorption spectra of complexes that contain substituents with one benzene cycle, the first three bands are caused by the transition of π electrons of the substituent to the LUMO of the chelate cycle. In complexes with two cycles in the substituent, the number of these transitions increases to five. As the π system becomes more extended, a bathochromic shift of the first absorption band and an increase in the transition probability are observed.

  1. 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.

  2. Quantitative fluorescence spectroscopy in turbid media: a practical solution to the problem of scattering and absorption.

    PubMed

    Chen, Yao; Chen, Zeng-Ping; Yang, Jing; Jin, Jing-Wen; Zhang, Juan; Yu, Ru-Qin

    2013-02-19

    The presence of practically unavoidable scatterers and background absorbers in turbid media such as biological tissue or cell suspensions can significantly distort the shape and intensity of fluorescence spectra of fluorophores and, hence, greatly hinder the in situ quantitative determination of fluorophores in turbid media. In this contribution, a quantitative fluorescence model (QFM) was proposed to explicitly model the effects of the scattering and absorption on fluorescence measurements. On the basis of the proposed model, a calibration strategy was developed to remove the detrimental effects of scattering and absorption and, hence, realize accurate quantitative analysis of fluorophores in turbid media. A proof-of-concept model system, the determination of free Ca(2+) in turbid media using Fura-2, was utilized to evaluate the performance of the proposed method. Experimental results showed that QFM can provide quite precise concentration predictions for free Ca(2+) in turbid media with an average relative error of about 7%, probably the best results ever achieved for turbid media without the use of advanced optical technologies. QFM has not only good performance but also simplicity of implementation. It does not require characterization of the light scattering properties of turbid media, provided that the light scattering and absorption properties of the test samples are reasonably close to those of the calibration samples. QFM can be developed and extended in many application areas such as ratiometric fluorescent sensors for quantitative live cell imaging.

  3. In situ phytoplankton absorption, fluorescence emission, and particulate backscattering spectra determined from reflectance

    NASA Technical Reports Server (NTRS)

    Roesler, Collin S.; Pery, Mary Jane

    1995-01-01

    An inverse model was developed to extract the absortion and scattering (elastic and inelastic) properties of oceanic constituents from surface spectral reflectance measurements. In particular, phytoplankton spectral absorption coefficients, solar-stimulated chlorophyll a fluorescence spectra, and particle backscattering spectra were modeled. The model was tested on 35 reflectance spectra obtained from irradiance measurements in optically diverse ocean waters (0.07 to 25.35 mg/cu m range in surface chlorophyll a concentrations). The universality of the model was demonstrated by the accurate estimation of the spectral phytoplankton absorption coefficents over a range of 3 orders of magnitude (rho = 0.94 at 500 nm). Under most oceanic conditions (chlorophyll a less than 3 mg/cu m) the percent difference between measured and modeled phytoplankton absorption coefficents was less than 35%. Spectral variations in measured phytoplankton absorption spectra were well predicted by the inverse model. Modeled volume fluorescence was weakly correlated with measured chl a; fluorescence quantum yield varied from 0.008 to 0.09 as a function of environment and incident irradiance. Modeled particle backscattering coefficients were linearly related to total particle cross section over a twentyfold range in backscattering coefficents (rho = 0.996, n = 12).

  4. 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

  5. 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

  6. 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

  7. Study of cadmium-induced cytotoxicity using two-photon excitation endogenous fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Li, Dong; Yang, Mildred S.; Lin, Tao; Zheng, Wei; Qu, Jianan Y.

    2009-09-01

    We demonstrate that using time-resolved two-photon excitation endogenous fluorescence microscopy, the cadmium (Cd)-induced cellular toxic level can be assessed by the free-to protein-bound reduced nicotinamide adenine dinucleotide (free/bound NADH) ratio in a living cell. NADH fluorescence excited at 730 nm is captured at different times following exposure to cadmium at a variety of concentrations. The temporal characteristics of NADH fluorescence from mitochondrial and nuclear compartments are analyzed, respectively. The results show that cadmium induces a significant increase of the free/bound NADH ratio in mitochondria and nucleus, caused by the inhibition effect on the electron transport chain (ETC) and the stimulating effect on the glycolysis pathway, respectively. It is found that induction of metallothionein (MT) in cells occurs after 4 h of exposure to a sublethal concentration of Cd and reaches a peak at 6 h. More importantly, the increase in MT level can effectively suppress the elevation of the free/bound NADH ratio caused by a subsequent exposure to a higher concentration of Cd, indicating that MT plays a key role in protecting cells from Cd-induced toxicity. Our findings show that the free/bound NADH ratio can potentially be used as a sensitive indicator of toxic and carcinogenic actions induced by Cd.

  8. Adapting a compact confocal microscope system to a two-photon excitation fluorescence imaging architecture.

    PubMed

    Diaspro, A; Corosu, M; Ramoino, P; Robello, M

    1999-11-01

    Within the framework of a national National Institute of Physics of Matter (INFM) project, we have realised a two-photon excitation (TPE) fluorescence microscope based on a new generation commercial confocal scanning head. The core of the architecture is a mode-locked Ti:Sapphire laser (Tsunami 3960, Spectra Physics Inc., Mountain View, CA) pumped by a high-power (5 W, 532 nm) laser (Millennia V, Spectra Physics Inc.) and an ultracompact confocal scanning head, Nikon PCM2000 (Nikon Instruments, Florence, Italy) using a single-pinhole design. Three-dimensional point-spread function has been measured to define spatial resolution performances. The TPE microscope has been used with a wide range of excitable fluorescent molecules (DAPI, Fura-2, Indo-1, DiOC(6)(3), fluoresceine, Texas red) covering a single photon spectral range from UV to green. An example is reported on 3D imaging of the helical structure of the sperm head of the Octopus Eledone cirrhosa labelled with an UV excitable dye, i.e., DAPI. The system can be easily switched for operating both in conventional and two-photon mode.

  9. Electronically excited states of membrane fluorescent probe 4-dimethylaminochalcone. Results of quantum chemical calculations.

    PubMed

    Romanov, Alexey N; Gularyan, Samvel K; Polyak, Boris M; Sakovich, Ruslan A; Dobretsov, Gennady E; Sarkisov, Oleg M

    2011-05-28

    Quantum-chemical calculations of ground and excited states for membrane fluorescent probe 4-dimethylaminochalcone (DMAC) in vacuum were performed. Optimized geometries and dipole moments for lowest-lying singlet and triplet states were obtained. The nature of these electronic transitions and the relaxation path in the excited states were determined; changes in geometry and charge distribution were assessed. It was shown that in vacuum the lowest existed level is of (n, π*) nature, and the closest to it is the level of (π, π*) nature; the energy gap between them is narrow. This led to an effective (1)(π, π*) →(1)(n, π*) relaxation. After photoexcitation the molecule undergoes significant transformations, including changes in bond orders, pyramidalization angle of the dimethylamino group, and planarity of the molecule. Its dipole moment rises from 5.5 Debye in the ground state to 17.1 Debye in the (1)(π, π*) state, and then falls to 2 Debye in the (1)(n, π*) state. The excited (1)(n, π*) state is a short living state; it has a high probability of intersystem crossing into the (3)(π, π*) triplet state. This relaxation path explains the low quantum yield of DMAC fluorescence in non-polar media. It is possible that (3)(π, π*) is responsible for observed DMAC phosphorescence.

  10. 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.

  11. 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.

  12. Shock-tube measurements of excited oxygen atoms using cavity-enhanced absorption spectroscopy.

    PubMed

    Nations, Marcel; Wang, Shengkai; Goldenstein, Christopher S; Sun, Kai; Davidson, David F; Jeffries, Jay B; Hanson, Ronald K

    2015-10-10

    We report the use of cavity-enhanced absorption spectroscopy (CEAS) using two distributed feedback diode lasers near 777.2 and 844.6 nm for sensitive, time-resolved, in situ measurements of excited-state populations of atomic oxygen in a shock tube. Here, a 1% O2/Ar mixture was shock-heated to 5400-8000 K behind reflected shock waves. The combined use of a low-finesse cavity, fast wavelength scanning of the lasers, and an off-axis alignment enabled measurements with 10 μs time response and low cavity noise. The CEAS absorption gain factors of 104 and 142 for the P35←S520 (777.2 nm) and P0,1,23←S310 (844.6 nm) atomic oxygen transitions, respectively, significantly improved the detection sensitivity over conventional single-pass measurements. This work demonstrates the potential of using CEAS to improve shock-tube studies of nonequilibrium electronic-excitation processes at high temperatures.

  13. Dual-wavelength excitation to reduce background fluorescence for fluorescence spectroscopic quantitation of erythrocyte zinc protoporphyrin-IX and protoporphyrin-IX from whole blood and oral mucosa

    NASA Astrophysics Data System (ADS)

    Hennig, Georg; Vogeser, Michael; Holdt, Lesca M.; Homann, Christian; Großmann, Michael; Stepp, Herbert; Gruber, Christian; Erdogan, Ilknur; Hasmüller, Stephan; Hasbargen, Uwe; Brittenham, Gary M.

    2014-02-01

    Erythrocyte zinc protoporphyrin-IX (ZnPP) and protoporphyrin-IX (PPIX) accumulate in a variety of disorders that restrict or disrupt the biosynthesis of heme, including iron deficiency and various porphyrias. We describe a reagent-free spectroscopic method based on dual-wavelength excitation that can measure simultaneously both ZnPP and PPIX fluorescence from unwashed whole blood while virtually eliminating background fluorescence. We further aim to quantify ZnPP and PPIX non-invasively from the intact oral mucosa using dual-wavelength excitation to reduce the strong tissue background fluorescence while retaining the faint porphyrin fluorescence signal originating from erythrocytes. Fluorescence spectroscopic measurements were made on 35 diluted EDTA blood samples using a custom front-face fluorometer. The difference spectrum between fluorescence at 425 nm and 407 nm excitation effectively eliminated background autofluorescence while retaining the characteristic porphyrin peaks. These peaks were evaluated quantitatively and the results compared to a reference HPLC-kit method. A modified instrument using a single 1000 μm fiber for light delivery and detection was used to record fluorescence spectra from oral mucosa. For blood measurements, the ZnPP and PPIX fluorescence intensities from the difference spectra correlated well with the reference method (ZnPP: Spearman's rho rs = 0.943, p < 0.0001; PPIX: rs = 0.959, p < 0.0001). In difference spectra from oral mucosa, background fluorescence was reduced significantly, while porphyrin signals remained observable. The dual-wavelength excitation method evaluates quantitatively the ZnPP/heme and PPIX/heme ratios from unwashed whole blood, simplifying clinical laboratory measurements. The difference technique reduces the background fluorescence from measurements on oral mucosa, allowing for future non-invasive quantitation of erythrocyte ZnPP and PPIX.

  14. Two-photon two-focus fluorescence correlation spectroscopy with a tunable distance between the excitation volumes.

    PubMed

    Didier, Pascal; Godet, Julien; Mély, Yves

    2009-05-01

    In the present work, a Michelson interferometer was combined with a two-photon excitation microscope to perform two-focus Fluorescence Correlation Spectroscopy. This simple and original approach allows us to tune the distance between the two excitation volumes and determine absolute diffusion constants. The technique was validated on different model systems that demonstrate the sensitivity of the approach.

  15. 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

  16. 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

  17. Interactions of hypericin with a model mutagen - Acridine orange analyzed by light absorption and fluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Pietrzak, Monika; Szabelski, Mariusz; Kasparek, Adam; Wieczorek, Zbigniew

    2017-02-01

    The present study was designed to estimate the ability of hypericin to interact with a model mutagen - acridine orange. The hetero-association of hypericin and acridine orange was investigated with absorption and fluorescence spectroscopy methods in aqueous solution of DMSO. The data indicate that hypericin forms complexes with acridine orange and that the association constants are relatively high and depend on DMSO concentration. The absorption spectra of the hypericin - acridine orange complexes were examined as well. Owing to its ability to interact with flat aromatic compounds, hypericin may potentially be used as an interceptor molecule.

  18. Photobleaching response of different sources of chromophoric dissolved organic matter exposed to natural solar radiation using absorption and excitation-emission matrix spectra.

    PubMed

    Zhang, Yunlin; Liu, Xiaohan; Osburn, Christopher L; Wang, Mingzhu; Qin, Boqiang; Zhou, Yongqiang

    2013-01-01

    CDOM biogeochemical cycle is driven by several physical and biological processes such as river input, biogeneration and photobleaching that act as primary sinks and sources of CDOM. Watershed-derived allochthonous (WDA) and phytoplankton-derived autochthonous (PDA) CDOM were exposed to 9 days of natural solar radiation to assess the photobleaching response of different CDOM sources, using absorption and fluorescence (excitation-emission matrix) spectroscopy. Our results showed a marked decrease in total dissolved nitrogen (TDN) concentration under natural sunlight exposure for both WDA and PDA CDOM, indicating photoproduction of ammonium from TDN. In contrast, photobleaching caused a marked increase in total dissolved phosphorus (TDP) concentration for both WDA and PDA CDOM. Thus TDN:TDP ratios decreased significantly both for WDA and PDA CDOM, which partially explained the seasonal dynamic of TDN:TDP ratio in Lake Taihu. Photobleaching rate of CDOM absorption a(254), was 0.032 m/MJ for WDA CDOM and 0.051 m/MJ for PDA CDOM from days 0-9, indicating that phototransformations were initially more rapid for the newly produced CDOM from phytoplankton than for the river CDOM. Extrapolation of these values to the field indicated that 3.9%-5.1% CDOM at the water surface was photobleached and mineralized every day in summer in Lake Taihu. Photobleaching caused the increase of spectral slope, spectral slope ratio and molecular size, indicating the CDOM mean molecular weight decrease which was favorable to further microbial degradation of mineralization. Three fluorescent components were validated in parallel factor analysis models calculated separately for WDA and PDA CDOM. Our study suggests that the humic-like fluorescence materials could be rapidly and easily photobleached for WDA and PDA CDOM, but the protein-like fluorescence materials was not photobleached and even increased from the transformation of the humic-like fluorescence substance to the protein

  19. Development of UV-excitable red and near-infrared fluorescent labels and their application for simultaneous multicolor bioimaging by single-wavelength excitation.

    PubMed

    Mizuno, Tetsuya; Umezawa, Keitaro; Shindo, Yutaka; Citterio, Daniel; Oka, Kotaro; Suzuki, Koji

    2013-09-01

    We report a new type of UV-excitable red/NIR-emissive fluorescent dyads (PKF series). Conjugation of a pyrene and a novel bright red/near-infrared (NIR) fluorophore resulted in large quasi-Stokes shift while retaining intense fluorescence emission and sharp spectral bands. Labeling of PKF dyads to biomolecules was performed by means of introduction of a succinimidyl ester. Simultaneous Ca(2+)/albumin dual-color intracellular imaging by PKF in combination with fura-2 (UV-excitable/VIS-emissive Ca(2+) indicator) reveals its usefulness as a new bioimaging tool.

  20. 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.

  1. High-resolution fluorescence excitation spectra of jet-cooled benzyl and p-methylbenzyl radicals

    NASA Astrophysics Data System (ADS)

    Lin, Tai-Yuan David; Tan, Xue-Qing; Cerny, Timothy M.; Williamson, James M.; Cullin, David W.; Miller, Terry A.

    1992-11-01

    High-resolution, rotationally resolved, laser-induced, fluorescence excitation spectra of the A 1 and 6a 10 bands of benzyl and the 0 00 band of p-methylbenzyl radicals were obtained in supersonic expansions. All three spectra were assigned and fit, using the rigid rotor Hamiltonian as well as methyl group internal rotation theory. The results of the rotational analysis provide good rotation constants for benzyl and p-methylbenzyl and establish unambiguously that the symmetry of the excited electronic state in this transition of p-methylbenzyl is 2A 2 (in C 2v). The heights of torsional barriers that hinder the internal rotation of the methyl group in p-methylbenzyl also are determined. The torsional results are compared to those obtained previously for this radical in a vibrational analysis and to other open shell radicals.

  2. Full elastic characterization of absorptive rubber using laser excited guided ultrasonic waves

    NASA Astrophysics Data System (ADS)

    Verstraeten, Bert; Xu, Xiadong; Martinez, Loïc; Glorieux, Christ

    2012-05-01

    Because of the highly damping nature of rubber, it is difficult to characterize its dynamic elastic properties using classical methods. In this paper, an experimental approach employing laser excited guided acoustic waves is proposed to accurately determine the real and imaginary part of the longitudinal and shear elastic modulus of a rubber layer. From the spatiotemporal evolution of a propagating laser excited Lamb wave measured by a laser Doppler vibrometer, which is scanning along a line perpendicular to a line of excitation, the phase velocity dispersion curves in the wave number - frequency domain are obtained. The results are interpreted in the framework of a detailed semianalytical study, analyzing the influence of elastic damping on the Lamb dispersion curves. This analysis is exploited to adequately fit the experimental dispersion curves and thus extract information about the elastic moduli and absorption coefficients of the rubber plate. The results are validated by a pulse-echo measurement, and by guided wave propagation results with the rubber layer connected in a bi-layer plate configuration to non-damping plates.

  3. 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.

  4. 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.

  5. Fluorescent excitation of photospheric Fe K-alpha emission during solar flares

    NASA Technical Reports Server (NTRS)

    Parmar, A. N.; Culhane, J. L.; Rapley, C. G.; Phillips, K. J. H.; Wolfson, C. J.; Acton, L. W.; Dennis, B. R.

    1982-01-01

    The Bent Crystal Spectrometer on the NASA Solar Maximum Mission satellite provides high spectral and temporal resolution observations of the Fe K-alpha lines. Analyses have been conducted of spectra from almost 50 solar flares that occurred during 1980. These data strongly support fluorescent excitation of photospheric iron by photons of E greater than 7.11 keV emitted by the hot coronal plasma produced during the flare. After comparison of the data with a model, the observed K-alpha line widths are discussed along with estimates of the size of the emitting region, the height of the coronal source and the photospheric iron abundance.

  6. Solvatochromic Shifts on Absorption and Fluorescence Bands of N,N-Dimethylaniline.

    PubMed

    Fdez Galván, Ignacio; Elena Martín, M; Muñoz-Losa, Aurora; Aguilar, Manuel A

    2009-02-10

    A theoretical study of the absorption and fluorescence UV/vis spectra of N,N-dimethylaniline in different solvents has been performed, using a method combining quantum mechanics, molecular mechanics, and the mean field approximation. The transitions between the three lowest-lying states have been calculated in vacuum as well as in cyclohexane, tetrahydrofuran, and water. The apparent anomalies experimentally found in water (a blue shift in the absorption bands with respect to the trend in other solvents, and an abnormally high red shift for the fluorescence band) are well reproduced and explained in view of the electronic structure of the solute and the solvent distribution around it. Additional calculations were done with a mixture of cyclohexane and tetrahydrofuran as solvent, which displays a nonlinear solvatochromic shift. Results, although not conclusive, are consistent with experiment and provide a possible explanation for the nonlinear behavior in the solvent mixture.

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

    NASA Astrophysics Data System (ADS)

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

    2013-05-01

    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.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. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr00308f

  8. Effects of solvent polarity on the absorption and fluorescence spectra of chlorogenic acid and caffeic acid compounds: determination of the dipole moments.

    PubMed

    Belay, Abebe; Libnedengel, Ermias; Kim, Hyung Kook; Hwang, Yoon-Hwae

    2016-02-01

    The effects of solvent polarity on absorption and fluorescence spectra of biologically active compounds (chlorogenic acid (CGA) and caffeic acids (CA)) have been investigated. In both spectra pronounced solvatochromic effects were observed with shift of emission peaks larger than the corresponding UV-vis electronic absorption spectra. From solvatochromic theory the ground and excited-state dipole moments were determined experimentally and theoretically. The differences between the excited and ground state dipole moment determined by Bakhshiev, Kawski-Chamma-Viallet and Reichardt equations are quite similar. The ground and excited-state dipole moments were determined by theoretical quantum chemical calculation using density function theory (DFT) method (Gaussian 09) and were also similar to the experimental results. The HOMO-LUMO energy band gaps for CGA and CFA were calculated and found to be 4.1119 and 1.8732 eV respectively. The results also indicated the CGA molecule is more stable than that of CFA. It was also observed that in both compounds the excited state possesses a higher dipole moment than that of the ground state. This confirms that the excited state of the hydroxycinnamic compounds is more polarized than that of the ground state and therefore is more sensitive to the solvent.

  9. Internal energy distributions from nitrogen dioxide fluorescence. 2. Collisional energy transfer from excited nitrogen dioxide

    SciTech Connect

    Patten, K.O. Jr.; Johnston, H.S. Lawrence Berkeley Lab., CA )

    1993-09-30

    We follow the collisional deactivation of laser-excited nitrogen dioxide through its dispersed fluorescence. The energy acceptor gases are NO[sub 2] at four excitation energies ranging from 18828 to 24989 cm[sup [minus]1] and five monatomic gases, four diatomic gases, and three polyatomic gases with 18828-cm[sup [minus]1] excitation energy. The nominal products are the shapes of the internal energy distributions, which are obtained and plotted for several representative cases. From these distributions, the first three moments of the internal energy distributions are derived as a function of molecular collisions and tabulated as (i) the average internal energy, (ii) energy spread, and (iii) skewness. These quantities are plotted against c(M)t, the product of buffer gas concentration c(M) and delay time after laser excitation t(0.5-2 [mu]s), which is a quantity proportional to number of collisions. The negative slope of average energy vs c(M)t is the macroscopic energy-transfer rate constant, k[sub [epsilon

  10. Time-resolved and steady-state fluorescence studies of excited-state proton-transfer reactions of proflavine

    NASA Astrophysics Data System (ADS)

    De Silvestri, S.; Laporta, P.

    1984-01-01

    Time-resolved and steady-state fluorescence studies of proflavine in aqueous solution are presented. The observation of a monoexponential fluorescence decay with a time constant decreasing with increasing pH and the presence of an anomalous red-shift in the fluorescence spectrum as a function of pH indicate the existence of a complex proton-transfer mechanism in the excited state. A reaction scheme is proposed and the corresponding proton-transfer rates are evaluated. An excited-state pK value of 12.85 is obtained for the equilibrium between the cationic form of proflavine and the same form dissociated at an amino group.

  11. 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.

  12. X-ray absorption and soft x-ray fluorescence analysis of KDP optics

    SciTech Connect

    Nelson, A J; van Buuren, T; Miller, E; Land, T A; Bostedt, C; Franco, N; Whitman, P K; Baisden, P A; Terminello, L J; Callcott, T A

    2000-08-09

    Potassium Dihydrogen Phosphate (KDP) is a non-linear optical material used for laser frequency conversion and optical switches. Unfortunately, when KDP crystals are coated with a porous silica anti-reflection coating [1] and then exposed to ambient humidity, they develop dissolution pits [2,3]. Previous investigations [2] have shown that thermal annealing renders KDP optics less susceptible to pitting suggesting that a modification of surface chemistry has occurred. X-ray absorption and fluorescence were used to characterize changes in the composition and structure of KDP optics as a function of process parameters. KDP native crystals were also analyzed to provide a standard basis for interpretation. Surface sensitive total electron yield and bulk sensitive fluorescence yield from the K 2p, P 2p (L{sub 2,3}-edge) and O 1s (K-edge) absorption edges were measured at each process step. Soft X-ray fluorescence was also used to observe changes associated with spectral differences noted in the absorption measurements. Results indicate that annealing at 160 C dehydrates the surface of KDP resulting in a metaphosphate surface composition with K:P:O = 1:1:3.

  13. 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.

  14. 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.

  15. Fluorenyl porphyrins for combined two-photon excited fluorescence and photosensitization

    NASA Astrophysics Data System (ADS)

    Mongin, Olivier; Hugues, Vincent; Blanchard-Desce, Mireille; Merhi, Areej; Drouet, Samuel; Yao, Dandan; Paul-Roth, Christine

    2015-04-01

    The two-photon absorption (2PA), the luminescence and the photosensitization properties of porphyrin-cored fluorenyl dendrimers and meso-substituted fluorenylporphyrin monomer, dimer and trimer are described. In comparison with model tetraphenylporphyrin, these compounds combine enhanced (non-resonant) 2PA cross-sections in the near infrared and enhanced fluorescence quantum yields, together with maintained singlet oxygen generation quantum yields. 'Semi-disconnection' between fluorenyl groups and porphyrins (i.e. direct meso substitution) proved to be more efficient than non-conjugated systems (based on efficient FRET between fluorenyl antennae and porphyrins). These results are of interest for combined two-photon imaging and photodynamic therapy.

  16. 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.

  17. 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.

  18. Study of preferential solvation of 2,6-diaminoanthraquinone in binary mixtures by absorption and fluorescence studies

    NASA Astrophysics Data System (ADS)

    Sasirekha, V.; Ramakrishnan, V.

    2008-08-01

    The role of solute-solvent and solvent-solvent interaction on the preferential solvation characteristics of 2,6-diaminoanthraquinone (DAAQ) has been analysed by monitoring the optical absorption and fluorescence emission spectra. Binary mixtures consist of dimethylformamide (DMF)-ethanol (EtOH), DMF-dimelthylsulfoxide (DMSO), benzene (BZ)-DMF and acetonitrile (ACN)-DMF. The optical absorption spectra maximum and emission spectra maximum of DAAQ show the changes with varying the solvents and change in the composition in the case of binary mixtures. Non-ideal solvation characteristics are observed in all binary mixtures. It is found that at certain concentrations two mixed solvents interact to form a common structure with a ν12 (wave number in cm -1) value not always intermediate ( ν1 and ν2) between the values of the solvents mixed. Synergistic effect is observed in the case of DMF-EtOH mixtures. The preferential solvation parameters local mole fraction X2L, solvation index δS2, exchange constant K12 are calculated in all binary mixtures expect in the case of DMF-BZ mixture and DMF-EtOH mixture in the ground state. We have also monitored excitation wavelength effect on the probe molecule in aprotic polar and protic polar solvents.

  19. Laser-Excited Atomic Fluorescence and Ionization in a Graphite Furnace for the Determination of Metals and Nonmetals

    NASA Astrophysics Data System (ADS)

    Butcher, David James

    1990-01-01

    Here is reported novel instrumentation for atomic spectrometry that combined the use of a pulsed laser system as the light source and an electrothermal atomizer as the atom cell. The main goal of the research was to develop instrumentation that was more sensitive for elemental analysis than commercially available instruments and could be used to determine elements in real sample matrices. Laser excited atomic fluorescence spectrometry (LEAFS) in an electrothermal atomizer (ETA) was compared to ETA atomic absorption spectrometry (AAS) for the determination of thallium, manganese, and lead in food and agricultural standard reference materials (SRMs). Compared to ETA AAS, ETA LEAFS has a longer linear dynamic range (LDR) (5-7 orders of magnitude compared to 2-3 orders of magnitude) and higher sensitivity (10 ^{-16} to 10^{ -14} g as compared to 10^{ -13} to 10^{-11} g). Consequently, ETA LEAFS allows elemental analysis to be done over a wider range of concentrations with less dilution steps. Thallium was accurately determined in biological samples by ETA LEAFS at amounts five to one hundred times below the ETA AAS detection limit. ETA AAS and ETA LEAFS were compared for the determination of lead and manganese, and in general, the accuracies and precisions of ETA AAS were the same, with typical precisions between 3% and 6%. Fluorine was determined using laser excited molecular fluorescence spectrometry (LEMOFS) in an ETA. Molecular fluorescence from magnesium fluoride was collected, and the detection limit of 0.3 pg fluorine was two to six orders of magnitude more sensitive than other methods commonly used for the determination of fluorine. Significant interferences from ions were observed, but the sensitivity was high enough that fluorine could be determined in freeze dried urine SRMs by diluting the samples by a factor of one hundred to remove the interferences. Laser enhanced ionization (LEI) in an ETA was used for the determination of metals. For thallium, indium

  20. Fluorescence excitation-emission matrix spectroscopy analysis of landfill leachate DOM in coagulation-flocculation process.

    PubMed

    Zhu, Guocheng; Wang, Chuang; Dong, Xingwei

    2016-09-29

    Landfill leachate contains a variety of organic matters, some of which can be excited and emit fluorescence signal. In order to degrade these organic matters, the pretreatment of the leachate is needed, which can improve the degradation performance of post-treatment process. Coagulation-flocculation is one of the important pretreatment processes to treat landfill leachate. Assessing the chemical compositions of landfill leachate is helpful in the understanding of their sources and fates as well as the mechanistic behaviors in the water environment. The present work aimed to use fluorescence excitation-emission matrix spectroscopy (EEMs) to characterize the chemical fractions of landfill leachate dissolved organic matter (DOM) in conjunction with parallel factor analysis (PARAFAC). Results showed that the DOM of landfill leachate tested in this study was identified resulting from microbial input, which included five typical characteristic peaks and four kinds of PARAFAC fractions. These fractions were mainly composed of hydrophobic macromolecule humic acid-like (HM-HA), hydrophilic intermediate molecular fulvic acid-like (HIM-FA), and hydrophilic small molecule protein-like substances (HSM-PS). HM-HA and HIM-FA were found to be easier to remove than HSM-PS. Further research on HSM-PS removal by coagulation-flocculation still needs to be improved.

  1. Increased signals from short-wavelength-excited fluorescent molecules using sub-Ti:Sapphire wavelengths.

    PubMed

    Norris, G; Amor, R; Dempster, J; Amos, W B; McConnell, G

    2012-11-01

    We report the use of an all-solid-state ultrashort pulsed source specifically for two-photon microscopy at wavelengths shorter than those of the conventional Ti:Sapphire laser. Our approach involves sum-frequency mixing of the output from an optical parametric oscillator (λ= 1400-1640 nm) synchronously pumped by a Yb-doped fibre laser (λ= 1064 nm), with the residual pump radiation. This generated an fs-pulsed output tunable in the red spectral region (λ= 620-636 nm, ~150 mW, 405 fs, 80 MHz, M(2) ~ 1.3). We demonstrate the performance of our ultrashort pulsed system using fluorescently labelled and autofluorescent tissue, and compare with conventional Ti:Sapphire excitation. We observe a more than 3-fold increase in fluorescence signal intensity using our visible laser source in comparison with the Ti:Sapphire laser for two-photon excitation at equal illumination peak powers of 1.16 kW or less.

  2. 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.

  3. Time-Resolved Fluorescence Spectroscopy and Imaging of DNA Labeled with DAPI and Hoechst 33342 Using Three-Photon Excitation

    PubMed Central

    Lakowicz, Joseph R.; Gryczynski, Ignacy; Malak, Henryk; Schrader, Martin; Engelhardt, Peter; Kano, Hiroski; Hell, Stefan W.

    1997-01-01

    We examined the fluorescence spectral properties of the DNA stains DAPI (4′,6-diamidino-2-phenylindole, hydrochloride) and Hoechst 33342 (bis-benzimide, or 2,5′-bi-1H-benzimidazole2′-(4-ethoxyphenyl)-5-(4-methyl-1-piperazinyl)) with two-photon (2hν) and three-photon (3hν) excitation using femtosecond pulses from a Ti:sapphire laser from 830 to 885 nm. The mode of excitation of DAPI bound to DNA changed from two-photon at 830 nm to three-photon at 885 nm. In contrast, Hoechst 33342 displayed only two-photon excitation from 830 to 885 nm. DAPI-DNA displayed the same emission spectra and decay times for 2hν and 3hν excitation. Hoechst 33342-DNA displayed the same intensity decay for excitation at 830 and 885 nm. Both probes displayed higher anisotropies for multiphoton excitation as compared to one-photon excitation with ultraviolet wavelengths, and DAPI-DNA displays a higher anisotropy for 3hν at 885 nm than for 2hν at 830 nm. We used 970-nm excitation of DAPI-stained chromosomes to obtain the first three-dimensional images with three-photon excitation. Three-photon excitation of DAPI-stained chromosomes at 970 nm was demonstrated by the power dependence in the fluorescence microscope. ImagesFIGURE 10FIGURE 11FIGURE 12 PMID:9017187

  4. Time-resolved detection of the one- and two-photon excited fluorescence of single molecules of a folding enhanced green fluorescent protein

    NASA Astrophysics Data System (ADS)

    Cotlet, Mircea; Goodwin, Peter M.; Waldo, Geoffrey S.; Werner, James H.

    2006-02-01

    We use time-resolved single molecule fluorescence detection (MSMD) to investigate the fluorescence dynamics of a mutant of the wild-type Green Fluorescent Protein (GFP) from Aequorea victoria, the folding enhanced GFP (FEGFP). The folding enhanced GFP is a novel and robust variant designed for in vivo high-throughput screening of protein expression levels. This variant shows increased thermal stability and the ability to retain its fluorescence when fused to poorly folding proteins. Here we apply one- (OPE) and two- (TPE) photon excitation on freely diffusing FEGFP molecules. Under OPE, single FEGFP molecules undergo fluorescence flickering in the time scale of μs and tens of μs due to triplet formation and ground-state protonation-deprotonation, respectively. OPE fluorescence lifetimes of single FEGFP molecules show evidence for the presence of different emitting species, the I and B forms of FEGFP chromophore. TPE single FEGFP molecules flicker in fluorescence in the time scale of μs due to singlet-triplet transitions of the chromophore. Two-photon excitation of single FEGFP molecules results in the creation of a photoconverted species with a fluorescence lifetime of 2.5 ns, a species which is bright enough to be detected at the single molecule level. Our results indicate FEGFP is a promising fusion reporter for intracellular applications when using OPE and TPE microscopy with single molecule sensitivity.

  5. Optical limiting and excited-state absorption in fullerene solutions and doped glasses

    SciTech Connect

    McBranch, D.; Smilowitz, L.; Klimov, V.

    1995-09-01

    We report the ground state and excited state optical absorption spectra in the visible and near infrared for several substituted fullerenes and higher fullerenes in toluene solutions. Based on these measurements, broadband predictions of the optical limiting performance of these molecules can be deduced. These predictions are then tested at 532 to 700 nm in intensity-dependent transmission measurements. We observe optical limiting in all fullerenes measured; higher fullerenes show the greatest potential for limiting in the near infrared (650-1000 nm), while substituted C{sub 60} shows optimal limiting in the visible (450-700 nm). We observe dramatically reduced limiting for solid forms of C{sub 60} (thin films and C{sub 60}-doped porous glasses), indicating that efficient optical limiting in fullerenes requires true molecular solutions.

  6. Homogeneous competitive hybridization assay based on two-photon excitation fluorescence resonance energy transfer.

    PubMed

    Liu, Lingzhi; Dong, Xiaohu; Lian, Wenlong; Peng, Xiaoniu; Liu, Zhihong; He, Zhike; Wang, Ququan

    2010-02-15

    Recently, we have successfully developed a two-photon excitation fluorescence resonance energy transfer (TPE-FRET)-based homogeneous immunoassay using two-photon excitable small organic molecule as the energy donor. In the present work, the newly emerging TPE-FRET technique was extended to the determination of oligonucleotide. A new TPE molecule with favorable two-photon action cross section was synthesized [2-(2,5-bis(4-(dimethylamino)styryl)-1H-pyrrol-1-yl)acetic acid, abbreviated as TP-COOH], with the tagged reactive carboxyl group allowing facile conjugation with streptavidin (SA). Employing the TP-COOH molecule as energy donor and black hole quencher 1 (BHQ-1) as acceptor, a TPE-FRET-based homogeneous competitive hybridization model was constructed via a biotin-streptavidin bridge. Through the hybridization between a biotinylated single-stranded DNA (ssDNA) and a BHQ-1-linked ssDNA, and the subsequent capture of the as-formed hybrid by TP-COOH labeled SA, the donor fluorescence was quenched due to the FRET between TP-COOH and BHQ-1. Upon the competition between a target ssDNA and the quencher-linked ssDNA toward the biotinylated oligonucleotide, the donor fluorescence was recovered in a target-dependent manner. Good linearity was obtained with the target oligonucleotide ranging from 0.08 to 1.52 microM. The method was applied to spiked serum and urine samples with satisfying recoveries obtained. The results of this work verified the applicability of TPE-FRET technique in hybridization assay and confirmed the advantages of TPE-FRET in complicated matrix.

  7. 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}

  8. A ratiometric fluorescent probe for alkaline phosphatase via regulation of excited-state intramolecular proton transfer.

    PubMed

    Fan, Chunlei; Luo, Shengxu; Qi, Haiping

    2016-03-01

    A ratiometric fluorescent probe 2-(benzimidazol-2-yl)phenyl phosphoric acid (1) for alkaline phosphatase (ALP) is designed and synthesized. The method employs the modulation of the excited-state intramolecular proton transfer (ESIPT) process of 2-(2'-hydroxyphenyl)benzimidazole (HPBI) through the hydroxyl group protection/deprotection reaction. Upon phosphorylated with POCl3 , HPBI shows only an emission peak at 363 nm due to the blockage of ESIPT. However, once selective enzymatic hydrolysis with alkaline phosphatase (ALP) in Tris-HCl buffer occurs, the probe 1 is returned to HPBI and the ESIPT process is switched on, which results in a decrease in the emission band at 363 nm and an increase in a new fluorescence peak around 430 nm. The fluorescence intensity ratio at 430 and 360 nm (I430/I360) increases linearly with the activity of ALP up to 0.050 U/mL and the detection limit is 0.0013 U/mL. The proposed probe shows excellent specificity toward ALP.

  9. 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.

  10. 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.

  11. An excited-state intramolecular photon transfer fluorescence probe for localizable live cell imaging of cysteine

    NASA Astrophysics Data System (ADS)

    Liu, Wei; Chen, Wen; Liu, Si-Jia; Jiang, Jian-Hui

    2017-03-01

    Small molecule probes suitable for selective and specific fluorescence imaging of some important but low-concentration intracellular reactive sulfur species such as cysteine (Cys) pose a challenge in chemical biology. We present a readily available, fast-response fluorescence probe CHCQ-Ac, with 2-(5‧-chloro-2-hydroxyl-phenyl)-6-chloro-4(3 H)-quinazolinone (CHCQ) as the fluorophore and acrylate group as the functional moiety, that enables high-selectivity and high-sensitivity for detecting Cys in both solution and biological system. After specifically reacted with Cys, the probe undergoes a seven-membered intramolecular cyclization and released the fluorophore CHCQ with excited-state intramolecular photon transfer effect. A highly fluorescent, insoluble aggregate was then formed to facilitate high-sensitivity and high-resolution imaging. The results showed that probe CHCQ-Ac affords a remarkably large Stokes shift and can detect Cys under physiological pH condition with no interference from other analytes. Moreover, this probe was proved to have excellent chemical stability, low cytotoxicity and good cell permeability. Our design of this probe provides a novel potential tool to visualize and localize cysteine in bioimaging of live cells that would greatly help to explore various Cys-related physiological and pathological cellular processes in cell biology and diagnostics.

  12. 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.

  13. Two-parametric scaling law and figures of merit of excited-state absorption of organic dyes

    NASA Astrophysics Data System (ADS)

    Koldunov, Modest F.; Koldunov, Leonid M.

    2017-02-01

    Sequential theoretical analysis of the excited-state absorption of organic dyes is carried out. It is established that the transmission dependence on laser radiation intensity of optical elements activated by organic dyes is described by the scaling law. It contains two parameters (the critical intensity and the contrast parameter) which are figures of merit of the excited state absorption of dye. Requirements to experimental conditions needed for correctly determination of figures of merit of the excited-state absorption efficiency of organic dyes are formulated. It is shown that there is a good agreement between experimental data of the sample transmission dependence on laser radiation intensity with the result of its calculation based on the scaling law. Figures of merit for several organic dyes are obtained.

  14. Absorption spectra and spectral-kinetic characteristics of the fluorescence of Sanguinarine in complexes with polyelectrolytes and DNA

    NASA Astrophysics Data System (ADS)

    Motevich, I. G.; Strekal, N. D.; Nowicky, J. W.; Maskevich, S. A.

    2010-07-01

    The absorption spectra and stationary and time resolved fluorescence spectra of the isoquinoline alkaloid sanguinarine are studied in aqueous media and during interactions with synthetic polyelectrolytes (polystyrene sulfonate and polyallylamine) and a natural polyelectrolyte (DNA).

  15. Excited-state dynamics of dGMP measured by steady-state and femtosecond fluorescence spectroscopy.

    PubMed

    Miannay, Francois-Alexandre; Gustavsson, Thomas; Banyasz, Akos; Markovitsi, Dimitra

    2010-03-11

    The room-temperature fluorescence of 2'-deoxyguanosine 5'-monophosphate (dGMP) in aqueous solution is studied by steady-state and time-resolved fluorescence spectroscopy. The steady-state fluorescence spectrum of dGMP shows one band centered at 334 nm but has an extraordinary long red tail, extending beyond 700 nm. Both the fluorescence quantum yield and the relative weight of the 334 nm peak increase with the excitation wavelength. The initial fluorescence anisotropy after excitation at 267 nm is lower than 0.2 for all emission wavelengths, indicating an ultrafast S(2) --> S(1) internal conversion. The fluorescence decays depend strongly on the emission wavelength, getting longer with the wavelength. A rise time of 100-150 fs was observed for wavelengths longer than 450 nm, in accordance with a gradual red shift of the time-resolved spectra. The results are discussed in terms of a relaxation occurring mainly on the lowest excited (1)pi pi*-state surface toward a conical intersection with the ground state, in line with recent theoretical predictions. Our results show that the excited-state population undergoes a substantial "spreading out" before reaching the CI, explaining the complex dynamics observed.

  16. Dressing effects in the attosecond transient absorption spectra of doubly excited states in helium

    NASA Astrophysics Data System (ADS)

    Argenti, L.; Jiménez-Galán, Á.; Marante, C.; Ott, C.; Pfeifer, T.; Martín, F.

    2015-06-01

    Strong-field manipulation of autoionizing states is a crucial aspect of electronic quantum control. Recent measurements of the attosecond transient absorption spectrum (ATAS) of helium dressed by a few-cycle visible pulse [C. Ott et al., Nature (London) 516, 374 (2014), 10.1038/nature14026] provide evidence of the inversion of Fano profiles. With the support of accurate ab initio calculations that reproduce the results of the latter experiment, here we investigate the new physics that arise from ATAS when the laser intensity is increased. In particular, we show that (i) previously unnoticed signatures of the dark 2 p21S doubly excited state are observed in the experimental spectrum, (ii) inversion of Fano profiles is predicted to be periodic in the laser intensity, and (iii) the ac Stark shift of the higher terms in the s p2,n + autoionizing series exceeds the ponderomotive energy, which is the result of a genuine two-electron contribution to the polarization of the excited atom.

  17. Intermediate coupling for core-level excited states: Consequences for X-Ray absorption spectroscopy

    SciTech Connect

    Bagus, Paul S.; Sassi, Michel J.; Rosso, Kevin M.

    2015-04-01

    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.

  18. Transient absorption phenomena and related structural transformations in femtosecond laser-excited Si

    NASA Astrophysics Data System (ADS)

    Kudryashov, Sergey I.

    2004-09-01

    Analysis of processes affecting transient optical absorption and photogeneration of electron-hole plasma in silicon pumped by an intense NIR or visible femtosecond laser pulse has been performed taking into account the most important electron-photon, electron-electron and electron-phonon interactions and, as a result, two main regimes of such laser-matter interaction have been revealed. The first regime is concerned with indirect interband optical absorption in Si, enhanced by a coherent shrinkage of its smallest indirect bandgap due to dynamic Franz-Keldysh effect (DFKE). The second regime takes place due to the critical renormalization of the Si direct bandgap along Λ-axis of its first Brillouin zone because of DFKE and the deformation potential electron-phonon interaction and occurs as intense direct single-photon excitation of electrons into one of the quadruplet of equivalent Λ-valleys in the lowest conduction band, which is split down due to the electron-phonon interaction.

  19. 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.

  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. 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.

  2. 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.

  3. 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.

  4. Time-dependent excitation and ionization modelling of absorption-line variability due to GRB 080310

    NASA Astrophysics Data System (ADS)

    Vreeswijk, P. M.; Ledoux, C.; Raassen, A. J. J.; Smette, A.; De Cia, A.; Woźniak, P. R.; Fox, A. J.; Vestrand, W. T.; Jakobsson, P.

    2013-01-01

    We model the time-variable absorption of Fe II, Fe III, Si II, C II and Cr II detected in Ultraviolet and Visual Echelle Spectrograph (UVES) spectra of gamma-ray burst (GRB) 080310, with the afterglow radiation exciting and ionizing the interstellar medium in the host galaxy at a redshift of z = 2.42743. To estimate the rest-frame afterglow brightness as a function of time, we use a combination of the optical VRI photometry obtained by the RAPTOR-T telescope array, which is presented in this paper, and Swift's X-Ray Telescope (XRT) observations. Excitation alone, which has been successfully applied for a handful of other GRBs, fails to describe the observed column density evolution in the case of GRB 080310. Inclusion of ionization is required to explain the column density decrease of all observed Fe II levels (including the ground state 6D9/2) and increase of the Fe III 7S3 level. The large population of ions in this latter level (up to 10% of all Fe III) can only be explained through ionization of Fe II, as a large fraction of the ionized Fe II ions (we calculate 31% using the Flexible Atomic and Cowan codes) initially populate the 7S3 level of Fe III rather than the ground state. This channel for producing a significant Fe III 7S3 level population may be relevant for other objects in which absorption lines from this level, the UV34 triplet, are observed, such as broad absorption line (BAL) quasars and η Carinae. This provides conclusive evidence for time-variable ionization in the circumburst medium, which to date has not been convincingly detected. However, the best-fit distance of the neutral absorbing cloud to the GRB is 200-400 pc, i.e. similar to GRB-absorber distance estimates for GRBs without any evidence for ionization. We find that the presence of time-varying ionization in GRB 080310 is likely due to a combination of the super-solar iron abundance ([Fe/H] = +0.2) and the low H I column density (log N(H i) = 18.7) in the host of GRB 080310. Finally

  5. UV fluorescence excitation imaging of healing of wounds in skin: Evaluation of wound closure in organ culture model

    PubMed Central

    Wang, Ying; Gutierrez‐Herrera, Enoch; Ortega‐Martinez, Antonio; Anderson, Richard Rox

    2016-01-01

    Background and Objective Molecules native to tissue that fluoresce upon light excitation can serve as reporters of cellular activity and protein structure. In skin, the fluorescence ascribed to tryptophan is a marker of cellular proliferation, whereas the fluorescence ascribed to cross‐links of collagen is a structural marker. In this work, we introduce and demonstrate a simple but robust optical method to image the functional process of epithelialization and the exposed dermal collagen in wound healing of human skin in an organ culture model. Materials and Methods Non‐closing non‐grafted, partial closing non‐grafted, and grafted wounds were created in ex vivo human skin and kept in culture. A wide‐field UV fluorescence excitation imaging system was used to visualize epithelialization of the exposed dermis and quantitate wound area, closure, and gap. Histology (H&E staining) was also used to evaluate epithelialization. Results The endogenous fluorescence excitation of cross‐links of collagen at 335 nm clearly shows the dermis missing epithelium, while the endogenous fluorescence excitation of tryptophan at 295 nm shows keratinocytes in higher proliferating state. The size of the non‐closing wound was 11.4 ± 1.8 mm and remained constant during the observation period, while the partial‐close wound reached 65.5 ± 4.9% closure by day 16. Evaluations of wound gaps using fluorescence excitation images and histology images are in agreement. Conclusions We have established a fluorescence imaging method for studying epithelialization processes, evaluating keratinocyte proliferation, and quantitating closure during wound healing of skin in an organ culture model: the dermal fluorescence of pepsin‐digestible collagen cross‐links can be used to quantitate wound size, closure extents, and gaps; and, the epidermal fluorescence ascribed to tryptophan can be used to monitor and quantitate functional states of epithelialization. UV fluorescence

  6. 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.

  7. 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.

  8. 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.

  9. The use of ultraviolet excitation of native fluorescence for identifying biomarkers in halite crystals

    NASA Astrophysics Data System (ADS)

    Mormile, Melanie R.; Storrie-Lombardi, Michael

    2005-09-01

    Recent findings by the NASA's Mars Exploration Rovers and ESA's Mars Express indicate that during an earlier era in the planets' evolution, evaporation of surface water may have left behind vast evaporite deposits1,2,3. This makes the possibility of finding biological material preserved in halite inclusions most intriguing4. The retrieval and characterization of microorganisms from ancient halite crystals5,6 suggests that it might be possible to locate their remains as biomarkers or even living cells from evaporites sampled from extraterrestrial environments. However, before this is possible, techniques for the detection of bacterial cells or biomolecules in halite and other evaporite crystals need further refining. Specifically, contamination must be minimized and quantified during the microbial analysis of such crystals. Aseptic techniques that allow for the direct extraction of fluid brines from micron to millimeter size inclusions significantly reduce the possibility for contamination. However, even with extreme precautions, the possibility for contamination cannot be entirely eliminated, particularly when culture-based methods are employed. We have elicited native fluorescence from a variety of biomolecules, including the aromatic amino acids and nucleic acids, by laser excitation at 248 and 224 nm from haloarchaea and haloarchaea residues trapped in halite. Energy to each sample, (positive control crystals with Halobacteria salinarum and bacteria-free negative control crystals), was 80 microwatts at 224 nm and 25 microwatts at 248 nm. A 30 s exposure of the inclusions within the positive control elicited easily detectable fluorescence while there was no response from the negative control crystals during the same exposure. Analysis of halite crystals sampled from the Waste Isolation Pilot Plant, Carlsbad, New Mexico yielded similar results. To minimize microbial damage from the high-energy 224-248 nm beams and to make the technique more widely available to the

  10. 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.

  11. 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

    2016-02-09

    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.

  12. 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

  13. Do fluorescence and transient absorption probe the same intramolecular charge transfer state of 4-(dimethylamino)benzonitrile?

    SciTech Connect

    Gustavsson, Thomas; Fujiwara, Takashige; Lim, Edward C.

    2009-07-21

    We present here the results of time-resolved absorption and emission experiments for 4-(dimethylamino)benzonitrile in solution, which suggest that the fluorescent intramolecular charge transfer (ICT) state may differ from the twisted ICT (TICT) state observed in transient absorption.

  14. Dynamics of Excited States for Fluorescent Emitters with Hybridized Local and Charge-Transfer Excited State in Solid Phase: A QM/MM Study.

    PubMed

    Fan, Jianzhong; Cai, Lei; Lin, Lili; Wang, Chuan-Kui

    2016-12-01

    The highly efficient organic light-emitting diodes (OLEDS) based on fluorescent emitters with hybridized local and charge-transfer (HLCT) excited state have attracted great attention recently. The excited-state dynamics of the fluorescent molecule with consideration of molecular interaction are studied using the hybrid quantum mechanics/molecular mechanics method. The results show that, in solid state, the internal conversion rate (KIC) between the first singlet excited state (S1) and the ground state (S0) is smaller than the fluorescent rate (Kr), while in gas phase KIC is much larger than Kr. By analyzing the Huang-Rhys (HR) factor and reorganization energy (λ), we find that these two parameters in solid state are much smaller than those in gas phase due to the suppression of the vibration modes in low-frequency regions (<200 cm(-1)) related with dihedral angles between donor and acceptor groups. This is further demonstrated by the geometrical analysis that variation of the dihedral angle between geometries of S1 and S0 is smaller in solid state than that in gas phase. Moreover, combining the dynamics of the excited states and the adiabatic energy structures calculated in solid state, we illustrate the suggested "hot-exciton" mechanism of the HLCT emitters in OLEDs. Our work presents a rational explanation for the experimental results and demonstrates the importance of molecular interaction for theoretical simulation of the working principle of OLEDs.

  15. Truncated Newton's optimization scheme for absorption and fluorescence optical tomography: Part I theory and formulation.

    PubMed

    Roy, R; Sevick-Muraca, E

    1999-05-10

    The development of non-invasive, biomedical optical imaging from time-dependent measurements of near-infrared (NIR) light propagation in tissues depends upon two crucial advances: (i) the instrumental tools to enable photon "time-of-flight" measurement within rapid and clinically realistic times, and (ii) the computational tools enabling the reconstruction of interior tissue optical property maps from exterior measurements of photon "time-of-flight" or photon migration. In this contribution, the image reconstruction algorithm is formulated as an optimization problem in which an interior map of tissue optical properties of absorption and fluorescence lifetime is reconstructed from synthetically generated exterior measurements of frequency-domain photon migration (FDPM). The inverse solution is accomplished using a truncated Newtons method with trust region to match synthetic fluorescence FDPM measurements with that predicted by the finite element prediction. The computational overhead and error associated with computing the gradient numerically is minimized upon using modified techniques of reverse automatic differentiation.

  16. Observation of upconversion fluorescence and stimulated emission based on three-photon absorption

    NASA Astrophysics Data System (ADS)

    Yang, Q.; Lin, S.; Xu, L.; Yang, F.; Yang, Y.; Pan, L.; Sun, C.; Li, Y.; Sun, G.; Jiang, Z.

    2005-06-01

    The observations of three-photon-induced frequency-upconversion fluorescence and the highly directional stimulated visible emission in two dyes, 4-[p-(dicyanoethylamino) styryl]-N-methylpyridinium iodide (abbreviated as CEASP) and the complex of CEASP and Ce(NO3) (abbreviated as CEASP-Ce), are reported. The photographs of the forward amplified spontaneous emissions spots, pumped by an optical parametric oscillator idler with a pulse width of 8 ns and a wavelength of 1.3 μ m, are shown. The upconversion fluorescence produced both in dimethyl formamide solution and 2-hydroxyethyl methacrylate (HEMA) polymer spans from green to red, with a cubic dependence on the pump light intensity. The experimental results imply that the existence of the lanthanide ion Ce3 + sensitizes the nonlinear absorption and emission.

  17. Fluorescence excitation and emission spectra of 1,5-dihydroxyanthraquinone-d2 in n-hexane at 10 K

    NASA Astrophysics Data System (ADS)

    Smulevich, Giulietta; Foggi, Paolo

    1987-11-01

    The fluorescence excitation, between 430 and 505 nm, and emission, between 505 and 725 nm, spectra in n-hexane of 1,5-dihydroxyanthraquinone-d0 and -d2 at 10 K have been measured. Dual excitation and emission associated to excited state proton transfer were observed. Apart from the long wavelength emission, well resolved vibrational structures were obtained. A remarkable spectral shift (684 cm-1) of the origin of the high frequency transition was observed upon deuteration. The energy gaps between the transition origins both in excitation and emission as well as the isotopic shifts of the origins, were interpreted in terms of Lippincott-Schroeder asymmetric double minimum potential functions along the OH coordinate. An extra fluorescence occurs in the low frequency range, vanishing upon deuteration. It was explained as due to the ν(OH) stretching mode of the high frequency emission enhanced via vibronic coupling between the two ground states.

  18. Rational design, synthesis, and spectroscopic and photophysical properties of a visible-light-excitable, ratiometric, fluorescent near-neutral pH indicator based on BODIPY.

    PubMed

    Boens, Noël; Qin, Wenwu; Baruah, Mukulesh; De Borggraeve, Wim M; Filarowski, Aleksander; Smisdom, Nick; Ameloot, Marcel; Crovetto, Luis; Talavera, Eva M; Alvarez-Pez, Jose M

    2011-09-19

    A visible-light-excitable, ratiometric, brightly fluorescent pH indicator for measurements in the pH range 5-7 has been designed and synthesized by conjugatively linking the BODIPY fluorophore at the 3-position to the pH-sensitive ligand imidazole through an ethenyl bridge. The probe is available as cell membrane permeable methyl ester 8-(4-carbomethoxyphenyl)-4,4-difluoro-3-[2-(1H-imidazol-4-yl)ethenyl]-1,5,7-trimethyl-3a,4a-diaza-4-bora-s-indacene (I) and corresponding water-soluble sodium carboxylate, sodium 8-(4-carboxylatophenyl)-4,4-difluoro-3-[2-(1H-imidazol-4-yl)ethenyl]-1,5,7-trimethyl-3a,4a-diaza-4-bora-s-indacene (II). The fluorescence quantum yield Φ(f) of ester I is very high (0.8-1.0) in the organic solvents tested. The fluorescence lifetime (ca. 4 ns) of I in organic solvents with varying polarity/polarizability (from cyclohexane to acetonitrile) is independent of the solvent with a fluorescence rate constant k(f) of 2.4×10(8) s(-1). Probe I is readily loaded in the cytosol of live cells, where its high fluorescence intensity remains nearly constant over an extended time period. Water-soluble indicator II exhibits two acid-base equilibria in aqueous solution, characterized by pK(a) values of 6.0 and 12.6. The Φ(f) value of II in aqueous solution is high: 0.6 for the cationic and anionic forms of the imidazole ligand, and 0.8 for neutral imidazole. On protonation-deprotonation in the near-neutral pH range, UV/Vis absorption and fluorescence spectral shifts along with isosbestic and pseudo-isoemissive points are observed. This dual-excitation and dual-emission pH indicator emits intense green-yellow fluorescence at lower pH and intense orange fluorescence at higher pH. The influence of ionic strength and buffer concentration on the absorbance and steady-state fluorescence of II has also been investigated. The apparent pK(a) of the near-neutral acid-base equilibrium determined by spectrophotometric and fluorometric titration is nearly independent of

  19. Ultrabright fluorescent silica particles with a large number of complex spectra excited with a single wavelength for multiplex applications.

    PubMed

    Palantavida, S; Peng, B; Sokolov, I

    2017-02-08

    We report on a novel approach to synthesize ultrabright fluorescent silica particles capable of producing a large number of complex spectra. The spectra can be excited using a single wavelength which is paramount in quantitative fluorescence imaging, flow cytometry and sensing applications. The approach employs the physical encapsulation of organic fluorescent molecules inside a nanoporous silica matrix with no dye leakage. As was recently demonstrated, such an encapsulation allowed for the encapsulation of very high concentrations of organic dyes without quenching their fluorescent efficiency. As a result, dye molecules are distanced within ∼5 nm from each other; it theoretically allows for efficient exchange of excitation energy via Förster resonance energy transfer (FRET). Here we present the first experimental demonstration of the encapsulation of fluorescent dyes in the FRET sequence. Attaining a FRET sequence of up to five different dyes is presented. The number of distinguishable spectra can be further increased by using different relative concentrations of encapsulated dyes. Combining these approaches allows for creating a large number of ultrabright fluorescent particles with substantially different fluorescence spectra. We also demonstrate the utilization of these particles for potential multiplexing applications. Though fluorescence spectra of the obtained multiplex probes are typically overlapping, they can be distinguished by using standard linear decomposition algorithms.

  20. 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.

  1. Identification of calcifications in intracranial neoplasms using two photon excitation fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Lin, Peihua; Wang, Xingfu; Wu, Zanyi; Fang, Na; Li, Lianhuang; Kang, Dezhi; Chen, Jianxin

    2016-10-01

    Calcifications within brain tumors may be an indicator of a relatively long survival because a long time is required for the formation of calcium deposits, and may present a novel biomarker associated with response and improved outcome of therapy. In this paper, we describe the use of two-photon excitation fluorescent (TPEF) microscopy combined second harmonic generation (SHG) microscopy for high-resolution imaging that can be applied in identification of intratumoral calcifications. Our results demonstrate that the calcification has stronger TPEF signal than the area around it and the emission spectra shows the difference between the two areas clearly. The TPEF image of calcified region corresponds well with the corresponding H&E stained image. In this work, we present that the label-free imaging technique is able to distinguish the calcified mass lesions in intracranial neoplasms reliably.

  2. Quantitative optical biomarkers of lung cancer based intrinsic two-photon excited fluorescence signal

    NASA Astrophysics Data System (ADS)

    Li, Jingwen; Zhan, Zhenlin; Lin, Hongxin; Zuo, Ning; Zhu, Xiaoqin; Xie, Shusen; Chen, Jianxin; Zhuo, Shuangmu

    2016-10-01

    Alterations in the elastic fibers have been implicated in lung cancer. However, the label-free, microscopic imaging of elastic fibers in situ remains a major challenge. Here, we present the use of intrinsic two-photon excited fluorescence (TPEF) signal as a novel means for quantification of the elastic fibers in intact fresh human lung tissues. We obtained the TPEF images of elastic fibers from ex vivo the human lung tissues. We found that three features, including the elastic fibers area, the elastic fibers orientation, the elastic fibers structure, provide the quantitative identification of lung cancer and the direct visual cues for cancer versus non-cancer areas. These results suggest that the TPEF signal can be used as the label-free optical biomarkers for rapid clinical lung diagnosis and instant image-guided surgery.

  3. 4Pi microscopy of type A with 1-photon excitation in biological fluorescence imaging

    NASA Astrophysics Data System (ADS)

    Lang, Marion; Müller, Tobias; Engelhardt, Johann; Hell, Stefan W.

    2007-03-01

    We demonstrate that oil immersion lenses with a semi-aperture angle ≥ 74° enable 4Pi confocal fluorescence microscopy of type A with 1 photon excitation. The axial sidelobes amount to < 50 % of the main diffraction maximum, implying that lobe induced artifacts can be removed from the image data. The advancement reported herein enables a relative inexpensive implementation of 4Pi microscopy, providing axially superresolved 3D-imaging in transparent samples. As an example, we show dual-color 4Pi images of double stained Golgi stacks in a mammalian cell with 110 nm axial resolution. The resolution can be further enhanced to values slightly below 100 nm by image deconvolution.

  4. Optimization of native fluorescence detection of proteins using a pulsed nano laser excitation source

    PubMed Central

    Heywood, Matthew S.; Farnsworth, Paul B.

    2010-01-01

    We present a mathematical description of the S/N ratio in a fluorescence-based protein detector for capillary electrophoresis that uses a pulsed UV laser at 266 nm as an excitation source. The model accounts for photobleaching, detector volume, laser repetition rate, and analyte flow rate. We have experimentally characterized such a system, and present a comparison of the experimental data with the predictions of the model. Using the model, the system was optimized for test analytes tryptophan, tyrosine, BSA, and conalbumin, producing detection limits (3σ) of 0.67 nM, 5.7 nM, 0.9 nM, and 1.5 nM, respectively. Based on the photobleaching data, a photobleaching cross section of 1.4×10−18 cm2 at 266 nm was calculated for tryptophan. PMID:21073798

  5. 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.

  6. Determination of photophysical parameters of red fluorescent protein mRFP1 under ultraviolet excitation by methods of laser fluorimetry.

    PubMed

    Banishev, Alexandr A; Shirshin, Evgeny A; Fadeev, Victor V

    2010-12-01

    We investigate photophysical processes that take place in macromolecules of a fluorescent protein mRFP1 under UV excitation [when the energy transfer in a localized donor-acceptor (LDA) pair, which is presented in the molecules of the protein, becomes apparent]. We used a special approach based on the fluorescence laser spectroscopy technique. The energy transfer rates in LDA pairs and photophysical parameters of fluorophores (chromophores) of three spectral forms, which coexist in the ensemble of the macromolecules of this protein, were determined under pulse UV laser excitation.

  7. [Theoretical study of fluorescence of photosynthetic pigments at complex dependence of intensity of exciting light on time].

    PubMed

    Alekseev, A A; Belov, A A; Kirzhanov, D V; Kukushkin, A K

    2012-01-01

    Now the methods using the radiance with complex dependence of light intensity on time are applied to research of photosynthesis by means of fluorescence, exciting photosynthetic pigments. One of these methods is applied in PAM-fluorometers--the commercial devices currently widely used to investigate a state of photosynthesizing systems. However, if excitation is performed in this way, the question, what components of fluorescence are registered at an output of such devices, remains to be open-ended. In this work an attempt to analyse this task has been made.

  8. 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.

  9. 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.

  10. Donor-Acceptor Conjugated Linear Polyenes: A Study of Excited State Intramolecular Charge Transfer, Photoisomerization and Fluorescence Probe Properties.

    PubMed

    Hota, Prasanta Kumar; Singh, Anil Kumar

    2014-07-27

    Numerous studies of donor-acceptor conjugated linear polyenes have been carried out with the goal to understand the exact nature of the excited state electronic structure and dynamics. In this article we discuss our endeavours with regard to the excited state intramolecular charge transfer, photoisomerization and fluorescence probe properties of various donor-acceptor substituted compounds of diphenylpolyene [Ar(CH = CH) n Ar] series and ethenylindoles.

  11. Two-photon excited fluorescence detection at 420 nm for label-free detection of small aromatics and proteins in microchip electrophoresis.

    PubMed

    Schulze, Philipp; Schüttpelz, Mark; Sauer, Markus; Belder, Detlev

    2007-12-01

    Two photon excited (TPE) fluorescence detection was applied to native fluorescence detection of aromatics in microchip electrophoresis (MCE). This technique was evaluated as an alternative to common one photon excitation in the deep UV spectral range. TPE enables fluorescence detection of unlabeled aromatic compounds, even in non-deep UV-transparent microfluidic chips. In this study, we demonstrate the proof of concept of native TPE fluorescence detection of small aromatics in commercial microfluidic glass chips. Label-free TPE fluorescence detection of native proteins and small aromatics in MCE was achieved within the micromolar concentration range, utilising 420 nm excitation light.

  12. 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; ...

    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

  13. Imaging electronic trap states in perovskite thin films with combined fluorescence and femtosecond transient absorption microscopy

    SciTech Connect

    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. The 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.

  14. Substituted benzoxadiazoles as fluorogenic probes: a computational study of absorption and fluorescence.

    PubMed

    Brown, Alex; Ngai, Tsz Yan; Barnes, Marie A; Key, Jessie A; Cairo, Christopher W

    2012-01-12

    General chemical strategies which provide controlled changes in the emission or absorption properties of biologically compatible fluorophores remain elusive. One strategy employed is the conversion of a fluorophore-attached alkyne (or azide) to a triazole through a copper-catalyzed azide-alkyne coupling (CuAAC) reaction. In this study, we have computationally examined a series of structurally related 2,1,3-benzoxadiazole (benzofurazan) fluorophores and evaluated changes in their photophysical properties upon conversion from alkyne (or azide) to triazole forms. We have also determined the photophysical properties for a known set of benzoxadiazole compounds. The absorption and emission energies have been determined computationally using time-dependent density functional theory (TD-DFT) with the Perdew, Burke, and Ernzerhof exchange-correlation density functional (PBE0) and the 6-31+G(d) basis set. The TD-DFT results consistently agreed with the experimentally determined absorption and emission wavelengths except for certain compounds where charge-transfer excited states occurred. In addition to determining the absorption and emission wavelengths, simple methods for predicting relative quantum yields previously derived from semiempirical calculations were reevaluated on the basis of the new TD-DFT results and shown to be deficient. These results provide a necessary framework for the design of new substituted benzoxadiazole fluorophores.

  15. Large two-photon absorption cross sections of hemiporphyrazines in the excited state: the multiphoton absorption process of hemiporphyrazines with different central metals.

    PubMed

    Dini, Danilo; Calvete, Mario J F; Hanack, Michael; Amendola, Vincenzo; Meneghetti, Moreno

    2008-09-17

    A series of five hemiporphyrazines (Hps) with different coordinating central atoms (H2, GeCl2, InCl, Pt, Pb), and the acyclic derivative 1,3-bis-(6'-amino-4'-butoxy-2'-pyridylimino)-1,3-dihydroisoindoline have been synthesized and their multiphoton absorption properties examined at the second harmonic frequency of the Nd:YAG laser in the nanosecond time regime. Metal-free and platinum Hps display saturation of optical transmittance within incident fluence values of 6 J cm(-2). Comparison with other similar molecular structures like phthalocyanines and related molecules shows that Hps are strong nonlinear absorbers. The experimental curves of nonlinear transmission at 532 nm have been fitted by means of a three-level model with the occurrence of simultaneous two-photon absorption from an excited state. In the sole case of the InCl complex we found that a five-level model is needed because of the participation of triplet states. Contrary to phthalocyanines, naphthalocyanines, and porphyrins, a heavy central atom does not improve the nonlinear absorption properties since a different excited states dynamic is involved. The large nonlinear absorption of Hps combined with the very small absorption in the visible spectral range makes these molecules a very interesting class of molecules for nonlinear optical applications.

  16. Time-resolved measurements of short-wavelength fluorescence from x-ray-excited ions.

    PubMed

    Kapteyn, H C; Murnane, M M; Falcone, R W

    1987-09-01

    We demonstrate a novel technique for time-resolved spectroscopic studies of highly excited ions. The technique uses a laser-produced plasma as a short-pulse, soft-x-ray light source with a high repetition rate. A Nd:YAG laser with a pulse duration of 90 psec, a pulse energy of 70 microJ, and repetition rate of 10(4) pulses per second is focused onto a rotating metal target. Soft x rays from the resulting plasma photoionize a gas surrounding the target, and fluorescence from the gas is detected by using a spectrometer and a high-speed photodetector. Using the technique of time-correlated photon counting, we determined the radiative lifetime and collisional quenching rate of the Xe III 5s(0)5p(6)(1)S(0) state by observing its fluorescence at 108.9 nm. A time resolution of better than 400 psec was obtained. We also measured relative Auger decay yields of a core hole state in xenon using a higher-energy laser-produced plasma light source at a lower repetition rate.

  17. Fluorescence 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.

  18. Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Bukara, Katarina; Jovanić, Svetlana; Drvenica, Ivana T.; Stančić, Ana; Ilić, Vesna; Rabasović, Mihailo D.; Pantelić, Dejan; Jelenković, Branislav; Bugarski, Branko; Krmpot, Aleksandar J.

    2017-02-01

    The present study describes utilization of two photon excitation fluorescence (2PE) microscopy for visualization of the hemoglobin in human and porcine erythrocytes and their empty membranes (i.e., ghosts). High-quality, label- and fixation-free visualization of hemoglobin was achieved at excitation wavelength 730 nm by detecting visible autofluorescence. Localization in the suspension and spatial distribution (i.e., mapping) of residual hemoglobin in erythrocyte ghosts has been resolved by 2PE. Prior to the 2PE mapping, the presence of residual hemoglobin in the bulk suspension of erythrocyte ghosts was confirmed by cyanmethemoglobin assay. 2PE analysis revealed that the distribution of hemoglobin in intact erythrocytes follows the cells' shape. Two types of erythrocytes, human and porcine, characterized with discocyte and echinocyte morphology, respectively, showed significant differences in hemoglobin distribution. The 2PE images have revealed that despite an extensive washing out procedure after gradual hypotonic hemolysis, a certain amount of hemoglobin localized on the intracellular side always remains bound to the membrane and cannot be eliminated. The obtained results open the possibility to use 2PE microscopy to examine hemoglobin distribution in erythrocytes and estimate the purity level of erythrocyte ghosts in biotechnological processes.

  19. 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.

  20. In vitro characterization of Fluorescence by Unbound Excitation from Luminescence: broadening the scope of energy transfer.

    PubMed

    Holland, Alexandra D; Rückerl, Florian; Dragavon, Joseph M; Rekiki, Abdessalem; Tinevez, Jean-Yves; Tournebize, Régis; Shorte, Spencer L

    2014-03-15

    Energy transfer mechanisms represent the basis for an array of valuable tools to infer interactions in vitro and in vivo, enhance detection or resolve interspecies distances such as with resonance. Based upon our own previously published studies and new results shown here we present a novel framework describing for the first time a model giving a view of the biophysical relationship between Fluorescence by Unbound Excitation from Luminescence (FUEL), a conventional radiative excitation-emission process, and bioluminescence resonance energy transfer. We show here that in homogeneous solutions and in fluorophore-targeted bacteria, FUEL is the dominant mechanism responsible for the production of red-shifted photons. The minor resonance contribution was ascertained by comparing the intensity of the experimental signal to its theoretical resonance counterpart. Distinctive features of the in vitro FUEL signal include a macroscopic depth dependency, a lack of enhancement upon targeting at a constant fluorophore concentration cf and a non-square dependency on cf. Significantly, FUEL is an important, so far overlooked, component of all resonance phenomena which should guide the design of appropriate controls when elucidating interactions. Last, our results highlight the potential for FUEL as a means to enhance in vivo and in vitro detection through complex media while alleviating the need for targeting.

  1. Fluorescence imaging of local membrane electric fields during the excitation of single neurons in culture.

    PubMed Central

    Gogan, P; Schmiedel-Jakob, I; Chitti, Y; Tyc-Dumont, S

    1995-01-01

    The spatial distribution of depolarized patches of membrane during the excitation of single neurons in culture has been recorded with a high spatial resolution (1 micron2/pixel) imaging system based on a liquid-nitrogen-cooled astronomical camera mounted on an inverted microscope. Images were captured from rat nodose neurons stained with the voltage-sensitive dye RH237. Conventional intracellular microelectrode recordings were made in synchrony with the images. During an action potential the fluorescence changes occurred in localized, unevenly distributed membrane areas, which formed clusters of depolarized sites of different sizes and intensities. When fast conductances were blocked by the addition of tetrodotoxin, a reduction in the number and the intensities of the depolarized sites was observed. The blockade by tetrodotoxin of voltage-clamped neurons also reduced the number of depolarized sites, although the same depolarizing voltage step was applied. Similarly, when a voltage-clamped neuron was depolarized by a constant-amplitude voltage step, the number of depolarized sites varied according to the degree of activation of the voltage-sensitive channels, which was modified by changing the holding potential. These results suggest that the spatial patterns of depolarization observed during excitation are related to the operations of ionic channels in the membrane. Images FIGURE 1 FIGURE 2 FIGURE 4 FIGURE 5 FIGURE 6 FIGURE 7 PMID:8527643

  2. 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.

  3. Excited-state absorption and circular dichroism of ruthenium(II) tris(phenanthroline) in the ultraviolet region.

    PubMed

    Niezborala, Claire; Hache, François

    2007-08-16

    Excitation of ruthenium(II) tris(phenanthroline) in the visible region results in the tranfer of an electron from the central atom toward one of the ligands. To probe this excited state, we have performed pump-induced absorption and circular dichroism in the ultraviolet wavelengths, in the intraligand pi-pi* transition region. On top of the bleaching of the ground state transitions, new structures appear in the absorption and CD spectra. Thanks to a classical calculation based on the polarizability theory, we can interpret these features as the result of a strong reduction of the excitonic coupling due to a blue shift of the pi-pi* transition in the reduced ligand accompanied by the onset of new excited-state transitions.

  4. Structural determination of a short-lived excited iron(II) complex by picosecond x-ray absorption spectroscopy.

    PubMed

    Gawelda, Wojciech; Pham, Van-Thai; Benfatto, Maurizio; Zaushitsyn, Yuri; Kaiser, Maik; Grolimund, Daniel; Johnson, Steven L; Abela, Rafael; Hauser, Andreas; Bressler, Christian; Chergui, Majed

    2007-02-02

    Structural changes of the iron(II)-tris-bipyridine ([Fe(II)(bpy)(3)](2+)) complex induced by ultrashort pulse excitation and population of its short-lived (< or =0.6 ns) quintet high spin state have been detected by picosecond x-ray absorption spectroscopy. The structural relaxation from the high spin to the low spin state was followed over the entire lifetime of the excited state. A combined analysis of the x-ray-absorption near-edge structure and extended x-ray-absorption fine structure spectroscopy features delivers an Fe-N bond elongation of 0.2 A in the quintet state compared to the singlet ground state.

  5. 5-HT spatial distribution imaging with multiphoton excitation of 5-HT correlative visible fluorescence in live cells

    NASA Astrophysics Data System (ADS)

    Zhang, Zhihong; Zeng, Shaoqun; Liu, Yafeng; Zhou, Wei; Chen, Tongsheng; Luo, Qingming

    2002-04-01

    The autofluorescence of 5-Hydroxytryptamine (5-HT) loaded rat mucosal mast cells (RBL-2H3 cells) is imaged with multiphoton excitation laser scanning microscope (MPELSM). 5-HT correlative visible fluorescence (Fco-vis) excited with 740-nm multiphoton excitation is observed in live cells for the first time, and the generating mechanism of 5-HT Fco-vis is studied. The spatial distribution of 5-HT in live cells is imaged at high spatial resolution in our experiment, which provides a new way to study the correlation between 5-HT spatial distribution and content, and the cellular functional state in live tissue or cells.

  6. 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.

  7. 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.

  8. Fluorescence excitation-emission matrix spectra coupled with parallel factor and regional integration analysis to characterize organic matter humification.

    PubMed

    He, Xiao-Song; Xi, Bei-Dou; Li, Xiang; Pan, Hong-Wei; An, Da; Bai, Shuo-Guo; Li, Dan; Cui, Dong-Yu

    2013-11-01

    The present several humification indexes cannot provide the whole fluorescence information on organic matter composition and the evaluation results from them are inconsistent sometimes. In this study, fluorescence excitation-emission matrix spectra coupled with parallel factor analysis and fluorescence regional integration analysis were utilized to investigate organic matter humification, and the projection pursuit cluster (PPC) model was applied to form a suitable index for overcoming the difficulties in multi-index evaluation. The result showed that the ratio between the volume of humic- and fulvic-like fluorescence region and the volume of protein-like fluorescence region not only revealed the heterogeneity of organic matter, but also provided more accurate information on organic matter humification. In addition, the results showed that the PPC model could be used to characterize integrally the humification, and the projected characteristic value calculated from the PPC model could be used as the integrated humification evaluation index.

  9. Two-photon absorption laser induced fluorescence measurement of atomic oxygen density in an air atmospheric pressure plasma jet

    NASA Astrophysics Data System (ADS)

    Conway, Jim; Gogna, Gurusharan; Daniels, Stephen

    2016-09-01

    Two-photon Absorption Laser Induced Fluorescence (TALIF) is used to measure atomic oxygen number density [O] in an air Atmospheric Pressure Plasma Jet (APPJ). A novel technique based on photolysis of O2 is used to calibrate the TALIF system ensuring the same species (O) is probed during calibration and measurement. As a result, laser intensity can be increased outside the TALIF quadratic laser power region without affecting calibration reliability as any high intensity saturation effects will be identical for calibration and experiment. Higher laser intensity gives stronger TALIF signals helping overcome weak TALIF signals often experienced at atmospheric pressure due to collisional quenching. O2 photo-dissociation and two-photon excitation of the resulting [O] are both achieved within the same laser pulse. The photolysis [O] is spatially non-uniform and time varying. To allow valid comparison with [O] in a plasma, spatial and temporal correction factors are required. Knowledge of the laser pulse intensity I0(t), and wavelength allows correction factors to be found using a rate equation model. The air flow into the jet was fixed and the RF power coupled into the system varied. The resulting [O] was found to increase with RF power.

  10. Ion pairs of indobenzimidazolo cyanines: a structural study based on conductivity, absorption, fluorescence and 1H-NMR

    NASA Astrophysics Data System (ADS)

    Tatikolov, Aleksandr S.; Ishchenko, Aleksandr A.; Ghelli, Stefano; Ponterini, Glauco

    1998-11-01

    Asymmetric benzimidazolo carbo, di- and tricarbocyanines form ion pairs of the solvent-separated and contact types with different counterions in tetrahydrofuran, toluene and toluene-nitrile mixtures. The dissociation constants of the ion pairs in tetrahydrofuran, evaluated from conductivity data, do not depend on the length of the polymethine chain and show only a small decrease with decreasing counterion size. The absorption and fluorescence excitation spectra of the contact ion pairs exhibit a pronounced hypsochromic shift with respect to the solvated ions and the solvent-separated ion pairs. 1H-NMR experiments have provided information about the electronic structures of the ions of both the asymmetric dyes and the corresponding symmetric carbocyanines. They have also revealed different preferred anion locations in the contact ion pairs of the symmetric indocarbocyanine on one hand, and of the benzimidazolo carbocyanine and the asymmetric dyes on the other. This structural difference is suggested to be a cause of the observed opposite effects of ion pairing on the isomerization kinetics of the two groups of dyes.

  11. 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.

  12. One-colour control of activation, excitation and deactivation of a fluorescent diarylethene derivative in super-resolution microscopy.

    PubMed

    Arai, Yuhei; Ito, Syoji; Fujita, Hajime; Yoneda, Yusuke; Kaji, Takahiro; Takei, Satoshi; Kashihara, Ryota; Morimoto, Masakazu; Irie, Masahiro; Miyasaka, Hiroshi

    2017-02-28

    We demonstrated one-colour control of activation, excitation and deactivation of a fluorescent diarylethene derivative by using a 532 nm CW laser at a weak output power. This one-colour control method was applied to single-molecule tracking in polymer films over a total duration of a few hours at room temperature and PALM for the nanostructures of polymer systems.

  13. 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.

  14. Excitation and propagation of X-ray fluorescence through thin devices with hollowed ordered structures: comparison of experimental and theoretical spectra.

    PubMed

    Mazuritskiy, M I; Dabagov, S B; Marcelli, A; Lerer, A M; Dziedzic-Kocurek, K

    2016-01-01

    The lack of models describing the propagation of X-rays in waveguides and the interference mechanism between incident and reflected radiation waves hamper the understanding and the control of wave propagation phenomena occurring in many real systems. Here, experimental spectra collected at the exit of microchannel plates (MCPs) under the total X-ray reflection condition are presented. The results are discussed in the framework of a theoretical model in which the wave propagation is enhanced by the presence of a transition layer at the surface. The angular distributions of the propagating radiation at the exit of these MCPs with microchannels of ∼3 µm diameter will also be presented and discussed. These spectra show contributions associated with the reflection of the primary monochromatic beam and with the fluorescence radiation originating from the excitation of atoms composing the surface of the microchannel. The soft X-ray fluorescence spectra collected at the exit of microcapillaries were analyzed in the framework of a wave approximation while diffraction contributions observed at the exit of these hollow X-ray waveguides have been calculated using the Fraunhofer diffraction model for waves in the far-field domain. Data collected at the Si L-edge show that in glassy MCPs the fluorescence radiation can be detected only when the energy of the primary monochromatic radiation is above the absorption edge for grazing angles higher than half of the critical angle of the total reflection phenomenon. Experimental data and simulations of the propagating radiation represent a clear experimental confirmation of the channeling phenomenon of the excited fluorescence radiation inside a medium and point out that a high transmission can be obtained in waveguide optics for parameters relevant to X-ray imaging.

  15. 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.

  16. 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.

  17. 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.

  18. Excited state assisted three-photon absorption based optical limiting in nanocrystalline Cu2Se and FeSe2

    NASA Astrophysics Data System (ADS)

    Anand, Benoy; Molli, Muralikrishna; Aditha, Saikiran; Mimani Rattan, Tanu; Siva Sankara Sai, S.; Kamisetti, Venkataramaniah

    2013-09-01

    Transition metal selenides (FeSe2 and Cu2Se) are synthesized by the hydrothermal co-reduction method. XRD results revealed the crystalline nature of their single phase and the elemental compositions are obtained using EDS. TEM images of the as-prepared samples show the formation of nanorods of 10-20 nm diameter in case of iron selenide and nanoparticles of 10-35 nm diameter in case of copper selenide. The energy bandgap values are calculated using tauc plots obtained from UV-Visible absorption spectra. The open aperture Z-scan measurements carried out using 5 ns pulses at 532 nm revealed that the samples showed excellent optical limiting behavior owing to strong nonlinear absorption (NLA). Through numerical simulations, the mechanism of NLA is found to be effective three-photon absorption which has significant contribution from excited state absorption.

  19. New Fluorescence Parameters for the Determination of QA Redox State and Excitation Energy Fluxes.

    PubMed

    Kramer, David M; Johnson, Giles; Kiirats, Olavi; Edwards, Gerald E

    2004-02-01

    A number of useful photosynthetic parameters are commonly derived from saturation pulse-induced fluorescence analysis. We show, that qP, an estimate of the fraction of open centers, is based on a pure 'puddle' antenna model, where each Photosystem (PS) II center possesses its own independent antenna system. This parameter is incompatible with more realistic models of the photosynthetic unit, where reaction centers are connected by shared antenna, that is, the so-called 'lake' or 'connected units' models. We thus introduce a new parameter, qL, based on a Stern-Volmer approach using a lake model, which estimates the fraction of open PS II centers. We suggest that qL should be a useful parameter for terrestrial plants consistent with a high connectivity of PS II units, whereas some marine species with distinct antenna architecture, may require the use of more complex parameters based on intermediate models of the photosynthetic unit. Another useful parameter calculated from fluorescence analysis is ΦII, the yield of PS II. In contrast to qL, we show that the ΦII parameter can be derived from either a pure 'lake' or pure 'puddle' model, and is thus likely to be a robust parameter. The energy absorbed by PS II is divided between the fraction used in photochemistry, ΦII, and that lost non-photochemically. We introduce two additional parameters that can be used to estimate the flux of excitation energy into competing non-photochemical pathways, the yield induced by downregulatory processes, ΦNPQ, and the yield for other energy losses, ΦNO.

  20. Optimal Spectral Regions For Laser Excited Fluorescence Diagnostics For Point Of Care Application

    NASA Astrophysics Data System (ADS)

    Vaitkuviene, A.; Gėgžna, V.; Varanius, D.; Vaitkus, J.

    2011-09-01

    The tissue fluorescence gives the response of light emitting molecule signature, and characterizes the cell composition and peculiarities of metabolism. Both are useful for the biomedical diagnostics, as reported in previous our and others works. The present work demonstrates the results of application of laser excited autofluorescence for diagnostics of pathology in genital tissues, and the feasibility for the bedside at "point of care—off lab" application. A portable device using the USB spectrophotometer, micro laser (355 nm Nd:YAG, 0,5 ns pulse, repetition rate 10 kHz, output power 15 mW), three channel optical fiber and computer with diagnostic program was designed and ready for clinical trial to be used for cytology and biopsy specimen on site diagnostics, and for the endoscopy/puncture procedures. The biopsy and cytology samples, as well as intervertebral disc specimen were evaluated by pathology experts and the fluorescence spectra were investigated in the fresh and preserved specimens. The spectra were recorded in the spectral range 350-900 nm. At the initial stage the Gaussian components of spectra were found and the Mann-Whitney test was used for the groups' differentiation and the spectral regions for optimal diagnostics purpose were found. Then a formal dividing of spectra in the components or the definite width bands, where the main difference of the different group spectra was observed, was used to compare these groups. The ROC analysis based diagnostic algorithms were created for medical prognosis. The positive prognostic values and negative prediction values were determined for cervical Liquid PAP smear supernatant sediment diagnosis of being Cervicitis and Norma versus CIN2+. In a case of intervertebral disc the analysis allows to get the additional information about the disc degeneration status. All these results demonstrated an efficiency of the proposed procedure and the designed device could be tested at the point-of-care site or for

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. A X-Ray Absorption Study on Melting, Double Electron Excitation and Hemerythrin.

    NASA Astrophysics Data System (ADS)

    Zhang, Ke.

    In this study, X-ray absorption spectroscopy is applied to three systems: xenon atoms; an oxygen transport protein--hemerythrin; and mercury impurities in lead metal. An X-ray absorption experiment on xenon gas shows a clear evidence of the electron shaking up (off) from the valence shells when an L-shell electron is excited. A (DELTA)SCF (self consistent field) calculation in the dipole approximation was performed, which used a local exchange potential and the sudden approximation. The calculation shows that the (DELTA)SCF model is qualitatively and semi -quantitatively correct. The lowest lying shake up channel was found to be more than 70 percent of the total double excitation strength. The model was also used to investigate the many-body effect in the EXAFS. Various forms of hemerythrin (Hr) and their model compounds have been measured by EXAFS. The data analysis shows that the oxy-form of Hr is similar to the met-forms. On release of O(,2) in oxy-Hr, the (mu)-oxo bridge with Fe -O distance of 1.8 (ANGSTROM) is converted to a (mu)-OH bridge of 2.0 (ANGSTROM) and the iron-iron distance is changed from 3.24 (ANGSTROM) to 3.50 (ANGSTROM). Accompanying this change is a large increase in vibrational amplitude around the active site. The entropy increase associated with this vibrational change contributes most of the entropy change driving the transition. A new error analysis procedure is introduced to estimate the fitting errors introduced by modeling the unknown with the standards. It is found that the dominant part of the errors was introduced by the modeling itself, namely, the standards used are not quite transferable with the unknowns. The EXAFS on the samples of Hg impurities in Pb and pure Pb have been measured in the temperature range from 10 K to right below the melting temperatures. The measurement focuses on the local structure of both lead and mercury atoms, which is an advantage provided by the technique. The Debye-Waller factor (DWF) change exhibits

  6. 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

  7. Photoisomerization dynamics of azobenzene in solution with S1 excitation: a femtosecond fluorescence anisotropy study.

    PubMed

    Chang, Chih-Wei; Lu, Ying-Chih; Wang, Tsai-Te; Diau, Eric Wei-Guang

    2004-08-18

    Measurements of anisotropy of femtosecond fluorescence after direct excitation of the S1(n,pi*) state of azobenzene in hexane and ethylene glycol solutions have been carried out to address the controversy about inversion and rotation in the mechanism of photoisomerization. The observed anisotropies in hexane decay to a nonzero asymptotic level with a relaxation period the same as that for slow decay of the corresponding biexponential transient; this effect is attributed to involvement of the out-of-plane CNNC-torsional motion on approach to a twisted conical intersection along the "rotation channel" that depolarizes the original in-plane transition moment. In contrast, when the rotational channel becomes substantially hindered in ethylene glycol, the anisotropies show no discernible decay feature, but the corresponding transients show prominent decays attributed to involvement of in-plane symmetric motions; the latter approach a planar-sloped conical intersection along a "concerted inversion channel" for efficient internal conversion through vibronic coupling. The proposed mechanism is consistent with theoretical calculations and rationalizes both results on quantum yields and ultrafast observations.

  8. 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.

  9. Finding the Elusive Iodocarbene: Fluorescence Excitation and Single Vibronic Level Emission Spectroscopy of Chi

    NASA Astrophysics Data System (ADS)

    Tao, C.; Ebben, C.; Ko, H. T.; Reid, S. A.; Wang, Z.; Sears, T. J.

    2009-06-01

    Among the triatomic halocarbenes, only the iodocarbenes remain to be characterized. The search for these elusive species is motivated by a controversy regarding the multiplicity of the ground state. Photoelectron spectra of Lineberger and co-workers suggest a triplet ground state for CHI, at variance with recent ab initio studies, which suggest a singlet ground state with a singlet-triplet gap of around 4 kcal mol^{-1}. In this work, we have succeeded in finding the spectra of CHI and its deuterated isotopomer using pulsed discharge jet spectroscopy. Rotationally resolved fluorescence excitation spectra are consistent with a singlet-singlet transition, and the derived rotational constants are in good agreement with theoretical predictions. Single vibronic level emission spectra confirm a singlet multiplicity for the ground state, and reveal extensive mixing of the singlet and triplet levels at higher energy. We are able to set a lower limit on the singlet-triplet gap of 4.1 kcal mol^{-1}, in excellent agreement with theory. Extrapolation of the observed bending levels for CHI and CDI to a common origin suggests that the origin of the A^{1}A^'' state lies near 10 500 cm^{-1}, and we will report on high resolution measurements near the electronic origin made at Brookhaven National Laboratory. M. K. Gilles, K. M. Ervin, J. Ho, and W. C. Lineberger, J. Phys. Chem. 96, 1130 (1992).

  10. Two-photon excitation fluorescence imaging of the living juxtaglomerular apparatus.

    PubMed

    Peti-Peterdi, János; Morishima, Shigeru; Bell, P Darwin; Okada, Yasunobu

    2002-07-01

    Recently, multiphoton excitation fluorescence microscopy has been developed that offers important advantages over confocal imaging, particularly for in vivo visualization of thick tissue samples. We used this state-of-the-art technique to capture high-quality images and study the function of otherwise inaccessible cell types and complex cell structures of the juxtaglomerular apparatus (JGA) in living preparations of the kidney. This structure has multiple cell types that exhibit a complex array of functions, which regulate the process of filtrate formation and renal hemodynamics. We report, for the first time, on high-resolution three-dimensional morphology and Z-sectioning through isolated, perfused kidney glomeruli, tubules, and JGA. Time-series images show how alterations in tubular fluid composition cause striking changes in single-cell volume of the unique macula densa tubular epithelium in situ and how they also affect glomerular filtration through alterations in associated structures within the JGA. In addition, calcium imaging of the glomerulus and JGA demonstrates the utility of this system in capturing the complexity of events and effects that are exerted by the specific hypertensive autacoid angiotensin II. This imaging approach to the study of isolated, perfused live tissue with multiphoton microscopy may be applied to other biological systems in which multiple cell types form a functionally integrated syncytium.

  11. Absorption, fluorescence, and acid-base equilibria of rhodamines in micellar media of sodium dodecyl sulfate

    NASA Astrophysics Data System (ADS)

    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 pKaapp = 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.

  12. Advanced glycation end products in hemodialysates as fluorescent and optical absorption markers of patients mortality

    NASA Astrophysics Data System (ADS)

    Kuznetsov, A.; Frorip, A.; Maiste, A.; Ots-Rosenberg, M.; Sünter, A.; Sablonin, J.; Vasil'chenko, J.

    2014-10-01

    Hemodialysate (HD) samples collected from the end stage renal disease patients (ESRD Pts) were used for search for possible correlation between the intensity of HD visible auto-fluorescence (VF) detected at 420 nm as well as their optical absorption at 320 nm and the mortality events among the Pts. Previous but strongly promising correlations has been found in both cases which deserve further supplementation and examination. Investigation of possible influence of quenchers onto the VF intensity has been carried out. Endogenous inorganic ions present in biological fluids (serum, urine and HD) (Na, K, Ca, Mg and ammonia) do not affect the VF intensity remarkably but exogenous Al ions do that indirectly and specifically. Carbon based entities (nanoparticles of graphene type, humins) quench the VF effectively according to the Stern-Volmer law. The quenching phenomena and influence of aluminium must be taken into account by the further investigations, medical care and nutrition.

  13. 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.

  14. 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.

  15. Base pairing enhances fluorescence and favors cyclobutane dimer formation induced upon absorption of UVA radiation by DNA.

    PubMed

    Banyasz, Akos; Vayá, Ignacio; Changenet-Barret, Pascale; Gustavsson, Thomas; Douki, Thierry; Markovitsi, Dimitra

    2011-04-13

    The photochemical properties of the DNA duplex (dA)(20)·(dT)(20) are compared with those of the parent single strands. It is shown that base pairing increases the probability of absorbing UVA photons, probably due to the formation of charge-transfer states. UVA excitation induces fluorescence peaking at ∼420 nm and decaying on the nanosecond time scale. The fluorescence quantum yield, the fluorescence lifetime, and the quantum yield for cyclobutane dimer formation increase upon base pairing. Such behavior contrasts with that of the UVC-induced processes.

  16. 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.

  17. Calculation of Vibrational and Electronic Excited-State Absorption Spectra of Arsenic-Water Complexes Using Density Functional Theory

    DTIC Science & Technology

    2016-06-03

    of Arsenic- Water Complexes Using Density Functional Theory June 3, 2016 Approved for public release; distribution is unlimited. L. Huang S.g...NUMBER OF PAGES 17. LIMITATION OF ABSTRACT Calculation of Vibrational and Electronic Excited-State Absorption Spectra of Arsenic- Water Complexes Using... water molecules should be associated with response features that are intermediate between that of isolated molecules and that of a bulk system. DFT and

  18. Heavy Metals Effect on Cyanobacteria Synechocystis aquatilis Study Using Absorption, Fluorescence, Flow Cytometry, and Photothermal Measurements

    NASA Astrophysics Data System (ADS)

    Dudkowiak, A.; Olejarz, B.; Łukasiewicz, J.; Banaszek, J.; Sikora, J.; Wiktorowicz, K.

    2011-04-01

    The toxic effect of six heavy metals on cyanobacteria Synechocystis aquatilis was studied by absorption, fluorescence, flow cytometry, and photothermal measurements. This study indicates that at the concentration used, the cyanobacteria are more sensitive to silver, copper, and mercury than to cadmium, lead, and zinc metals. Disregarding the decrease in the yields of the related radiative processes caused by photochemical processes and/or damage to phycobilisomes, no changes were detected in the efficiency of thermal deactivation processes within a few microseconds, which can indicate the lack of disturbances in the photosynthetic light reaction and the lack of damage to the photosystem caused by the heavy metal ions in the concentrations used. The results demonstrate that the relative values of fluorescence yield as well as promptly generated heat calculated for the metal-affected and unaffected (reference) bacteria are sensitive indicators of environmental pollution with heavy metal ions, whereas the complementary methods proposed could be used as a noninvasive and fast procedure for in vivo assessment of their toxicity.

  19. 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.

  20. 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.

  1. [Three-dimensional excitation emission matrix fluorescence spectroscopic characterization of dissolved organic matter in water of coal-mining area].

    PubMed

    Yang, Ce; Zhong, Ning-Ning; Shui, Yu-Lei; Wang, Fei-Yu; Chen, Dang-Yi

    2008-01-01

    Three-dimensional excitation emission matrix was applied to characterize the fluorescence properties of dissolved organic matter in various waters of Shilong coal-mining area. Fluorescence peak I (fulvic-like) and peak II (humic-like) were strong, while peak IV and peak V (protein-like) were weak or even undetected in some samples. Fluorescence peaks in various waters and different zones showed great difference in intensities and the fluorescence peaks in underground water tended to be much lower than those of surface waters. Furthermore, the fluorescence peaks of rivers and lakes were higher than those of mine drainage, and also the fluorescence peaks in coking zone and coal mining zone were higher than those in sewage-irrigated zone, or even much higher than those in farming zone. The reason may be that coal mining activities and coal industry can bring plenty of organic matter from coal to surroundings. Meanwhile, surface water would accept mine drainage, waste water of coal-washing and sewage from daily life easier than underground water, so surface water can be polluted seriously. Fluorescence peaks in waters from coal mining area are little influenced by pH of the water but can be influenced by the content of Ca2+ to water in some extent.

  2. Multiphoton molecular spectroscopy and excited-state dynamics of enhanced green fluorescent protein (EGFP): acid base specificity

    NASA Astrophysics Data System (ADS)

    Heikal, Ahmed A.; Hess, Samuel T.; Webb, Watt W.

    2001-12-01

    Green fluorescent protein (GFP), isolated from Aequorea victoria jellyfish, has been used extensively as a noninvasive intracellular pH indicator and site-specific fluorescent marker in biochemistry, cell biology, and molecular genetics. Numerous mutations, aimed at optimizing spectroscopic and thermodynamic properties of GFP, have been created for different applications. Fluorescence correlation spectroscopy (FCS) reveals that the enhanced green fluorescent protein mutant (EGFP; S65T/F64L) undergoes external proton exchange with the buffer on ˜45-300 μs time scale with p Ka=5.8±0.1 [Proc. Natl. Acad. Sci. USA 95 (1998) 13573]. This contribution represents a comprehensive characterization of pH and excitation mode (wavelength, one and two photon (2P)) effects on the spectroscopy, excited-state dynamics, and rotational mobility of EGFP aiming at elucidating the significant electronic states of this molecular system. EGFP exhibits a large 2P action cross-section and, therefore, is well suited for intracellular imaging using 2P fluorescence microscopy.

  3. Triplet excited state distortions in a pyrazolate bridged platinum dimer measured by X-ray transient absorption spectroscopy.

    PubMed

    Lockard, Jenny V; Rachford, Aaron A; Smolentsev, Grigory; Stickrath, Andrew B; Wang, Xianghuai; Zhang, Xiaoyi; Atenkoffer, Klaus; Jennings, Guy; Soldatov, Alexander; Rheingold, Arnold L; Castellano, Felix N; Chen, Lin X

    2010-12-09

    The excited-state structure of a dinuclear platinum(II) complex with tert-butyl substituted pyrazolate bridging units, [Pt(ppy)(μ-(t)Bu(2)pz)](2) (ppy = 2-phenylpyridine; (t)Bu(2)pz = 3,5-di-tert-butylpyrazolate) is studied by X-ray transient absorption (XTA) spectroscopy to reveal the transient electronic and nuclear geometry. DFT calculations predict that the lowest energy triplet excited state, assigned to a metal-metal-to-ligand charge transfer (MMLCT) transition, has a contraction in the Pt-Pt distance. The Pt-Pt bond length and other structural parameters extracted from fitting the experimental XTA difference spectra from full multiple scattering (FMS) and multidimensional interpolation calculations indicates a metal-metal distance decrease by approximately 0.2 Å in the triplet excited state. The advantages and challenges of this approach in resolving dynamic transient structures of nonbonding or weak-bonding dinuclear metal complexes in solution are discussed.

  4. Femtosecond excited-state absorption dynamics and optical limiting in fullerene solutions, sol-gel glasses, and thin films

    SciTech Connect

    McBranch, D.; Klimov, V.; Smilowitz, L.; Wang, H.; Wudl, F.

    1996-11-01

    We compare detailed dynamics of the excited-state absorption for C{sub 60} in solution, thin films, and entrapped in an inorganic sol-gel glass matrix. Our results demonstrate that the microscopic morphology of the C{sub 60} molecule plays a crucial role in determining the relaxation dynamics. This is a key factor for applications in optical limiting for nanosecond pulses using reverse saturable absorption. We find that the dynamics of the C{sub 60}-glass composite occur on long (ns) timescales, comparable to that in solution; thin film samples, by contrast, show rapid decay (<20 picoseconds). These results demonstrate that the C{sub 60}-sol-gel glass composites contain C{sub 60} in a molecular dispersion, and are suitable candidates for solid-state optical limiting. Multispectral analysis of the decay dynamics in solution allows accurate determination of both the intersystem crossing time (600 {+-}100 ps) and the relative strengths of the singlet and triplet excited-state cross sections as a function of wavelength from 450-950 nm. The triplet excited-state cross section is greater than that for the singlet excited-state over the range from 620-810 nm.

  5. 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.

  6. 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.

  7. Kinetics of excited states of pigment clusters in solubilized light-harvesting complex II: photon density-dependent fluorescence yield and transmittance.

    PubMed Central

    Schödel, R; Hillmann, F; Schrötter, T; Voigt, J; Irrgang, K D; Renger, G

    1996-01-01

    Relative fluorescence yield, phi F, and transmittance, T, were measured in solubilized light-harvesting complex II (LHCII) as a function of photon density, Ip, of monochromatic 645-nm laser pulses (duration: approximately 2.5 ns). Special efforts were made in constructing an optical set-up that allows the accurate determination of the fluorescence from an area of constant Ip, phi F(Ip) starts to decline at approximately 10(14) and drops to values below 0.01% at maximum Ip (approximately 10(19) photons cm-2 pulse-1). T(Ip) decreases only slightly at photon densities of approximately 10(15) but increases steeply at values of > 10(17) photons cm-2 pulse-1. The interpretation of the phi F(Ip) data using the saturation limit of Mauzerall's multiple hit model leads to a unit size of about 10-15 chlorophyll molecules. One interpretation is to attribute this result to a very fast exciton-exciton annihilation of multiple excited states generated within this small domain. Alternatively, based on the assumption that delocalized cluster states within the monomeric/trimeric subunit of LHCII exist, the results can be consistently described by a kinetic model comprising ground, monoexcitonic, and biexcitonic states of clusters and a triplet state that is quenched by carotenoids in LHCII. Within the framework of this model the annihilation of multiple excitations is explained as ultrafast radiationless relaxation of higher excited cluster states. Comparative measurements in diluted acetonic Chl a solution are consistently described by the depletion of the ground state, taking the absorption cross section at the used wavelength. Images FIGURE 1 PMID:8968606

  8. 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.

  9. Bright blue-shifted fluorescent proteins with Cys in the GAF domain engineered from bacterial phytochromes: fluorescence mechanisms and excited-state dynamics

    PubMed Central

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

    2016-01-01

    Near-infrared fluorescent proteins (NIR FPs) engineered from bacterial phytochromes (BphPs) are of great interest for in vivo imaging. They utilize biliverdin (BV) as a chromophore, which is a heme degradation product, and therefore they are straightforward to use in mammalian tissues. Here, we report on fluorescence properties of NIR FPs with key alterations in their BV binding sites. BphP1-FP, iRFP670 and iRFP682 have Cys residues in both PAS and GAF domains, rather than in the PAS domain alone as in wild-type BphPs. We found that NIR FP variants with Cys in the GAF or with Cys in both PAS and GAF show blue-shifted emission with long fluorescence lifetimes. In contrast, mutants with Cys in the PAS only or no Cys residues at all exhibit red-shifted emission with shorter lifetimes. Combining these results with previous biochemical and BphP1-FP structural data, we conclude that BV adducts bound to Cys in the GAF are the origin of bright blue-shifted fluorescence. We propose that the long fluorescence lifetime follows from (i) a sterically more constrained thioether linkage, leaving less mobility for ring A than in canonical BphPs, and (ii) that π-electron conjugation does not extend on ring A, making excited-state deactivation less sensitive to ring A mobility. PMID:27857208

  10. Bright blue-shifted fluorescent proteins with Cys in the GAF domain engineered from bacterial phytochromes: fluorescence mechanisms and excited-state dynamics

    NASA Astrophysics Data System (ADS)

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

    2016-11-01

    Near-infrared fluorescent proteins (NIR FPs) engineered from bacterial phytochromes (BphPs) are of great interest for in vivo imaging. They utilize biliverdin (BV) as a chromophore, which is a heme degradation product, and therefore they are straightforward to use in mammalian tissues. Here, we report on fluorescence properties of NIR FPs with key alterations in their BV binding sites. BphP1-FP, iRFP670 and iRFP682 have Cys residues in both PAS and GAF domains, rather than in the PAS domain alone as in wild-type BphPs. We found that NIR FP variants with Cys in the GAF or with Cys in both PAS and GAF show blue-shifted emission with long fluorescence lifetimes. In contrast, mutants with Cys in the PAS only or no Cys residues at all exhibit red-shifted emission with shorter lifetimes. Combining these results with previous biochemical and BphP1-FP structural data, we conclude that BV adducts bound to Cys in the GAF are the origin of bright blue-shifted fluorescence. We propose that the long fluorescence lifetime follows from (i) a sterically more constrained thioether linkage, leaving less mobility for ring A than in canonical BphPs, and (ii) that π-electron conjugation does not extend on ring A, making excited-state deactivation less sensitive to ring A mobility.

  11. 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

  12. Minimally invasive imaging method based on second harmonic generation and multiphoton excitation fluorescence in translational respiratory research.

    PubMed

    Abraham, Thomas; Wadsworth, Samuel; Carthy, Jon M; Pechkovsky, Dmitri V; McManus, Bruce

    2011-01-01

    For translational respiratory research including in the development of clinical diagnostic tools, a minimally invasive imaging method, which can provide both cellular and extracellular structural details with sufficient specificity, sensitivity and spatial resolution, is particularly useful. Multiphoton microscopy causes excitation of endogenously fluorescent macromolecular systems and induces highly specific second harmonic generation signals from non-centrosymmetric macromolecules such as fibrillar collagens. Both these signals can be captured simultaneously to provide spatially resolved 3D structural organization of extracellular matrix as well as the cellular morphologies in their native states. Besides briefly discussing the fundamentals of multiphoton excitation fluorescence and harmonic generation signals and the instrumentation details, this review focuses on the specific applications of these imaging modalities in lung structural imaging, particularly morphological features of alveolar structures, visualizing and quantifying extracellular matrix remodelling accompanying emphysematous destructions as well as the IPF, detecting lung cancers and the potential use in the tissue engineering applications.

  13. 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.

  14. 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.

  15. Probing CuI in Homogeneous Catalysis using High-Energy-Resolution Fluorescence-Detected X-ray Absorption Spectroscopy

    PubMed Central

    Walroth, Richard C.; Uebler, Jacob W. H.

    2015-01-01

    Metal-to-ligand charge transfer excitations in CuI 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. PMID:25994112

  16. Combined system for high-time-resolution dual-excitation fluorescence photometry and fluorescence imaging of calcium transients in single normal and diseased skeletal muscle fibers

    NASA Astrophysics Data System (ADS)

    Uttenweiler, Dietmar; Wojciechowski, Reinhold; Makabe, Makoto; Veigel, Claudia; Fink, Rainer H.

    1994-12-01

    Fast photometric measurements and video-imaging of fluorescent indicators both are powerful tools in measuring the intracellular free calcium concentration of muscle and many other cells. as photometric systems yield a high temporal resolution, calcium imaging systems have high spatial but significantly reduced temporal resolution. Therefore we have developed an integrated system combining both methods and based mostly on standard components. As a common, sensitive Ca2+- indicator we used the fluorescent probe Fura-2, which is alternatingly excited for ratio measurements at 340/380 nm. We used a commercially available dual excitation photometric system (OSP-3; Olympus) for attaching a CCD-camera and a frame grabber board. To achieve the synchronization we had to design circuitries for external triggering, synchronization and accurate control of the filter changer, which we added to the system. Additionally, the software for a triggered image acquisition was developed. With this integrated setup one can easily switch between the fast photometric mode (ratio frequency 100 Hz) and the imaging mode (ratio frequency 4.17 Hz). The calcium images are correlated with the 25 times faster spot measurements and are analyzed by means of image processing. With this combined system we study release and uptake of calcium ions of normal and diseased skeletal muscle from mdx mice. Such a system will also be important for other cellular studies in which fluorescence indicators are used to monitor similar time dependent alterations as well as changes in cellular distributions of calcium.

  17. Intravital excitation increases detection sensitivity for pulmonary tuberculosis by whole-body imaging with β-lactamase reporter enzyme fluorescence.

    PubMed

    Nooshabadi, Fatemeh; Yang, Hee-Jeong; Cheng, Yunfeng; Durkee, Madeleine S; Xie, Hexin; Rao, Jianghong; Cirillo, Jeffrey D; Maitland, Kristen C

    2016-10-18

    Tuberculosis is a pulmonary disease with an especially high mortality rate in immuno-compromised populations, specifically children and HIV positive individuals. The causative agent, Mycobacterium tuberculosis (Mtb), is a very slow growing and difficult organism to work with, making both diagnosis and development of effective treatments cumbersome. We utilize a fiber-optic fluorescence microendoscope integrated with a whole-body imaging system for in vivo Mtb detection. The system exploits an endogenous enzyme of Mtb (β-lactamase, or BlaC) using a BlaC-specific NIR fluorogenic substrate. In the presence of BlaC, this substrate is cleaved and becomes fluorescent. Using intravital illumination of the lung to excite this probe, sensitivity of the optical system increases over trans- and epi-illumination methods of whole-body fluorescence imaging. We demonstrate that integration of these imaging technologies with BlaC-specific fluorescent reporter probe improves the level of detection to ∼100 colony forming units, a 100× increase in sensitivity in comparison to epi-illumination and a 10× increase in sensitivity in comparison to previous work in intravital excitation of tdTomato-expressing Mtb. This lower detection threshold enables the study of early stage bacterial infections with clinical strains of Mtb and longitudinal studies of disease pathogenesis and therapeutic efficacy with multiple time points in a single animal.

  18. Mechanism for optical enhancement and suppression of fluorescence.

    PubMed

    Bradshaw, David S; Andrews, David L

    2009-06-18

    When fluorescence from electronically excited states follows the absorption of radiation, the emission spectrum is often a key to identification of the excited species. It now emerges that passing off-resonant laser light through such an electronically excited system may enhance or suppress the fluorescent emission. This report establishes the mechanism and theory for this optical control of spontaneous fluorescence, derived by quantum electrodynamical analysis. Experimental techniques to detect the enhanced signal are also proposed.

  19. Simulating the absorption spectra of helium clusters (N = 70, 150, 231, 300) using a charge transfer correction to superposition of fragment single excitations.

    PubMed

    Ge, Qinghui; Mao, Yuezhi; White, Alec F; Epifanovsky, Evgeny; Closser, Kristina D; Head-Gordon, Martin

    2017-01-28

    Simulations of the n = 2 absorption spectra of HeN (N = 70, 150, 231, 300) clusters are reported, with nuclear configurations sampled by path integral molecular dynamics. The electronic structure is treated by a new approach, ALMO-CIS+CT, which is a formulation of configuration interaction singles (CIS) based on absolutely localized molecular orbitals (ALMOs). The method generalizes the previously reported ALMO-CIS model [K. D. Closser et al. J. Chem. Theory Comput. 11, 5791 (2015)] to include spatially localized charge transfer (CT) effects. It is designed to recover large numbers of excited states in atomic and molecular clusters, such as the entire n = 2 Rydberg band in helium clusters. ALMO-CIS+CT is shown to recover most of the error caused by neglecting charge transfer in ALMO-CIS and has comparable accuracy to standard CIS for helium clusters. For the n = 2 band, CT stabilizes states towards the blue edge by up to 0.5 eV. ALMO-CIS+CT retains the formal cubic scaling of ALMO-CIS with respect to system size. With improvements to the implementation over that originally reported for ALMO-CIS, ALMO-CIS+CT is able to treat helium clusters with hundreds of atoms using modest computing resources. A detailed simulation of the absorption spectra associated with the 2s and 2p bands of helium clusters up to 300 atoms is reported, using path integral molecular dynamics with a spherical boundary condition to generate atomic configurations at 3 K. The main features of experimentally reported fluorescence excitation spectra for helium clusters are reproduced.

  20. Simulating the absorption spectra of helium clusters (N = 70, 150, 231, 300) using a charge transfer correction to superposition of fragment single excitations

    NASA Astrophysics Data System (ADS)

    Ge, Qinghui; Mao, Yuezhi; White, Alec F.; Epifanovsky, Evgeny; Closser, Kristina D.; Head-Gordon, Martin

    2017-01-01

    Simulations of the n = 2 absorption spectra of HeN (N = 70, 150, 231, 300) clusters are reported, with nuclear configurations sampled by path integral molecular dynamics. The electronic structure is treated by a new approach, ALMO-CIS+CT, which is a formulation of configuration interaction singles (CIS) based on absolutely localized molecular orbitals (ALMOs). The method generalizes the previously reported ALMO-CIS model [K. D. Closser et al. J. Chem. Theory Comput. 11, 5791 (2015)] to include spatially localized charge transfer (CT) effects. It is designed to recover large numbers of excited states in atomic and molecular clusters, such as the entire n = 2 Rydberg band in helium clusters. ALMO-CIS+CT is shown to recover most of the error caused by neglecting charge transfer in ALMO-CIS and has comparable accuracy to standard CIS for helium clusters. For the n = 2 band, CT stabilizes states towards the blue edge by up to 0.5 eV. ALMO-CIS+CT retains the formal cubic scaling of ALMO-CIS with respect to system size. With improvements to the implementation over that originally reported for ALMO-CIS, ALMO-CIS+CT is able to treat helium clusters with hundreds of atoms using modest computing resources. A detailed simulation of the absorption spectra associated with the 2s and 2p bands of helium clusters up to 300 atoms is reported, using path integral molecular dynamics with a spherical boundary condition to generate atomic configurations at 3 K. The main features of experimentally reported fluorescence excitation spectra for helium clusters are reproduced.

  1. Application of the Kubelka-Munk correction for self-absorption of fluorescence emission in carmine lake paint layers.

    PubMed

    Clementi, Catia; Miliani, Costanza; Verri, Giovanni; Sotiropoulou, Sophia; Romani, Aldo; Brunetti, Brunetto G; Sgamellotti, A

    2009-12-01

    The variations of the fluorescence emission of carmine lake travelling through an absorbing and scattering medium, such as a paint layer, were investigated by ultraviolet (UV)-visible absorption, fluorescence spectroscopy, and imaging techniques. Samples of the lake were studied in dilute and saturated solutions, on a reference test panel and a real case study. Relevant spectral modifications have been observed as a function of the lake concentration mainly consisting of a fluorescence quenching, red shift of emission maxima, and deformation of emission band. The application of a correction factor based on the Kubelka-Munk model allowed fluorescence spectra obtained in solution and on painted samples of known composition to be compared and correlated, highlighting that the fluorescence of the lake within paint layers is affected by both self-absorption and aggregation phenomena. This approach has been successfully applied on a painting by G. Vasari for the noninvasive identification of carmine lake. The results reported here emphasize the necessity of taking physical phenomena into account in the interpretation of the fluorescence spectra for a proper and reliable characterization and identification of painting materials in works of art.

  2. Solvent-induced configuration mixing and triplet excited-state inversion: insights from transient absorption and transient dc photoconductivity measurements.

    PubMed

    She, Chunxing; Rachford, Aaron A; Wang, Xianghuai; Goeb, Sébastien; El-Ballouli, Ala'a O; Castellano, Felix N; Hupp, Joseph T

    2009-10-14

    Solvent-induced excited-state configuration mixing in a Pt(II) diimine chromophore with phenylene ethynylene containing acetylide ligands, [Pt((t)Bu2bpy)(PE3)2] (1), was characterized by nanosecond transient absorption spectroscopy and transient dc photoconductivity (TDCP). The mixing is a result of closely spaced triplet charge transfer (3CT) and intraligand-localized (3IL) triplet energy levels that are finely tuned with solvent polarity as ascertained by their parent model chromophores [Pt((t)Bu2bpy)(PE1)2] (2) and [Pt(P2)(PE3)2] (3), respectively. The absorption difference spectrum of the mixed triplet state is dramatically different from those of the 3CT and 3IL state model chromophores. The 3CT, 3IL and configuration-mixed triplet states led to distinct TDCP signals. The TDCP response is of negative polarity for 3CT excited states but of positive polarity for 3IL excited states. TDCP transients for 1 in mixed solvents are a combination of signals from the 3IL and 3CT states, with the signal magnitude depending on the polarity of solvent composition. The fraction of 3CT state character in the configurationally mixed excited state was quantified by TDCP to be approximately 0.24 in pure benzene, while it decreased to approximately 0.05 in 20 : 80 (v : v) benzene-CH2Cl2. The charge transfer fraction appears to increase slightly to approximately 0.11 in the lower polarity 20 : 80 n-hexane-CH2Cl2 medium. TDCP is shown to be a useful tool for the identification of the lowest excited state in electrically neutral metal-organic chromophores.

  3. Dual-wavelength excitation for fluorescence-based quantification of zinc protoporphyrin IX and protoporphyrin IX in whole blood.

    PubMed

    Hennig, Georg; Gruber, Christian; Vogeser, Michael; Stepp, Herbert; Dittmar, Stephan; Sroka, Ronald; Brittenham, Gary M

    2014-07-01

    Quantification of erythrocyte zinc protoporphyrin IX (ZnPP) and protoporphyrin IX (PPIX), individually or jointly, is useful for the diagnostic evaluation of iron deficiency, iron-restricted erythropoiesis, lead exposure, and porphyrias. A method for simultaneous quantification of ZnPP and PPIX in unwashed blood samples is described, using dual-wavelength excitation to effectively eliminate background fluorescence from other blood constituents. In blood samples from 35 subjects, the results of the dual-wavelength excitation method and a reference high performance liquid chromatography (HPLC) assay were closely correlated both for ZnPP (rs = 0.943, p < 0.0001; range 37-689 μmol ZnPP/mol heme, 84-1238 nmol/L) and for PPIX (rs = 0.959, p < 0.0001; range 42-4212 μmol PPIX/mol heme, 93-5394 nmol/L). In addition, for ZnPP, the proposed method is compared with conventional single-wavelength excitation and with commercial front-face fluorimetry of washed erythrocytes and whole blood. We hypothesize that dual-wavelength excitation fluorimetry will provide a new approach to the suppression of background fluorescence in blood and tissue measurements of ZnPP and PPIX.

  4. Excited-state intramolecular proton transfer as a fluorescence probe for protein binding-site static polarity.

    PubMed Central

    Sytnik, A; Kasha, M

    1994-01-01

    A fluorescence probe is introduced for protein conformation and binding-site monitoring as the proton-transfer (PT) tautomer fluorescence by using 4-hydroxy-5-azaphenanthrene (HAP) as a prototype. A typical grossly-wavelength-shifted PT fluorescence for HAP is observed in the 600-nm spectral region for this UV-absorbing molecule (absorption onset, 400 nm), for which case PT occurs even in protic solvents. It is shown that PT fluorescence of HAP can serve as a protein-binding-site static-polarity calibrator, shifting from a lambda max of 612 nm in cyclohexane to 585 nm in ethanol at 298 K, contrary to the usual dispersion red shift. A small mechanical solvent-cage effect is noted in ethanol at 77 K, but solvent dielectric relaxation is not apparent from the fluorescence spectrum. Thus, HAP serves to distinguish static solvent-cage polarity from dynamical solvent dielectric relaxation and other solvent-cage effects (mechanical restriction of molecular conformation). HAP as a PT-fluorescence probe is applied to human serum albumin (HSA) and beaver apomyoglobin. PMID:8078934

  5. 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.

  6. Mini-Sized Carbon Nitride Nanosheets with Double Excitation- and pH-Dependent Fluorescence Behaviors for Two-Photon Cell Imaging.

    PubMed

    Liu, Jinhua; Zhao, Duoduo; Li, Lin; Weng, Minrui; Zhang, Chengwu; Zhang, Shiyu; Zhu, Jixin; Feng, Yong; Shih, Kaimin; Huang, Wei

    2017-02-26

    Synthesis of mini-sized carbon nitride nanosheets (CNNSs) by traditional methods remains a challenge. Herein, size-tunable and uniform mini-sized CNNSs are synthesized by hydrothermal carbonization of a single polyethyleneimine (PEI) precursor. The as-obtained mini-sized CNNSs possess uniform size, good hydrophilicity and abundant nitrogen active sites, which not only exhibit double excitation- and pH-dependent fluorescence behaviors, but also two-photon excitation fluorescence. áThe resulting CNNSs display low toxicity and can be efficiently delivered into live cells for two-photon fluorescence imaging, offering great potential as fluorescence probes in biochemical applications.

  7. The equipment for time-resolved measurements of excitation-emission matrix of seawater fluorescence in natural conditions

    NASA Astrophysics Data System (ADS)

    Salyuk, Pavel A.; Nagorny, Ivan G.; Mayor, Alexander A.; Krikun, Vladimir A.; Shmirko, Konstantin A.

    2016-10-01

    Excitation-emission matrix (EEM) technique is widely used to study the properties of organic matter in seawater. Additional variations of the exciting radiation parameters (in addition to the wavelengths), such as duration and intensity, allow to get more spectral information about the object of research, and allow to identify some individual components or to study the dynamics of the molecular changes in the sample of analyzed liquid. One of the main limitations of the EEM is that usually necessary to carry out research in the laboratory on the specialized equipment. The paper describes the hardware system allowing carry out measurements in the field studies. The hardware system includes a set of light-emitted diodes (LEDs) working in the range from 245 to 600 nm, which produce sequential excitation fluorescence of the sample liquid, which is detected using a 32-channel photomultiplier tube (PMT). Number of LEDs can vary from 7 to 16. Through the use of multi-channel photomultiplier reached the required measurement sensitivity and efficiency measurement is less than 1 minute for full cycle. Minimum 4 channel in excitation range of 350-600 nm support the work of variable duration excitation pulses from microseconds to seconds. It makes possible to measure the fluorescence of the chlorophyll-a with closed or open reaction centers of phytoplankton cells. The use of multiple excitation radiation allows to activate various mechanisms of energy transfer in the photosynthetic apparatus of phytoplankton cells, improves measurement accuracy, reduces dependence on the variation in species composition and functional state of phytoplankton.

  8. 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.

  9. Acceptor and Excitation Density Dependence of the Ultrafast Polaron Absorption Signal in Donor-Acceptor Organic Solar Cell Blends.

    PubMed

    Zarrabi, Nasim; Burn, Paul L; Meredith, Paul; Shaw, Paul E

    2016-07-21

    Transient absorption spectroscopy on organic semiconductor blends for solar cells typically shows efficient charge generation within ∼100 fs, accounting for the majority of the charge carriers. In this Letter, we show using transient absorption spectroscopy on blends containing a broad range of acceptor content (0.01-50% by weight) that the rise of the polaron signal is dependent on the acceptor concentration. For low acceptor content (<10% by weight), the polaron signal rises gradually over ∼1 ps with most polarons generated after 200 fs, while for higher acceptor concentrations (>10%) most polarons are generated within 200 fs. The rise time in blends with low acceptor content was also found to be sensitive to the pump fluence, decreasing with increasing excitation density. These results indicate that the sub-100 fs rise of the polaron signal is a natural consequence of both the high acceptor concentrations in many donor-acceptor blends and the high excitation densities needed for transient absorption spectroscopy, which results in a short average distance between the exciton and the donor-acceptor interface.

  10. 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.

  11. 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.

  12. Atomic resolution mapping of the excited-state electronic structure of Cu2O with time-resolved x-ray absorption spectroscopy

    NASA Astrophysics Data System (ADS)

    Hillyard, P. W.; Kuchibhatla, S. V. N. T.; Glover, T. E.; Hertlein, M. P.; Huse, N.; Nachimuthu, P.; Saraf, L. V.; Thevuthasan, S.; Gaffney, K. J.

    2009-09-01

    We have used time-resolved soft x-ray spectroscopy to investigate the electronic structure of optically excited cuprous oxide at the OK edge and the CuL3 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 Cu3d and O2p orbital contributions. The 2.1 eV optical band gap of Cu2O significantly exceeds the one eV shift in the CuL3 - and OK -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.

  13. 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.

  14. 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