WorldWideScience

Sample records for two-photon induced fluorescence

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

    NARCIS (Netherlands)

    Marinov, D.; Drag, C.; Blondel, C.; Guaitella, O.; Golda, J.; Klarenaar, B.L.M.; Engeln, R.A.H.; Schulz-von der Gathen, V.; Booth, J.-P.

    2016-01-01

    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

  2. Two-photon laser-induced fluorescence studies of HS radicals, DS radicals, and I atoms

    Energy Technology Data Exchange (ETDEWEB)

    Tiee, J J; Ferris, M J; Loge, G W; Wampler, F B

    1983-04-15

    A two-photon laser-induced excitation and fluorescence technique has been used to study the A /sup 2/..sigma../sup +/ - X/sup 2/PI transition of HS and DS radicals and various high-lying /sup 4/P/sup 0/, /sup 2/D/sup 0/, and /sup 4/D/sup 0/ states of the I atom. The two-photon excitation cross sections and detection sensitivity are discussed. 13 references, 5 figures.

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

    Energy Technology Data Exchange (ETDEWEB)

    Elliott, Drew; Scime, Earl; Short, Zachary, E-mail: zdshort@mix.wvu.edu [Department of Physics and Astronomy, West Virginia University, Morgantown, West Virginia 26056 (United States)

    2016-11-15

    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.

  4. Direct measurements of neutral density depletion by two-photon absorption laser-induced fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Aanesland, A.; Liard, L.; Leray, G.; Jolly, J.; Chabert, P.

    2007-01-01

    The ground state density of xenon atoms has been measured by spatially resolved laser-induced fluorescence spectroscopy with two-photon excitation in the diffusion chamber of a magnetized Helicon plasma. This technique allows the authors to directly measure the relative variations of the xenon atom density without any assumptions. A significant neutral gas density depletion was measured in the core of the magnetized plasma, in agreement with previous theoretical and experimental works. It was also found that the neutral gas density was depleted near the radial walls

  5. Two-photon induced fluorescence and other optical effects in irradiated and doped fused silica

    International Nuclear Information System (INIS)

    Kramer, S.D.

    1986-07-01

    The objective of this program was to assess and identify irradiation techniques which could be used to modify the optical charactistics of doped fused silica. Primary emphasis was placed on determining if gamma ray or neutron bombardment of the glass would enhance certain Raman and nonlinear optical effects. In particular, the effect of irradiation on optical two photon induced fluorescence was studied in detail. The maximum radiation exposures used were 10 6 rads (Si) of gamma rays and neutron fluences of 1 x 10 14 neutrons/cm 2 . The optical measurements were made at room temperature between one and four months after irradiation. The maximum input light intensity was 10 9 watts/cm 2 at a near infrared (1.06 μ) input wavelength which was chosen to lie in a transparent spectral region of the glass. Under these experimental conditions a careful search revealed no detectable two-photon induced fluorescence in the region from 550 to 900 nm. The upper limit for the photon efficiency of this process was determined to be less than 1 x 10 -10 %. 89 refs., 12 figs

  6. Femtosecond two-photon laser-induced fluorescence of krypton for high-speed flow imaging.

    Science.gov (United States)

    Wang, Yejun; Capps, Cade; Kulatilaka, Waruna D

    2017-02-15

    Ultrashort-pulse (femtosecond-duration) two-photon laser-induced fluorescence (fs-TPLIF) of an inert gas tracer krypton (Kr) is investigated. A detailed spectroscopic study of fluorescence channels followed by the 5p'←←4p excitation of Kr at 204.1 nm is reported. The experimental line positions in the 750-840 nm emission region agree well with the NIST Atomic Spectra Database. The present work provides an accurate listing of relative line strengths in this spectral region. In the range of laser pulse energies investigated, a quadratic dependence was observed between the Kr-TPLIF signal and the laser pulse energy. The single-laser-shot 2D TPLIF images recorded in an unsteady jet demonstrate the potential of using fs excitation at 204.1 nm for mixing and flow diagnostic studies using Kr as an inert gas tracer.

  7. Two-photon induced fluorescence of Cy5-DNA in buffer solution and on silver island films

    International Nuclear Information System (INIS)

    Lukomska, Joanna; Gryczynski, Ignacy; Malicka, Joanna; Makowiec, Slawomir; Lakowicz, Joseph R.; Gryczynski, Zygmunt

    2005-01-01

    We report the observation of a strong two-photon induced fluorescence emission of Cy5-DNA within the tunable range of a Ti:Sapphire laser. The estimated two-photon cross-section for Cy5-DNA of 400 GM is about 3.5-fold higher than it was reported for rhodamine B. The fundamental anisotropies of Cy5-DNA are close to the theoretical limits of 2/5 and 4/7 for one- and two-photon excitation, respectively. We also observed an enhanced two-photon induced fluorescence (TPIF) of Cy5-DNA deposited on silver island films (SIFs). In the presence of SIFs, the TPIF is about 100-fold brighter. The brightness increase of Cy5-DNA TPIF near SIFs is mostly due to enhanced local field

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

    Energy Technology Data Exchange (ETDEWEB)

    Scime, Earl E. [West Virginia Univ., Morgantown, WV (United States)

    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

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

    International Nuclear Information System (INIS)

    Scime, Earl E.

    2016-01-01

    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 H α 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 installation on their

  10. Imaging hydrogen flames by two-photon, laser-induced fluorescence

    Science.gov (United States)

    Miles, R.; Lempert, W.; Kumar, V.; Diskin, G.

    1991-01-01

    A nonintrusive multicomponent imaging system is developed which can image hydrogen, hot oxygen, and air simultaneously. An Ar-F excimer laser is injection-locked to cover the Q1 two-photon transition in molecular hydrogen which allows the observation of both hot oxygen and cold hydrogen. Rayleigh scattering from the water molecules occurs at the same frequency as the illuminating laser allowing analysis of the air density. Images of ignited and nonignited hydrogen jets are recorded with a high-sensitivity gated video camera. The images permit the analysis of turbulent hydrogen-core jet, the combustion zone, and the surrounding air, and two-dimensional spatial correlations can be made to study the turbulent structure and couplings between different regions of the flow field. The method is of interest to the study of practical combustion systems which employ hydrogen-air diffusion flames.

  11. Femtosecond, two-photon laser-induced-fluorescence imaging of atomic oxygen in an atmospheric-pressure plasma jet

    Science.gov (United States)

    Schmidt, Jacob B.; Sands, Brian L.; Kulatilaka, Waruna D.; Roy, Sukesh; Scofield, James; Gord, James R.

    2015-06-01

    Femtosecond, two-photon-absorption laser-induced-fluorescence (fs-TALIF) spectroscopy is employed to measure space- and time-resolved atomic-oxygen distributions in a nanosecond, repetitively pulsed, externally grounded, atmospheric-pressure plasma jet flowing helium with a variable oxygen admixture. The high-peak-intensity, low-average-energy femtosecond pulses result in increased TALIF signal with reduced photolytic inferences. This allows 2D imaging of absolute atomic-oxygen number densities ranging from 5.8   ×   1015 to 2.0   ×   1012cm-3 using a cooled CCD with an external intensifier. Xenon is used for signal and imaging-system calibrations to quantify the atomic-oxygen fluorescence signal. Initial results highlight a transition in discharge morphology from annular to filamentary, corresponding with a change in plasma chemistry from ozone to atomic oxygen production, as the concentration of oxygen in the feed gas is changed at a fixed voltage-pulse-repetition rate. In this configuration, significant concentrations of reactive oxygen species may be remotely generated by sustaining an active discharge beyond the confines of the dielectric capillary, which may benefit applications that require large concentrations of reactive oxygen species such as material processing or biomedical devices.

  12. Two-photon absorption laser-induced fluorescence of atomic oxygen in the afterglow of pulsed positive corona discharge

    Science.gov (United States)

    Ono, Ryo; Takezawa, Kei; Oda, Tetsuji

    2009-08-01

    Atomic oxygen is measured in the afterglow of pulsed positive corona discharge using time-resolved two-photon absorption laser-induced fluorescence. The discharge occurs in a 14 mm point-to-plane gap in dry air. After the discharge pulse, the atomic oxygen density decreases at a rate of 5×104 s-1. Simultaneously, ozone density increases at almost the same rate, where the ozone density is measured using laser absorption method. This agreement between the increasing rate of atomic oxygen and decreasing rate of ozone proves that ozone is mainly produced by the well-known three-body reaction, O+O2+M→O3+M. No other process for ozone production such as O2(v)+O2→O3+O is observed. The spatial distribution of atomic oxygen density is in agreement with that of the secondary streamer luminous intensity. This agreement indicates that atomic oxygen is mainly produced in the secondary streamer channels, not in the primary streamer channels.

  13. Spatially and Temporally Resolved Atomic Oxygen Measurements in Short Pulse Discharges by Two Photon Laser Induced Fluorescence

    Science.gov (United States)

    Lempert, Walter; Uddi, Mruthunjaya; Mintusov, Eugene; Jiang, Naibo; Adamovich, Igor

    2007-10-01

    Two Photon Laser Induced Fluorescence (TALIF) is used to measure time-dependent absolute oxygen atom concentrations in O2/He, O2/N2, and CH4/air plasmas produced with a 20 nanosecond duration, 20 kV pulsed discharge at 10 Hz repetition rate. Xenon calibrated spectra show that a single discharge pulse creates initial oxygen dissociation fraction of ˜0.0005 for air like mixtures at 40-60 torr total pressure. Peak O atom concentration is a factor of approximately two lower in fuel lean (φ=0.5) methane/air mixtures. In helium buffer, the initially formed atomic oxygen decays monotonically, with decay time consistent with formation of ozone. In all nitrogen containing mixtures, atomic oxygen concentrations are found to initially increase, for time scales on the order of 10-100 microseconds, due presumably to additional O2 dissociation caused by collisions with electronically excited nitrogen. Further evidence of the role of metastable N2 is demonstrated from time-dependent N2 2^nd Positive and NO Gamma band emission spectroscopy. Comparisons with modeling predictions show qualitative, but not quantitative, agreement with the experimental data.

  14. Holographic Two-Photon Induced Photopolymerization

    Data.gov (United States)

    Federal Laboratory Consortium — Holographic two-photon-induced photopolymerization (HTPIP) offers distinct advantages over conventional one-photon-induced photopolymerization and current techniques...

  15. A bistriphenylamine-substituted spirobifluorene derivative exhibiting excellent nonlinearity/transparency/thermal stability trade-off and strong two-photon induced blue fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Yin, Hongyao [Department of Chemistry, Shanghai Normal University, Shanghai 200234 (China); Xiao, Haibo, E-mail: xiaohb@shnu.edu.cn [Department of Chemistry, Shanghai Normal University, Shanghai 200234 (China); Ding, Lei [Department of Chemistry, Shanghai Normal University, Shanghai 200234 (China); Zhang, Chun; Ren, Aiming [State Key Laboratory of Theoretical and Computational Chemistry, Institute of Theoretical Chemistry, Jilin University, Changchun 130023 (China); Li, Bo [Key Laboratory of Polar Materials and Devices, Ministry of Education, East China Normal University, Shanghai 200241 (China)

    2015-02-01

    A spirobifluorene-bridged donor/donor chromophore, 2,7-bis-(4-(N,N-diphenylamino)phen-1-yl)-9,9′-spirobifluorene (SPF-TP), was found to combine excellent transparency in the near UV–visible region (λ{sub cut-off} ≤ 420 nm), large two-photon absorption cross-section (4.5 × 10{sup 3}GM) and high thermal stability (T{sub d} = 501 °C). In comparison to the reported two-photon absorption molecules, SPF-TP represents the best thermal stability so far described in the literature. The main electronic factors explaining the high two-photon absorption activities of SPF-TP were analyzed by theoretical calculations. Cyclic voltammograms were employed to explore the causes of the excellent transparency of SPF-TP. It was found that the spiroconjugation effect is responsible for the excellent nonlinearity/transparency/thermal stability trade-off in SPF-TP. In addition, SPF-TP is also a good two-photon induced blue fluorescent material with high fluorescence quantum yield (Φ = 0.90, in THF). - Highlights: • We report a molecule exhibiting excellent transparency. • The two-photon absorption cross-section is as large as 4.5 × 10{sup 3}GM. • The molecule exhibits excellent thermal stability. • The molecule is a good two-photon induced blue fluorescent material. • The spiroconjugation effect explains the excellent properties.

  16. A bistriphenylamine-substituted spirobifluorene derivative exhibiting excellent nonlinearity/transparency/thermal stability trade-off and strong two-photon induced blue fluorescence

    International Nuclear Information System (INIS)

    Yin, Hongyao; Xiao, Haibo; Ding, Lei; Zhang, Chun; Ren, Aiming; Li, Bo

    2015-01-01

    A spirobifluorene-bridged donor/donor chromophore, 2,7-bis-(4-(N,N-diphenylamino)phen-1-yl)-9,9′-spirobifluorene (SPF-TP), was found to combine excellent transparency in the near UV–visible region (λ cut-off  ≤ 420 nm), large two-photon absorption cross-section (4.5 × 10 3 GM) and high thermal stability (T d  = 501 °C). In comparison to the reported two-photon absorption molecules, SPF-TP represents the best thermal stability so far described in the literature. The main electronic factors explaining the high two-photon absorption activities of SPF-TP were analyzed by theoretical calculations. Cyclic voltammograms were employed to explore the causes of the excellent transparency of SPF-TP. It was found that the spiroconjugation effect is responsible for the excellent nonlinearity/transparency/thermal stability trade-off in SPF-TP. In addition, SPF-TP is also a good two-photon induced blue fluorescent material with high fluorescence quantum yield (Φ = 0.90, in THF). - Highlights: • We report a molecule exhibiting excellent transparency. • The two-photon absorption cross-section is as large as 4.5 × 10 3 GM. • The molecule exhibits excellent thermal stability. • The molecule is a good two-photon induced blue fluorescent material. • The spiroconjugation effect explains the excellent properties

  17. Two-Photon Fluorescence Microscope for Microgravity Research

    Science.gov (United States)

    Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

    2005-01-01

    A two-photon fluorescence microscope has been developed for the study of biophysical phenomena. Two-photon microscopy is a novel form of laser-based scanning microscopy that enables three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon optical microscopy, two-photon microscopy utilizes the simultaneous nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption, so an ultra-fast pulsed laser source is typically employed. On the other hand, the critical energy threshold for two-photon absorption leads to fluorophore excitation that is intrinsically localized to the focal volume. Consequently, two-photon microscopy enables optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction (relative to one-photon optical microscopy) in photon-induced damage because of the longer excitation wavelength. This reduction is especially advantageous for in vivo studies. Relative to confocal microscopy, there is also a reduction in background fluorescence, and, because of a reduction in Rayleigh scattering, there is a 4 increase of penetration depth. The prohibitive cost of a commercial two-photon fluorescence-microscope system, as well as a need for modularity, has led to the construction of a custom-built system (see Figure 1). This system includes a coherent mode-locked titanium: sapphire laser emitting 120-fs-duration pulses at a repetition rate of 80 MHz. The pulsed laser has an average output power of 800 mW and a wavelength tuning range of 700 to 980 nm, enabling the excitation of a variety of targeted fluorophores. The output from the laser is attenuated, spatially filtered, and then directed into a confocal scanning head that has been modified to provide for side entry of the laser beam. The laser output coupler has been replaced with a dichroic filter that reflects the

  18. Electromagnetically induced two-photon transparency in rubidium atoms

    International Nuclear Information System (INIS)

    Wang, D.; Gao, J.Y.; Xu, J.H.; Bassani, F.; La Rocca, G.C.; Salerno Univ.

    2001-01-01

    We present an experimental demonstration of electromagnetically induced two-photon transparency (EITT) in room temperature rubidium vapor. The 8S 1/2 to 5P 1/2 fluorescence is used to monitor the 5S 1/2 (F = 3) to 8S 1/2 (F = 3) two-photon absorption near resonance with the intermediate state 5P 3/2 . A controlling pump laser beam is employed to coherently couple the 5P 3/2 and 5D 5/2 states, thus producing two dressed intermediate states which give rise to destructive interference in the two-photon transition. An induced two-photon transparency of about 80% has been obtained at resonance; our experimental findings are in good agreement with the general theory of Agarwal et al. (1996), when the appropriate spectroscopic parameters are used. (orig.)

  19. Uniform silica nanoparticles encapsulating two-photon absorbing fluorescent dye

    International Nuclear Information System (INIS)

    Wu Weibing; Liu Chang; Wang Mingliang; Huang Wei; Zhou Shengrui; Jiang Wei; Sun Yueming; Cui Yiping; Xu Chunxinag

    2009-01-01

    We have prepared uniform silica nanoparticles (NPs) doped with a two-photon absorbing zwitterionic hemicyanine dye by reverse microemulsion method. Obvious solvatochromism on the absorption spectra of dye-doped NPs indicates that solvents can partly penetrate into the silica matrix and then affect the ground and excited state of dye molecules. For dye-doped NP suspensions, both one-photon and two-photon excited fluorescence are much stronger and recorded at shorter wavelength compared to those of free dye solutions with comparative overall dye concentration. This behavior is possibly attributed to the restricted twisted intramolecular charge transfer (TICT), which reduces fluorescence quenching when dye molecules are trapped in the silica matrix. Images from two-photon laser scanning fluorescence microscopy demonstrate that the dye-doped silica NPs can be actively uptaken by Hela cells with low cytotoxicity. - Graphical abstract: Water-soluble silica NPs doped with a two-photon absorbing zwitterionic hemicyanine dye were prepared. They were found of enhanced one-photon and two-photon excited fluorescence compared to free dye solutions. Images from two-photon laser scanning fluorescence microscopy demonstrate that the dye-doped silica NPs can be actively uptaken by Hela cells.

  20. Electrically Induced Two-Photon Transparency in Semiconductor Quantum Wells

    International Nuclear Information System (INIS)

    Hayat, Alex; Nevet, Amir; Orenstein, Meir

    2009-01-01

    We demonstrate experimentally two-photon transparency, achieved by current injection into a semiconductor quantum-well structure which exhibits two-photon emission. The two-photon induced luminescence is progressively reduced by the injected current, reaching the point of two-photon transparency - a necessary condition for semiconductor two-photon gain and lasing. These results agree with our calculations.

  1. Detection of carbon monoxide (CO) in sooting hydrocarbon flames using femtosecond two-photon laser-induced fluorescence (fs-TPLIF)

    Science.gov (United States)

    Wang, Yejun; Kulatilaka, Waruna D.

    2018-01-01

    Ultrashort-pulse, femtosecond (fs)-duration, two-photon laser-induced fluorescence (fs-TPLIF) measurements of carbon monoxide (CO) are reported in rich, sooting hydrocarbon flames. CO-TPLIF detection using conventional nanosecond or picosecond lasers are often plagued by photochemical interferences, specifically under fuel-rich flames conditions. In the current study, we investigate the commonly used CO two-photon excitation scheme of the B1Σ+ ← X1Σ+ electronic transition, using approximately 100-fs-duration excitation pulses. Fluorescence emission was observed in the Ångström band originating from directly populated B1Σ+ upper state, as well as, in the third positive band from collisionally populated b3Σ+ upper state. The current work was focused on the Ångström band emission. Interference from nascent C2 emissions originating from hot soot particles in the flame could be reduced to a negligible level using a narrower detection gate width. In contrast, avoiding interferences from laser-generated C2 Swan-band emissions required specific narrowband spectral filtering in sooting flame conditions. The observed less than quadratic laser pulse energy dependence of the TPLIF signal suggests the presence of strong three-photon ionization and stimulated emission processes. In a range of CH4/air and C2H4/air premixed flames investigated, the measured CO fluorescence signals agree well with the calculated equilibrium CO number densities. Reduced-interference CO-TPLIF imaging in premixed C2H4/O2/N2 jet flames is also reported.

  2. Determination of the D/T fuel mixture using two-photon laser induced fluorescence in combination with neutral beam injection

    International Nuclear Information System (INIS)

    Voslamber, D.; Mandl, W.

    1997-08-01

    Doppler-free two-photon induced fluorescence in the Lyman-α lines of H, D and T has been suggested previously as a local and isotope-selective diagnostic of the intrinsic neutral hydrogen densities in magnetically confined fusion plasmas. In the present paper it is shown that the diagnostic potential of this method is significantly increased if it is combined with neutral atom beams whose characteristics are such that efficient production of thermal ground state atoms via charge exchange reactions is achieved. Considerably deeper plasma regions than just the plasma edge can thus be probed and local, isotope-selective information is obtained on the more relevant ions rather than on the neutrals. Additional diagnostic possibilities, e.g. those arising from the spectroscopic investigation of the beam particles themselves, are also discussed. (author)

  3. Two-photon excited fluorescence emission from hemoglobin

    Science.gov (United States)

    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.

  4. Quantitative 3-dimensional imaging of auxin and cytokinin levels in transgenic soybean and medicago truncatula roots via two-photon induced fluorescence imaging

    Science.gov (United States)

    Fisher, Jon; Gaillard, Paul; Nurmalasari, Ni Putu Dewi; Fellbaum, Carl; Subramaniam, Sen; Smith, Steve

    2018-02-01

    Industrial nitrogen fertilizers account for nearly 50% of the fossil fuel costs in modern agriculture and contribute to soil and water pollution. Therefore, significant interest exists in understanding and characterizing the efficiency of nitrogen fixation, and the biochemical signaling pathways which orchestrate the plant-microbial symbiosis through which plants fix nitrogen. Legume plant species exhibit a particularly efficient nitrogen uptake mechanism, using root nodules which house nitrogen-fixing rhizobial bacteria. While nodule development has been widely studied, there remain significant gaps in understanding the regulatory hormones' role in plant development. In this work, we produce 3-dimensional maps of auxin (AX) and cytokinin (CK) hormone concentrations within model plant root tips and nodules with respect to root architecture and cell type. Soybean and Medicago plants were transfected with a two-color fluorescent vector with AXsensitive green fluorescent protein (GFP) and CK-sensitive TdTomato (TdT). 3D images of soybean root nodules were captured using two-photon induced fluorescence microscopy. The resulting images were computationally analyzed using the localization code first developed by Weeks and later adapted by Kilfoil, and analyzed in the context of the root architecture. Statistical analysis of the resulting 3D hormone level maps reproduce-well the known roles of AX and CK in developing plant roots, and are the first quantitative description of these regulatory hormones tied to specific plant architecture. The analytical methods used, and the spatial distribution of these key regulatory hormones in plant roots, nodule primordia and root nodules, and their statistical interpretation are presented.

  5. Production mechanism of atomic nitrogen in atmospheric pressure pulsed corona discharge measured using two-photon absorption laser-induced fluorescence

    International Nuclear Information System (INIS)

    Teramoto, Yoshiyuki; Ono, Ryo; Oda, Tetsuji

    2012-01-01

    To study the production mechanism of atomic nitrogen, the temporal profile and spatial distribution of atomic nitrogen are measured in atmospheric pressure pulsed positive corona discharge using two-photon absorption laser-induced fluorescence. The absolute atomic nitrogen density in the streamer filaments is estimated from decay rate of atomic nitrogen in N 2 discharge. The results indicate that the absolute atomic nitrogen density is approximately constant against discharge energy. When the discharge voltage is 21.5 kV, production yield of atomic nitrogen produced by an N 2 discharge pulse is estimated to be 2.9 - 9.8 × 10 13 atoms and the energy efficiency of atomic nitrogen production is estimated to be about 1.8 - 6.1 × 10 16 atoms/J. The energy efficiency of atomic nitrogen production in N 2 discharge is constant against the discharge energy, while that in N 2 /O 2 discharge increases with discharge energy. In the N 2 /O 2 discharge, two-step process of N 2 dissociation plays significant role for atomic nitrogen production.

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

    International Nuclear Information System (INIS)

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

    2011-01-01

    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

  7. Fluorescent Pluronic nanodots for in vivo two-photon imaging

    International Nuclear Information System (INIS)

    Maurin, Mathieu; Vurth, Laeticia; Vial, Jean-Claude; Baldeck, Patrice; Stephan, Olivier; Marder, Seth R; Sanden, Boudewijn Van der

    2009-01-01

    We report the synthesis of new nanosized fluorescent probes based on bio-compatible polyethylene-polypropylene glycol (Pluronic) materials. In aqueous solution, mini-emulsification of Pluronic with a high fluorescent di-stryl benzene-modified derivative, exhibiting a two-photon absorption cross section as high as 2500 Goeppert-Mayer units at 800 nm, leads to nanoparticles exhibiting a hydrodynamic radius below 100 nm. We have demonstrated that these new probes with luminescence located in the spectral region of interest for bio-imaging (the yellow part of the visible spectrum) allow deep (500 μm) bio-imaging of the mice brain vasculature. The dose injected during our experiments is ten times lower when compared to the classical commercial rhodamine-B isothicyanate-Dextran system but gives similar results to homogeneous blood plasma staining. The mean fluorescent signal intensity stayed constant during more than 1 h.

  8. Imaging Live Drosophila Brain with Two-Photon Fluorescence Microscopy

    Science.gov (United States)

    Ahmed, Syeed Ehsan

    Two-photon fluorescence microscopy is an imaging technique which delivers distinct benefits for in vivo cellular and molecular imaging. Cyclic adenosine monophosphate (cAMP), a second messenger molecule, is responsible for triggering many physiological changes in neural system. However, the mechanism by which this molecule regulates responses in neuron cells is not yet clearly understood. When cAMP binds to a target protein, it changes the structure of that protein. Therefore, studying this molecular structure change with fluorescence resonance energy transfer (FRET) imaging can shed light on the cAMP functioning mechanism. FRET is a non-radiative dipole-dipole coupling which is sensitive to small distance change in nanometer scale. In this study we have investigated the effect of dopamine in cAMP dynamics in vivo. In our study two-photon fluorescence microscope was used for imaging mushroom bodies inside live Drosophila melanogaster brain and we developed a method for studying the change in cyclic AMP level.

  9. Enhanced-locality fiber-optic two-photon-fluorescence live-brain interrogation

    Energy Technology Data Exchange (ETDEWEB)

    Fedotov, I. V.; Doronina-Amitonova, L. V. [International Laser Center, Physics Department, M.V. Lomonosov Moscow State University, Moscow 119992 (Russian Federation); Russian Quantum Center, ul. Novaya 100, Skolkovo, Moscow Region 1430125 (Russian Federation); Kurchatov Institute National Research Center, Moscow (Russian Federation); Sidorov-Biryukov, D. A.; Fedotov, A. B. [International Laser Center, Physics Department, M.V. Lomonosov Moscow State University, Moscow 119992 (Russian Federation); Russian Quantum Center, ul. Novaya 100, Skolkovo, Moscow Region 1430125 (Russian Federation); Anokhin, K. V. [Kurchatov Institute National Research Center, Moscow (Russian Federation); P.K. Anokhin Institute of Normal Physiology, Russian Academy of Medical Sciences, Moscow (Russian Federation); Kilin, S. Ya. [B.I. Stepanov Institute of Physics, National Academy of Sciences of Belarus, Minsk (Belarus); Sakoda, K. [National Institute for Materials Science, 1-1 Namiki, Tsukuba 305-0044 (Japan); Zheltikov, A. M. [International Laser Center, Physics Department, M.V. Lomonosov Moscow State University, Moscow 119992 (Russian Federation); Russian Quantum Center, ul. Novaya 100, Skolkovo, Moscow Region 1430125 (Russian Federation); Department of Physics and Astronomy, Texas A and M University, College Station, Texas 77843 (United States); Center of Photochemistry, Russian Academy of Sciences, ul. Novatorov 7a, Moscow 117421 (Russian Federation)

    2014-02-24

    Two-photon excitation is shown to substantially enhance the locality of fiber-based optical interrogation of strongly scattering biotissues. In our experiments, a high-numerical-aperture, large-core-are fiber probe is used to deliver the 200-fs output of a 100-MHz mode-locked ytterbium fiber laser to samples of live mouse brain, induce two-photon fluorescence of nitrogen–vacancy centers in diamond markers in brain sample. Fiber probes with a high numerical aperture and a large core area are shown to enable locality enhancement in fiber-laser–fiber-probe two-photon brain excitation and interrogation without sacrificing the efficiency of fluorescence response collection.

  10. Two-Photon Fluorescence Microscopy Developed for Microgravity Fluid Physics

    Science.gov (United States)

    Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

    2004-01-01

    Recent research efforts within the Microgravity Fluid Physics Branch of the NASA Glenn Research Center have necessitated the development of a microscope capable of high-resolution, three-dimensional imaging of intracellular structure and tissue morphology. Standard optical microscopy works well for thin samples, but it does not allow the imaging of thick samples because of severe degradation caused by out-of-focus object structure. Confocal microscopy, which is a laser-based scanning microscopy, provides improved three-dimensional imaging and true optical sectioning by excluding the out-of-focus light. However, in confocal microscopy, out-of-focus object structure is still illuminated by the incoming beam, which can lead to substantial photo-bleaching. In addition, confocal microscopy is plagued by limited penetration depth, signal loss due to the presence of a confocal pinhole, and the possibility of live-cell damage. Two-photon microscopy is a novel form of laser-based scanning microscopy that allows three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon microscopy, it utilizes the nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption because of the nonlinear (i.e., quadratic) electric field dependence, so an ultrafast pulsed laser source must typically be employed. On the other hand, this stringent energy density requirement effectively localizes fluorophore excitation to the focal volume. Consequently, two-photon microscopy provides optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction in photo-damage because of the longer excitation wavelength, a reduction in background fluorescence, and a 4 increase in penetration depth over confocal methods because of the reduction in Rayleigh scattering.

  11. A spirobifluorene-based two-photon fluorescence probe for mercury ions and its applications in living cells

    Energy Technology Data Exchange (ETDEWEB)

    Xiao, Haibo, E-mail: xiaohb@shnu.edu.cn; Zhang, Yanzhen; Zhang, Wu; Li, Shaozhi; Tan, Jingjing; Han, Zhongying

    2017-05-01

    A novel spirobifluorene derivative SPF-TMS, which containing dithioacetal groups and triphenylamine units, was synthesized. The probing behaviors toward various metal ions were investigated via UV/Vis absorption spectra as well as one-photon fluorescence changes. The results indicated that SPF-TMS exhibits high sensitivity and selectivity for mercury ions. The detection limit was at least 8.6 × 10{sup −8}M, which is excellent comparing with other optical sensors for Hg{sup 2+}. When measured by two-photon excited fluorescence technique in THF at 800 nm, the two-photon cross-section of SPF-TMS is 272 GM. Especially, upon reaction with mercury species, SPF-TMS yielded another two-photon dye SPF-DA. Both SPF-TMS and SPF-DA emit strong two-photon induced fluorescence and can be applied in cell imaging by two-photon microscopy. - Highlights: • We report a spirobifluorene-based molecule as two-photon fluorescent probe with large two-photon cross-section. • The molecule has exclusive selectivity and sensitivity for mercury species. • The molecule has large two-photon emission changes before and after addition of Hg{sup 2+}. • Both the probe and the mercury ion-promoted reaction product can be applied in cell imaging by two-photon microscopy.

  12. Sensing for intracellular thiols by water-insoluble two-photon fluorescent probe incorporating nanogel

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Xudong; Zhang, Xin; Wang, Shuangqing; Li, Shayu [Beijing National Laboratory for Molecular Sciences, Key laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Hu, Rui, E-mail: hurui@iccas.ac.cn [Beijing National Laboratory for Molecular Sciences, Key laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Li, Yi, E-mail: yili@mail.ipc.ac.cn [Key Laboratory of Photochemical Conversion and Optoelectronic Materials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Yang, Guoqiang, E-mail: gqyang@iccas.ac.cn [Beijing National Laboratory for Molecular Sciences, Key laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China)

    2015-04-15

    Highlights: • A novel “turn-on” two-photon fluorescent probe based on a π-conjugated triarylboron luminogen was designed and synthesized. • Fast, selective and sensitive detection of biothiols in 100% aqueous solution by simply loaded on a nanogel. • Single-photon and two-photon fluorescent bioimaging of biothiols in NIH/3T3 fibroblasts. - Abstract: A novel “turn-on” two-photon fluorescent probe containing a π-conjugated triarylboron luminogen and a maleimide moiety DMDP-M based on the photo-induced electron transfer (PET) mechanism for biothiol detection was designed and synthesized. By simply loading the hydrophobic DMDP-M on a cross-linked Pluronic{sup ®} F127 nanogel (CL-F127), a probing system DMDP-M/CL-F127 was established, which shows quick response, high selectivity and sensitivity to cysteine (Cys), homocysteine (Hcy) and glutathione (GSH) in aqueous phase. The DMDP-M/CL-F127 system presented the fastest response to Cys with a rate constant of 0.56 min{sup −1}, and the detection limit to Cys was calculated to be as low as 0.18 μM. The DMDP-M/CL-F127 system has been successfully applied to the fluorescence imaging of biothiols in NIH/3T3 fibroblasts either with single-photon or two-photon excitation because of its high biocompatibility and cell-membrane permeability. The present work provides a general, simple and efficient strategy for the application of hydrophobic molecules to sensing biothiols in aqueous phase, and a novel sensing system for intracellular biothiols fitted for both single-photon and two-photon fluorescence imaging.

  13. Diagnostics of MCF plasmas using Lyman-α fluorescence excited by one or two photons

    International Nuclear Information System (INIS)

    Voslamber, D.

    1998-11-01

    Laser-induced Lyman-α fluorescence of the hydrogen isotopes is investigated with regard to diagnostic applications in magnetically confined fusion plasmas. A formal analysis is presented for two excitation schemes: one-photon and Doppler-free two-photon excitation. The analysis includes estimates of the expected experimental errors arising from the photon noise and from the sensitivity of the observed fluorescence signals to variations of the plasma and laser parameters. Both excitation schemes are suitable primarily for application in the plasma edge, but even in the plasma bulk of large machines they can still be applied in combination with a diagnostic neutral beam. The two-photon excitation scheme is particularly attractive because it involves absorption spectra that are resolved within the Doppler width. This implies a large diagnostic potential and in particular offers a way to measure the deuterium-tritium fuel mix in fusion reactors. (author)

  14. Time gated fluorescence lifetime imaging and micro-volume spectroscopy using two-photon excitation

    NARCIS (Netherlands)

    Sytsma, J.; Vroom, J.M.; de Grauw, C.J.; Gerritsen, H.C.

    A scanning microscope utilizing two-photon excitation in combination with fluorescence lifetime contrast is presented. The microscope makes use of a tunable femtosecond titanium:sapphire laser enabling the two-photon excitation of a broad range of fluorescent molecules, including UV probes.

  15. Two-photon fluorescence and fluorescence imaging of two styryl heterocyclic dyes combined with DNA.

    Science.gov (United States)

    Gao, Chao; Liu, Shu-yao; Zhang, Xian; Liu, Ying-kai; Qiao, Cong-de; Liu, Zhao-e

    2016-03-05

    Two new styryl heterocyclic two-photon (TP) materials, 4-[4-(N-methyl)styrene]-imidazo [4,5-f][1,10] phenanthroline-benzene iodated salt (probe-1) and 4,4-[4-(N-methyl)styrene]-benzene iodated salt (probe-2) were successfully synthesized and studied as potential fluorescent probes of DNA detection. The linear and nonlinear photophysical properties of two compounds in different solvents were investigated. The absorption, one- and two-photon fluorescent spectra of the free dye and dye-DNA complex were also examined to evaluate their photophysical properties. The binding constants of dye-DNA were obtained according to Scatchard equation with good values. The results showed that two probes could be used as fluorescent DNA probes by two-photon excitation, and TP fluorescent properties of probe-1 are superior to that of probe-2. The fluorescent method date indicated that the mechanisms of dye-DNA complex interaction may be groove binding for probe-1 and electrostatic interaction for probe-2, respectively. The MTT assay experiments showed two probes are low toxicity. Moreover, the TP fluorescence imaging of DNA detection in living cells at 800 nm indicated that the ability to locate in cell nuclei of probe-1 is better than that of probe-2. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. A new approach to dual-color two-photon microscopy with fluorescent proteins

    Directory of Open Access Journals (Sweden)

    Rebane Aleks

    2010-02-01

    Full Text Available Abstract Background Two-photon dual-color imaging of tissues and cells labeled with fluorescent proteins (FPs is challenging because most two-photon microscopes only provide one laser excitation wavelength at a time. At present, methods for two-photon dual-color imaging are limited due to the requirement of large differences in Stokes shifts between the FPs used and their low two-photon absorption (2PA efficiency. Results Here we present a new method of dual-color two-photon microscopy that uses the simultaneous excitation of the lowest-energy electronic transition of a blue fluorescent protein and a higher-energy electronic transition of a red fluorescent protein. Conclusion Our method does not require large differences in Stokes shifts and can be extended to a variety of FP pairs with larger 2PA efficiency and more optimal imaging properties.

  17. Folate receptor targeting silica nanoparticle probe for two-photon fluorescence bioimaging

    Science.gov (United States)

    Wang, Xuhua; Yao, Sheng; Ahn, Hyo-Yang; Zhang, Yuanwei; Bondar, Mykhailo V.; Torres, Joseph A.; Belfield, Kevin D.

    2010-01-01

    Narrow dispersity organically modified silica nanoparticles (SiNPs), diameter ~30 nm, entrapping a hydrophobic two-photon absorbing fluorenyl dye, were synthesized by hydrolysis of triethoxyvinylsilane and (3-aminopropyl)triethoxysilane in the nonpolar core of Aerosol-OT micelles. The surface of the SiNPs were functionalized with folic acid, to specifically deliver the probe to folate receptor (FR) over-expressing Hela cells, making these folate two-photon dye-doped SiNPs potential candidates as probes for two-photon fluorescence microscopy (2PFM) bioimaging. In vitro studies using FR over-expressing Hela cells and low FR expressing MG63 cells demonstrated specific cellular uptake of the functionalized nanoparticles. One-photon fluorescence microscopy (1PFM) imaging, 2PFM imaging, and two-photon fluorescence lifetime microscopy (2P-FLIM) imaging of Hela cells incubated with folate-modified two-photon dye-doped SiNPs were demonstrated. PMID:21258480

  18. Thermally activated delayed fluorescence organic dots for two-photon fluorescence lifetime imaging

    Science.gov (United States)

    He, Tingchao; Ren, Can; Li, Zhuohua; Xiao, Shuyu; Li, Junzi; Lin, Xiaodong; Ye, Chuanxiang; Zhang, Junmin; Guo, Lihong; Hu, Wenbo; Chen, Rui

    2018-05-01

    Autofluorescence is a major challenge in complex tissue imaging when molecules present in the biological tissue compete with the fluorophore. This issue may be resolved by designing organic molecules with long fluorescence lifetimes. The present work reports the two-photon absorption (TPA) properties of a thermally activated delayed fluorescence (TADF) molecule with carbazole as the electron donor and dicyanobenzene as the electron acceptor (i.e., 4CzIPN). The results indicate that 4CzIPN exhibits a moderate TPA cross-section (˜9 × 10-50 cm4 s photon-1), high fluorescence quantum yield, and a long fluorescence lifetime (˜1.47 μs). 4CzIPN was compactly encapsulated into an amphiphilic copolymer via nanoprecipitation to achieve water-soluble organic dots. Interestingly, 4CzIPN organic dots have been utilized in applications involving two-photon fluorescence lifetime imaging (FLIM). Our work aptly demonstrates that TADF molecules are promising candidates of nonlinear optical probes for developing next-generation multiphoton FLIM applications.

  19. Highly sensitive measurement in two-photon absorption cross section and investigation of the mechanism of two-photon-induced polymerization

    Energy Technology Data Exchange (ETDEWEB)

    Lu Youmei E-mail: luym19@cc.tuat.ac.jp; Hasegawa, Fuyuki; Goto, Takamichi; Ohkuma, Satoshi; Fukuhara, Setsuko; Kawazu, Yukie; Totani, Kenro; Yamashita, Takashi; Watanabe, Toshiyuki E-mail: toshi@cc.tuat.ac.jp

    2004-10-01

    A novel two-photon initiator, 4,4'-bis[4-(di-n-butylamino)styryl]-benzene with the side-group methyl (Me) (abbreviated as Chromophore 1), was synthesized in comparison with the chromophore with the side group methoxy (MeO) (abbreviated as Chromophore 2). Femtosecond laser-induced fluorescence intensity was used to evaluate two-photon absorption (TPA) cross section, {delta}, by means of a charge-coupled device, USB-2000 (abbreviated as CCD). Results showed that changing the side group from Me to MeO led to a significant red-shift of the two-photon absorption ({sup 2}{lambda}{sub max}). However, the microstructures obtained by two-photon-induced polymerization (TPIP) demonstrated that the sensitivities of Chromophore 1 increased despite a two-fold decrease in the two-photon cross section {delta}{sub max,} relative to Chromophore 2. Correlated with the appearance that the long-lived charge transfer emission of the chromophore in the monomer bulk, we suggest that the intramolecular charge transfer (intra-CT) takes place within the excited dye. Then intermolecular charge transfer was successive as a result of the formation of an exciplex between the dye and the monomer. The Me group was favorable for the intra-CT, relative to MeO, which contributed to the enhancement of the sensitivity of TPIP.

  20. Near-IR Two-Photon Fluorescent Sensor for K(+) Imaging in Live Cells.

    Science.gov (United States)

    Sui, Binglin; Yue, Xiling; Kim, Bosung; Belfield, Kevin D

    2015-08-19

    A new two-photon excited fluorescent K(+) sensor is reported. The sensor comprises three moieties, a highly selective K(+) chelator as the K(+) recognition unit, a boron-dipyrromethene (BODIPY) derivative modified with phenylethynyl groups as the fluorophore, and two polyethylene glycol chains to afford water solubility. The sensor displays very high selectivity (>52-fold) in detecting K(+) over other physiological metal cations. Upon binding K(+), the sensor switches from nonfluorescent to highly fluorescent, emitting red to near-IR (NIR) fluorescence. The sensor exhibited a good two-photon absorption cross section, 500 GM at 940 nm. Moreover, it is not sensitive to pH in the physiological pH range. Time-dependent cell imaging studies via both one- and two-photon fluorescence microscopy demonstrate that the sensor is suitable for dynamic K(+) sensing in living cells.

  1. Phosphorus ligand imaging with two-photon fluorescence spectroscopy: towards rational catalyst immobilization

    NARCIS (Netherlands)

    Marras, F.; Kluwer, A.M.; Siekierzycka, J.R.; Vozza, A.; Brouwer, A.M.; Reek, J.N.H.

    2010-01-01

    Spotless catalysts: Ligand immobilization was studied by two-photon fluorescence microscopy with a fluorescent nixantphos ligand as probe (see picture). In the immobilization process ligand aggregates form in solution and are deposited on the support, where they appear as bright spots in

  2. Combined Raman and continuous-wave-excited two-photon fluorescence cell imaging

    NARCIS (Netherlands)

    Uzunbajakava, N.; Otto, Cornelis

    2003-01-01

    We demonstrate a confocal optical microscope that combines cw two-photon-excited fluorescence microscopy with confocal Raman microscopy. With this microscope fast image acquisition with fluorescence imaging can be used to select areas of interest for subsequent chemical analysis with spontaneous

  3. Molecular engineering of two-photon fluorescent probes for bioimaging applications

    Science.gov (United States)

    Liu, Hong-Wen; Liu, Yongchao; Wang, Peng; Zhang, Xiao-Bing

    2017-03-01

    During the past two decades, two-photon microscopy (TPM), which utilizes two near-infrared photons as the excitation source, has emerged as a novel, attractive imaging tool for biological research. Compared with one-photon microscopy, TPM offers several advantages, such as lowering background fluorescence in living cells and tissues, reducing photodamage to biosamples, and a photobleaching phenomenon, offering better 3D spatial localization, and increasing penetration depth. Small-molecule-based two-photon fluorescent probes have been well developed for the detection and imaging of various analytes in biological systems. In this review, we will give a general introduction of molecular engineering of two-photon fluorescent probes based on different fluorescence response mechanisms for bioimaging applications during the past decade. Inspired by the desired advantages of small-molecule two-photon fluorescent probes in biological imaging applications, we expect that more attention will be devoted to the development of new two-photon fluorophores and applications of TPM in areas of bioanalysis and disease diagnosis.

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

    Science.gov (United States)

    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.

  5. Fluorenyl benzothiadiazole and benzoselenadiazole near-IR fluorescent probes for two-photon fluorescence imaging (Conference Presentation)

    Science.gov (United States)

    Belfield, Kevin D.; Yao, Sheng; Kim, Bosung; Yue, Xiling

    2016-03-01

    Imaging biological samples with two-photon fluorescence (2PF) microscopy has the unique advantage of resulting high contrast 3D resolution subcellular image that can reach up to several millimeters depth. 2PF probes that absorb and emit at near IR region need to be developed. Two-photon excitation (2PE) wavelengths are less concerned as 2PE uses wavelengths doubles the absorption wavelength of the probe, which means 2PE wavelengths for probes even with absorption at visible wavelength will fall into NIR region. Therefore, probes that fluoresce at near IR region with high quantum yields are needed. A series of dyes based on 5-thienyl-2, 1, 3-benzothiadiazole and 5-thienyl-2, 1, 3-benzoselenadiazole core were synthesized as near infrared two-photon fluorophores. Fluorescence maxima wavelengths as long as 714 nm and fluorescence quantum yields as high as 0.67 were achieved. The fluorescence quantum yields of the dyes were nearly constant, regardless of solvents polarity. These diazoles exhibited large Stokes shift (GM), and high two-photon fluorescence figure of merit (FM , 1.04×10-2 GM). Cells incubated on a 3D scaffold with one of the new probes (encapsulated in Pluronic micelles) exhibited bright fluorescence, enabling 3D two-photon fluorescence imaging to a depth of 100 µm.

  6. Circular dichroism probed by two-photon fluorescence microscopy in enantiopure chiral polyfluorene thin films

    NARCIS (Netherlands)

    Savoini, M.; Wu, Xiaofei; Celebrano, M.; Ziegler, J.; Biagioni, P.; Meskers, S.C.J.; Duo, L.; Hecht, B.; Finazzi, M.

    2012-01-01

    Two-photon fluorescence scanning confocal microscopy sensitive to circular dichroism with a diffraction-limited resolution well below 500 nm is demonstrated. With this method, the spatial variation of the circular dichroism of thermally annealed chiral polyfluorene thin films has been imaged. We

  7. Applications of two-photon fluorescence microscopy in deep-tissue imaging

    Science.gov (United States)

    Dong, Chen-Yuan; Yu, Betty; Hsu, Lily L.; Kaplan, Peter D.; Blankschstein, D.; Langer, Robert; So, Peter T. C.

    2000-07-01

    Based on the non-linear excitation of fluorescence molecules, two-photon fluorescence microscopy has become a significant new tool for biological imaging. The point-like excitation characteristic of this technique enhances image quality by the virtual elimination of off-focal fluorescence. Furthermore, sample photodamage is greatly reduced because fluorescence excitation is limited to the focal region. For deep tissue imaging, two-photon microscopy has the additional benefit in the greatly improved imaging depth penetration. Since the near- infrared laser sources used in two-photon microscopy scatter less than their UV/glue-green counterparts, in-depth imaging of highly scattering specimen can be greatly improved. In this work, we will present data characterizing both the imaging characteristics (point-spread-functions) and tissue samples (skin) images using this novel technology. In particular, we will demonstrate how blind deconvolution can be used further improve two-photon image quality and how this technique can be used to study mechanisms of chemically-enhanced, transdermal drug delivery.

  8. Spectral, energy, and time parameters of two-photon fluorescence of 2,5-diphenyloxazole polycrystals

    International Nuclear Information System (INIS)

    Agal'tsov, A.M.; Gorelik, V.S.; Rakhmatullaev, I.A.

    1995-01-01

    Two-photon fluorescence (TPF) spectra of 2,5-diphenyloxazole polycrystals (known in the literature as PPO) were obtained and studied as a function of the pump power and time delay. The fluorescence spectrum shape observed upon two-photon excitation is shown to be distinctly different from that observed upon electron-beam excitation. It is shown that high pump powers result in stimulated fluorescence. PPO exhibits a high TPF quantum yield, the integrated conversion efficiency of exciting radiation to TPF being 40%. The TPF decay time is measured to be 20 ns. The spectral data obtained for PPO polycrystals can be used in the development of new TPF light sources tunable in the UV region. 10 refs., 4 figs., 1 tab

  9. Two-photon excited fluorescence microscopy application for ex vivo investigation of ocular fundus samples

    Science.gov (United States)

    Peters, Sven; Hammer, Martin; Schweitzer, Dietrich

    2011-07-01

    Two-photon excited fluorescence (TPEF) imaging of ocular tissue has recently become a promising tool in ophthalmology for diagnostic and research purposes. The feasibility and the advantages of TPEF imaging, namely deeper tissue penetration and improved high-resolution imaging of microstructures, have been demonstrated lately using human ocular samples. The autofluorescence properties of endogenous fluorophores in ocular fundus tissue are well known from spectrophotometric analysis. But fluorophores, especially when it comes to fluorescence lifetime, typically display a dependence of their fluorescence properties on local environmental parameters. Hence, a more detailed investigation of ocular fundus autofluorescence ideally in vivo is of utmost interest. The aim of this study is to determine space-resolved the stationary and time-resolved fluorescence properties of endogenous fluorophores in ex vivo porcine ocular fundus samples by means of two-photon excited fluorescence spectrum and lifetime imaging microscopy (FSIM/FLIM). By our first results, we characterized the autofluorescence of individual anatomical structures of porcine retina samples excited at 760 nm. The fluorescence properties of almost all investigated retinal layers are relatively homogenous. But as previously unknown, ganglion cell bodies show a significantly shorter fluorescence lifetime compared to the adjacent mueller cells. Since all retinal layers exhibit bi-exponential autofluorescence decays, we were able to achieve a more precise characterization of fluorescence properties of endogenous fluorophores compared to a present in vivo FLIM approach by confocal scanning laser ophthalmoscope (cSLO).

  10. Bulky Counterions: Enhancing the Two-Photon Excited Fluorescence of Gold Nanoclusters.

    Science.gov (United States)

    Bertorelle, Franck; Moulin, Christophe; Soleilhac, Antonin; Comby-Zerbino, Clothilde; Dugourd, Philippe; Russier-Antoine, Isabelle; Brevet, Pierre-François; Antoine, Rodolphe

    2018-01-19

    Increasing fluorescence quantum yields of ligand-protected gold nanoclusters has attracted wide research interest. The strategy consisting in using bulky counterions has been found to dramatically enhance the fluorescence. In this Communication, we push forward this concept to the nonlinear optical regime. We show that by an appropriate choice of bulky counterions and of solvent, a 30-fold increase in two-photon excited fluorescence (TPEF) signal at ≈600 nm for gold nanoclusters can be obtained. This would correspond to a TPEF cross-section in the range of 0.1 to 1 GM. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Supramolecular assembly affording a ratiometric two-photon fluorescent nanoprobe for quantitative detection and bioimaging.

    Science.gov (United States)

    Wang, Peng; Zhang, Cheng; Liu, Hong-Wen; Xiong, Mengyi; Yin, Sheng-Yan; Yang, Yue; Hu, Xiao-Xiao; Yin, Xia; Zhang, Xiao-Bing; Tan, Weihong

    2017-12-01

    Fluorescence quantitative analyses for vital biomolecules are in great demand in biomedical science owing to their unique detection advantages with rapid, sensitive, non-damaging and specific identification. However, available fluorescence strategies for quantitative detection are usually hard to design and achieve. Inspired by supramolecular chemistry, a two-photon-excited fluorescent supramolecular nanoplatform ( TPSNP ) was designed for quantitative analysis with three parts: host molecules (β-CD polymers), a guest fluorophore of sensing probes (Np-Ad) and a guest internal reference (NpRh-Ad). In this strategy, the TPSNP possesses the merits of (i) improved water-solubility and biocompatibility; (ii) increased tissue penetration depth for bioimaging by two-photon excitation; (iii) quantitative and tunable assembly of functional guest molecules to obtain optimized detection conditions; (iv) a common approach to avoid the limitation of complicated design by adjustment of sensing probes; and (v) accurate quantitative analysis by virtue of reference molecules. As a proof-of-concept, we utilized the two-photon fluorescent probe NHS-Ad-based TPSNP-1 to realize accurate quantitative analysis of hydrogen sulfide (H 2 S), with high sensitivity and good selectivity in live cells, deep tissues and ex vivo -dissected organs, suggesting that the TPSNP is an ideal quantitative indicator for clinical samples. What's more, TPSNP will pave the way for designing and preparing advanced supramolecular sensors for biosensing and biomedicine.

  12. Diagnosis of basal cell carcinoma by two photon excited fluorescence combined with lifetime imaging

    Science.gov (United States)

    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. Concentration of atomic hydrogen in a dielectric barrier discharge measured by two-photon absorption fluorescence

    Science.gov (United States)

    Dvořák, P.; Talába, M.; Obrusník, A.; Kratzer, J.; Dědina, J.

    2017-08-01

    Two-photon absorption laser-induced fluorescence (TALIF) was utilized for measuring the concentration of atomic hydrogen in a volume dielectric barrier discharge (DBD) ignited in mixtures of Ar, H2 and O2 at atmospheric pressure. The method was calibrated by TALIF of krypton diluted in argon at atmospheric pressure, proving that three-body collisions had a negligible effect on quenching of excited krypton atoms. The diagnostic study was complemented with a 3D numerical model of the gas flow and a zero-dimensional model of the chemistry in order to better understand the reaction kinetics and identify the key pathways leading to the production and destruction of atomic hydrogen. It was determined that the density of atomic hydrogen in Ar-H2 mixtures was in the order of 1021 m-3 and decreased when oxygen was added into the gas mixture. Spatially resolved measurements and simulations revealed a sharply bordered region with low atomic hydrogen concentration when oxygen was added to the gas mixture. At substoichiometric oxygen/hydrogen ratios, this H-poor region is confined to an area close to the gas inlet and it is shown that the size of this region is not only influenced by the chemistry but also by the gas flow patterns. Experimentally, it was observed that a decrease in H2 concentration in the feeding Ar-H2 mixture led to an increase in H production in the DBD.

  14. In vivo imaging of cerebral energy metabolism with two-photon fluorescence lifetime microscopy of NADH.

    Science.gov (United States)

    Yaseen, Mohammad A; Sakadžić, Sava; Wu, Weicheng; Becker, Wolfgang; Kasischke, Karl A; Boas, David A

    2013-02-01

    Minimally invasive, specific measurement of cellular energy metabolism is crucial for understanding cerebral pathophysiology. Here, we present high-resolution, in vivo observations of autofluorescence lifetime as a biomarker of cerebral energy metabolism in exposed rat cortices. We describe a customized two-photon imaging system with time correlated single photon counting detection and specialized software for modeling multiple-component fits of fluorescence decay and monitoring their transient behaviors. In vivo cerebral NADH fluorescence suggests the presence of four distinct components, which respond differently to brief periods of anoxia and likely indicate different enzymatic formulations. Individual components show potential as indicators of specific molecular pathways involved in oxidative metabolism.

  15. CdSe/AsS core-shell quantum dots: preparation and two-photon fluorescence.

    Science.gov (United States)

    Wang, Junzhong; Lin, Ming; Yan, Yongli; Wang, Zhe; Ho, Paul C; Loh, Kian Ping

    2009-08-19

    Arsenic(II) sulfide (AsS)-coated CdSe core-shell nanocrystals can be prepared by a cluster-complex deposition approach under mild conditions. At 60 degrees C, growth of an AsS shell onto a CdSe nanocrystal can be realized through the crystallization of a cluster complex of AsS/butylamine in a mixed solvent of isopropanol/chloroform. The new, type I core-shell nanocrystal exhibits markedly enhanced one-photon fluorescence as well two-photon upconversion fluorescence. The nanocrystals can be used for infrared-excited upconversion cellular labeling.

  16. Selective labeling of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein

    Science.gov (United States)

    Watanabe, Wataru; Shimada, Tomoko; Matsunaga, Sachihiro; Kurihara, Daisuke; Arimura, Shin-ichi; Tsutsumi, Nobuhiro; Fukui, Kiichi; Itoh, Kazuyoshi

    2008-02-01

    We present space-selective labeling of organelles by using two-photon conversion of a photoconvertible fluorescent protein with near-infrared femtosecond laser pulses. Two-photon excitation of photoconvertible fluorescent-protein, Kaede, enables space-selective labeling of organelles. We alter the fluorescence of target mitochondria in a tobacco BY-2 cell from green to red by focusing femtosecond laser pulses with a wavelength of 750 nm.

  17. Deep brain two-photon NIR fluorescence imaging for study of Alzheimer's disease

    Science.gov (United States)

    Chen, Congping; Liang, Zhuoyi; Zhou, Biao; Ip, Nancy Y.; Qu, Jianan Y.

    2018-02-01

    Amyloid depositions in the brain represent the characteristic hallmarks of Alzheimer's disease (AD) pathology. The abnormal accumulation of extracellular amyloid-beta (Aβ) and resulting toxic amyloid plaques are considered to be responsible for the clinical deficits including cognitive decline and memory loss. In vivo two-photon fluorescence imaging of amyloid plaques in live AD mouse model through a chronic imaging window (thinned skull or craniotomy) provides a mean to greatly facilitate the study of the pathological mechanism of AD owing to its high spatial resolution and long-term continuous monitoring. However, the imaging depth for amyloid plaques is largely limited to upper cortical layers due to the short-wavelength fluorescence emission of commonly used amyloid probes. In this work, we reported that CRANAD-3, a near-infrared (NIR) probe for amyloid species with excitation wavelength at 900 nm and emission wavelength around 650 nm, has great advantages over conventionally used probes and is well suited for twophoton deep imaging of amyloid plaques in AD mouse brain. Compared with a commonly used MeO-X04 probe, the imaging depth of CRANAD-3 is largely extended for open skull cranial window. Furthermore, by using two-photon excited fluorescence spectroscopic imaging, we characterized the intrinsic fluorescence of the "aging pigment" lipofuscin in vivo, which has distinct spectra from CRANAD-3 labeled plaques. This study reveals the unique potential of NIR probes for in vivo, high-resolution and deep imaging of brain amyloid in Alzheimer's disease.

  18. Two photon versus one photon fluorescence excitation in whispering gallery mode microresonators

    International Nuclear Information System (INIS)

    Pastells, Carme; Marco, M.-Pilar; Merino, David; Loza-Alvarez, Pablo; Pasquardini, Laura; Lunelli, Lorenzo; Pederzolli, Cecilia; Daldosso, Nicola; Farnesi, Daniele; Berneschi, Simone; Righini, Giancarlo C.; Quercioli, Franco; Nunzi Conti, Gualtiero; Soria, Silvia

    2016-01-01

    We investigate the feasibility of both one photon and two photon fluorescence excitation using whispering gallery mode microresonators. We report the linear and non linear fluorescence real-time detection of labeled IgG covalently bonded to the surface of a silica whispering gallery mode resonator (WGMR). The immunoreagents have been immobilized onto the surface of the WGMR sensor after being activated with an epoxy silane and an orienting layer. The developed immunosensor presents great potential as a robust sensing device for fast and early detection of immunoreactions. We also investigate the potential of microbubbles as nonlinear enhancement platform. The dyes used in these studies are dylight800, tetramethyl rhodamine isothiocyanate, rhodamine 6G and fluorescein. All measurements were performed in a modified confocal microscope. - Highlights: • One photon fluorescence overlaps with the semiconductor pump laser gain bandwidth. • We report on the feasibility to excite two photon fluorescence in microbubble resonators. • Our functionalization process maintains a good quality factor of the microresonator.

  19. Two photon versus one photon fluorescence excitation in whispering gallery mode microresonators

    Energy Technology Data Exchange (ETDEWEB)

    Pastells, Carme; Marco, M.-Pilar [Nanobiotechnology for Diagnostics Group (Nb4Dg), IQAC-CSIC, 08034 Barcelona (Spain); CIBER de Bioingeniería, Biomateriales y Nanomedicina, 08034 Barcelona (Spain); Merino, David; Loza-Alvarez, Pablo [ICFO-Institut de Ciències Fotòniques, Castelldefels, 08860 Barcelona (Spain); Pasquardini, Laura [Fondazione Bruno Kessler, 38123 Povo, TN (Italy); Lunelli, Lorenzo [Fondazione Bruno Kessler, 38123 Povo, TN (Italy); IBF-CNR, 38123 Povo, TN (Italy); Pederzolli, Cecilia [Fondazione Bruno Kessler, 38123 Povo, TN (Italy); Daldosso, Nicola [Department of Computer Science, University of Verona, Strada le Grazie 15, 37134 Verona (Italy); Farnesi, Daniele [CNR-IFAC “Nello Carrara” Institute of Applied Physics, 50019 Sesto Fiorentino, FI (Italy); Museo Storico della Fisica e Centro Studi e Ricerche “E. Fermi”, 00184 Roma (Italy); Berneschi, Simone [CNR-IFAC “Nello Carrara” Institute of Applied Physics, 50019 Sesto Fiorentino, FI (Italy); Righini, Giancarlo C. [CNR-IFAC “Nello Carrara” Institute of Applied Physics, 50019 Sesto Fiorentino, FI (Italy); Museo Storico della Fisica e Centro Studi e Ricerche “E. Fermi”, 00184 Roma (Italy); Quercioli, Franco [CNR-INO National Institute of Optics, Sesto Fiorentino, FI (Italy); Nunzi Conti, Gualtiero [CNR-IFAC “Nello Carrara” Institute of Applied Physics, 50019 Sesto Fiorentino, FI (Italy); Soria, Silvia, E-mail: s.soria@ifac.cnr.it [CNR-IFAC “Nello Carrara” Institute of Applied Physics, 50019 Sesto Fiorentino, FI (Italy)

    2016-02-15

    We investigate the feasibility of both one photon and two photon fluorescence excitation using whispering gallery mode microresonators. We report the linear and non linear fluorescence real-time detection of labeled IgG covalently bonded to the surface of a silica whispering gallery mode resonator (WGMR). The immunoreagents have been immobilized onto the surface of the WGMR sensor after being activated with an epoxy silane and an orienting layer. The developed immunosensor presents great potential as a robust sensing device for fast and early detection of immunoreactions. We also investigate the potential of microbubbles as nonlinear enhancement platform. The dyes used in these studies are dylight800, tetramethyl rhodamine isothiocyanate, rhodamine 6G and fluorescein. All measurements were performed in a modified confocal microscope. - Highlights: • One photon fluorescence overlaps with the semiconductor pump laser gain bandwidth. • We report on the feasibility to excite two photon fluorescence in microbubble resonators. • Our functionalization process maintains a good quality factor of the microresonator.

  20. Non-invasive imaging of skin cancer with fluorescence lifetime imaging using two photon tomography

    Science.gov (United States)

    Patalay, Rakesh; Talbot, Clifford; Alexandrov, Yuriy; Munro, Ian; Breunig, Hans Georg; König, Karsten; Warren, Sean; Neil, Mark A. A.; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Christopher

    2011-07-01

    Multispectral fluorescence lifetime imaging (FLIM) using two photon microscopy as a non-invasive technique for the diagnosis of skin lesions is described. Skin contains fluorophores including elastin, keratin, collagen, FAD and NADH. This endogenous contrast allows tissue to be imaged without the addition of exogenous agents and allows the in vivo state of cells and tissues to be studied. A modified DermaInspect® multiphoton tomography system was used to excite autofluorescence at 760 nm in vivo and on freshly excised ex vivo tissue. This instrument simultaneously acquires fluorescence lifetime images in four spectral channels between 360-655 nm using time-correlated single photon counting and can also provide hyperspectral images. The multispectral fluorescence lifetime images were spatially segmented and binned to determine lifetimes for each cell by fitting to a double exponential lifetime model. A comparative analysis between the cellular lifetimes from different diagnoses demonstrates significant diagnostic potential.

  1. Whispering gallery modes in two-photon fluorescence from spherical DCM dye microresonators

    Science.gov (United States)

    Mamonov, Evgeniy A.; Maydykovskiy, Anton I.; Mitetelo, Nikolai V.; Venkatakrishnarao, Dasari; Chandrasekar, Rajadurai; Murzina, Tatyana V.

    2018-03-01

    Organic microstructures are well known for their resonator properties, which bring about whispering gallery mode (WGM) excitation. Here we report on experimental evidence of the WGM in the two-photon fluorescence (TPF) of DCM dye microspheres made using the self-assembly method. The WGM excitation accompanying the overall TPF in the spectral range from 530\\div640 nm demonstrated a quality factor of approximately 102 for spheres that were several microns in diameter. The power dependence of the TPF intensity proved the second order nature of the interaction process involved.

  2. Simple fibre based dispersion management for two-photon excited fluorescence imaging through an endoscope

    DEFF Research Database (Denmark)

    Dimopoulos, Konstantinos; Marti, Dominik; Andersen, Peter E.

    2018-01-01

    We want to implement two-photon excitation fluorescence microscopy (TPEFM) into endoscopes, since TPEFM can provide relevant biomarkers for cancer staging and grading in hollow organs, endoscopically accessible through natural orifices. However, many obstacles must be overcome, among others...... the delivery of short laser pulses to the distal end of the endoscope. To this avail, we present imaging results using an all-fibre dispersion management scheme in a TPEFM setup. The scheme has been conceived by Jespersen et al. in 20101 and relies on the combination of a single mode fibre with normal...

  3. Photonic crystal fibre enables short-wavelength two-photon laser scanning fluorescence microscopy with fura-2

    International Nuclear Information System (INIS)

    McConnell, Gail; Riis, Erling

    2004-01-01

    We report on a novel and compact reliable laser source capable of short-wavelength two-photon laser scanning fluorescence microscopy based on soliton self-frequency shift effects in photonic crystal fibre. We demonstrate the function of the system by performing two-photon microscopy of smooth muscle cells and cardiac myocytes from the rat pulmonary vein and Chinese hamster ovary cells loaded with the fluorescent calcium indicator fura-2/AM

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

    Science.gov (United States)

    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. Copyright 1999 Wiley-Liss, Inc.

  5. Two-photon induced collagen cross-linking in bioartificial cardiac tissue

    Science.gov (United States)

    Kuetemeyer, Kai; Kensah, George; Heidrich, Marko; Meyer, Heiko; Martin, Ulrich; Gruh, Ina; Heisterkamp, Alexander

    2011-08-01

    Cardiac tissue engineering is a promising strategy for regenerative therapies to overcome the shortage of donor organs for transplantation. Besides contractile function, the stiffness of tissue engineered constructs is crucial to generate transplantable tissue surrogates with sufficient mechanical stability to withstand the high pressure present in the heart. Although several collagen cross-linking techniques have proven to be efficient in stabilizing biomaterials, they cannot be applied to cardiac tissue engineering, as cell death occurs in the treated area. Here, we present a novel method using femtosecond (fs) laser pulses to increase the stiffness of collagen-based tissue constructs without impairing cell viability. Raster scanning of the fs laser beam over riboflavin-treated tissue induced collagen cross-linking by two-photon photosensitized singlet oxygen production. One day post-irradiation, stress-strain measurements revealed increased tissue stiffness by around 40% being dependent on the fibroblast content in the tissue. At the same time, cells remained viable and fully functional as demonstrated by fluorescence imaging of cardiomyocyte mitochondrial activity and preservation of active contraction force. Our results indicate that two-photon induced collagen cross-linking has great potential for studying and improving artificially engineered tissue for regenerative therapies.

  6. Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate.

    Science.gov (United States)

    Eibl, Matthias; Karpf, Sebastian; Weng, Daniel; Hakert, Hubertus; Pfeiffer, Tom; Kolb, Jan Philip; Huber, Robert

    2017-07-01

    Two-photon-excited fluorescence lifetime imaging microscopy (FLIM) is a chemically specific 3-D sensing modality providing valuable information about the microstructure, composition and function of a sample. However, a more widespread application of this technique is hindered by the need for a sophisticated ultra-short pulse laser source and by speed limitations of current FLIM detection systems. To overcome these limitations, we combined a robust sub-nanosecond fiber laser as the excitation source with high analog bandwidth detection. Due to the long pulse length in our configuration, more fluorescence photons are generated per pulse, which allows us to derive the lifetime with a single excitation pulse only. In this paper, we show high quality FLIM images acquired at a pixel rate of 1 MHz. This approach is a promising candidate for an easy-to-use and benchtop FLIM system to make this technique available to a wider research community.

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

    Science.gov (United States)

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

    2017-07-01

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

  8. N-doped carbon dots derived from bovine serum albumin and formic acid with one- and two-photon fluorescence for live cell nuclear imaging.

    Science.gov (United States)

    Tan, Mingqian; Li, Xintong; Wu, Hao; Wang, Beibei; Wu, Jing

    2015-12-01

    Carbon dots with both one- and two-photon fluorescence have drawn great attention for biomedical imaging. Herein, nitrogen-doped carbon dots were facilely developed by one-pot hydrothermal method using bovine serum albumin and formic acid as carbon sources. They are highly water-soluble with strong fluorescence when excited with ultraviolet or near infrared light. The carbon dots have a diameter of ~8.32 nm and can emit strong two-photon induced fluorescence upon excitation at 750 nm with a femtosecond laser. X-ray photoelectron spectrometer analysis revealed that the carbon dots contained three components, C, N and O, corresponding to the peak at 285, 398 and 532 eV, respectively. The Fourier-transform infrared spectroscopy analysis revealed that there are carboxyl and carboxylic groups on the surface, which allowed further linking of functional molecules. pH stability study demonstrated that the carbon dots are able to be used in a wide range of pH values. The fluorescence mechanism is also discussed in this study. Importantly, these carbon dots are biocompatible and highly photostable, which can be directly applied for both one- and two-photon living cell imaging. After proper surface functionalization with TAT peptide, they can be used as fluorescent probes for live cell nuclear-targeted imaging. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Stratum corneum lipid organization as observed by atomic force, confocal and two-photon excitation fluorescence microscopy

    DEFF Research Database (Denmark)

    Norlén, Lars; Plasencia Gil, Maria Inés; Bagatolli, Luis

    2008-01-01

    -related biophysical techniques (e.g. atomic force microscopy and confocal/two-photon excitation fluorescence microscopy), it was recently shown that reconstituted membranes composed of extracted decontaminated human stratum corneum lipids do not form a fluid phase, but exclusively a single-gel phase that segregates...

  10. Two-photon excited fluorescence spectroscopy and imaging of melanin in vitro and in vivo

    Science.gov (United States)

    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.

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

    Science.gov (United States)

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

    2016-10-04

    Three water-soluble tetracationic quadrupolar chromophores comprising two three-coordinate boron π-acceptor groups bridged by thiophene-containing moieties were synthesised for biological imaging applications. Compound 3 containing the bulkier 5-(3,5-Me2 C6 H2 )-2,2'-(C4 H2 S)2 -5'-(3,5-Me2 C6 H2 ) bridge is stable over a long period of time, exhibits a high fluorescence quantum yield and strong one- and two-photon absorption (TPA), and has a TPA cross section of 268 GM at 800 nm in water. Confocal laser scanning fluorescence microscopy studies in live cells indicated localisation of the chromophore at the mitochondria; moreover, cytotoxicity measurements proved biocompatibility. Thus, chromophore 3 has excellent potential for one- and two-photon-excited fluorescence imaging of mitochondrial function in cells. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Imaging of Fluoride Ion in Living Cells and Tissues with a Two-Photon Ratiometric Fluorescence Probe

    Directory of Open Access Journals (Sweden)

    Xinyue Zhu

    2015-01-01

    Full Text Available A reaction-based two-photon (TP ratiometric fluorescence probe Z2 has been developed and successfully applied to detect and image fluoride ion in living cells and tissues. The Z2 probe was designed designed to utilize an ICT mechanism between n-butylnaphthalimide as a fluorophore and tert-butyldiphenylsilane (TBDPS as a response group. Upon addition of fluoride ion, the Si-O bond in the Z2 would be cleaved, and then a stronger electron-donating group was released. The fluorescent changes at 450 and 540 nm, respectively, made it possible to achieve ratiometric fluorescence detection. The results indicated that the Z2 could ratiometrically detect and image fluoride ion in living cells and tissues in a depth of 250 μm by two-photon microscopy (TPM.

  13. Silole-Based Red Fluorescent Organic Dots for Bright Two-Photon Fluorescence In vitro Cell and In vivo Blood Vessel Imaging.

    Science.gov (United States)

    Chen, Bin; Feng, Guangxue; He, Bairong; Goh, Chiching; Xu, Shidang; Ramos-Ortiz, Gabriel; Aparicio-Ixta, Laura; Zhou, Jian; Ng, Laiguan; Zhao, Zujin; Liu, Bin; Tang, Ben Zhong

    2016-02-10

    Robust luminescent dyes with efficient two-photon fluorescence are highly desirable for biological imaging applications, but those suitable for organic dots fabrication are still rare because of aggregation-caused quenching. In this work, a red fluorescent silole, 2,5-bis[5-(dimesitylboranyl)thiophen-2-yl]-1-methyl-1,3,4-triphenylsilole ((MesB)2 DTTPS), is synthesized and characterized. (MesB)2 DTTPS exhibits enhanced fluorescence efficiency in nanoaggregates, indicative of aggregation-enhanced emission (AEE). The organic dots fabricated by encapsulating (MesB)2 DTTPS within lipid-PEG show red fluorescence peaking at 598 nm and a high fluorescence quantum yield of 32%. Upon excitation at 820 nm, the dots show a large two-photon absorption cross section of 3.43 × 10(5) GM, which yields a two-photon action cross section of 1.09 × 10(5) GM. These (MesB)2 DTTPS dots show good biocompatibility and are successfully applied to one-photon and two-photon fluorescence imaging of MCF-7 cells and two-photon in vivo visualization of the blood vascular of mouse muscle in a high-contrast and noninvasive manner. Moreover, the 3D blood vasculature located at the mouse ear skin with a depth of over 100 μm can also be visualized clearly, providing the spatiotemporal information about the whole blood vascular network. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Dynamic characterization of hydrophobic and hydrophilic solutes in oleic-acid enhanced transdermal delivery using two-photon fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tseng, Te-Yu; Yang, Chiu-Sheng; Chen, Yang-Fang [Department of Physics, National Taiwan University, Taipei, Taiwan (China); Tsai, Tsung-Hua [Department of Dermatology, Far Eastern Memorial Hospital, New Taipei City, Taiwan (China); Dong, Chen-Yuan, E-mail: cydong@phys.ntu.edu.tw [Department of Physics, National Taiwan University, Taipei, Taiwan (China); Center for Quantum Science and Engineering, National Taiwan University, Taipei, Taiwan (China); Center for Optoelectronic Biomedicine, National Taiwan University, Taipei, Taiwan (China)

    2014-10-20

    In this letter, we propose an efficient methodology of investigating dynamic properties of sulforhodamine B and rhodamine B hexyl ester molecules transporting across ex-vivo human stratum corneum with and without oleic acid enhancement. Three-dimensional, time-lapse fluorescence images of the stratum corneum can be obtained using two-photon fluorescence microscopy. Furthermore, temporal quantifications of transport enhancements in diffusion parameters can be achieved with the use of Fick's second law. Dynamic characterization of solutes transporting across the stratum corneum is an effective method for understanding transient phenomena in transdermal delivery of probe molecules, leading to improved delivery strategies of molecular species for therapeutic purposes.

  15. Direct Vpr-Vpr Interaction in Cells monitored by two Photon Fluorescence Correlation Spectroscopy and Fluorescence Lifetime Imaging

    Directory of Open Access Journals (Sweden)

    Mély Yves

    2008-09-01

    Full Text Available Abstract Background The human immunodeficiency virus type 1 (HIV-1 encodes several regulatory proteins, notably Vpr which influences the survival of the infected cells by causing a G2/M arrest and apoptosis. Such an important role of Vpr in HIV-1 disease progression has fuelled a large number of studies, from its 3D structure to the characterization of specific cellular partners. However, no direct imaging and quantification of Vpr-Vpr interaction in living cells has yet been reported. To address this issue, eGFP- and mCherry proteins were tagged by Vpr, expressed in HeLa cells and their interaction was studied by two photon fluorescence lifetime imaging microscopy and fluorescence correlation spectroscopy. Results Results show that Vpr forms homo-oligomers at or close to the nuclear envelope. Moreover, Vpr dimers and trimers were found in the cytoplasm and in the nucleus. Point mutations in the three α helices of Vpr drastically impaired Vpr oligomerization and localization at the nuclear envelope while point mutations outside the helical regions had no effect. Theoretical structures of Vpr mutants reveal that mutations within the α-helices could perturb the leucine zipper like motifs. The ΔQ44 mutation has the most drastic effect since it likely disrupts the second helix. Finally, all Vpr point mutants caused cell apoptosis suggesting that Vpr-mediated apoptosis functions independently from Vpr oligomerization. Conclusion We report that Vpr oligomerization in HeLa cells relies on the hydrophobic core formed by the three α helices. This oligomerization is required for Vpr localization at the nuclear envelope but not for Vpr-mediated apoptosis.

  16. Sensitive and rapid detection of endogenous hydrogen sulfide distributing in different mouse viscera via a two-photon fluorescent probe

    International Nuclear Information System (INIS)

    Chen, Qian; Yang, Jinfeng; Li, Yinhui; Zheng, Jing; Yang, Ronghua

    2015-01-01

    Development of efficient methods for detection of endogenous H 2 S in living cells and tissues is of considerable significance for better understanding the biological and pathological functions of H 2 S. Two-photon (TP) fluorescent probes are favorable as powerful molecular tools for studying physiological process due to its non-invasiveness, high spatiotemporal resolution and deep-tissues imaging. Up to date, several TP probes for intracellular H 2 S imaging have been designed, but real-time imaging of endogenous H 2 S-related biological processes in tissues is hampered due to low sensitivity, long response time and interference from other biothiols. To address this issue, we herein report a novel two-photon fluorescent probe (TPP-H 2 S) for highly sensitive and fast monitoring and imaging H 2 S levels in living cells and tissues. In the presence of H 2 S, it exhibits obviously improved sensitivity (LOD: 0.12 μM) and fast response time (about 2 min) compared with the reported two-photon H 2 S probes. With two-photon excitation, TPP-H 2 S displays high signal-to-noise ratio and sensitivity even no interference in cell growth media. As further application, TPP-H 2 S is applied for fast imaging of H 2 S in living cells and different fresh tissues by two-photon confocal microscope. Most importantly we first measured the endogenous H 2 S level in different viscera by vivisection and found that the distribution of endogenous H 2 S mostly in brain, liver and lung. The excellent sensing properties of TPP-H 2 S make it a practically useful tool for further studying biological roles of H 2 S. - Highlights: • This two-photon probe exhibits an improved sensitivity and response time to H 2 S. • This probe shows excellent membrane permeability and fast visualization of H 2 S in living cells and tissues. • This probe is successfully applied to measure the endogenously produced H 2 S levels in different viscera of mouse.

  17. Two-photon excitation with pico-second fluorescence lifetime imaging to detect nuclear association of flavanols

    Energy Technology Data Exchange (ETDEWEB)

    Mueller-Harvey, Irene, E-mail: i.mueller-harvey@reading.ac.uk [Chemistry and Biochemistry Laboratory, Food Production and Quality Research Division, School of Agriculture, Policy and Development, University of Reading, P O Box 236, Reading RG6 6AT (United Kingdom); Feucht, Walter, E-mail: walter.feucht@gmail.com [Department of Plant Sciences, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Polster, Juergen, E-mail: j.polster@wzw.tum.de [Department of Physical Biochemistry, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Trnkova, Lucie, E-mail: lucie.trnkova@uhk.cz [University of Hradec Kralove, Faculty of Science, Department of Chemistry, Rokitanskeho 62, 50003 Hradec Kralove (Czech Republic); Burgos, Pierre, E-mail: pierre.burgos@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Parker, Anthony W., E-mail: tony.parker@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Botchway, Stanley W., E-mail: stan.botchway@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom)

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer This fluorescence lifetime imaging microscopy (FLIM) technique for flavanols overcomes autofluorescence interference in cells. Black-Right-Pointing-Pointer Plant flavanols differed in their lifetimes. Black-Right-Pointing-Pointer Dissolved and bound flavanols revealed contrasting lifetime changes. Black-Right-Pointing-Pointer This technique will allow studying of flavanol trafficking in live cells. - Abstract: Two-photon excitation enabled for the first time the observation and measurement of excited state fluorescence lifetimes from three flavanols in solution, which were {approx}1.0 ns for catechin and epicatechin, but <45 ps for epigallocatechin gallate (EGCG). The shorter lifetime for EGCG is in line with a lower fluorescence quantum yield of 0.003 compared to catechin (0.015) and epicatechin (0.018). In vivo experiments with onion cells demonstrated that tryptophan and quercetin, which tend to be major contributors of background fluorescence in plant cells, have sufficiently low cross sections for two-photon excitation at 630 nm and therefore do not interfere with detection of externally added or endogenous flavanols in Allium cepa or Taxus baccata cells. Applying two-photon excitation to flavanols enabled 3-D fluorescence lifetime imaging microscopy and showed that added EGCG penetrated the whole nucleus of onion cells. Interestingly, EGCG and catechin showed different lifetime behaviour when bound to the nucleus: EGCG lifetime increased from <45 to 200 ps, whilst catechin lifetime decreased from 1.0 ns to 500 ps. Semi-quantitative measurements revealed that the relative ratios of EGCG concentrations in nucleoli associated vesicles: nucleus: cytoplasm were ca. 100:10:1. Solution experiments with catechin, epicatechin and histone proteins provided preliminary evidence, via the appearance of a second lifetime ({tau}{sub 2} = 1.9-3.1 ns), that both flavanols may be interacting with histone proteins. We conclude that there

  18. Construction of an efficient two-photon fluorescent probe for imaging nitroreductase in live cells and tissues

    Science.gov (United States)

    Zhou, Liyi; Gong, Liang; Hu, Shunqin

    2018-06-01

    Compared with traditional confocal microscopy, two-photon fluorescence microscopy (TPFM), which excites a two-photon (TP) fluorophore by near-infrared light, provides improved three-dimensional image resolution with increased tissue-image depth (>500 μm) and an extended observation time. Therefore, the development of novel functional TP fluorophores has attracted great attention in recent years. Herein, a novel TP fluorophore CM-NH2, which have the donor-π-acceptor (D-π-A)-structure, was designed and synthesized. We further used this dye developed a new type of TP fluorescent probe CM-NO2 for detecting nitroreductase (NTR). Upon incubated with NTR for 15 min, CM-NO2 displayed a 90-fold fluorescence enhancement at 505 nm and the maximal TP action cross-section value after reaction was detected and calculated to be 200 GM at 760 nm. The probe exhibited excellent properties such as high sensitivity, high selectivity, low cytotoxicity, and high photostability. Moreover, the probe was utilized to image the tumor hypoxia in live HeLa cells. Finally, using the CM-NO2 to image NTR in tissues was demonstrated.

  19. Application of laser fluorescence spectroscopy by two-photon excitation into atomic hydrogen density measurement in reactive plasmas

    International Nuclear Information System (INIS)

    Kajiwara, Toshinori; Takeda, Kazuyuki; Kim, Hee Je; Park, Won Zoo; Muraoka, Katsunori; Akazaki, Masanori; Okada, Tatsuo; Maeda, Mitsuo.

    1990-01-01

    Density profiles of hydrogen atoms in reactive plasmas of hydrogen and methane gases were measured, for the first time, using the laser fluorescence spectroscopy by two-photon excitation of Lyman beta transition and observation at the Balmer alpha radiation. Absolute density determinations showed atomic densities of around 3 x 10 17 m -3 , or the degree of dissociation to be 10 -4 . Densities along the axis perpendicular to the RF electrode showed peaked profiles, which were due to the balance of atomic hydrogen production by electron impact on molecules against diffusion loss to the walls. (author)

  20. Resonant two-photon absorption and electromagnetically induced transparency in open ladder-type atomic system.

    Science.gov (United States)

    Moon, Han Seb; Noh, Heung-Ryoul

    2013-03-25

    We have experimentally and theoretically studied resonant two-photon absorption (TPA) and electromagnetically induced transparency (EIT) in the open ladder-type atomic system of the 5S(1/2) (F = 1)-5P(3/2) (F' = 0, 1, 2)-5D(5/2) (F″ = 1, 2, 3) transitions in (87)Rb atoms. As the coupling laser intensity was increased, the resonant TPA was transformed to EIT for the 5S(1/2) (F = 1)-5P(3/2) (F' = 2)-5D(5/2) (F″ = 3) transition. The transformation of resonant TPA into EIT was numerically calculated for various coupling laser intensities, considering all the degenerate magnetic sublevels of the 5S(1/2)-5P(3/2)-5D(5/2) transition. From the numerical results, the crossover from TPA to EIT could be understood by the decomposition of the spectrum into an EIT component owing to the pure two-photon coherence and a TPA component caused by the mixed term.

  1. 3D imaging of intrinsic crystalline defects in zinc oxide by spectrally resolved two-photon fluorescence microscopy

    Science.gov (United States)

    Al-Tabich, A.; Inami, W.; Kawata, Y.; Jablonski, R.; Worasawat, S.; Mimura, H.

    2017-05-01

    We present a method for three-dimensional intrinsic defect imaging in zinc oxide (ZnO) by spectrally resolved two-photon fluorescence microscopy, based on the previously presented method of observing a photoluminescence distribution in wide-gap semiconductor crystals [Noor et al., Appl. Phys. Lett. 92(16), 161106 (2008)]. A tightly focused light beam radiated by a titanium-sapphire laser is used to obtain a two-photon excitation of selected area of the ZnO sample. Photoluminescence intensity of a specific spectral range is then selected by optical band pass filters and measured by a photomultiplier tube. Reconstruction of the specimen image is done by scanning the volume of interest by a piezoelectric positioning stage and measuring the spectrally resolved photoluminescence intensity at each point. The method has been proved to be effective at locating intrinsic defects of the ZnO crystalline structure in the volume of the crystal. The method was compared with other defect imaging and 3D imaging techniques like scanning tunneling microscopy and confocal microscopy. In both cases, our method shows superior penetration abilities and, as the only method, allows location of the defects of the chosen type in 3D. In this paper, we present the results of oxygen vacancies and zinc antisites imaging in ZnO nanorods.

  2. Preparation of a Two-Photon Fluorescent Probe for Imaging H2O2 in Lysosomes in Living Cells and Tissues.

    Science.gov (United States)

    Ren, Mingguang; Deng, Beibei; Kong, Xiuqi; Tang, Yonghe; Lin, Weiying

    2017-01-01

    Hydrogen peroxide (H 2 O 2 ) plays important roles in many physiological and pathological processes. At the cellular organelle level, the abnormal concentrations of H 2 O 2 in the lysosomes may cause redox imbalance and the loss of the critical functions of the lysosomes. Herein, we describe the preparation of a potent lysosome-targeted two-photon fluorescent probe (Lyso-HP) for the detection of H 2 O 2 in the lysosomes in the living cells. This unique fluorescent probe can also be employed to effectively detect H 2 O 2 in the living tissues using two-photon fluorescence microscopy.

  3. A bioaccumulative cyclometalated platinum(II) complex with two-photon-induced emission for live cell imaging.

    Science.gov (United States)

    Koo, Chi-Kin; Wong, Ka-Leung; Man, Cornelia Wing-Yin; Lam, Yun-Wah; So, Leo King-Yan; Tam, Hoi-Lam; Tsao, Sai-Wah; Cheah, Kok-Wai; Lau, Kai-Chung; Yang, Yang-Yi; Chen, Jin-Can; Lam, Michael Hon-Wah

    2009-02-02

    The cyclometalated platinum(II) complex [Pt(L)Cl], where HL is a new cyclometalating ligand 2-phenyl-6-(1H-pyrazol-3-yl)pyridine containing C(phenyl), N(pyridyl), and N(pyrazolyl) donor moieties, was found to possess two-photon-induced luminescent properties. The two-photon-absorption cross section of the complex in N,N-dimethylformamide at room temperature was measured to be 20.8 GM. Upon two-photon excitation at 730 nm from a Ti:sapphire laser, bright-green emission was observed. Besides its two-photon-induced luminescent properties, [Pt(L)Cl] was able to be rapidly accumulated in live HeLa and NIH3T3 cells. The two-photon-induced luminescence of the complex was retained after live cell internalization and can be observed by two-photon confocal microscopy. Its bioaccumulation properties enabled time-lapse imaging of the internalization process of the dye into living cells. Cytotoxicity of [Pt(L)Cl] to both tested cell lines was low, according to MTT assays, even at loadings as high as 20 times the dose concentration for imaging for 6 h.

  4. Two-Photon Absorption Properties of Gold Fluorescent Protein: A Combined Molecular Dynamics and Quantum Chemistry Study.

    Science.gov (United States)

    Simsek, Yusuf; Brown, Alex

    2018-05-09

    Molecular dynamic (MD) simulations were carried out to obtain the conformational changes of the chromophore in the gold fluorescent protein (PDB ID: 1OXF). To obtain two-photon absorption (TPA) cross-sections, time dependent density functional theory (TD-DFT) computations were performed for chromophore geometries sampled along the trajectory. The TD-DFT computations used the CAM-B3LYP functional and 6-31+G(d) basis set with the conductor-like polarizable continuum model (PCM) with parameters for water. Results showed that two dihedral angles change remarkably over the simulation time. TPA cross-sections were found to average 20 GM for the excitation to S1 between 430 and 460 nm; however, the maximal and minimal values were 35GM and 5GM, respectively. Besides the effects of the dihedrals on the spectroscopic properties, some bond lengths affected the excitation energies and the TPA cross-sections significantly (up to ±25-30%) while the effects of bond angles were smaller (±5%). Overall the present results provide insight in the effects of conformational exibility on TPA (with gold fluorescent protein as a specific example) and suggest that further experimental measurements of TPA for gold fluorescent protein should be undertaken.

  5. Mercury effects on Thalassiosira weissflogii: Applications of two-photon excitation chlorophyll fluorescence lifetime imaging and flow cytometry

    International Nuclear Information System (INIS)

    Wu Yun; Zeng Yan; Qu, Jianan Y.; Wang Wenxiong

    2012-01-01

    The toxic effects of inorganic mercury [Hg(II)] and methylmercury (MeHg) on the photosynthesis and population growth in a marine diatom Thalassiosira weissflogii were investigated using two methods: two-photon excitation fluorescence lifetime imaging (FLIM) and flow cytometry (FCM). For photosynthesis, Hg(II) exposure increased the average chlorophyll fluorescence lifetime, whereas such increment was not found under MeHg stress. This may be caused by the inhibitory effect of Hg(II) instead of MeHg on the electron transport chain. For population growth, modeled specific growth rate data showed that the reduction in population growth by Hg(II) mainly resulted from an increased number of injured cells, while the live cells divided at the normal rates. However, MeHg inhibitory effects on population growth were contributed by the reduced division rates of all cells. Furthermore, the cell images and the FCM data reflected the morphological changes of diatom cells under Hg(II)/MeHg exposure vividly and quantitatively. Our results demonstrated that the toxigenicity mechanisms between Hg(II) and MeHg were different in the algal cells.

  6. Photonic devices based on patterning by two photon induced polymerization techniques

    Science.gov (United States)

    Fortunati, I.; Dainese, T.; Signorini, R.; Bozio, R.; Tagliazucca, V.; Dirè, S.; Lemercier, G.; Mulatier, J.-C.; Andraud, C.; Schiavuta, P.; Rinaldi, A.; Licoccia, S.; Bottazzo, J.; Franco Perez, A.; Guglielmi, M.; Brusatin, G.

    2008-04-01

    Two and three dimensional structures with micron and submicron resolution have been achieved in commercial resists, polymeric materials and sol-gel materials by several lithographic techniques. In this context, silicon-based sol-gel materials are particularly interesting because of their versatility, chemical and thermal stability, amount of embeddable active compounds. Compared with other micro- and nano-fabrication schemes, the Two Photon Induced Polymerization is unique in its 3D processing capability. The photopolymerization is performed with laser beam in the near-IR region, where samples show less absorption and less scattering, giving rise to a deeper penetration of the light. The use of ultrashort laser pulses allows the starting of nonlinear processes like multiphoton absorption at relatively low average power without thermally damaging the samples. In this work we report results on the photopolymerization process in hybrid organic-inorganic films based photopolymerizable methacrylate-containing Si-nanobuilding blocks. Films, obtained through sol-gel synthesis, are doped with a photo-initiator allowing a radical polymerization of methacrylic groups. The photo-initiator is activated by femtosecond laser source, at different input energies. The development of the unexposed regions is performed with a suitable solvent and the photopolymerized structures are characterized by microscopy techniques.

  7. Biexcitonic photocurrent induced by two-photon process at a telecommunication band

    International Nuclear Information System (INIS)

    Kodera, Tetsuo; Miyazawa, Toshiyuki; Kumagai, Naoto; Watanabe, Katsuyuki; Suzuki, Ayako; Takagi, Hiroyuki; Nakaoka, Toshihiro; Arakawa, Yasuhiko

    2009-01-01

    We report on photocurrent (PC) measurements of biexciton in a single self-assembled InAs quantum dot (QD) at a telecommunication wavelength of 1.3μm. We use shadow mask technique on an n-i Schottky photodiode structure with QDs to excite a single QD resonantly. Coherent pulse excitation is realized in two types of setups utilizing (i) an optical parametric oscillator and (ii) a stable semiconductor laser diode. In both setups we observe the biexcitonic PC peaks induced by a coherent two-photon process. Especially in the latter setups, the narrower pulse linewidth in energy provides a clearer biexcitonic PC peak because of reduced unwanted excitation. We estimate the binding energy ΔE B of our telecom-band biexciton to be 0.9 meV from the splitting between excitonic and biexcitonic resonances. The result suggests our telecom-band exciton-biexciton system is a good candidate for the building block of fiber-based controlled-rotation quantum logic operation. (copyright 2009 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  8. Integrated femtosecond stimulated Raman scattering and two-photon fluorescence imaging of subcellular lipid and vesicular structures

    Science.gov (United States)

    Li, Xuesong; Lam, Wen Jiun; Cao, Zhe; Hao, Yan; Sun, Qiqi; He, Sicong; Mak, Ho Yi; Qu, Jianan Y.

    2015-11-01

    The primary goal of this study is to demonstrate that stimulated Raman scattering (SRS) as a new imaging modality can be integrated into a femtosecond (fs) nonlinear optical (NLO) microscope system. The fs sources of high pulse peak power are routinely used in multimodal nonlinear microscopy to enable efficient excitation of multiple NLO signals. However, with fs excitations, the SRS imaging of subcellular lipid and vesicular structures encounters significant interference from proteins due to poor spectral resolution and a lack of chemical specificity, respectively. We developed a unique NLO microscope of fs excitation that enables rapid acquisition of SRS and multiple two-photon excited fluorescence (TPEF) signals. In the in vivo imaging of transgenic C. elegans animals, we discovered that by cross-filtering false positive lipid signals based on the TPEF signals from tryptophan-bearing endogenous proteins and lysosome-related organelles, the imaging system produced highly accurate assignment of SRS signals to lipid. Furthermore, we demonstrated that the multimodal NLO microscope system could sequentially image lipid structure/content and organelles, such as mitochondria, lysosomes, and the endoplasmic reticulum, which are intricately linked to lipid metabolism.

  9. Volumetric label-free imaging and 3D reconstruction of mammalian cochlea based on two-photon excitation fluorescence microscopy

    International Nuclear Information System (INIS)

    Zhang, Xianzeng; Zhan, Zhenlin; Xie, Shusen; Geng, Yang; Ye, Qing

    2013-01-01

    The visualization of the delicate structure and spatial relationship of intracochlear sensory cells has relied on the laborious procedures of tissue excision, fixation, sectioning and staining for light and electron microscopy. Confocal microscopy is advantageous for its high resolution and deep penetration depth, yet disadvantageous due to the necessity of exogenous labeling. In this study, we present the volumetric imaging of rat cochlea without exogenous dyes using a near-infrared femtosecond laser as the excitation mechanism and endogenous two-photon excitation fluorescence (TPEF) as the contrast mechanism. We find that TPEF exhibits strong contrast, allowing cellular and even subcellular resolution imaging of the cochlea, differentiating cell types, visualizing delicate structures and the radial nerve fiber. Our results further demonstrate that 3D reconstruction rendered with z-stacks of optical sections enables better revealment of fine structures and spatial relationships, and easily performed morphometric analysis. The TPEF-based optical biopsy technique provides great potential for new and sensitive diagnostic tools for hearing loss or hearing disorders, especially when combined with fiber-based microendoscopy. (paper)

  10. Endogenous Two-Photon Excited Fluorescence Provides Label-Free Visualization of the Inflammatory Response in the Rodent Spinal Cord

    Directory of Open Access Journals (Sweden)

    Ortrud Uckermann

    2015-01-01

    Full Text Available Activation of CNS resident microglia and invasion of external macrophages plays a central role in spinal cord injuries and diseases. Multiphoton microscopy based on intrinsic tissue properties offers the possibility of label-free imaging and has the potential to be applied in vivo. In this work, we analyzed cellular structures displaying endogenous two-photon excited fluorescence (TPEF in the pathologic spinal cord. It was compared qualitatively and quantitatively to Iba1 and CD68 immunohistochemical staining in two models: rat spinal cord injury and mouse encephalomyelitis. The extent of tissue damage was retrieved by coherent anti-Stokes Raman scattering (CARS and second harmonic generation imaging. The pattern of CD68-positive cells representing postinjury activated microglia/macrophages was colocalized to the TPEF signal. Iba1-positive microglia were found in areas lacking any TPEF signal. In peripheral areas of inflammation, we found similar numbers of CD68-positive microglia/macrophages and TPEF-positive structures while the number of Iba1-positive cells was significantly higher. Therefore, we conclude that multiphoton imaging of unstained spinal cord tissue enables retrieving the extent of microglia activation by acquisition of endogenous TPEF. Future application of this technique in vivo will enable monitoring inflammatory responses of the nervous system allowing new insights into degenerative and regenerative processes.

  11. Endogenous Two-Photon Excited Fluorescence Provides Label-Free Visualization of the Inflammatory Response in the Rodent Spinal Cord

    Science.gov (United States)

    Uckermann, Ortrud; Galli, Roberta; Beiermeister, Rudolf; Sitoci-Ficici, Kerim-Hakan; Later, Robert; Leipnitz, Elke; Chavakis, Triantafyllos; Koch, Edmund; Schackert, Gabriele; Steiner, Gerald; Kirsch, Matthias

    2015-01-01

    Activation of CNS resident microglia and invasion of external macrophages plays a central role in spinal cord injuries and diseases. Multiphoton microscopy based on intrinsic tissue properties offers the possibility of label-free imaging and has the potential to be applied in vivo. In this work, we analyzed cellular structures displaying endogenous two-photon excited fluorescence (TPEF) in the pathologic spinal cord. It was compared qualitatively and quantitatively to Iba1 and CD68 immunohistochemical staining in two models: rat spinal cord injury and mouse encephalomyelitis. The extent of tissue damage was retrieved by coherent anti-Stokes Raman scattering (CARS) and second harmonic generation imaging. The pattern of CD68-positive cells representing postinjury activated microglia/macrophages was colocalized to the TPEF signal. Iba1-positive microglia were found in areas lacking any TPEF signal. In peripheral areas of inflammation, we found similar numbers of CD68-positive microglia/macrophages and TPEF-positive structures while the number of Iba1-positive cells was significantly higher. Therefore, we conclude that multiphoton imaging of unstained spinal cord tissue enables retrieving the extent of microglia activation by acquisition of endogenous TPEF. Future application of this technique in vivo will enable monitoring inflammatory responses of the nervous system allowing new insights into degenerative and regenerative processes. PMID:26355949

  12. Fs-transient absorption and fluorescence upconversion after two- photon excitation of carotenoids in solution and in LHC II

    CERN Document Server

    Wall, P J; Fleming, G R

    2000-01-01

    With time resolved two-photon techniques we determined the lifetime and two-photon spectrum of the forbidden S/sub 1/ state of beta - carotene (9+or-0.2 ps), lutein (15+or-0.5 ps) and the energy transferring carotenoids in LHC II (250+or-50 fs). (7 refs).

  13. Two-photon imaging of formaldehyde in live cells and animals utilizing a lysosome-targetable and acidic pH-activatable fluorescent probe.

    Science.gov (United States)

    Xie, Xilei; Tang, Fuyan; Shangguan, Xiaoyan; Che, Shiyi; Niu, Jinye; Xiao, Yongsheng; Wang, Xu; Tang, Bo

    2017-06-13

    Lyso-TPFP presents lysosomal targetability and an acidic pH-activatable response toward formaldehyde. Thus, it exclusively visualizes lysosomal formaldehyde and is immune against it in neutral cytosol and other organelles. In addition, two-photon fluorescence imaging endows Lyso-TPFP with the capability of in situ tracking formaldehyde in live cells and animals.

  14. Enhanced Emission from Single Isolated Gold Quantum Dots Investigated Using Two-Photon-Excited Fluorescence Near-Field Scanning Optical Microscopy.

    Science.gov (United States)

    Abeyasinghe, Neranga; Kumar, Santosh; Sun, Kai; Mansfield, John F; Jin, Rongchao; Goodson, Theodore

    2016-12-21

    New approaches in molecular nanoscopy are greatly desired for interrogation of biological, organic, and inorganic objects with sizes below the diffraction limit. Our current work investigates emergent monolayer-protected gold quantum dots (nanoclusters, NCs) composed of 25 Au atoms by utilizing two-photon-excited fluorescence (TPEF) near-field scanning optical microscopy (NSOM) at single NC concentrations. Here, we demonstrate an approach to synthesize and isolate single NCs on solid glass substrates. Subsequent investigation of the NCs using TPEF NSOM reveals that, even when they are separated by distances of several tens of nanometers, we can excite and interrogate single NCs individually. Interestingly, we observe an enhanced two-photon absorption (TPA) cross section for single Au 25 NCs that can be attributed to few-atom local field effects and to local field-induced microscopic cascading, indicating their potential for use in ultrasensitive sensing, disease diagnostics, cancer cell therapy, and molecular computers. Finally, we report room-temperature aperture-based TPEF NSOM imaging of these NCs for the first time at 30 nm point resolution, which is a ∼5-fold improvement compared to the previous best result for the same technique. This report unveils the unique combination of an unusually large TPA cross section and the high photostability of Au NCs to (non-destructively) investigate stable isolated single NCs using TPEF NSOM. This is the first reported optical study of monolayer-protected single quantum clusters, opening some very promising opportunities in spectroscopy of nanosized objects, bioimaging, ultrasensitive sensing, molecular computers, and high-density data storage.

  15. A fast-response two-photon fluorescent probe for imaging endogenous H2O2 in living cells and tissues

    Science.gov (United States)

    Lu, Yanan; Shi, Xiaomin; Fan, Wenlong; Black, Cory A.; Lu, Zhengliang; Fan, Chunhua

    2018-02-01

    As a second messenger, hydrogen peroxide plays significant roles in numerous physiological and pathological processes and is related to various diseases including inflammatory disease, diabetes, neurodegenerative disorders, cardiovascular disease and Alzheimer's disease. Two-photon (TP) fluorescent probes reported for the detection of endogenous H2O2 are rare and most have drawbacks such as slow response and low sensitivity. In this report, we demonstrate a simple H2O2-specific TP fluorescent probe (TX-HP) containing a two-photon dye 6-hydroxy-2,3,4,4a-tetrahydro-1H-xanthen-1-one (TX) on the modulation of the ICT process. The probe exhibits a rapid fluorescent response to H2O2 in 9 min with both high sensitivity and selectivity. The probe can detect exogenous H2O2 in living cells. Furthermore, the probe is successfully utilized for imaging H2O2 in liver tissues.

  16. An ESIPT-based two-photon fluorescent probe detection of hydrogen peroxide in live cells and tissues.

    Science.gov (United States)

    Zhou, Liyi; Peng, Yongbo; Wang, Qianqian; Lin, Qinlu

    2017-02-01

    A variety of diseases associated with human aging, which have a strong oxidative stress, but connecting age-related diseases and oxidative stress of the basic molecular mechanisms still insufficiently understood. Oxidative stress origins from the unregulated production of reactive oxygen species (ROS), and oxidative damaging to tissues and organs from subsequent oxidation-reduction chemistry by cellular mismanagement. In particular, H 2 O 2 is a major by-product of ROS in live organisms and a common marker for oxidative stress, and its dynamic equilibrium can have various physiological and pathological consequences. H 2 O 2 is a small molecule, but it is an essential oxygen metabolite in living systems and acts as an important compound in cellular signal transduction by reversible oxidation of proteins. To quantitatively detect of H 2 O 2 in biosystems, herein, we adopted a 2-(2'-hydroxyphenyl)-4(3H)-quinazolinone (HPQ), a small organic fluorophore known for its luminescence mechanism through excited-state intramolecular proton transfer (ESIPT). HPQ was employed as a precursor to develop a turn-on probe (HPQ-H) for bioimaging applications. After cleavaging the boronic ester moiety by H 2 O 2 , HPQ-H releases a HPQ fluorophore which shows a 45-fold fluorescence intensity enhancement with high sensitivity and selectivity over other reactive oxygen species (ROS), and a high resolution imaging and large tissue-imaging depth (70-170μm) in living cells and tissues images under two-photon excitation (720nm). Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Two Photon Induced Lasing in 1550 nm Quantum Dash Optical Gain Media

    DEFF Research Database (Denmark)

    Capua, Amir; Saal, Abigael; Reithmaier, Johann Peter

    2011-01-01

    We report on a unique lasing mechanism observed in quantum dash Gain media. While the gain media is electrically pumped below lasing threshold, a strong optical pulse excites carriers by two photon absorption into high energy states of the quantum dashes and wetting layer. Fast inter band carrier...

  18. Measurement Induced Enhancement of Squeezing in Nondegenerate Two-Photon Jaynes-Cummings Model

    International Nuclear Information System (INIS)

    Ye Saiyun

    2006-01-01

    Squeezing properties in the nondegenerate two-photon Jaynes-Cummings model are investigated. The effects of direct selective atomic measurement and the application of the classical field followed by atomic measurement are analyzed. Different values of the parameters of the classical field are taken into account. It is found that the field squeezing can be enhanced by measurement.

  19. Two-photon fluorescent polysiloxane-based films with thermally responsive self switching properties achieved by a unique reversible spirocyclization mechanism.

    Science.gov (United States)

    Zuo, Yujing; Yang, Tingxin; Zhang, Yu; Gou, Zhiming; Tian, Minggang; Kong, Xiuqi; Lin, Weiying

    2018-03-14

    Responsiveness and reversibility are present in nature, and are ubiquitous in biological systems. The realization of reversibility and responsiveness is of great importance in the development of properties and the design of new materials. However, two-photon fluorescent thermal-responsive materials have not been reported to date. Herein, we engineered thermally responsive polysiloxane materials ( Dns-non ) that exhibited unique two-photon luminescence, and this is the first report about thermally responsive luminescent materials with two-photon fluorescence. The fluorescence of Dns-non could switch from the "on" to "off" state through a facile heating and cooling process, which could be observed by the naked eye. Monitoring the temperature of the CPU in situ was achieved by easily coating D1-non onto the CPU surface, which verified the potential application in devices of Dns-non . A unique alkaline tuned reversible transition mechanism of rhodamine-B from its spirocyclic to its ring-open state was proposed. Furthermore, Dns-non appeared to be a useful cell adhesive for the culture of cells on the surface. We believe that the constructed thermally responsive silicon films which have promising utilization as a new type of functional fluorescent material, may show broad applications in materials chemistry or bioscience.

  20. Fluorescence excitation analysis by two-photon confocal laser scanning microscopy: a new method to identify fluorescent nanoparticles on histological tissue sections

    Directory of Open Access Journals (Sweden)

    Kahn E

    2012-10-01

    Full Text Available Edmond Kahn,1 Nicolas Tissot,3 Perrine Frere,3 Aurélien Dauphin,3 Mohamed Boumhras,2,4 Claude-Marie Bachelet,3 Frédérique Frouin,1 Gérard Lizard21Institut National de la Santé et de la Recherche Médicale (INSERM U678/UMR-S UPMC, CHU Pitié-Salpêtrière, Paris, France; 2Equipe Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique EA7270, Faculté des Sciences Gabriel, Université de Bourgogne-INSERM Dijon, France; 3Plateforme d'Imagerie cellulaire, UPMC, Paris, France; 4Laboratory of Biochemistry and Neuroscience, Applied Toxicology Group, Faculty of Science and Technology, Settat, MoroccoAbstract: In the present study, we make use of the ability of two-photon confocal laser scanning microscopes (CLSMs equipped with tunable lasers to produce spectral excitation image sequences. Furthermore, unmixing, which is usually performed on emission image sequences, is performed on these excitation image sequences. We use factor analysis of medical image sequences (FAMIS, which produces factor images, to unmix spectral image sequences of stained structures in tissue sections to provide images of characterized stained cellular structures. This new approach is applied to histological tissue sections of mouse aorta containing labeled iron nanoparticles stained with Texas Red and counterstained with SYTO13, to obtain visual information about the accumulation of these nanoparticles in the arterial wall. The possible presence of Texas Red is determined using a two-photon CLSM associated with FAMIS via the excitation spectra. Texas Red and SYTO13 are thus differentiated, and corresponding factor images specify their possible presence and cellular localization. In conclusion, the designed protocol shows that sequences of images obtained by excitation in a two-photon CLSM enables characterization of Texas Red-stained nanoparticles and other markers. This methodology offers an alternative and complementary solution to the conventional use of emission

  1. pH-Induced Modulation of One- and Two-Photon Absorption Properties in a Naphthalene-Based Molecular Probe.

    Science.gov (United States)

    Murugan, N Arul; Kongsted, Jacob; Ågren, Hans

    2013-08-13

    Presently, there is a great demand for small probe molecules that can be used for two-photon excitation microscopy (TPM)-based monitoring of intracellular and intraorganelle activity and pH. The candidate molecules should ideally possess a large two-photon absorption cross section with optical properties sensitive to pH changes. In the present work, we investigate the potential of a methoxy napthalene (MONAP) derivative for its suitability to serve as a pH sensor using TPM. Using an integrated approach rooted in hybrid quantum mechanics/molecular mechanics, the structures, dynamics, and the one- and two-photon properties of the probe in dimethylformamide solvent are studied. It is found that the protonated form is responsible for the optical property of MONAP at moderately low pH, for which the calculated pH-induced red shift is in good agreement with experiments. A 2-fold increase in the two-photon absorption cross section in the IR region of the spectrum is predicted for the moderately low pH form of the probe, suggesting that this can be a potential probe for pH monitoring of living cells. We also propose some design principles aimed at obtaining control of the absorption spectral range of the probe by structural tuning. Our work indicates that the integrated approach employed is capable of capturing the pH-induced changes in structure and optical properties of organic molecular probes and that such in silico tools can be used to draw structure-property relationships to design novel molecular probes suitable for a specific application.

  2. Details of the Collagen and Elastin Architecture in the Human Limbal Conjunctiva, Tenon's Capsule and Sclera Revealed by Two-Photon Excited Fluorescence Microscopy.

    Science.gov (United States)

    Park, Choul Yong; Marando, Catherine M; Liao, Jason A; Lee, Jimmy K; Kwon, Jiwon; Chuck, Roy S

    2016-10-01

    To investigate the architecture and distribution of collagen and elastin in human limbal conjunctiva, Tenon's capsule, and sclera. The limbal conjunctiva, Tenon's capsule, and sclera of human donor corneal buttons were imaged with an inverted two-photon excited fluorescence microscope. No fixation process was necessary. The laser (Ti:sapphire) was tuned at 850 nm for two-photon excitation. Backscatter signals of second harmonic generation (SHG) and autofluorescence (AF) were collected through a 425/30-nm and a 525/45-nm emission filter, respectively. Multiple, consecutive, and overlapping (z-stack) images were acquired. Collagen signals were collected with SHG, whereas elastin signals were collected with AF. The size and density of collagen bundles varied widely depending on depth: increasing from conjunctiva to sclera. In superficial image planes, collagen bundles were image planes (episclera and superficial sclera), collagen bundles were thicker (near 100 μm in width) and densely packed. Comparatively, elastin fibers were thinner and sparse. The orientation of elastin fibers was independent of collagen fibers in superficial layers; but in deep sclera, elastin fibers wove through collagen interbundle gaps. At the limbus, both collagen and elastin fibers were relatively compact and were distributed perpendicular to the limbal annulus. Two-photon excited fluorescence microscopy has enabled us to understand in greater detail the collagen and elastin architecture of the human limbal conjunctiva, Tenon's capsule, and sclera.

  3. Band structure of cavity-type hypersonic phononic crystals fabricated by femtosecond laser-induced two-photon polymerization

    Energy Technology Data Exchange (ETDEWEB)

    Rakhymzhanov, A. M.; Utegulov, Z. N., E-mail: zhutegulov@nu.edu.kz, E-mail: fytas@mpip-mainz.mpg.de [Department of Physics, School of Science and Technology, Nazarbayev University, Astana 010000 (Kazakhstan); Optics Laboratory, National Laboratory Astana, Nazarbayev University, Astana 10000 (Kazakhstan); Gueddida, A. [Institut d' Electronique, Microélectronique et Nanotechnologie, 59650 Villeneuve d' Ascq (France); LPMR, Département de Physique, Faculté des Sciences, Université Mohamed I, 60000 Oujda (Morocco); Alonso-Redondo, E. [Max Planck Institute of Polymer Research, Ackermannweg 10, 55128 Mainz (Germany); Perevoznik, D.; Kurselis, K. [Laser Zentrum Hannover e.V., 30419 Hannover (Germany); Chichkov, B. N. [Laser Zentrum Hannover e.V., 30419 Hannover (Germany); Institute of Laser and Information Technologies RAS, Moscow, 142092 Troitsk (Russian Federation); El Boudouti, E. H. [LPMR, Département de Physique, Faculté des Sciences, Université Mohamed I, 60000 Oujda (Morocco); Djafari-Rouhani, B. [Institut d' Electronique, Microélectronique et Nanotechnologie, 59650 Villeneuve d' Ascq (France); Fytas, G., E-mail: zhutegulov@nu.edu.kz, E-mail: fytas@mpip-mainz.mpg.de [Max Planck Institute of Polymer Research, Ackermannweg 10, 55128 Mainz (Germany); Department of Materials Science, University of Crete and FORTH, 71110 Heraklion (Greece)

    2016-05-16

    The phononic band diagram of a periodic square structure fabricated by femtosecond laser pulse-induced two photon polymerization is recorded by Brillouin light scattering (BLS) at hypersonic (GHz) frequencies and computed by finite element method. The theoretical calculations along the two main symmetry directions quantitatively capture the band diagrams of the air- and liquid-filled structure and moreover represent the BLS intensities. The theory helps identify the observed modes, reveals the origin of the observed bandgaps at the Brillouin zone boundaries, and unravels direction dependent effective medium behavior.

  4. Effect of detergents on the physico-chemical properties of skin stratum corneum: A two-photon excitation fluorescence microscopy study

    DEFF Research Database (Denmark)

    Bloksgaard, Maria; Brewer, Jonathan R.; Pashkovski, Eugene

    2014-01-01

    OBJECTIVE: Understanding the structural and dynamical features of skin is critical for advancing innovation in personal care and drug discovery. Synthetic detergent mixtures used in commercially available body wash products are thought to be less aggressive towards the skin barrier when compared...... to conventional detergents. The aim of this work is to comparatively characterize the effect of a mild synthetic cleanser mixture (SCM) and sodium dodecyl sulphate (SDS) on the hydration state of the intercellular lipid matrix and on proton activity of excised skin stratum corneum (SC). METHOD: Experiments were...... performed using two-photon excitation fluorescence microscopy. Fluorescent images of fluorescence reporters sensitive to proton activity and hydration of SC were obtained in excised skin and examined in presence and absence of SCM and SDS detergents. RESULTS: Hydration of the intercellular lipid matrix...

  5. Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf.

    Science.gov (United States)

    Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; Nguyen, Thien An; Alfano, Robert R

    2014-06-01

    Two-photon (2P) excitation of the second singlet (S₂) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S₂ state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S₂ state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

  6. Water soluble two-photon fluorescent organic probes for long-term imaging of lysosomes in live cells and tumor spheroids.

    Science.gov (United States)

    Kumari, Pratibha; Verma, Sanjay K; Mobin, Shaikh M

    2018-01-11

    The morphological alteration of lysosomes is a powerful indicator of various pathological disorders. In this regard, we have designed and synthesized a new water soluble fluorescent Schiff-base ligand (L-lyso) containing two hydroxyl groups. L-lyso exhibits excellent two-photon properties with tracking of lysosomes in live cells as well as in 3D tumor spheroids. Furthermore, it can label lysosomes for more than 3 days. Thus, L-lyso has an edge over the commercially available expensive LysoTracker probes and also over other reported probes in terms of its long-term imaging, water solubility and facile synthesis.

  7. Dipolar versus octupolar triphenylamine-based fluorescent organic nanoparticles as brilliant one- and two-photon emitters for (bio)imaging.

    Science.gov (United States)

    Parthasarathy, Venkatakrishnan; Fery-Forgues, Suzanne; Campioli, Elisa; Recher, Gaëlle; Terenziani, Francesca; Blanchard-Desce, Mireille

    2011-11-18

    Two related triphenylamine-based dipolar and octupolar fluorophores are used to prepare aqueous suspensions of fluorescent organic nanoparticles (FONs) via the reprecipitation method. The obtained spherical nanoparticles (30-40 nm in diameter) are fluorescent in aqueous solution (up to 15% fluorescence quantum yield) and exhibit extremely high one- and two-photon brightness, superior to those obtained for quantum dots. Despite the two chromophores showing similar fluorescence in solution, the fluorescence of FONs made from the octupolar derivative is significantly red-shifted compared to that generated by the dipolar FONs. In addition, the maximum two-photon absorption cross section of the FONs made from the octupolar derivative is 55% larger than that of the dipolar derivative FONs. The experimental observations provide evidence that the different molecular shape (rodlike versus three-branched) and charge distribution (dipolar versus octupolar) of the two chromophores strongly affect the packing inside the nanoparticles as well as their spectroscopic properties and colloidal stability in pure water. The use of these FONs as probes for biphotonic in-vivo imaging is investigated on Xenopus laevis tadpoles to test their utilization for angiography. When using FONs made from the octupolar dye, the formation of microagglomerates (2-5 μm scale) is observed in vivo, with subsequent lethal occlusion of the blood vessels. Conversely, the nanoparticles of the dipolar dye allow acute imaging of blood vessels thanks to their suitable size and brightness, while no toxic effect is observed. Such a goal cannot be achieved with the dissolved dye, which permeates the vessel walls. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Two-photon excited fluorescence from higher electronic states of chlorophylls in photosynthetic antenna complexes a new approach to detect strong excitonic chlorophyll a/b coupling

    CERN Document Server

    Leupold, D; Ehlert, J; Irrgang, K D; Renger, G; Lokstein, H

    2002-01-01

    Stepwise two-photon excitation of chlorophyll a and b in the higher plant main light-harvesting complex (LHC II) and the minor complex CP29 (as well as in organic solution) with 100-fs pulses in the Q/sub y/ region results in a weak blue fluorescence. The dependence of the spectral shape of the blue fluorescence on excitation wavelength offers a new approach to elucidate the long-standing problem of the origin of spectral "chlorophyll forms" in pigment-protein complexes, in particular the characterization of chlorophyll a/b-heterodimers. As a first result we present evidence for the existence of strong chlorophyll a/b-interactions (excitonically coupled transitions at 650 and 680 nm) in LHC II at ambient temperature. In comparison with LHC II, the experiments with CP29 provide further evidence that the lowest energy chlorophyll a transition (at ~680 nm) is not excitonically coupled to chlorophyll b. (22 refs).

  9. Two-Photon Fluorescence Microscopy for Determination of the Riboflavin Concentration in the Anterior Corneal Stroma When Using the Dresden Protocol.

    Science.gov (United States)

    Seiler, Theo G; Ehmke, Tobias; Fischinger, Isaak; Zapp, Daniel; Stachs, Oliver; Seiler, Theo; Heisterkamp, Alexander

    2015-10-01

    To determine the riboflavin concentration gradient in the anterior corneal stroma when using the Dresden protocol with different dextran solutions. Three different groups of porcine corneas, five each, were compared regarding the riboflavin concentration in the anterior stroma. Before all experiments, stable hydration conditions were established for the corresponding solution. All groups were treated with 0.1% riboflavin in different dextran solutions (15%, 16%, 20%). After imbibition, two-photon microscopy was used to determine fluorescence intensity. For signal attenuation and concentration determination corneas were saturated and measured a second time by two-photon microscopy. Additionally, the distribution was calculated mathematically and compared to the empiric results. Riboflavin concentration is decreasing with depth for all dextran solutions. A nearly constant concentration could be determined over the first 75 μm. Analysis of the fit functions leads to diffusion coefficients of D = 2.97 × 10-7 cm2/s for the 15% dextran solution, D = 2.34 × 10-7 cm2/s for the 16% dextran solution, and D = 1.28 × 10-7 cm2/s for the 20% dextran solution. The riboflavin gradients of the 20% dextran group were statistically significantly different from 15% dextran starting at a depth of 220 μm and deeper (P = 0.047). The 16% dextran group differed statistically at a depth of 250 μm and deeper (P = 0.047). These results show a significant difference to those published previously. With correct settings two-photon microscopy is a precise way to determine the concentration of riboflavin in cornea. The measured gradient is excellently fit by a Gaussian distribution, which comes out as a solution of Fick's second law.

  10. An organic dye with very large Stokes-shift and broad tunability of fluorescence: Potential two-photon probe for bioimaging and ultra-sensitive solid-state gas sensor

    Energy Technology Data Exchange (ETDEWEB)

    He, Tingchao; Tian, Xiaoqing; Lin, Xiaodong, E-mail: linxd@szu.edu.cn, E-mail: hdsun@ntu.edu.sg [College of Physics Science and Technology, Shenzhen University, Shenzhen 518060 (China); Wang, Yue; Zhao, Xin; Sun, Handong, E-mail: linxd@szu.edu.cn, E-mail: hdsun@ntu.edu.sg [Division of Physics and Applied Physics, and Centre for Disruptive Photonic Technologies (CDPT), School of Physical and Mathematical Sciences, Nanyang Technological University, 21 Nanyang Link, Singapore 637371 (Singapore); Gao, Yang; Grimsdale, Andrew C. [School of Materials Science and Engineering, Nanyang Technological University, Singapore 639798 (Singapore)

    2016-01-04

    Light-emitting nonlinear optical molecules, especially those with large Stokes shifts and broad tunability of their emission wavelength, have attracted considerable attention for various applications including biomedical imaging and fluorescent sensors. However, most fluorescent chromophores have only limited potential for such applications due to small Stokes shifts, narrow tunability of fluorescence emissions, and small optical nonlinearity in highly polar solvents. In this work, we demonstrate that a two-photon absorbing stilbene chromophore exhibits a large two-photon absorption action cross-section (ηδ = 320 GM) in dimethylsulfoxide (DMSO) and shows broad fluorescence tunability (125 nm) by manipulating the polarity of the surrounding medium. Importantly, a very large Stokes shift of up to 227 nm is achieved in DMSO. Thanks to these features, this chromophore can be utilized as a two-photon probe for bioimaging applications and in an ultrasensitive solid-state gas detector.

  11. Nonlinear spectral imaging of human normal skin, basal cell carcinoma and squamous cell carcinoma based on two-photon excited fluorescence and second-harmonic generation

    Science.gov (United States)

    Xiong, S. Y.; Yang, J. G.; Zhuang, J.

    2011-10-01

    In this work, we use nonlinear spectral imaging based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) for analyzing the morphology of collagen and elastin and their biochemical variations in basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and normal skin tissue. It was found in this work that there existed apparent differences among BCC, SCC and normal skin in terms of their thickness of the keratin and epithelial layers, their size of elastic fibers, as well as their distribution and spectral characteristics of collagen. These differences can potentially be used to distinguish BCC and SCC from normal skin, and to discriminate between BCC and SCC, as well as to evaluate treatment responses.

  12. Laser fluorescence spectroscopy by two-photon excitation for detection of hydrogen atoms in a periphery region of high temperature plasmas

    International Nuclear Information System (INIS)

    Kim, Hee-Je; Kajiwara, Toshinori; Motoyama, Sumio; Muraoka, Katsunori; Akazaki, Masanori; Okada, Tatsuo; Maeda, Mitsuo

    1989-01-01

    For measurements of atomic hydrogen density in the periphery region of high temperature plasmas, laser fluorescence spectroscopy (LFS) by two-photon excitation (1s-3s, 3d) was developed. Based upon the theoretical estimates for laser source requirements, which indicated the laser energy and spectral width to be more than 10 mJ (assuming the pulse duration of 10 ns) and several tens of picometers around the wavelength of 205.1 nm, respectively, the first Stokes generation in deuterium gas of ArF laser output was adopted and shown to have the necessary performance. Through the LFS experiment employing the laser source, the minimum detectable limit of atomic hydrogen, normalized by a laser power and an observing solid angle, was demonstrated to be 1 x 10 14 [m -3 · MW · sr], which is usually sufficient for the above purpose, and the accuracy of the density determination was shown to be within a factor 2. (author)

  13. A novel mitochondria-targeted two-photon fluorescent probe for dynamic and reversible detection of the redox cycles between peroxynitrite and glutathione.

    Science.gov (United States)

    Sun, Chunlong; Du, Wen; Wang, Peng; Wu, Yang; Wang, Baoqin; Wang, Jun; Xie, Wenjun

    2017-12-16

    Redox homeostasis is important for maintenance of normal physiological functions within cells. Redox state of cells is primarily a consequence of precise balance between levels of reducing equivalents and reactive oxygen species. Redox homeostasis between peroxynitrite (ONOO - ) and glutathione (GSH) is closely associated with physiological and pathological processes, such as prolonged relaxation in vascular tissues and smooth muscle preparations, attenuation of hepatic necrosis, and activation of matrix metalloproteinase-2. We report a two-photon fluorescent probe (TP-Se) based on water-soluble carbazole-based compound, which integrates with organic selenium, to monitor changes in ONOO - /GSH levels in cells. This probe can reversibly respond to ONOO - and GSH and exhibits high selectivity, sensitivity, and mitochondrial targeting. The probe was successfully applied to visualize changes in redox cycles during ONOO - outbreak and antioxidant GSH repair in cells. The probe will lead to significant development on redox events involved in cellular redox regulation. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Label-free imaging of brain and brain tumor specimens with combined two-photon excited fluorescence and second harmonic generation microscopy

    Science.gov (United States)

    Jiang, Liwei; Wang, Xingfu; Wu, Zanyi; Du, Huiping; Wang, Shu; Li, Lianhuang; Fang, Na; Lin, Peihua; Chen, Jianxin; Kang, Dezhi; Zhuo, Shuangmu

    2017-10-01

    Label-free imaging techniques are gaining acceptance within the medical imaging field, including brain imaging, because they have the potential to be applied to intraoperative in situ identifications of pathological conditions. In this paper, we describe the use of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) microscopy in combination for the label-free detection of brain and brain tumor specimens; gliomas. Two independently detecting channels were chosen to subsequently collect TPEF/SHG signals from the specimen to increase TPEF/SHG image contrasts. Our results indicate that the combined TPEF/SHG microscopic techniques can provide similar rat brain structural information and produce a similar resolution like conventional H&E staining in neuropathology; including meninges, cerebral cortex, white-matter structure corpus callosum, choroid plexus, hippocampus, striatum, and cerebellar cortex. It can simultaneously detect infiltrating human brain tumor cells, the extracellular matrix collagen fiber of connective stroma within brain vessels and collagen depostion in tumor microenvironments. The nuclear-to-cytoplasmic ratio and collagen content can be extracted as quantitative indicators for differentiating brain gliomas from healthy brain tissues. With the development of two-photon fiberscopes and microendoscope probes and their clinical applications, the combined TPEF and SHG microcopy may become an important multimodal, nonlinear optical imaging approach for real-time intraoperative histological diagnostics of residual brain tumors. These occur in various brain regions during ongoing surgeries through the method of simultaneously identifying tumor cells, and the change of tumor microenvironments, without the need for the removal biopsies and without the need for tissue labelling or fluorescent markers.

  15. The translated conceptual survey of physics / stablization of the focal plane in two photon excitation fluorescence microscopy

    Science.gov (United States)

    Wada, Asma

    As a reflection of my career to be an effective college physics teacher, my thesis is in two parts. The first is in education research, the focus of this part is to have a tool to evaluate pedagogies I have learned at the school and plan to apply in my classrooms back home. Consequently, this resulted in the development of the translated conceptual survey of physics ( TCSP). (TCSP) was designed by combining some questions from the Force Conceptual Inventory (FCI), and the Conceptual Survey of Electricity and Magnetism (CSEM) to assess student's understanding of basic concepts of Newtonian mechanics and electricity and magnetism in introductory physics. The idea of developing this questionnaire is to use it in classrooms back home as a part of a long term objective to implement what has been realized in the area of education research to improve the quality of teaching physics there. The survey was initially written in English, validated with interviews with native English speakers, translated into Arabic, and then validated via an interview with a native Arabic speaker. We then administered the survey to two different English-speaking intro physics courses and analyzed the results for consistency. The objective of the second part in my thesis is to expand my knowledge in an area of physics that I have interest in, and getting involved in a scientific research to develop skills I need as a teacher. My research is in optical physics, in particular, I am working on one of the challenges in implementing two photon excitation luorescence (TPEF) microscopy in imaging living systems. (TPEF) microscopy has been shown to be an invaluable tool for investigating biological structure and function in living organisms. The utility of (TPEF) imaging for this application arises from several important factors including it's ability to image deep within tissue, and to do so without harming the organism. Both of these advantages arise from the fact that (TPEF) imaging is done with

  16. Two-photon physics

    International Nuclear Information System (INIS)

    Bardeen, W.A.

    1981-10-01

    A new experimental frontier has recently been opened to the study of two photon processes. The first results of many aspects of these reactions are being presented at this conference. In contrast, the theoretical development of research ito two photon processes has a much longer history. This talk reviews the many different theoretical ideas which provide a detailed framework for our understanding of two photon processes

  17. Optical beam induced current measurements based on two-photon absorption process in 4H-SiC bipolar diodes

    Energy Technology Data Exchange (ETDEWEB)

    Hamad, H.; Raynaud, C.; Bevilacqua, P.; Tournier, D.; Planson, D. [Ampère Laboratory - UMR 5005, 21, Avenue Jean Capelle, 69621 Villeurbanne Cedex (France); Vergne, B. [Franco-Allemand Institute of Saint-Louis ISL, 5, Rue du Général Cassagnou, 68300 Saint-Louis (France)

    2014-02-24

    Using a pulsed green laser with a wavelength of 532 nm, a duration pulse of ∼1 ns, and a mean power varying between 1 and 100 mW, induced photocurrents have been measured in 4H-SiC bipolar diodes. Considering the photon energy (2.33 eV) and the bandgap of 4H-SiC (3.2 eV), the generation of electron-hole pair by the conventional single photon absorption process should be negligible. The intensity of the measured photocurrents depends quadratically on the power beam intensity. This clearly shows that they are generated using two-photon absorption process. As in conventional OBIC (Optical Beam Induced Current), the measurements give an image of the electric field distribution in the structure under test, and the minority carrier lifetime can be extracted from the decrease of the photocurrent at the edge of the structure. The extracted minority carrier lifetime of 210 ns is consistent with results obtained in case of single photon absorption.

  18. Optical beam induced current measurements based on two-photon absorption process in 4H-SiC bipolar diodes

    International Nuclear Information System (INIS)

    Hamad, H.; Raynaud, C.; Bevilacqua, P.; Tournier, D.; Planson, D.; Vergne, B.

    2014-01-01

    Using a pulsed green laser with a wavelength of 532 nm, a duration pulse of ∼1 ns, and a mean power varying between 1 and 100 mW, induced photocurrents have been measured in 4H-SiC bipolar diodes. Considering the photon energy (2.33 eV) and the bandgap of 4H-SiC (3.2 eV), the generation of electron-hole pair by the conventional single photon absorption process should be negligible. The intensity of the measured photocurrents depends quadratically on the power beam intensity. This clearly shows that they are generated using two-photon absorption process. As in conventional OBIC (Optical Beam Induced Current), the measurements give an image of the electric field distribution in the structure under test, and the minority carrier lifetime can be extracted from the decrease of the photocurrent at the edge of the structure. The extracted minority carrier lifetime of 210 ns is consistent with results obtained in case of single photon absorption

  19. A multifunctional magnetic nanocarrier bearing fluorescent dye for targeted drug delivery by enhanced two-photon triggered release

    International Nuclear Information System (INIS)

    Banerjee, Shashwat S; Chen, D-H

    2009-01-01

    We report a novel nanoformulation for targeted drug delivery which utilizes nanophotonics through the fusion of nanotechnology with biomedical application. The approach involves an energy-transferring magnetic nanoscopic co-assembly fabricated of rhodamine B (RDB) fluorescent dye grafted gum arabic modified Fe 3 O 4 magnetic nanoparticle and photosensitive linker by which dexamethasone drug is conjugated to the magnetic nano-assembly. The advantage offered by this nanoformulation is the indirect photo-triggered-on-demand drug release by efficient up-converting energy of the near-IR (NIR) light to higher energy and intraparticle energy transfer from the dye grafted magnetic nanoparticle to the linker for drug release by cleavage. The synthesized nanoparticles were found to be of ultra-small size (13.33 nm) and are monodispersed in an aqueous suspension. Dexamethasone (Dexa) drug conjugated to RDB-GAMNP by photosensitive linker showed appreciable release of Dexa by photo-triggered response on exposure to radiation having a wavelength in the NIR region whereas no detectable release was observed in the dark. Photo-triggered response for the nanoformulation not bearing the rhodamine B dye was drastically less as less Dexa was released on exposure to NIR radiation which suggest that the photo-cleavage of linker and release of Dexa mainly originated from the indirect excitation through the uphill energy conversions based on donor-acceptor model FRET. The promising pathway of nanophotonics for the on-demand release of the drug makes this nanocarrier very promising for applications in nanomedicine.

  20. Cardiac muscle organization revealed in 3-D by imaging whole-mount mouse hearts using two-photon fluorescence and confocal microscopy.

    Science.gov (United States)

    Sivaguru, Mayandi; Fried, Glenn; Sivaguru, Barghav S; Sivaguru, Vignesh A; Lu, Xiaochen; Choi, Kyung Hwa; Saif, M Taher A; Lin, Brian; Sadayappan, Sakthivel

    2015-11-01

    The ability to image the entire adult mouse heart at high resolution in 3-D would provide enormous advantages in the study of heart disease. However, a technique for imaging nuclear/cellular detail as well as the overall structure of the entire heart in 3-D with minimal effort is lacking. To solve this problem, we modified the benzyl alcohol:benzyl benzoate (BABB) clearing technique by labeling mouse hearts with periodic acid Schiff (PAS) stain. We then imaged the hearts with a combination of two-photon fluorescence microscopy and automated tile-scan imaging/stitching. Utilizing the differential spectral properties of PAS, we could identify muscle and nuclear compartments in the heart. We were also able to visualize the differences between a 3-month-old normal mouse heart and a mouse heart that had undergone heart failure due to the expression of cardiac myosin binding protein-C (cMyBP-C) gene mutation (t/t). Using 2-D and 3-D morphometric analysis, we found that the t/t heart had anomalous ventricular shape, volume, and wall thickness, as well as a disrupted sarcomere pattern. We further validated our approach using decellularized hearts that had been cultured with 3T3 fibroblasts, which were tracked using a nuclear label. We were able to detect the 3T3 cells inside the decellularized intact heart tissue, achieving nuclear/cellular resolution in 3-D. The combination of labeling, clearing, and two-photon microscopy together with tiling eliminates laborious and time-consuming physical sectioning, alignment, and 3-D reconstruction.

  1. Correlating two-photon excited fluorescence imaging of breast cancer cellular redox state with seahorse flux analysis of normalized cellular oxygen consumption

    Science.gov (United States)

    Hou, Jue; Wright, Heather J.; Chan, Nicole; Tran, Richard; Razorenova, Olga V.; Potma, Eric O.; Tromberg, Bruce J.

    2016-06-01

    Two-photon excited fluorescence (TPEF) imaging of the cellular cofactors nicotinamide adenine dinucleotide and oxidized flavin adenine dinucleotide is widely used to measure cellular metabolism, both in normal and pathological cells and tissues. When dual-wavelength excitation is used, ratiometric TPEF imaging of the intrinsic cofactor fluorescence provides a metabolic index of cells-the "optical redox ratio" (ORR). With increased interest in understanding and controlling cellular metabolism in cancer, there is a need to evaluate the performance of ORR in malignant cells. We compare TPEF metabolic imaging with seahorse flux analysis of cellular oxygen consumption in two different breast cancer cell lines (MCF-7 and MDA-MB-231). We monitor metabolic index in living cells under both normal culture conditions and, for MCF-7, in response to cell respiration inhibitors and uncouplers. We observe a significant correlation between the TPEF-derived ORR and the flux analyzer measurements (R=0.7901, p<0.001). Our results confirm that the ORR is a valid dynamic index of cell metabolism under a range of oxygen consumption conditions relevant for cancer imaging.

  2. Interface-Targeting Strategy Enables Two-Photon Fluorescent Lipid Droplet Probes for High-Fidelity Imaging of Turbid Tissues and Detecting Fatty Liver.

    Science.gov (United States)

    Guo, Lifang; Tian, Minggang; Feng, Ruiqing; Zhang, Ge; Zhang, Ruoyao; Li, Xuechen; Liu, Zhiqiang; He, Xiuquan; Sun, Jing Zhi; Yu, Xiaoqiang

    2018-04-04

    Lipid droplets (LDs) with unique interfacial architecture not only play crucial roles in protecting a cell from lipotoxicity and lipoapoptosis but also closely relate with many diseases such as fatty liver and diabetes. Thus, as one of the important applied biomaterials, fluorescent probes with ultrahigh selectivity for in situ and high-fidelity imaging of LDs in living cells and tissues are critical to elucidate relevant physiological and pathological events as well as detect related diseases. However, available probes only utilizing LDs' waterless neutral cores but ignoring the unique phospholipid monolayer interfaces exhibit low selectivity. They cannot differentiate neutral cores of LDs from intracellular other lipophilic microenvironments, which results in extensively cloud-like background noise and severely limited their bioapplications. Herein, to design LD probes with ultrahigh selectivity, the exceptional interfacial architecture of LDs is considered adequately and thus an interface-targeting strategy is proposed for the first time. According to the novel strategy, we have developed two amphipathic fluorescent probes (N-Cy and N-Py) by introducing different cations into a lipophilic fluorophore (nitrobenzoxadiazole (NBD)). Consequently, their cationic moiety precisely locates the interfaces through electrostatic interaction and simultaneously NBD entirely embeds into the waterless core via hydrophobic interaction. Thus, high-fidelity and background-free fluorescence imaging of LDs are expectably realized in living cells in situ. Moreover, LDs in turbid tissues like skeletal muscle slices have been clearly imaged (up to 82 μm depth) by a two-photon microscope. Importantly, using N-Cy, we not only intuitively monitored the variations of LDs in number, size, and morphology but also clearly revealed their abnormity in hepatic tissues resulting from fatty liver. Therefore, these unique probes provide excellent imaging tools for elucidating LD

  3. Broadband two-photon absorption cross sections of benzothiazole derivatives and benzobisthiazolium salts

    Science.gov (United States)

    Noskovičova, Eva; Lorenc, Dušan; Magdolen, Peter; Sigmundová, Ivica; Zahradník, Pavol; Velič, Dušan

    2018-05-01

    Two-photon absorption (TPA) cross sections of conjugated donor-π-acceptor dipolar structures containing benzothiazole or benzobisthiazolium moieties are determined in a broad spectral range from 700 nm to 1000 nm using two-photon induced fluorescence technique. The TPA cross section values range from 150 GM to 4600 GM. The largest values are observed in near-infrared region. The dipolar derivative of benzothiazole has the largest TPA cross section of 4600 GM at wavelength of 890 nm. A combination of the large TPA in the near-infrared region and the high emission quantum yield makes these compounds excellent candidates for two-photon fluorescence microscopy.

  4. Stimulated emission depletion following two photon excitation

    OpenAIRE

    Marsh, R. J.; Armoogum, D. A.; Bain, A. J.

    2002-01-01

    The technique of stimulated emission depletion of fluorescence (STED) from a two photon excited molecular population is demonstrated in the S, excited state of fluorescein in ethylene glycol and methanol. Two photon excitation (pump) is achieved using the partial output of a regeneratively amplified Ti:Sapphire laser in conjunction with an optical parametric amplifier whose tuneable output provides a synchronous depletion (dump) pulse. Time resolved fluorescence intensity and anisotropy measu...

  5. Extreme nonlinearities in InAs/InP nanowire gain media: the two-photon induced laser

    DEFF Research Database (Denmark)

    Capua, Amir; Kami, Ouri; Eisenstein, Gadi

    2012-01-01

    We demonstrate a novel laser oscillation scheme in an InAs / InP wire-like quantum dash gain medium. A short optical pulse excites carriers by two photon absorption which relax to the energy levels providing gain thereby enabling laser oscillations. The nonlinear dynamic interaction is analyzed a...

  6. Tale of two photons

    International Nuclear Information System (INIS)

    Anon.

    1994-01-01

    A very profitable spinoff from electron- positron collisions is two-photon physics. Rather than the electron and positron interacting directly via an exchanged photon, two virtual (transient) photons, one from each particle, get tangled up. With new electron-positron colliders appearing on the scene, a topical meeting on two-photon physics - 'From DAPHNE to LEP 200 and beyond' - held from 2-4 February in Paris, in the premises of the Ministry of Higher Education and Research, was particularly timely. Some 60 physicists, both experimentalists and theorists, participated, with some thirty speakers

  7. Two Photon Distribution Amplitudes

    International Nuclear Information System (INIS)

    El Beiyad, M.; Pire, B.; Szymanowski, L.; Wallon, S.

    2008-01-01

    The factorization of the amplitude of the process γ*γ→γγ in the low energy and high photon virtuality region is demonstrated at the Born order and in the leading logarithmic approximation. The leading order two photon (generalized) distribution amplitudes exhibit a characteristic ln Q 2 behaviour and obey new inhomogeneous evolution equations

  8. A two photon absorption laser induced fluorescence diagnostic for fusion plasmas

    Energy Technology Data Exchange (ETDEWEB)

    Magee, R. M.; Galante, M. E.; McCarren, D.; Scime, E. E. [Physics Department, West Virginia University, Morgantown, West Virginia 26506 (United States); Boivin, R. L.; Brooks, N. H.; Groebner, R. J.; Hill, D. N. [General Atomics, San Diego, California 92121 (United States); Porter, G. D. [Lawrence Livermore National Laboratory, Livermore, California 94550 (United States)

    2012-10-15

    The quality of plasma produced in a magnetic confinement fusion device is influenced to a large extent by the neutral gas surrounding the plasma. The plasma is fueled by the ionization of neutrals, and charge exchange interactions between edge neutrals and plasma ions are a sink of energy and momentum. Here we describe a diagnostic capable of measuring the spatial distribution of neutral gas in a magnetically confined fusion plasma. A high intensity (5 MW/cm{sup 2}), narrow bandwidth (0.1 cm{sup -1}) laser is injected into a hydrogen plasma to excite the Lyman {beta} transition via the simultaneous absorption of two 205 nm photons. The absorption rate, determined by measurement of subsequent Balmer {alpha} emission, is proportional to the number of particles with a given velocity. Calibration is performed in situ by filling the chamber to a known pressure of neutral krypton and exciting a transition close in wavelength to that used in hydrogen. We present details of the calibration procedure, including a technique for identifying saturation broadening, measurements of the neutral density profile in a hydrogen helicon plasma, and discuss the application of the diagnostic to plasmas in the DIII-D tokamak.

  9. A two photon absorption laser induced fluorescence diagnostic for fusion plasmas.

    Science.gov (United States)

    Magee, R M; Galante, M E; McCarren, D; Scime, E E; Boivin, R L; Brooks, N H; Groebner, R J; Hill, D N; Porter, G D

    2012-10-01

    The quality of plasma produced in a magnetic confinement fusion device is influenced to a large extent by the neutral gas surrounding the plasma. The plasma is fueled by the ionization of neutrals, and charge exchange interactions between edge neutrals and plasma ions are a sink of energy and momentum. Here we describe a diagnostic capable of measuring the spatial distribution of neutral gas in a magnetically confined fusion plasma. A high intensity (5 MW/cm(2)), narrow bandwidth (0.1 cm(-1)) laser is injected into a hydrogen plasma to excite the Lyman β transition via the simultaneous absorption of two 205 nm photons. The absorption rate, determined by measurement of subsequent Balmer α emission, is proportional to the number of particles with a given velocity. Calibration is performed in situ by filling the chamber to a known pressure of neutral krypton and exciting a transition close in wavelength to that used in hydrogen. We present details of the calibration procedure, including a technique for identifying saturation broadening, measurements of the neutral density profile in a hydrogen helicon plasma, and discuss the application of the diagnostic to plasmas in the DIII-D tokamak.

  10. Automated evaluation of liver fibrosis in thioacetamide, carbon tetrachloride, and bile duct ligation rodent models using second-harmonic generation/two-photon excited fluorescence microscopy.

    Science.gov (United States)

    Liu, Feng; Chen, Long; Rao, Hui-Ying; Teng, Xiao; Ren, Ya-Yun; Lu, Yan-Qiang; Zhang, Wei; Wu, Nan; Liu, Fang-Fang; Wei, Lai

    2017-01-01

    Animal models provide a useful platform for developing and testing new drugs to treat liver fibrosis. Accordingly, we developed a novel automated system to evaluate liver fibrosis in rodent models. This system uses second-harmonic generation (SHG)/two-photon excited fluorescence (TPEF) microscopy to assess a total of four mouse and rat models, using chemical treatment with either thioacetamide (TAA) or carbon tetrachloride (CCl 4 ), and a surgical method, bile duct ligation (BDL). The results obtained by the new technique were compared with that using Ishak fibrosis scores and two currently used quantitative methods for determining liver fibrosis: the collagen proportionate area (CPA) and measurement of hydroxyproline (HYP) content. We show that 11 shared morphological parameters faithfully recapitulate Ishak fibrosis scores in the models, with high area under the receiver operating characteristic (ROC) curve (AUC) performance. The AUC values of 11 shared parameters were greater than that of the CPA (TAA: 0.758-0.922 vs 0.752-0.908; BDL: 0.874-0.989 vs 0.678-0.966) in the TAA mice and BDL rat models and similar to that of the CPA in the TAA rat and CCl 4 mouse models. Similarly, based on the trends in these parameters at different time points, 9, 10, 7, and 2 model-specific parameters were selected for the TAA rats, TAA mice, CCl 4 mice, and BDL rats, respectively. These parameters identified differences among the time points in the four models, with high AUC accuracy, and the corresponding AUC values of these parameters were greater compared with those of the CPA in the TAA rat and mouse models (rats: 0.769-0.894 vs 0.64-0.799; mice: 0.87-0.93 vs 0.739-0.836) and similar to those of the CPA in the CCl 4 mouse and BDL rat models. Similarly, the AUC values of 11 shared parameters and model-specific parameters were greater than those of HYP in the TAA rats, TAA mice, and CCl 4 mouse models and were similar to those of HYP in the BDL rat models. The automated

  11. Two-photon-induced hot-electron transfer to a single molecule in a scanning tunneling microscope

    International Nuclear Information System (INIS)

    Wu, S. W.; Ho, W.

    2010-01-01

    The junction of a scanning tunneling microscope (STM) operating in the tunneling regime was irradiated with femtosecond laser pulses. A photoexcited hot electron in the STM tip resonantly tunnels into an excited state of a single molecule on the surface, converting it from the neutral to the anion. The electron-transfer rate depends quadratically on the incident laser power, suggesting a two-photon excitation process. This nonlinear optical process is further confirmed by the polarization measurement. Spatial dependence of the electron-transfer rate exhibits atomic-scale variations. A two-pulse correlation experiment reveals the ultrafast dynamic nature of photoinduced charging process in the STM junction. Results from these experiments are important for understanding photoinduced interfacial charge transfer in many nanoscale inorganic-organic structures.

  12. High contrast two-photon imaging of fingermarks

    Science.gov (United States)

    Stoltzfus, Caleb R.; Rebane, Aleksander

    2016-04-01

    Optically-acquired fingermarks are widely used as evidence across law enforcement agencies as well as in the courts of law. A common technique for visualizing latent fingermarks on nonporous surfaces consists of cyanoacrylate fuming of the fingerprint material, followed by impregnation with a fluorescent dye, which under ultra violet (UV) illumination makes the fingermarks visible and thus accessible for digital recording. However, there exist critical circumstances, when the image quality is compromised due to high background scattering, high auto-fluorescence of the substrate material, or other detrimental photo-physical and photo-chemical effects such as light-induced damage to the sample. Here we present a novel near-infrared (NIR), two-photon induced fluorescence imaging modality, which significantly enhances the quality of the fingermark images, especially when obtained from highly reflective and/or scattering surfaces, while at the same time reducing photo-damage to sensitive forensic samples.

  13. Air-Stable Surface-Passivated Perovskite Quantum Dots for Ultra-Robust, Single- and Two-Photon-Induced Amplified Spontaneous Emission

    KAUST Repository

    Pan, Jun

    2015-12-01

    We demonstrate ultra-air- and photostable CsPbBr3 quantum dots (QDs) by using an inorganic–organic hybrid ion pair as the capping ligand. This passivation approach to perovskite QDs yields high photoluminescence quantum yield with unprecedented operational stability in ambient conditions (60 ± 5% lab humidity) and high pump fluences, thus overcoming one of the greatest challenges impeding the development of perovskite-based applications. Due to the robustness of passivated perovskite QDs, we were able to induce ultrastable amplified spontaneous emission (ASE) in solution processed QD films not only through one photon but also through two-photon absorption processes. The latter has not been observed before in the family of perovskite materials. More importantly, passivated perovskite QD films showed remarkable photostability under continuous pulsed laser excitation in ambient conditions for at least 34 h (corresponds to 1.2 × 108 laser shots), substantially exceeding the stability of other colloidal QD systems in which ASE has been observed.

  14. Two-photon excitation in chip electrophoresis enabling label-free fluorescence detection in non-UV transparent full-body polymer chips.

    Science.gov (United States)

    Geissler, David; Belder, Detlev

    2015-12-01

    One of the most commonly employed detection methods in microfluidic research is fluorescence detection, due to its ease of integration and excellent sensitivity. Many analytes though do not show luminescence when excited in the visible light spectrum, require suitable dyes. Deep-ultraviolet (UV) excitation (electrophoresis of small aromatic compounds. Various polymers, such as poly(methyl methacrylate), cyclic olefin polymer, and copolymer as well as poly(dimethylsiloxane) were investigated and compared with respect to achievable LOD and ruggedness against photodamage. To demonstrate the applicability of the technique, the method was also applied to the determination of serotonin and tryptamine in fruit samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Single-organelle tracking by two-photon conversion

    Science.gov (United States)

    Watanabe, Wataru; Shimada, Tomoko; Matsunaga, Sachihiro; Kurihara, Daisuke; Fukui, Kiichi; Shin-Ichi Arimura, Shin-Ichi; Tsutsumi, Nobuhiro; Isobe, Keisuke; Itoh, Kazuyoshi

    2007-03-01

    Spatial and temporal information about intracellular objects and their dynamics within a living cell are essential for dynamic analysis of such objects in cell biology. A specific intracellular object can be discriminated by photoactivatable fluorescent proteins that exhibit pronounced light-induced spectral changes. Here, we report on selective labeling and tracking of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein with near-infrared femtosecond laser pulses. We performed selective labeling of a single mitochondrion in a living tobacco BY-2 cell using two-photon photoconversion of Kaede. Using this technique, we demonstrated that, in plants, the directed movement of individual mitochondria along the cytoskeletons was mediated by actin filaments, whereas microtubules were not required for the movement of mitochondria. This single-organelle labeling technique enabled us to track the dynamics of a single organelle, revealing the mechanisms involved in organelle dynamics. The technique has potential application in direct tracking of selective cellular and intracellular structures.

  16. Utilizing two-photon fluorescence and second harmonic generation microscopy to study human bone marrow mesenchymal stem cell morphogenesis in chitosan scaffold

    Science.gov (United States)

    Su, Ping-Jung; Huang, Chi-Hsiu; Huang, Yi-You; Lee, Hsuan-Sue; Dong, Chen-Yuan

    2008-02-01

    A major goal of tissue engineering is to cultivate the cartilage in vitro. One approach is to implant the human bone marrow mesenchymal stem cells into the three dimensional biocompatible and biodegradable material. Through the action of the chondrogenic factor TGF-β3, the stem cells can be induced to secrete collagen. In this study, mesenchymal stem cells are implanted on the chitosan scaffold and TGF-β3 was added to produce the cartilage tissue and TP autofluorescence and SHG microscopy was used to image the process of chondrogenesis. With additional development, multiphoton microscopy can be developed into an effective tool for evaluating the quality of tissue engineering products.

  17. Molecular photosensitisers for two-photon photodynamic therapy.

    Science.gov (United States)

    Bolze, F; Jenni, S; Sour, A; Heitz, V

    2017-11-30

    Two-photon excitation has attracted the attention of biologists, especially after the development of two-photon excited microscopy in the nineties. Since then, new applications have rapidly emerged such as the release of biologically active molecules and photodynamic therapy (PDT) using two-photon excitation. PDT, which requires a light-activated drug (photosensitiser), is a clinically approved and minimally invasive treatment for cancer and for non-malignant diseases. This feature article focuses on the engineering of molecular two-photon photosensitisers for PDT, which should bring important benefits to the treatment, increase the treatment penetration depth with near-infrared light excitation, improve the spatial selectivity and reduce the photodamage to healthy tissues. After an overview of the two-photon absorption phenomenon and the methods to evaluate two-photon induced phototoxicity on cell cultures, the different classes of photosensitisers described in the literature are discussed. The two-photon PDT performed with historical one-photon sensitisers are briefly presented, followed by specifically engineered cyclic tetrapyrrole photosensitisers, purely organic photosensitisers and transition metal complexes. Finally, targeted two-photon photosensitisers and theranostic agents that should enhance the selectivity and efficiency of the treatment are discussed.

  18. COMPARISON OF FEMTOSECOND AND NANOSECOND TWO PHOTON ABSORPTION LASER INDUCED FLUORESCENCE (TALIF) OF ATOMIC OXYGEN IN ATMOSPHERIC PRESSURE PLASMAS

    Science.gov (United States)

    2016-08-01

    were also collected with the CDBDs in an ozone production regime, as shown below in Figure 12. For the ns-TALIF setup, the discharge was operated in...these plasma systems and facilitate the development of model -based predictive capabilities. Diagnostic techniques used to quantify intermediate...quenching rate is defined as ii q q q Q k n=∑ , i.e. the sum of the product of the number density of the quenching species, nq, and its corresponding

  19. Polarized two-photon photoselection in EGFP: Theory and experiment.

    Science.gov (United States)

    Masters, T A; Marsh, R J; Blacker, T S; Armoogum, D A; Larijani, B; Bain, A J

    2018-04-07

    In this work, we present a complete theoretical description of the excited state order created by two-photon photoselection from an isotropic ground state; this encompasses both the conventionally measured quadrupolar (K = 2) and the "hidden" degree of hexadecapolar (K = 4) transition dipole alignment, their dependence on the two-photon transition tensor and emission transition dipole moment orientation. Linearly and circularly polarized two-photon absorption (TPA) and time-resolved single- and two-photon fluorescence anisotropy measurements are used to determine the structure of the transition tensor in the deprotonated form of enhanced green fluorescent protein. For excitation wavelengths between 800 nm and 900 nm, TPA is best described by a single element, almost completely diagonal, two-dimensional (planar) transition tensor whose principal axis is collinear to that of the single-photon S 0 → S 1 transition moment. These observations are in accordance with assignments of the near-infrared two-photon absorption band in fluorescent proteins to a vibronically enhanced S 0 → S 1 transition.

  20. Polarized two-photon photoselection in EGFP: Theory and experiment

    Science.gov (United States)

    Masters, T. A.; Marsh, R. J.; Blacker, T. S.; Armoogum, D. A.; Larijani, B.; Bain, A. J.

    2018-04-01

    In this work, we present a complete theoretical description of the excited state order created by two-photon photoselection from an isotropic ground state; this encompasses both the conventionally measured quadrupolar (K = 2) and the "hidden" degree of hexadecapolar (K = 4) transition dipole alignment, their dependence on the two-photon transition tensor and emission transition dipole moment orientation. Linearly and circularly polarized two-photon absorption (TPA) and time-resolved single- and two-photon fluorescence anisotropy measurements are used to determine the structure of the transition tensor in the deprotonated form of enhanced green fluorescent protein. For excitation wavelengths between 800 nm and 900 nm, TPA is best described by a single element, almost completely diagonal, two-dimensional (planar) transition tensor whose principal axis is collinear to that of the single-photon S0 → S1 transition moment. These observations are in accordance with assignments of the near-infrared two-photon absorption band in fluorescent proteins to a vibronically enhanced S0 → S1 transition.

  1. A series of fluorene-based two-photon absorbing molecules: synthesis, linear and nonlinear characterization, and bioimaging

    Science.gov (United States)

    Andrade, Carolina D.; Yanez, Ciceron O.; Rodriguez, Luis; Belfield, Kevin D.

    2010-01-01

    The synthesis, structural, and photophysical characterization of a series of new fluorescent donor–acceptor and acceptor-acceptor molecules, based on the fluorenyl ring system, with two-photon absorbing properties is described. These new compounds exhibited large Stokes shifts, high fluorescent quantum yields, and, significantly, high two-photon absorption cross sections, making them well suited for two-photon fluorescence microscopy (2PFM) imaging. Confocal and two-photon fluorescence microscopy imaging of COS-7 and HCT 116 cells incubated with probe I showed endosomal selectivity, demonstrating the potential of this class of fluorescent probes in multiphoton fluorescence microscopy. PMID:20481596

  2. Synthesis, singlet-oxygen photogeneration, two-photon absorption, photo-induced DNA cleavage and cytotoxic properties of an amphiphilic β-Schiff-base linked Ru(II) polypyridyl–porphyrin conjugate

    International Nuclear Information System (INIS)

    Ke, Hanzhong; Ma, Wanpeng; Wang, Hongda; Cheng, Guoe; Yuan, Han; Wong, Wai-Kwok; Kwong, Daniel W.J.; Tam, Hoi-Lam; Cheah, Kok-Wai; Chan, Chi-Fai; Wong, Ka-Leung

    2014-01-01

    A novel porphyrin–polypyridyl ruthenium(II) conjugate (TPP–Ru), in which the ruthenium(II) polypyridyl moiety is linked to the β-position of the tetraphenylporphyrin via a Schiff base linkage, has been synthesized and characterized by 1 H NMR, HRMS and UV–visible spectroscopy. The relative singlet oxygen quantum yield and two-photon absorption cross-section of this conjugate, together with its photo-induced DNA cleavage and cytotoxic activities were measured. The results show that the amphiphilic ruthenium(II) polypyridyl–porphyrin conjugate is an effective DNA photocleavage agent, with potential application in one- and two-photon absorption anti-cancer photodynamic therapy. - Highlights: • New porphyrin–ruthenium(II) polypyridyl complexes (TTP–Ru) have been synthesized. • The TTP–Ru shows substantial two-photon absorption cross-section (σ 2 =391 GM). • The TTP–Ru exhibits a substantial 1 O 2 quantum yield (0.64±0.13). • The TTP–Ru exhibits a strong DNA cleavage activity upon photo-excitation. • The TTP–Ru is available for in vitro imaging and as a photodynamic therapy agent

  3. Synthesis, singlet-oxygen photogeneration, two-photon absorption, photo-induced DNA cleavage and cytotoxic properties of an amphiphilic β-Schiff-base linked Ru(II) polypyridyl–porphyrin conjugate

    Energy Technology Data Exchange (ETDEWEB)

    Ke, Hanzhong, E-mail: kehanz@163.com [Faculty of Material Science and Chemistry, China University of Geosciences, Wuhan, Hubei 430074 (China); Ma, Wanpeng; Wang, Hongda; Cheng, Guoe [Faculty of Material Science and Chemistry, China University of Geosciences, Wuhan, Hubei 430074 (China); Yuan, Han [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong SAR (China); Wong, Wai-Kwok, E-mail: wkwong@hkbu.edu.hk [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong SAR (China); Institute of Advanced Materials, Hong Kong Baptist University, Kowloon Tong, Hong Kong SAR (China); Kwong, Daniel W.J. [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong SAR (China); Tam, Hoi-Lam; Cheah, Kok-Wai [Department of Physics, Hong Kong Baptist University, Kowloon Tong, Hong Kong SAR (China); Institute of Advanced Materials, Hong Kong Baptist University, Kowloon Tong, Hong Kong SAR (China); Chan, Chi-Fai; Wong, Ka-Leung [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong SAR (China)

    2014-10-15

    A novel porphyrin–polypyridyl ruthenium(II) conjugate (TPP–Ru), in which the ruthenium(II) polypyridyl moiety is linked to the β-position of the tetraphenylporphyrin via a Schiff base linkage, has been synthesized and characterized by {sup 1}H NMR, HRMS and UV–visible spectroscopy. The relative singlet oxygen quantum yield and two-photon absorption cross-section of this conjugate, together with its photo-induced DNA cleavage and cytotoxic activities were measured. The results show that the amphiphilic ruthenium(II) polypyridyl–porphyrin conjugate is an effective DNA photocleavage agent, with potential application in one- and two-photon absorption anti-cancer photodynamic therapy. - Highlights: • New porphyrin–ruthenium(II) polypyridyl complexes (TTP–Ru) have been synthesized. • The TTP–Ru shows substantial two-photon absorption cross-section (σ{sub 2}=391 GM). • The TTP–Ru exhibits a substantial {sup 1}O{sub 2} quantum yield (0.64±0.13). • The TTP–Ru exhibits a strong DNA cleavage activity upon photo-excitation. • The TTP–Ru is available for in vitro imaging and as a photodynamic therapy agent.

  4. Selective two-photon collagen crosslinking in situ measured by Brillouin microscopy (Conference Presentation)

    Science.gov (United States)

    Kwok, Sheldon J. J.; Kuznetsov, Ivan A.; Kim, Moonseok; Choi, Myunghwan; Scarcelli, Giuliano; Yun, Seok-Hyun

    2017-02-01

    Two-photon polymerization and crosslinking are commonly used methods for microfabrication of three-dimensional structures with applications spanning from photonic microdevices, drug delivery systems, to cellular scaffolds. However, the use of two-photon processes for precise, internal modification of biological tissues has not yet been reported. One of the major challenges has been a lack of appropriate tools to monitor and characterize crosslinked regions nondestructively. Here, we demonstrate spatially selective two-photon collagen crosslinking (2P-CXL) in intact tissue for the first time. Using riboflavin photosensitizer and femtosecond laser irradiation, we crosslinked a small volume of tissue within animal corneas. Collagen fiber orientations and photobleaching were characterized by second harmonic generation and two-photon fluorescence imaging, respectively. Using confocal Brillouin microscopy, we measured local changes in longitudinal mechanical moduli and visualized the cross-linked pattern without perturbing surrounding non-irradiated regions. 2P-CXL-induced tissue stiffening was comparable to that achieved with conventional one-photon CXL. Our results demonstrate the ability to selectively stiffen biological tissue in situ at high spatial resolution, with broad implications in ophthalmology, laser surgery, and tissue engineering.

  5. Doppler-free two-photon excitation of 238U

    International Nuclear Information System (INIS)

    Hodgkinson, D.P.; Wort, D.J.H.

    1981-04-01

    A theory of resonantly enhanced two-photon absorption is presented and tested in a number of experiments in which 238 U vapour is excited by two continuous wave dye lasers. Good quantitative agreement between theory and experiment is found. In particular the central prediction of the theory, that antiparallel laser beams of modest intensity can pump an appreciable fraction of the Maxwell velocity distribution, has been checked directly by measuring the spectral width of the fluorescence from the two-photon excited level. (author)

  6. Scanless two-photon excitation of channelrhodopsin-2

    DEFF Research Database (Denmark)

    Papagiakoumou, E.; Anselmi, F.; Bègue, A.

    2010-01-01

    developed a method that combines generalized phase contrast with temporal focusing (TF-GPC) to shape two-photon excitation for this purpose. The illumination patterns are generated automatically from fluorescence images of neurons and shaped to cover the cell body or dendrites, or distributed groups...... of cells. The TF-GPC two-photon excitation patterns generated large photocurrents in Channelrhodopsin-2–expressing cultured cells and neurons and in mouse acute cortical slices. The amplitudes of the photocurrents can be precisely modulated by controlling the size and shape of the excitation volume and...

  7. Linear photophysics, two-photon absorption and femtosecond transient absorption spectroscopy of styryl dye bases

    Energy Technology Data Exchange (ETDEWEB)

    Shaydyuk, Ye.O. [Institute of Physics, Prospect Nauki, 46, Kyiv-28 03028 Ukraine (Ukraine); Levchenko, S.M. [Institute of Molecular Biology and Genetics, 150, Akademika Zabolotnoho Str., Kyiv 036803 (Ukraine); Kurhuzenkau, S.A. [Department of Chemistry, University of Parma, Parco Area delle Scienze 17/A, Parma 43124 (Italy); Anderson, D. [NanoScienece Technology Center, University of Central Florida, 12424 Research Parkway, PAV400, Orlando, FL 32826 (United States); Department of Chemistry, University of Central Florida, 4111 Libra Drive, PSB225, Orlando, FL 32816 (United States); Masunov, A.E. [NanoScienece Technology Center, University of Central Florida, 12424 Research Parkway, PAV400, Orlando, FL 32826 (United States); Department of Chemistry, University of Central Florida, 4111 Libra Drive, PSB225, Orlando, FL 32816 (United States); South Ural State University, Lenin pr. 76, Chelyabinsk 454080 (Russian Federation); Department of Condensed Matter Physics, National Research Nuclear University MEPhI, Kashirskoye shosse 31, Moscow 115409 (Russian Federation); Photochemistry Center RAS, ul. Novatorov 7a, Moscow 119421 (Russian Federation); Kachkovsky, O.D.; Slominsky, Yu.L.; Bricks, J.L. [Insitute of Organic Chemistry, Murmanskaya Street, 5, Kyiv 03094 (Ukraine); Belfield, K.D. [College of Science and Liberal Arts, New Jersey Institute of Technology, University Heights, Newark, NJ 07102 (United States); School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi’an, 710062 (China); Bondar, M.V., E-mail: mbondar@mail.ucf.edu [Institute of Physics, Prospect Nauki, 46, Kyiv-28 03028 Ukraine (Ukraine)

    2017-03-15

    The steady-state and time-resolved linear spectral properties, two-photon absorption spectra and fast relaxation processes in the excited states of styryl base-type derivatives were investigated. The nature of linear absorption, fluorescence and excitation anisotropy spectra were analyzed in solvents of different polarity at room temperature and specific dependence of the solvatochromic behavior on the donor-acceptor strength of the terminal substituents was shown. Two-photon absorption (2PA) efficiency of styryl dye bases was determined in a broad spectral range using two-photon induced fluorescence technique, and cross-sections maxima of ~ 100 GM were found. The excited state absorption (ESA) and fast relaxation processes in the molecular structures were investigated by transient absorption femtosecond pump-probe methodology. The role of twisted intramolecular charge transfer (TICT) effect in the excited state of styryl dye base with dimethylamino substituent was shown. The experimental spectroscopic data were also verified by quantum chemical calculations at the Time Dependent Density Functional Theory level, combined with a polarizable continuum model.

  8. Calculation of the spatial resolution in two-photon absorption spectroscopy applied to plasma diagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Garcia-Lechuga, M. [Departamento de Física Teórica, Atómica y Óptica, Universidad de Valladolid, 47011-Valladolid (Spain); Laser Processing Group, Instituto de Óptica “Daza de Valdés,” CSIC, 28006-Madrid (Spain); Fuentes, L. M. [Departamento de Física Aplicada, Universidad de Valladolid, 47011-Valladolid (Spain); Grützmacher, K.; Pérez, C., E-mail: concha@opt.uva.es; Rosa, M. I. de la [Departamento de Física Teórica, Atómica y Óptica, Universidad de Valladolid, 47011-Valladolid (Spain)

    2014-10-07

    We report a detailed characterization of the spatial resolution provided by two-photon absorption spectroscopy suited for plasma diagnosis via the 1S-2S transition of atomic hydrogen for optogalvanic detection and laser induced fluorescence (LIF). A precise knowledge of the spatial resolution is crucial for a correct interpretation of measurements, if the plasma parameters to be analysed undergo strong spatial variations. The present study is based on a novel approach which provides a reliable and realistic determination of the spatial resolution. Measured irradiance distribution of laser beam waists in the overlap volume, provided by a high resolution UV camera, are employed to resolve coupled rate equations accounting for two-photon excitation, fluorescence decay and ionization. The resulting three-dimensional yield distributions reveal in detail the spatial resolution for optogalvanic and LIF detection and related saturation due to depletion. Two-photon absorption profiles broader than the Fourier transform-limited laser bandwidth are also incorporated in the calculations. The approach allows an accurate analysis of the spatial resolution present in recent and future measurements.

  9. Calculation of the spatial resolution in two-photon absorption spectroscopy applied to plasma diagnosis

    International Nuclear Information System (INIS)

    Garcia-Lechuga, M.; Fuentes, L. M.; Grützmacher, K.; Pérez, C.; Rosa, M. I. de la

    2014-01-01

    We report a detailed characterization of the spatial resolution provided by two-photon absorption spectroscopy suited for plasma diagnosis via the 1S-2S transition of atomic hydrogen for optogalvanic detection and laser induced fluorescence (LIF). A precise knowledge of the spatial resolution is crucial for a correct interpretation of measurements, if the plasma parameters to be analysed undergo strong spatial variations. The present study is based on a novel approach which provides a reliable and realistic determination of the spatial resolution. Measured irradiance distribution of laser beam waists in the overlap volume, provided by a high resolution UV camera, are employed to resolve coupled rate equations accounting for two-photon excitation, fluorescence decay and ionization. The resulting three-dimensional yield distributions reveal in detail the spatial resolution for optogalvanic and LIF detection and related saturation due to depletion. Two-photon absorption profiles broader than the Fourier transform-limited laser bandwidth are also incorporated in the calculations. The approach allows an accurate analysis of the spatial resolution present in recent and future measurements.

  10. Broadband high-resolution two-photon spectroscopy with laser frequency combs

    OpenAIRE

    Hipke, Arthur; Meek, Samuel A.; Ideguchi, Takuro; Hänsch, Theodor W.; Picqué, Nathalie

    2013-01-01

    Two-photon excitation spectroscopy with broad spectral span is demonstrated at Doppler-limited resolution. We describe first Fourier transform two-photon spectroscopy of an atomic sample with two mode-locked laser oscillators in a dual-comb technique. Each transition is uniquely identified by the modulation imparted by the interfering comb excitations. The temporal modulation of the spontaneous two-photon fluorescence is monitored with a single photodetector, and the spectrum is revealed by a...

  11. Broadband Doppler-limited two-photon and stepwise excitation spectroscopy with laser frequency combs

    Science.gov (United States)

    Hipke, Arthur; Meek, Samuel A.; Ideguchi, Takuro; Hänsch, Theodor W.; Picqué, Nathalie

    2014-07-01

    Multiplex two-photon excitation spectroscopy is demonstrated at Doppler-limited resolution. We describe first Fourier-transform two-photon spectroscopy of an atomic sample with two mode-locked laser oscillators in a dual-comb technique. Each transition is uniquely identified by the modulation imparted by the interfering comb excitations. The temporal modulation of the spontaneous two-photon fluorescence is monitored with a single photodetector, and the spectrum of all excited transitions is revealed by a Fourier transform.

  12. Synthesis, structure and two-photon absorption properties of a new multi-branched compound, 1,2,4,5-tetrakis(4-pyridylvinyl)benzene

    International Nuclear Information System (INIS)

    Wang Lei; Tao Xutang; Yang Jiaxiang; Yu Wentao; Ren Yan; Xin Qian; Liu Zhi; Jiang Minhua

    2004-01-01

    A conjugated and symmetric multi-branched compound, 1,2,4,5-tetrakis(4-pyridylvinyl)benzene (TKPVB), has been synthesized and the crystal structures of TKPVB and its intermediate, 1,2,4,5-tetrakis(dimethoxyphosphorylmethyl)benzene, were determined by diffraction method. TKPVB with four units of 4-vinylpyridine moieties attached to the central benzene core presents an A-π-A general framework, where A is a π-deficient pyridine ring. The single-photon and two-photon absorption and fluorescence properties in different solvents of varying polarity have been investigated. It is also found that the one- and two-photon-induced fluorescence spectra are quite similar, which indicate that the one- and two-photon allowed-excited states are the same

  13. Quarkonium two-photon decays in QCD

    International Nuclear Information System (INIS)

    Dulyan, L.S.; Khodjamirian, A.Yu.; Magakian, A.D.

    1989-01-01

    The two-photon decay of tensor charmonium χ c2 → 2 γ is calculated with account of gluon condensate effects. The result is in good agreement with experiment. The two-photon width of pseudoscalar b-quarkonium η b → 2 γ is estimated. 19 refs.; 1 fig.; 1 tab

  14. Comparison of confocal microscopy and two-photon microscopy in mouse cornea in vivo.

    Science.gov (United States)

    Lee, Jun Ho; Lee, Seunghun; Gho, Yong Song; Song, In Seok; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

    2015-03-01

    High-resolution imaging of the cornea is important for studying corneal diseases at cellular levels. Confocal microscopy (CM) has been widely used in the clinic, and two-photon microscopy (TPM) has recently been introduced in various pre-clinical studies. We compared the performance of CM and TPM in normal mouse corneas and neovascularized mouse corneas induced by suturing. Balb/C mice and C57BL/6 mice expressing green fluorescent protein (GFP) were used to compare modalities based on intrinsic contrast and extrinsic fluorescence contrast. CM based on reflection (CMR), CM based on fluorescence (CMF), and TPM based on intrinsic/extrinsic fluorescence and second harmonic generation (SHG) were compared by imaging the same sections of mouse corneas sequentially in vivo. In normal mouse corneas, CMR visualized corneal cell morphologies with some background noise, and CMF visualized GFP expressing corneal cells clearly. TPM visualized corneal cells and collagen in the stroma based on fluorescence and SHG, respectively. However, in neovascularized mouse corneas, CMR could not resolve cells deep inside the cornea due to high background noise from the effects of increased structural irregularity induced by suturing. CMF and TPM visualized cells and induced vasculature better than CMR because both collect signals from fluorescent cells only. Both CMF and TPM had signal decays with depth due to the structural irregularity, with CMF having faster signal decay than TPM. CMR, CMF, and TPM showed different degrees of image degradation in neovascularized mouse corneas. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Two-Photon Vibrational Spectroscopy using local optical fields of gold and silver nanostructures

    Science.gov (United States)

    Kneipp, Katrin; Kneipp, Janina; Kneipp, Harald

    2007-03-01

    Spectroscopic effects can be strongly affected when they take place in the immediate vicinity of metal nanostructures due to coupling to surface plasmons. We introduce a new approach that suggests highly efficient two-photon labels as well as two-photon vibrational spectroscopy for non-destructive chemical probing. The underlying spectroscopic effect is the incoherent inelastic scattering of two photons on the vibrational quantum states performed in the enhanced local optical fields of gold nanoparticles, surface enhanced hyper Raman scattering (SEHRS). We infer effective two-photon cross sections for SEHRS on the order of 10^5 GM, similar or higher than the best known cross sections for two-photon fluorescence. SEHRS combines the advantages of two-photon spectroscopy with the structural information of vibrational spectroscopy, and the high sensitivity and nanometer-scale local confinement of plasmonics-based spectroscopy.

  16. Laser induced fluorescence of some plant leaves

    International Nuclear Information System (INIS)

    Helmi, M.S.; Mohamed, M.M.; Amer, R.; Elshazly, O.; Elraey, M.

    1992-01-01

    Laser induced fluorescence (LIF) is successfully used as a technique for remote detection of spectral characteristics of some plants. A pulsed nitrogen laser at 337.1 nm is used to excite cotton, corn and rice leaves. The fluorescence spectrum is detected in the range from 340 nm to 820 nm. It is found that, these plant leaves have common fluorescence maxima at 440 nm, 685 nm and 740 nm. plant leaves are also found to be identifiable by the ratio of the fluorescence intensity at 440 nm to that at 685 nm. The present technique can be further used as a means of assessing, remotely, plant stresses. 5 fig

  17. Photo-induced processes in collagen-hypericin system revealed by fluorescence spectroscopy and multiphoton microscopy.

    Science.gov (United States)

    Hovhannisyan, V; Guo, H W; Hovhannisyan, A; Ghukasyan, V; Buryakina, T; Chen, Y F; Dong, C Y

    2014-05-01

    Collagen is the main structural protein and the key determinant of mechanical and functional properties of tissues and organs. Proper balance between synthesis and degradation of collagen molecules is critical for maintaining normal physiological functions. In addition, collagen influences tumor development and drug delivery, which makes it a potential cancer therapy target. Using second harmonic generation, two-photon excited fluorescence microscopy, and spectrofluorimetry, we show that the natural pigment hypericin induces photosensitized destruction of collagen-based tissues. We demonstrate that hypericin-mediated processes in collagen fibers are irreversible and may be used for the treatment of cancer and collagen-related disorders.

  18. Measurement of isotope shift of recycled uranium by laser induced fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Oba, Masaki; Wakaida, Ikuo; Akaoka, Katsuaki; Miyabe, Masabumi

    1999-07-01

    Isotope shift of the recycled uranium atoms including the 236 U was measured by laser induced fluorescence method. Eight even levels at 2 eV and three odd levels at 4 eV were measured with isotope shifts among 238 U, 236 U and 235 U obtained. As for the measurement of the 4 eV levels, the Doppler free two photon absorption method was used, and the hyperfine structure of the 235 U was analyzed simultaneously. The isotope shift of 234 U was also observed in the three transition. (J.P.N.)

  19. Charm production in two-photon collisions

    International Nuclear Information System (INIS)

    Linde, F.L.

    1988-01-01

    The analysis focuses on the production of charmonium resonances η c , χ 0 and χ 2 in two-photon interactions. The measurement of the inclusive production of charged D* mesons is described. 97 refs.; 54 figs.; 15 tabs

  20. Exclusive hadron production in two photon reactions

    International Nuclear Information System (INIS)

    Poppe, M.

    1986-02-01

    This paper summarises experimental results on exclusive hadron production in two photon collisions at electron positron storage rings and attempts some interpretation. Experimental know how is described and new suggestions are made for future analyses. New model calculations on resonance form factors and pair production amplitudes are presented. The two photon vertex is decomposed such that experiments can be parameterised with the minimal number of free parameters. Selection rules for off shell photon collisions are given in addition to Yang's theorems. (orig.)

  1. Single molecule photodynamics by means of one- and two-photon approach

    International Nuclear Information System (INIS)

    Chirico, Giuseppe; Cannone, Fabio; Diaspro, Alberto

    2003-01-01

    Single molecule spectroscopy allows to investigate heterogeneous behaviours on photochemical and structural grounds. We report on studies of the effect of the excitation intensity on the internal photodynamics of simple dyes immobilized on chemically etched glass slides. The use of the excitation intensity needed for two-photon excitation induces local heating, structural changes and transitions to dark states. Similar behaviour is found on single green fluorescent proteins immobilized on glass slides or embedded in silica gels upon single-photon excitation. However, by sampling the images with sufficiently low frequency, we are able to follow relevant biological events, such as the unfolding kinetics. We find that the glass slides are preferable in terms of the signal-to-noise ratio but the protein is not preserved in its native state, while evidence for the native conformation of the single proteins in the silica gels is found in the uniformity of the fluorescence emission

  2. Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy

    Science.gov (United States)

    Hunter, Jennifer J.; Masella, Benjamin; Dubra, Alfredo; Sharma, Robin; Yin, Lu; Merigan, William H.; Palczewska, Grazyna; Palczewski, Krzysztof; Williams, David R.

    2011-01-01

    In vivo two-photon imaging through the pupil of the primate eye has the potential to become a useful tool for functional imaging of the retina. Two-photon excited fluorescence images of the macaque cone mosaic were obtained using a fluorescence adaptive optics scanning laser ophthalmoscope, overcoming the challenges of a low numerical aperture, imperfect optics of the eye, high required light levels, and eye motion. Although the specific fluorophores are as yet unknown, strong in vivo intrinsic fluorescence allowed images of the cone mosaic. Imaging intact ex vivo retina revealed that the strongest two-photon excited fluorescence signal comes from the cone inner segments. The fluorescence response increased following light stimulation, which could provide a functional measure of the effects of light on photoreceptors. PMID:21326644

  3. Intravital two-photon microscopy of immune cell dynamics in corneal lymphatic vessels.

    Directory of Open Access Journals (Sweden)

    Philipp Steven

    Full Text Available BACKGROUND: The role of lymphatic vessels in tissue and organ transplantation as well as in tumor growth and metastasis has drawn great attention in recent years. METHODOLOGY/PRINCIPAL FINDINGS: We now developed a novel method using non-invasive two-photon microscopy to simultaneously visualize and track specifically stained lymphatic vessels and autofluorescent adjacent tissues such as collagen fibrils, blood vessels and immune cells in the mouse model of corneal neovascularization in vivo. The mouse cornea serves as an ideal tissue for this technique due to its easy accessibility and its inducible and modifiable state of pathological hem- and lymphvascularization. Neovascularization was induced by suture placement in corneas of Balb/C mice. Two weeks after treatment, lymphatic vessels were stained intravital by intrastromal injection of a fluorescently labeled LYVE-1 antibody and the corneas were evaluated in vivo by two-photon microscopy (TPM. Intravital TPM was performed at 710 nm and 826 nm excitation wavelengths to detect immunofluorescence and tissue autofluorescence using a custom made animal holder. Corneas were then harvested, fixed and analyzed by histology. Time lapse imaging demonstrated the first in vivo evidence of immune cell migration into lymphatic vessels and luminal transport of individual cells. Cells immigrated within 1-5.5 min into the vessel lumen. Mean velocities of intrastromal corneal immune cells were around 9 µm/min and therefore comparable to those of T-cells and macrophages in other mucosal surfaces. CONCLUSIONS: To our knowledge we here demonstrate for the first time the intravital real-time transmigration of immune cells into lymphatic vessels. Overall this study demonstrates the valuable use of intravital autofluorescence two-photon microscopy in the model of suture-induced corneal vascularizations to study interactions of immune and subsequently tumor cells with lymphatic vessels under close as possible

  4. Two-photon superradiance in extended medium

    International Nuclear Information System (INIS)

    Branzan, V.; Enache, N.

    1993-01-01

    The possibility of collectivization of an ensemble of atoms of an extended system (the distance between atoms is larger or equal to the wave-length of a spontaneous emitted radiation) during two-photon spontaneous decay is theoretically investigated. It is demonstrated that such systems of inverted atoms should emit phase-correlated pairs of photons. The time-space correlation among atoms is realized due to the two-photon exchanging through the electromagnetic field's vacuum. An increase of the spontaneous decay rate of the two-atom inverted ensemble is demonstrated. The dependence of two-photon superradiance on the sample geometry is investigated. A non-equilibrium method of the elimination of the atoms level Fermi-operators is proposed. (Author)

  5. Squeezing via two-photon transitions

    Science.gov (United States)

    Savage, C. M.; Walls, D. F.

    1986-05-01

    The squeezing spectrum for a cavity field mode interacting with an ensemble of three-level 'Lambda-configuration' atoms by an effective two-photon transition is calculated. The advantage of the three-level Lambda system as a squeezing medium, that is, optical nonlinearity without atomic saturation, has recently been pointed out by Reid, Walls, and Dalton. Perfect squeezing is predicted at the turning points for dispersive optical bistability and good squeezing for a range of other cases. Three-level ladder atoms interacting by an effective two-photon transition are also shown to give perfect squeezing in the dispersive limit.

  6. Two-photon excitation of argon

    International Nuclear Information System (INIS)

    Pindzola, P.S.; Payne, M.C.

    1982-01-01

    The authors calculate two photon excitation parameters for various excited states of argon assuming the absorption of near resonance broad-bandwidth laser radiation. Results are given for the case of two photons absorbed for the same laser beam as well as the case of absorbing photons of different frequency from each of two laser beams. The authors use multiconfiguration Hartree-Fock wave functions to evaluate the second-order sums over matrix elements. Various experimental laser schemes are suggested for the efficient excitation and subsequent ionization of argon

  7. A review of two photon physics

    International Nuclear Information System (INIS)

    Cooper, S.

    1982-08-01

    This talk is intended as an introduction for those not yet expert in two-photon physics, especially those e + e - one-photon physicists who still think of two-photon events as background. I concentrate on the physics questions involved, especially emphasizing the areas where I feel progress can be made in the near future, and of necessity leaving most experimental details to be found in the references. After a quick survey of the field and a few words about kinematics, I discuss in detail two major fields: the photon structure function and resonance production. (orig.)

  8. Effects of Depilation-Induced Skin Pigmentation and Diet-Induced Fluorescence on In Vivo Fluorescence Imaging

    OpenAIRE

    Kwon, Sunkuk; Sevick-Muraca, Eva M.

    2017-01-01

    Near-infrared fluorescence imaging (NIRFI) and far-red fluorescence imaging (FRFI) were used to investigate effects of depilation-induced skin pigmentation and diet-induced background fluorescence on fluorescent signal amplitude and lymphatic contraction frequency in C57BL6 mice. Far-red fluorescent signal amplitude, but not frequency, was affected by diet-induced fluorescence, which was removed by feeding the mice an alfalfa-free diet, and skin pigmentation further impacted the amplitude mea...

  9. Identification of CW two-photon transitions in Na2 and NaK

    International Nuclear Information System (INIS)

    Morgan, G.P.

    1983-01-01

    This thesis reports on the two-photon visible excitation spectra of sodium and potassium vapors. In the past, similar work has been performed on sodium and many atomic two-photon transitions have been characterized. However, many extra signals exist which do not possess the ground, 3S, state hyperfine splitting. These extra transitions are due to the sodium dimer Na 2 . 79 such transitions, from 5800A - 6500A, which lie within the resolution of the apparatus have been studied. The molecules are excited with a lowpower narrow band counterpropagating cw dye laser beam and two-photon fluorescence. The fluorescence intensities of many of these transitions are greater than the 3S to 5S and 3S to 4D atomic signals, where the 3P enhancing state lies 300 cm -1 from resonance. By comparing the number density of the atomic with any molecular ground state and also the two-photon transition rates to excited states, the intermediate enhancing state for a two-photon transition in Na 2 can be predicted to be less than 1 cm -1 from resonance with the two-photon transition. This observation, along with published Dunham coefficients, is used to identify the states involved in the two-photon transitions

  10. Laser-induced fluorescence imaging of bacteria

    Science.gov (United States)

    Hilton, Peter J.

    1998-12-01

    This paper outlines a method for optically detecting bacteria on various backgrounds, such as meat, by imaging their laser induced auto-fluorescence response. This method can potentially operate in real-time, which is many times faster than current bacterial detection methods, which require culturing of bacterial samples. This paper describes the imaging technique employed whereby a laser spot is scanned across an object while capturing, filtering, and digitizing the returned light. Preliminary results of the bacterial auto-fluorescence are reported and plans for future research are discussed. The results to date are encouraging with six of the eight bacterial strains investigated exhibiting auto-fluorescence when excited at 488 nm. Discrimination of these bacterial strains against red meat is shown and techniques for reducing background fluorescence discussed.

  11. Review of Two-Photon Interactions

    International Nuclear Information System (INIS)

    Urner, David

    2004-01-01

    Presented are recent results of two-photon interactions. Topics include photon structure functions, inclusive hadron production, differential cross sections derived from tagged γγ fusion events and results in exclusive hadron production, particularly the observations of the η'c

  12. Two-photon transitions to exciton polaritons

    International Nuclear Information System (INIS)

    Hassan, A.R.

    1979-08-01

    A semiclassical theory for the creation of excitonic polariton states by two-photon absorption, via an intermediate exciton state, is given. A band model has been introduced which gives the dominant contribution to this process. A numerical calculation is found to be in good agreement with a recent observation in CuCl. (author)

  13. Pion-pair production by two photons

    International Nuclear Information System (INIS)

    Terazawa, Hidezumi.

    1994-07-01

    The cross section for pion-pair production by two photons is calculated approximately by using the low energy theorem previously derived from partially-conserved-axial-vector-current hypothesis and current algebra, and found to agree very well with the experimental data recently obtained by the Mark II, TPC/Two-Gamma and CLEO Collaborations. (author)

  14. Two-photon excitation spectroscopy of carotenoid-containing and carotenoid-depleted LH2 complexes from purple bacteria.

    Science.gov (United States)

    Stepanenko, Ilya; Kompanetz, Viktor; Makhneva, Zoya; Chekalin, Sergey; Moskalenko, Andrei; Razjivin, Andrei

    2009-08-27

    We applied two-photon fluorescence excitation spectroscopy to LH2 complex from purple bacteria Allochromatium minutissimum and Rhodobacter sphaeroides . Bacteriochlorophyll fluorescence was measured under two-photon excitation of the samples within the 1200-1500 nm region. Spectra were obtained for both carotenoid-containing and -depleted complexes of each bacterium to allow their direct comparison. The depletion of carotenoids did not alter the two-photon excitation spectra of either bacteria. The spectra featured a wide excitation band around 1350 nm (2x675 nm, 14,800 cm(-1)) which strongly resembled two-photon fluorescence excitation spectra of similar complexes published by other authors. We consider obtained experimental data to be evidence of direct two-photon excitation of bacteriochlorophyll excitonic states in this spectral region.

  15. Two-photon imaging and analysis of neural network dynamics

    International Nuclear Information System (INIS)

    Luetcke, Henry; Helmchen, Fritjof

    2011-01-01

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  16. Two-photon imaging and analysis of neural network dynamics

    Science.gov (United States)

    Lütcke, Henry; Helmchen, Fritjof

    2011-08-01

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  17. Two-photon imaging and analysis of neural network dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Luetcke, Henry; Helmchen, Fritjof [Brain Research Institute, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich (Switzerland)

    2011-08-15

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  18. Can Two-Photon Interference be Considered the Interference of Two Photons?

    International Nuclear Information System (INIS)

    Pittman, T.B.; Strekalov, D.V.; Migdall, A.; Rubin, M.H.; Sergienko, A.V.; Shih, Y.H.

    1996-01-01

    We report on a open-quote open-quote postponed compensation close-quote close-quote experiment in which the observed two-photon entangled state interference cannot be pictured in terms of the overlap of the two individual photon wave packets of a parametric down-conversion pair on a beam splitter. In the sense of a quantum eraser, the distinguishability of the different two-photon Feynman amplitudes leading to a coincidence detection is removed by delaying the compensation until after the output of an unbalanced two-photon interferometer. copyright 1996 The American Physical Society

  19. Probing two-photon exchange with OLYMPUS

    International Nuclear Information System (INIS)

    Kohl, M.

    2014-01-01

    Two-photon exchange is believed to be responsible for the discrepancies in the proton electric to magnetic form factor ratio found with the Rosenbluth and polarization transfer methods. If this explanation is correct, one expects significant differences in the lepton-proton cross sections between positrons and electrons. The OLYMPUS experiment at DESY in Hamburg, Germany was designed to measure the ratio of unpolarized positron-proton and electron-proton elastic scattering cross sections over a wide kinematic range with high precision, in order to quantify the effect of two-photon exchange. The experiment used intense beams of electrons and positrons stored in the DORIS ring at 2.0 GeV interacting with an internal windowless hydrogen gas target. The current status of OLYMPUS will be discussed. (authors)

  20. Symmetry adaptation in two-photon spectroscopy

    International Nuclear Information System (INIS)

    Kibler, M.

    1991-11-01

    Symmetry adaptation techniques are applied to the determination of the intensity of two-photon transitions for transition ions in finite symmetry environments. The case of intra-configurational transitions are discussed with some details and some results on inter-configurational transitions are briefly reported. In particular, for intra-configurational transitions, a model is described which takes into account the following ingredients: (symmetry, second- plus third-order mechanisms, S-, L- and J-mixings). (author) 20 refs

  1. Two-photon Absorption In Quantum Dots,quantum Dashes And Related Materials

    Energy Technology Data Exchange (ETDEWEB)

    Jain, Ravinder

    2009-08-31

    We have proposed the use of USQDs for various deep-tissue biological imaging applications, notably wavelength-multiplexed multicolor imaging and intra-nuclear studies such as those involving cell apoptosis, and have studied the issue of maximizing two-photon absorption-induced fluorescence (TPAF) signals from CdSe/ZnS USQDs to be used for this application. In particular, using 2 nm USQDs, we have shown that the TPAF signal at 780 nm is ~ 8 times that at 850 nm and 68 times that at 900 nm, two wavelengths that have been used in previous studies using CdSe/ZnS SQDs for deep-tissue imaging of biological studies via TPAF .

  2. Two-photon spin generation and detection

    Energy Technology Data Exchange (ETDEWEB)

    Miah, M Idrish, E-mail: m.miah@griffith.edu.a [Nanoscale Science and Technology Centre, Griffith University, Nathan, Brisbane, QLD 4111 (Australia)

    2009-02-21

    A time- and polarization-resolved two-photon pump-probe investigation is performed in lightly doped GaAs. We generate spin-polarized electrons in bulk GaAs at various temperatures using right-circularly polarized two-photon excitation and detect them by probing the spin-dependent transmission of the sample. The spin polarization (P) of conduction band electrons, as measured using probe pulses with the same (right) and opposite (left) circular polarization, is measured in dependences of pump-probe delay ({Delta}t), lattice temperature (T{sub L}), doping density (n) as well as of the excess photon energy {Delta}E{sub 2{omega}}= {h_bar}2{omega} - E{sub g}, where E{sub g} is the band gap energy. P is found to be decayed with {Delta}t and enhanced with the decrease in T{sub L} or the increase in n. It is also found that P decreases with the increase in {Delta}E{sub 2{omega}}and depolarizes rapidly for {Delta}E{sub 2{omega}}> {Delta}E{sub SO}, where {Delta}E{sub SO} is the spin-orbit splitting energy. The results demonstrate that due to a much longer absorption depth highly polarized spins can be generated optically by two-photon pumping of bulk semiconductors.

  3. Two-photon spin generation and detection

    International Nuclear Information System (INIS)

    Miah, M Idrish

    2009-01-01

    A time- and polarization-resolved two-photon pump-probe investigation is performed in lightly doped GaAs. We generate spin-polarized electrons in bulk GaAs at various temperatures using right-circularly polarized two-photon excitation and detect them by probing the spin-dependent transmission of the sample. The spin polarization (P) of conduction band electrons, as measured using probe pulses with the same (right) and opposite (left) circular polarization, is measured in dependences of pump-probe delay (Δt), lattice temperature (T L ), doping density (n) as well as of the excess photon energy ΔE 2ω = ℎ2ω - E g , where E g is the band gap energy. P is found to be decayed with Δt and enhanced with the decrease in T L or the increase in n. It is also found that P decreases with the increase in ΔE 2ω and depolarizes rapidly for ΔE 2ω > ΔE SO , where ΔE SO is the spin-orbit splitting energy. The results demonstrate that due to a much longer absorption depth highly polarized spins can be generated optically by two-photon pumping of bulk semiconductors.

  4. Inhibition of two-photon absorption in a three-level system with a pair of bichromatic fields

    International Nuclear Information System (INIS)

    Zou Jinhua; Hu Xiangming; Cheng Guangling; Li Xing; Du Dan

    2005-01-01

    We study two-photon absorption in a three-level ladder atomic system driven by a pair of bichromatic fields of equal frequency differences. The high-frequency component of one bichromatic field and the low-frequency component of the other are on two-photon resonance. The transition probability is calculated by employing the method of harmonic expansion and matrix inversion. Unexpectedly, when the sums of the phases of the different pairs of field components on the two-photon resonance are equal to each other, two-photon absorption is dramatically suppressed and the atomic system becomes transparent against two-photon absorption. Physically, due to dynamical Stark splitting, the two-photon transitions induced by the different pairs of field components experience different dressed states with phase difference of π. As a result, destructive interference occurs between the two pathways and leads to the inhibition of two-photon absorption

  5. Two-Photon Probes for Lysosomes and Mitochondria: Simultaneous Detection of Lysosomes and Mitochondria in Live Tissues by Dual-Color Two-Photon Microscopy Imaging.

    Science.gov (United States)

    Lim, Chang Su; Hong, Seung Taek; Ryu, Seong Shick; Kang, Dong Eun; Cho, Bong Rae

    2015-10-01

    Novel two-photon (TP) probes were developed for lysosomes (PLT-yellow) and mitochondria (BMT-blue and PMT-yellow). These probes emitted strong TP-excited fluorescence in cells at widely separated wavelength regions and displayed high organelle selectivity, good cell permeability, low cytotoxicity, and pH insensitivity. The BMT-blue and PLT-yellow probes could be utilized to detect lysosomes and mitochondria simultaneously in live tissues by using dual-color two-photon microscopy, with minimum interference from each other. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Laser-induced fluorescence for medical diagnostics

    International Nuclear Information System (INIS)

    Andersson Engels, S.

    1989-12-01

    Laser-induced fluorescence as a tool for tissue diagnostics is discussed. Both spectrally and time-resolved fluorescence signals are studied to optimize the demarcation of diseased lesions from normal tissue. The presentation is focused on two fields of application: the identification of malignant tumours and atherosclerotic plaques. Tissue autofluorescence as well as fluorescence from administered drugs have been utilized in diseased tissue diagnosis. The fluorescence criterion for tissue diagnosis is, as far as possible, chosen to be independent of unknown fluorescence parameters, which are not correlated to the type of tissue investigated. Both a dependence on biological parameters, such as light absorption in blood, and instrumental characteristics, such as excitation pulse fluctuations and detection geometry, can be minimized. Several chemical compounds have been studied in animal experiments after intraveneous injection to verify their capacity as malignant tumour marking drugs under laser excitation and fluorescence detection. Another objective of these studies was to improve our understanding of the mechanism and chemistry behind the retention of the various drugs in tissue. The properties of a chemical which maximize its selective retention in tumours are discussed. In order to utilize this diagnostic modality, three different clinically adapted sets of instrumentation have been developed and are presented. Two of the systems are nitrogen-laser-based fluorosensors; one is a point-monitoring system with full spectral resolution and the other one is an imaging system with up to four simultaneously recorded images in different spectral bands. The third system is a low-cost point-monitoring mercury-lamp-based fluoroscence emission as well as reflection characteristics of tissue. (author)

  7. In vivo two-photon imaging of retina in rabbits and rats.

    Science.gov (United States)

    Jayabalan, Gopal Swamy; Wu, Yi-Kai; Bille, Josef F; Kim, Samuel; Mao, Xiao Wen; Gimbel, Howard V; Rauser, Michael E; Fan, Joseph T

    2018-01-01

    The purpose of this study was to evaluate the retina using near-infrared (NIR) two-photon scanning laser ophthalmoscopy. New Zealand white rabbits, albino rats, and brown Norway rats were used in this study. An autofluorescence image of the retina, including the retinal cells and its associated vasculatures was obtained by a real-time scan using the ophthalmoscope. Furthermore, the retinal vessels, nerve fiber layers and the non-pigmented retina were recorded with two-photon fluorescein angiography (FA); and the choroidal vasculatures were recorded using two-photon indocyanine green angiography (ICGA). Two-photon ICGA was achieved by exciting a second singlet state at ∼398 nm. Simultaneous two-photon FA and two-photon ICGA were performed to characterize the retinal and choroidal vessels with a single injection. The minimum laser power threshold required to elicit two-photon fluorescence was determined. The two-photon ophthalmoscope could serve as a promising tool to detect and monitor the disease progression in animal models. Moreover, these high-resolution images of retinal and choroidal vessels can be acquired in a real-time scan with a single light source, requiring no additional filters for FA or ICGA. The combination of FA and ICGA using the two-photon ophthalmoscope will help researchers to characterize the retinal diseases in animal models, and also to classify the types (classic, occult or mixed) of choroidal neovascularization (CNV) in macular degeneration. Furthermore, the prototype can be adapted to image the retina of rodents and rabbits. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Preparation, one- and two-photon properties of carbazole derivatives containing nitrogen heterocyclic ring

    Science.gov (United States)

    Zhang, Yichi; Wang, Ping; Li, Liang; Chen, Zhimin; He, Chunying; Wu, Yiqun

    Preparation of recording materials with high two-photon absorption activities is one of the important issues to superhigh- density two-photon absorption (TPA) three-dimensional (3D) optical data storage. In this paper, three new carbazole derivatives containing nitrogen heterocyclic ring with symmetric and asymmetric structures are prepared using ethylene as the π bridge between the carbazole unit and nitrogen heterocyclic ring, namely, 9-butyl-3-(2-(1,8- naphthyridin)vinyl)-carbazole (material 1), 9-butyl-3,6-bis(2-(1,8-naphthyl)vinyl)-carbazole (material 2) and 9-butyl-3,6- bis(2-(quinolin)vinyl)-carbazole (material 3). Their one photon properties including linear absorption spectra, fluorescence emission spectra, and fluorescence quantum yields are studied. The fluorescence excited by 120 fs pulse at 800 nm Ti: sapphire laser operating at 1 kHz repetition rate with different incident powers of 9-butyl-3-(2-(quinolin) vinyl)-carbazole (material 3) was investigated, and two-photon absorption cross-sections has been obtained. It is shown that material 3 containing quinoline rings as electron acceptor with symmetric structure exhibit high two-photon absorption activity. The result implies that material 3 (9-butyl-3-(2-(quinolin) vinyl)-carbazole) is a good candidate as a promising recording material for super-high-density two-photon absorption (TPA) three-dimensional (3D) optical data storage. The influence of chemical structure of the materials on the optical properties is discussed.

  9. Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye.

    Science.gov (United States)

    Sharma, Robin; Williams, David R; Palczewska, Grazyna; Palczewski, Krzysztof; Hunter, Jennifer J

    2016-02-01

    Although extrinsic fluorophores can be introduced to label specific cell types in the retina, endogenous fluorophores, such as NAD(P)H, FAD, collagen, and others, are present in all retinal layers. These molecules are a potential source of optical contrast and can enable noninvasive visualization of all cellular layers. We used a two-photon fluorescence adaptive optics scanning light ophthalmoscope (TPF-AOSLO) to explore the native autofluorescence of various cell classes spanning several layers in the unlabeled retina of a living primate eye. Three macaques were imaged on separate occasions using a custom TPF-AOSLO. Two-photon fluorescence was evoked by pulsed light at 730 and 920 nm excitation wavelengths, while fluorescence emission was collected in the visible range from several retinal layers and different locations. Backscattered light was recorded simultaneously in confocal modality and images were postprocessed to remove eye motion. All retinal layers yielded two-photon signals and the heterogeneous distribution of fluorophores provided optical contrast. Several structural features were observed, such as autofluorescence from vessel walls, Müller cell processes in the nerve fibers, mosaics of cells in the ganglion cell and other nuclear layers of the inner retina, as well as photoreceptor and RPE layers in the outer retina. This in vivo survey of two-photon autofluorescence throughout the primate retina demonstrates a wider variety of structural detail in the living eye than is available through conventional imaging methods, and broadens the use of two-photon imaging of normal and diseased eyes.

  10. Radiative corrections to two photon physics

    International Nuclear Information System (INIS)

    Neervan, W.L. van; Vermaseren, J.A.M.

    1983-06-01

    The authors develop a method to calculate radiative corrections to two photon reactions of the type e + e - →e + e - X where X is an arbitrary final state. To illustrate this they take the example where X stands for a point-like pseudoscalar. It will be shown that the method is an improvement on the standard way in adding real and virtual photon contributions to the (differential) cross-sections. This makes it possible to compute differential and total cross-sections to a very high precision and with a minimum of computer time which is not possible by using more conventional methods. (Auth.)

  11. Inclusive two-photon reactions at TRISTAN

    International Nuclear Information System (INIS)

    Drees, M.

    1995-01-01

    After briefly reviewing past accomplishments of TRISTAN experiments in the field of inclusive two-photon reactions, I discuss open problems in the Monte Carlo simulation of such reactions. The main emphasis is on multiple scattering, i.e. events where at least two pairs of partons scatter within the same γγ collision to form at least four (mini)jets. The cross section for such events might just be observable at TRISTAN. While theoretical arguments for the existence of such events are strong, they have not yet been directly observed experimentally, thereby potentially opening a new opportunity for TRISTAN experiments. (author)

  12. Two-photon physics at LEP

    International Nuclear Information System (INIS)

    Ginzburg, I.F.

    1988-01-01

    The two-photon production of hadrons in e + e - collisions e + e - →e + e - h from which the γγ→h cross sections are extracted is discussed. The common features of these processes are: hadrons move, as a rule, along e + e - beam axis, their total transverse momentum K perpendicular or perpendicular to is small; the total hadron energy is usually less than √S/2. Physical problems of soft processes, exotics, hard processes, semihard processes are considered. New possibilities of LEP, the most interesting and real are presented

  13. Two-photon cooling of magnesium atoms

    DEFF Research Database (Denmark)

    Malossi, N.; Damkjær, S.; Hansen, P. L.

    2005-01-01

    A two-photon mechanism for cooling atoms below the Doppler temperature is analyzed. We consider the magnesium ladder system (3s2)S01¿(3s3p)P11 at 285.2nm followed by the (3s3p)P11¿(3s3d)D21 transition at 880.7nm . For the ladder system quantum coherence effects may become important. Combined with...... and experiment is excellent. In addition, by properly choosing the Rabi frequencies of the two optical transitions a velocity independent atomic dark state is observed....

  14. Mixture-fraction imaging at 1  kHz using femtosecond laser-induced fluorescence of krypton.

    Science.gov (United States)

    Richardson, Daniel R; Jiang, Naibo; Stauffer, Hans U; Kearney, Sean P; Roy, Sukesh; Gord, James R

    2017-09-01

    Femtosecond, two-photon-absorption laser-induced-fluorescence (TALIF) imaging measurements of krypton (Kr) are demonstrated to study mixing in gaseous flows. A measurement approach is presented in which observed Kr TALIF signals are 7 times stronger than the current state-of-the-art methodology. Fluorescence emission is compared for different gas pressures and excitation wavelengths, and the strongest fluorescence signals were observed when the excitation wavelength was tuned to 212.56 nm. Using this optimized excitation scheme, 1-kHz, single-laser-shot visualizations of unsteady flows and two-dimensional measurements of mixture fraction and scalar dissipation rate of a Kr-seeded jet are demonstrated.

  15. Laser induced fluorescence of dental caries

    Science.gov (United States)

    Albin, S.; Byvik, C. E.; Buoncristiani, A. M.

    1988-01-01

    Significant differences between the optical spectra taken from sound regions of teeth and carious regions have been observed. These differences appear both in absorption and in laser induced fluorescence spectra. Excitation by the 488 nm line of an argon ion laser beam showed a peak in the emission intensity around 553 nm for the sound dental material while the emission peak from the carious region was red-shifted by approximately 40 nm. The relative absorption of carious region was significantly higher at 488 nm; however its fluorescence intensity peak was lower by an order of magnitude compared to the sound tooth. Implications of these results for a safe, reliable and early detection of dental caries are discussed.

  16. Resonant production in two photon collisions

    International Nuclear Information System (INIS)

    Butler, F.

    1988-12-01

    Using 220 picobarn/sup /minus/1/ of data collected by the Mark II detector at PEP, we have measured the width of the /eta/' for quasi real photons. The width is separately measured in the three reactions e + e/sup /minus// → e + e/sup /minus///eta/ 0 π + π/sup /minus//; /eta/ 0 → γγ, e + e/sup /minus// → e + e/sup /minus///eta/ 0 π + π/sup /minus//; /eta/ 0 → π + π/sup /minus//(π 0 ,γ) and e + e/sup /minus// → e + e/sup /minus///rho/ 0 γ, giving a statistically weighted average two-photon width of Γ/sub /eta/'→γγ/ = 5. 21+-0.28 keV. As a by-product of the measurement of Γ/sub /eta/'→γγ/ using the decay mode /eta/' → /rho/γ, we also measured a two-photon decay width for the a 2 (1320) of Γ/sub a 2(1320)→γγ/ = 1.17+-0. 15+-0.25 keV. 47 refs., 50 figs., 8 tabs

  17. Behaviour of atomic oxygen in a pulsed dielectric barrier discharge measured by laser-induced fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Ono, Ryo [High Temperature Plasma Center, University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa, Chiba, 227-8568 (Japan); Yamashita, Youta [Department of Electrical Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656 (Japan); Takezawa, Kei [Department of Electrical Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656 (Japan); Oda, Tetsuji [Department of Electrical Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656 (Japan)

    2005-08-21

    Atomic oxygen is measured in a pulsed dielectric barrier discharge (DBD) using two-photon absorption laser-induced fluorescence (TALIF). The ground-level atomic oxygen is excited to the 3p {sup 3}P state by two-photon absorption at 226 nm. Negative (-40 kV) or positive (+30 kV) pulsed DBD occurs in an O{sub 2}-N{sub 2} mixture at atmospheric pressure. The pulse width of the DBD current is approximately 50 ns. The TALIF experiment shows that the decay rate of atomic oxygen increases linearly with O{sub 2} concentration. This result proves that atomic oxygen decays mainly by the third-body reaction, O + O{sub 2} + M {yields} O{sub 3} + M. The rate coefficient of the third-body reaction is estimated to be 2.2 x 10{sup -34} cm{sup 6} s{sup -1} in the negative DBD and 0.89 x 10{sup -34} cm{sup 6} s{sup -1} in the positive DBD. It is shown that the decay rate of atomic oxygen increases linearly with humidity. This can explain the well-known fact that ozone production in DBD is suppressed by increasing humidity.

  18. Behaviour of atomic oxygen in a pulsed dielectric barrier discharge measured by laser-induced fluorescence

    International Nuclear Information System (INIS)

    Ono, Ryo; Yamashita, Youta; Takezawa, Kei; Oda, Tetsuji

    2005-01-01

    Atomic oxygen is measured in a pulsed dielectric barrier discharge (DBD) using two-photon absorption laser-induced fluorescence (TALIF). The ground-level atomic oxygen is excited to the 3p 3 P state by two-photon absorption at 226 nm. Negative (-40 kV) or positive (+30 kV) pulsed DBD occurs in an O 2 -N 2 mixture at atmospheric pressure. The pulse width of the DBD current is approximately 50 ns. The TALIF experiment shows that the decay rate of atomic oxygen increases linearly with O 2 concentration. This result proves that atomic oxygen decays mainly by the third-body reaction, O + O 2 + M → O 3 + M. The rate coefficient of the third-body reaction is estimated to be 2.2 x 10 -34 cm 6 s -1 in the negative DBD and 0.89 x 10 -34 cm 6 s -1 in the positive DBD. It is shown that the decay rate of atomic oxygen increases linearly with humidity. This can explain the well-known fact that ozone production in DBD is suppressed by increasing humidity

  19. Ultratrace analysis of transuranic actinides by laser-induced fluorescence

    Science.gov (United States)

    Miller, S.M.

    1983-10-31

    Ultratrace quantities of transuranic actinides are detected indirectly by their effect on the fluorescent emissions of a preselected fluorescent species. Transuranic actinides in a sample are coprecipitated with a host lattice material containing at least one preselected fluorescent species. The actinide either quenches or enhances the laser-induced fluorescence of the preselected fluorescent species. The degree of enhancement or quenching is quantitatively related to the concentration of actinide in the sample.

  20. Laser induced fluorescence of trapped molecular ions

    International Nuclear Information System (INIS)

    Grieman, F.J.

    1979-10-01

    An experimental apparatus for obtaining the optical spectra of molecular ions is described. The experimental technique includes the use of three dimensional ion trapping, laser induced fluorescence, and gated photon counting methods. The ions, which are produced by electron impact, are confined in a radio-frequency quadrupole ion trap of cylindrical design. Because the quadrupole ion trap allows mass selection of the molecular ion desired for study, the analysis of the spectra obtained is greatly simplified. The ion trap also confines the ions to a region easily probed by a laser beam. 18 references

  1. Laser induced fluorescence of trapped molecular ions

    Energy Technology Data Exchange (ETDEWEB)

    Grieman, F.J.

    1979-10-01

    An experimental apparatus for obtaining the optical spectra of molecular ions is described. The experimental technique includes the use of three dimensional ion trapping, laser induced fluorescence, and gated photon counting methods. The ions, which are produced by electron impact, are confined in a radio-frequency quadrupole ion trap of cylindrical design. Because the quadrupole ion trap allows mass selection of the molecular ion desired for study, the analysis of the spectra obtained is greatly simplified. The ion trap also confines the ions to a region easily probed by a laser beam. 18 references.

  2. Laser induced fluorescence spectroscopy for FTU

    International Nuclear Information System (INIS)

    Hughes, T.P.

    1995-07-01

    Laser induced fluorescence spectroscopy (LIFS) is based on the absorption of a short pulse of tuned laser light by a group of atoms and the observation of the resulting fluorescence radiation from the excited state. Because the excitation is resonant it is very efficient, and the fluorescence can be many times brighter than the normal spontaneous emission, so low number densities of the selected atoms can be detected and measured. Good spatial resolution can be achieved by using a narrow laser beam. If the laser is sufficiently monochromatic, and it can be tuned over the absorption line profile of the selected atoms, information can also be obtained about the velocities of the atoms from the Doppler effect which can broaden and shift the line. In this report two topics are examined in detail. The first is the effect of high laser irradiance, which can cause 'power broadening' of the apparent absorption line profile. The second is the effect of the high magnetic field in FTU. Detailed calculations are given for LIFS of neutral iron and molybdenum atoms, including the Zeeman effect, and the implementation of LIFS for these atoms on FTU is discussed

  3. Microsphere imaging with confocal microscopy and two photon microscopy

    International Nuclear Information System (INIS)

    Chun, Hyung Su; An, Kyung Won; Lee, Jai Hyung

    2002-01-01

    We have acquired images of polystyrene and fused-silica microsphere by using conventional optical microscopy, confocal microscopy and two-photon microscopy, and performed comparative analysis of these images. Different from conventional optical microscopy, confocal and two-photon microscopy had good optical sectioning capability. In addition, confocal microscopy and two-photon microscopy had better lateral resolution than conventional optical microscopy. These results are attributed to confocality and nonlinearity of confocal microscopy and two photon microscopy, respectively.

  4. On the kinematics of the two-photon Cherenkov effect

    International Nuclear Information System (INIS)

    Afanas'ev, G.N.; Stepanovskij, Yu.P.

    2003-01-01

    We study the kinematics of the two-photon Cherenkov effect. In the general case, the emission angles of two photons satisfy certain inequalities and the corresponding radiation intensities are rather diffused. In special cases, when the above inequalities reduce to equalities, the emission angles of two photons are fixed and the corresponding radiation intensities should have sharp maxima at these angles. This makes easier the experimental study of the two-photon Cherenkov effect

  5. A vacuum-UV laser-induced fluorescence experiment for measurement of rotationally and vibrationally excited H2

    International Nuclear Information System (INIS)

    Vankan, P.; Heil, S.B.S.; Mazouffre, S.; Engeln, R.; Schram, D.C.; Doebele, H.F.

    2004-01-01

    An experimental setup is built to detect spatially resolved rovibrationally excited hydrogen molecules via laser-induced fluorescence. To excite the hydrogen molecules, laser radiation is produced in the vacuum UV part of the spectrum. The laser radiation is tunable between 120 nm and 230 nm and has a bandwith of 0.15 cm -1 . The wavelength of the laser radiation is calibrated by simultaneous recording of the two-photon laser induced fluorescence spectrum of nitric oxide. The excited hydrogen populations are calibrated on the basis of coherent anti-Stokes Raman scattering measurements. A population distribution is measured in the shock region of a pure hydrogen plasma expansion. The higher rotational levels (J>5) show overpopulation compared to a Boltzmann distribution determined from the lower rotational levels (J≤5)

  6. Novel triphenylamine-cored two-photon absorbing dyes for labeling of biomolecules

    International Nuclear Information System (INIS)

    Xiao Haibo; Mei Chong; Wang Yaochuan; Li, Hui; Qian Shixiong; Yin Hongyao; Xu Zhisong

    2011-01-01

    Highlights: → Two novel triphenylamine-cored chromophores were synthesized. → These two dyes have sizable two-photon absorption cross-section at 800 nm. → They possess reasonable water solubility and are suitable as labels in aqueous biological environments. → These dyes have strong chelating ability. → They display a large set of reactivity for coupling to biomolecules. - Abstract: Two novel, V-shaped and Y-shaped dipicolinate derivatives branched from triphenylamine, {4-[(E)-2-(2,6-dimethoxycarbonylpyridin-4-yl)vinyl]}-N-phenyl-N-{4- [(E)-2-(2,6-dimethoxycarbonylpyridin-4-yl)vinylphenyl]}aniline (1) and {4-[(E)-2-(2,6-dimethoxycarbonylpyridin-4-yl) vinyl]}-N,N-bis {4-[(E)-2-(2,6-dimethoxycarbonyl pyridin-4-yl)vinylphenyl]}aniline (2) were synthesized. These compounds were designed for large two-photon absorption and in particular for labeling of biomolecules. Their linear absorption, fluorescence properties and their two-photon absorption properties as well as two-photon fluorescence cell imaging were examined. When excited at 800 nm, the two-photon absorption cross-section values of chromophores 1 and 2 in THF were 208 GM, 376 GM, respectively. These two-photon absorbing dyes possess reasonable water solubility, strong chelating ability and display a large set of reactivity for coupling to biomolecules, which are apparently due to the two methoxycarbonyl groups in pyridine ring. This work suggests that chromophores 1 and 2 are promising labels potentially applicable for the tracking of biomolecules using two-photon scanning microscopy.

  7. Two-photon interference : spatial aspects of two-photon entanglement, diffraction, and scattering

    NARCIS (Netherlands)

    Peeters, Wouter Herman

    2010-01-01

    This dissertation contains scientific research within the realm of quantum optics, which is a branch of physics. An experimental and theoretical study is made of two-photon interference phenomena in various optical systems. Spatially entangled photon pairs are produced via the nonlinear optical

  8. Two-photon NADH imaging exposes boundaries of oxygen diffusion in cortical vascular supply regions

    OpenAIRE

    Kasischke, Karl A; Lambert, Elton M; Panepento, Ben; Sun, Anita; Gelbard, Harris A; Burgess, Robert W; Foster, Thomas H; Nedergaard, Maiken

    2010-01-01

    Oxygen transport imposes a possible constraint on the brain's ability to sustain variable metabolic demands, but oxygen diffusion in the cerebral cortex has not yet been observed directly. We show that concurrent two-photon fluorescence imaging of endogenous nicotinamide adenine dinucleotide (NADH) and the cortical microcirculation exposes well-defined boundaries of tissue oxygen diffusion in the mouse cortex. The NADH fluorescence increases rapidly over a narrow, very low pO2 range with a p ...

  9. Synthesis of novel fluorene-based two-photon absorbing molecules and their applications in optical data storage, microfabrication, and stimulated emission depletion

    Science.gov (United States)

    Yanez, Ciceron

    2009-12-01

    Two-photon absorption (2PA) has been used for a number of scientific and technological applications, exploiting the fact that the 2PA probability is directly proportional to the square of the incident light intensity (while one-photon absorption bears a linear relation to the incident light intensity). This intrinsic property of 2PA leads to 3D spatial localization, important in fields such as optical data storage, fluorescence microscopy, and 3D microfabrication. The spatial confinement that 2PA enables has been used to induce photochemical and photophysical events in increasingly smaller volumes and allowed nonlinear, 2PA-based, technologies to reach sub-diffraction limit resolutions. The primary focus of this dissertation is the development of novel, efficient 2PA, fluorene-based molecules to be used either as photoacid generators (PAGs) or fluorophores. A second aim is to develop more effective methods of synthesizing these compounds. As a third and final objective, the new molecules were used to develop a write-once-read many (WORM) optical data storage system, and stimulated emission depletion probes for bioimaging. In Chapter I, the microwave-assisted synthesis of triarylsulfonium salt photoacid generators (PAGs) from their diphenyliodonium counterparts is reported. The microwave-assisted synthesis of these novel sulfonium salts afforded reaction times 90 to 420 times faster than conventional thermal conditions, with photoacid quantum yields of new sulfonium PAGs ranging from 0.01 to 0.4. These PAGs were used to develop a fluorescence readout-based, nonlinear three-dimensional (3D) optical data storage system (Chapter II). In this system, writing was achieved by acid generation upon two-photon absorption (2PA) of a PAG (at 710 or 730 nm). Readout was then performed by interrogating two-photon absorbing dyes, after protonation, at 860 nm. Two-photon recording and readout of voxels was demonstrated in five and eight consecutive, crosstalk-free layers within a

  10. Arduino Due based tool to facilitate in vivo two-photon excitation microscopy.

    Science.gov (United States)

    Artoni, Pietro; Landi, Silvia; Sato, Sebastian Sulis; Luin, Stefano; Ratto, Gian Michele

    2016-04-01

    Two-photon excitation spectroscopy is a powerful technique for the characterization of the optical properties of genetically encoded and synthetic fluorescent molecules. Excitation spectroscopy requires tuning the wavelength of the Ti:sapphire laser while carefully monitoring the delivered power. To assist laser tuning and the control of delivered power, we developed an Arduino Due based tool for the automatic acquisition of high quality spectra. This tool is portable, fast, affordable and precise. It allowed studying the impact of scattering and of blood absorption on two-photon excitation light. In this way, we determined the wavelength-dependent deformation of excitation spectra occurring in deep tissues in vivo.

  11. Insights into esophagus tissue architecture using two-photon confocal microscopy

    Science.gov (United States)

    Liu, Nenrong; Wang, Yue; Feng, Shangyuan; Chen, Rong

    2013-08-01

    In this paper, microstructures of human esophageal mucosa were evaluated using the two-photon laser scanning confocal microscopy (TPLSCM), based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). The distribution of epithelial cells, muscle fibers of muscularis mucosae has been distinctly obtained. Furthermore, esophageal submucosa characteristics with cancer cells invading into were detected. The variation of collagen, elastin and cancer cells is very relevant to the pathology in esophagus, especially early esophageal cancer. Our experimental results indicate that the MPM technique has the much more advantages for label-free imaging, and has the potential application in vivo in the clinical diagnosis and monitoring of early esophageal cancer.

  12. Two-photon-based photoactivation in live zebrafish embryos.

    Science.gov (United States)

    Russek-Blum, Niva; Nabel-Rosen, Helit; Levkowitz, Gil

    2010-12-24

    Photoactivation of target compounds in a living organism has proven a valuable approach to investigate various biological processes such as embryonic development, cellular signaling and adult physiology. In this respect, the use of multi-photon microscopy enables quantitative photoactivation of a given light responsive agent in deep tissues at a single cell resolution. As zebrafish embryos are optically transparent, their development can be monitored in vivo. These traits make the zebrafish a perfect model organism for controlling the activity of a variety of chemical agents and proteins by focused light. Here we describe the use of two-photon microscopy to induce the activation of chemically caged fluorescein, which in turn allows us to follow cell's destiny in live zebrafish embryos. We use embryos expressing a live genetic landmark (GFP) to locate and precisely target any cells of interest. This procedure can be similarly used for precise light induced activation of proteins, hormones, small molecules and other caged compounds.

  13. Laser induced fluorescence of trapped molecular ions

    International Nuclear Information System (INIS)

    Winn, J.S.

    1980-10-01

    Laser induced fluoresence (LIF) spectra (laser excitation spectra) are conceptually among the most simple spectra to obtain. One need only confine a gaseous sample in a suitable container, direct a laser along one axis of the container, and monitor the sample's fluorescence at a right angle to the laser beam. As the laser wavelength is changed, the changes in fluorescence intensity map the absorption spectrum of the sample. (More precisely, only absorption to states which have a significant radiative decay component are monitored.) For ion spectroscopy, one could benefit in many ways by such an experiment. Most optical ion spectra have been observed by emission techniques, and, aside from the problems of spectral analysis, discharge emission methods often produce the spectra of many species, some of which may be unknown or uncertain. Implicit in the description of LIF given above is certainty as to the chemical identity of the carrier of the spectrum. This article describes a method by which the simplifying aspects of LIF can be extended to molecular ions

  14. Two photon processes in surface photovoltage spectroscopy

    International Nuclear Information System (INIS)

    Craig, R.P.; Thurgate, S.M.

    1996-01-01

    Full text: A significant mid-gap effect has been found in Surface Photovoltage Spectroscopy measurements of cleaved GaAs, InP and Si wafer which is normally interpreted as arising from transitions between surface states and band edges. This large mid-gap effect common to various materials is puzzling as such a high proportion of mid-gap states seems unlikely. Most theories of surface states predict states that tail from the band edges into the gap or states that have a well defined energy in the gap. None propose a large state exactly at E G /2. We recently investigated the variation in SPS spectra with flux density. We find a non-linear correlation in the magnitude of Band-Bending arising from mid-gap and band-gap photon energies. We suggest that the mid-gap feature is due to a two photon absorption process leading to carrier pair generation mediated by mid-gap states in the continuum of band-gap surface states

  15. Enhanced two-photon emission from a dressed biexciton

    International Nuclear Information System (INIS)

    Sánchez Muñoz, Carlos; Laussy, Fabrice P; Tejedor, Carlos; Valle, Elena del

    2015-01-01

    Radiative two-photon cascades from biexcitons in semiconductor quantum dots under resonant two-photon excitation are promising candidates for the generation of photon pairs. In this work, we propose a scheme to obtain two-photon emission that allows us to operate under very intense driving fields. This approach relies on the Purcell enhancement of two-photon virtual transitions between states of the biexciton dressed by the laser. The richness provided by the biexcitonic level structure allows to reach a variety of regimes, from antibunched and bunched photon pairs with polarization orthogonal to the driving field, to polarization entangled two-photon emission. This provides evidence that the general paradigm of two-photon emission from a ladder of dressed states can find interesting, particular implementations in a variety of systems. (paper)

  16. Development of laser-induced fluorescence for precombustion diagnostics in spark-ignition engines

    Energy Technology Data Exchange (ETDEWEB)

    Neij, H.

    1998-11-01

    ) - was investigated with respect to optical properties relevant to laser-based combustion diagnostics, by flame emission, optical absorption, laser-induced fluorescence, spontaneous Raman scattering, and rotational CARS. The potential for LIF detection of water vapor in combustion processes was evaluated. Water molecules were excited in a two-photon process at 248 nm yielding fluorescence around 400-500 nm. Spectrally interfering species at flame conditions were identified as hot O{sub 2}, and laser-generated C{sub 2} and N{sub 2}{sup +}. The detection limit for two-dimensional single-shot detection of water vapor at atmospheric conditions was estimated to 0.2%. Extrapolations to flame conditions were presented. A pressure-dependent process was identified, which decreased the signal intensity, broadened the linewidths, and degraded the spectral-excitation feature as the ambient pressure was increased. Two-photon water vapor LIF was applied to a research engine for residual gas visualization. The accuracy and precision of both two-dimensional and spatially averaged data were discussed. The LIF data was used to explain the engine behavior on a cycle-by-cycle basis. A significant correlation was identified between the combustion event and the spatially averaged water signal around the spark gap at the time of ignition 181 refs, 26 figs, 7 tabs

  17. Two-photon decay in heavy atoms and ions

    International Nuclear Information System (INIS)

    Mokler, P.H.; Dunford, R.W

    2003-08-01

    We review the status of and comment on current developments in the field of two-photon decay in atomic physics research. Recent work has focused on two-photon decays in highly-charged ions and two-photon decay of inner-shell vacancies in heavy neutral atoms. We emphasize the importance of measuring the shape of the continuum emission in two-photon decay as a probe of relativistic effects in the strong central fields found in heavy atomic systems. New experimental approaches and their consequences will be discussed. (orig.)

  18. Simulating fluorescence light-canopy interaction in support of laser-induced fluorescence measurements

    International Nuclear Information System (INIS)

    Rosema, A.; Verhoef, W.; Schroote, J.; Snel, J.F.H.

    1991-01-01

    In the Netherlands an operational field instrument for the measurement of laser induced fluorescence of vegetation (LEAF) is developed. In addition, plant physiological and remote sensing research is done to support this new remote sensing instrument. This paper presents a general introduction on the subject of laser-induced fluorescence, including the relation between chlorophyll fluorescence and photosynthesis, spectral characteristics, and previous research. Also the LEAF system is briefly described. Subsequently, the development of a leaf fluorescence model (KMF) and a canopy fluorescence model (FLSAIL) are reported. With these simulation models a sensitivity study is carried out. Fluorescence of 685 nm appears to be most suitable to obtain information on photosynthesis and stress, but is also influenced by canopy structure. Separation of these two effects is studied

  19. Resonance production in two-photon interactions

    International Nuclear Information System (INIS)

    Roe, N.A.

    1989-02-01

    Resonance production in two-photon interactions is studied using data collected with the ASP detector at the PEP e + e/sup /minus// storage ring located at the Stanford Linear Accelerator Center. The ASP detector is a non-magnetic lead-glass calorimeter constructed from 632 lead-glass bars. It covers 94% of 4π in solid angle, extending to within 20/degree/ of the beamline. Lead-scintillator calorimeters extend the coverage to within 21 mr of the beamline on both sides. Energy resolution of √E/10%, where E is the energy is GeV, is achieved for electrons and photons in the lead-glass calorimeter, and particle trajectories are reconstructed with high efficiency. A total luminosity of 108 pb/sup /minus/1/ was collected with the ASP detector at a center-of-mass energy of 29 GeV. The observed process is e + e/sup /minus// → e + e/sup /minus//γ*γ* → e + e/sup /minus//X, is a pseudoscalar resonance (J/sup PC/ = 0/sup /minus/+/) and γ* is a virtual (mass /ne/ 0) photon. The outgoing electrons scatter down the beampipe and are not detected. The observed resonances are the /eta/ and /eta/' mesons, with masses of 549 and 958 MeV, respectively. They are detected in the γγ decay mode; a total of 2380 +- 49 /eta/ → γγ and 568 +- 26 /eta/' → γγ events are observed. From the number of events, the detection efficiency, and the calculated production cross sections the radiative widths, Γ/sub γγ/, of the /eta/ and /eta/' were measured and found to be: Γ/sub γγ/(/eta/) = .481 +- .010 +- .047keV and Γ/sub γγ/(/eta/') = 4.71 +- .22 +- .70keV. These results are in good agreement with the world average values. 67 refs., 42 figs., 20 tabs

  20. Resonance production in two-photon interactions

    Energy Technology Data Exchange (ETDEWEB)

    Roe, N.A.

    1989-02-01

    Resonance production in two-photon interactions is studied using data collected with the ASP detector at the PEP e/sup +/e/sup /minus// storage ring located at the Stanford Linear Accelerator Center. The ASP detector is a non-magnetic lead-glass calorimeter constructed from 632 lead-glass bars. It covers 94% of 4..pi.. in solid angle, extending to within 20/degree/ of the beamline. Lead-scintillator calorimeters extend the coverage to within 21 mr of the beamline on both sides. Energy resolution of ..sqrt..E/10%, where E is the energy is GeV, is achieved for electrons and photons in the lead-glass calorimeter, and particle trajectories are reconstructed with high efficiency. A total luminosity of 108 pb/sup /minus/1/ was collected with the ASP detector at a center-of-mass energy of 29 GeV. The observed process is e/sup +/e/sup /minus// ..-->.. e/sup +/e/sup /minus//..gamma..*..gamma..* ..-->.. e/sup +/e/sup /minus//X, is a pseudoscalar resonance (J/sup PC/ = 0/sup /minus/+/) and ..gamma..* is a virtual (mass /ne/ 0) photon. The outgoing electrons scatter down the beampipe and are not detected. The observed resonances are the /eta/ and /eta/' mesons, with masses of 549 and 958 MeV, respectively. They are detected in the ..gamma gamma.. decay mode; a total of 2380 +- 49 /eta/ ..-->.. ..gamma gamma.. and 568 +- 26 /eta/' ..-->.. ..gamma gamma.. events are observed. From the number of events, the detection efficiency, and the calculated production cross sections the radiative widths, GAMMA/sub ..gamma gamma../, of the /eta/ and /eta/' were measured and found to be: GAMMA/sub ..gamma gamma../(/eta/) = .481 +- .010 +- .047keV and GAMMA/sub ..gamma gamma../(/eta/') = 4.71 +- .22 +- .70keV. These results are in good agreement with the world average values. 67 refs., 42 figs., 20 tabs.

  1. Peroxy Radical Measurements via Laser Induced Fluorescence

    Science.gov (United States)

    Trawny, Katrin; Tatum Ernest, Cheryl; Novelli, Anna; Elste, Thomas; Plaß-Dülmer, Christian; Rudolf, Markus; Martinez, Monica; Harder, Hartwig; Lelieveld, Jos

    2013-04-01

    We present a newly built Laser Induced Fluorescence (LIF) system to measure the sum of all peroxy radicals (RO2) utilizing chemical conversion to OH. This instrument operates in two different modes: the ROx mode (sum of OH, HO2, and RO2) and the HOx mode (sum of OH and HO2). The HOx mode is used to derive the RO2 data from the ROx measurements. A model approach was used during instrumental development to identify the key parameters needed for the conversion process in front of the detection area and to optimize sensitivity. The instrument was then carefully characterized in various lab experiments, where it could be shown that the wall losses for HO2 are negligible and that nearly all HO2 is converted to OH in front of the detection zone. The pressure and temperature dependencies were also analyzed and assured that the instrument does not show any photolytical interference. As the instrument is calibrated with only one kind of peroxy radicals it was very important that the differences in sensitivity for different peroxy radicals are acceptable. Lab experiments as well as first results from the HOPE 2012 intensive field campaign, which took place in summer 2012 at the Global Atmosphere Watch (GAW) station of the German Weather Service, will be discussed.

  2. Laser induced fluorescence in atmospheric pressure discharges

    International Nuclear Information System (INIS)

    Dilecce, G; De Benedictis, S; Martini, L M; Tosi, P; Scotoni, M

    2015-01-01

    This paper offers an outline of laser induced fluorescence (LIF) diagnostics and practical recommendations for its use in atmospheric pressure discharges. LIF principles, technical requirements and rationalization of experimental outcomes by modelling are addressed. Important issues that are particularly relevant to small scale, spatially inhomogeneous discharges, like plasma-jets, are emphasized. For the first time, all collision processes and the spatial non-homogeneity of the laser beam are together accounted for in the LIF model. Saturation characteristics are discussed and used for the assessment of model parameters. A calibration procedure is discussed and implemented. Gas temperature measurements by LIF are also addressed. The whole description of the technique is given, without loss of generality, through the example of its application to the OH radical. Notes on other diatomic radicals, CH, NO and CN, are given along the paper. Some results in a RF plasma-jet are presented as an example of application in a discharge system where all the concepts developed in the paper are applied. (paper)

  3. Studies of photoionization in liquids using a laser two-photon ionization conductivity technique

    International Nuclear Information System (INIS)

    Siomos, K.; Christophorou, L.G.

    1981-01-01

    One-photon ionization studies of solute molecules in a liquid medium are limited by the absorption of the host medium. A laser two-photon ionization (TPI) technique using a frequency tunable dye laser has been developed, whereby the photoionization threshold of a solute molecule was determined from the induced conductivity in the liquid medium under study due to electron-ion pair formation via two-photon ionization of the solute. The two-photon induced electron-ion current is measured as a function of the laser wavelength, lambda/sub laser/. In this paper, results are reported and discussed on the photoionization of N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD), pyrene and fluoranthene in liquid n-pentane

  4. Robotic Automation of In Vivo Two-Photon Targeted Whole-Cell Patch-Clamp Electrophysiology.

    Science.gov (United States)

    Annecchino, Luca A; Morris, Alexander R; Copeland, Caroline S; Agabi, Oshiorenoya E; Chadderton, Paul; Schultz, Simon R

    2017-08-30

    Whole-cell patch-clamp electrophysiological recording is a powerful technique for studying cellular function. While in vivo patch-clamp recording has recently benefited from automation, it is normally performed "blind," meaning that throughput for sampling some genetically or morphologically defined cell types is unacceptably low. One solution to this problem is to use two-photon microscopy to target fluorescently labeled neurons. Combining this with robotic automation is difficult, however, as micropipette penetration induces tissue deformation, moving target cells from their initial location. Here we describe a platform for automated two-photon targeted patch-clamp recording, which solves this problem by making use of a closed loop visual servo algorithm. Our system keeps the target cell in focus while iteratively adjusting the pipette approach trajectory to compensate for tissue motion. We demonstrate platform validation with patch-clamp recordings from a variety of cells in the mouse neocortex and cerebellum. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  5. Two-photon stimulated emission and pulse amplification

    International Nuclear Information System (INIS)

    Yuen, H.P.

    1975-01-01

    Threshold conditions are given for the sustained operation of standing-wave and long-pulse traveling-wave two-photon lasers. Pulse shortening in long-pulse two-photon amplification, a behavior absent in the one-photon case, is also demonstrated analytically. (U.S.)

  6. Two-photon LIF on the HIT-SI3 experiment: Absolute density and temperature measurements of deuterium neutrals

    Energy Technology Data Exchange (ETDEWEB)

    Elliott, Drew, E-mail: dbelliott@mix.wvu.edu; Siddiqui, Umair; Scime, Earl [Department of Physics and Astronomy, West Virginia University, Morgantown, West Virginia 26056 (United States); Sutherland, Derek; Everson, Chris; Morgan, Kyle; Hossack, Aaron; Nelson, Brian; Jarboe, Tom [Department of Aerospace Engineering, University of Washington, Seattle, Washington 98105 (United States)

    2016-11-15

    Two-photon laser-induced fluorescence measurements were performed on the helicity injected torus (HIT-SI3) device to determine the density and temperature of the background neutral deuterium population. Measurements were taken in 2 ms long pulsed plasmas after the inductive helicity injectors were turned off. Attempts to measure neutrals during the main phase of the plasma were unsuccessful, likely due to the density of neutrals being below the detection threshold of the diagnostic. An unexpectedly low density of atomic deuterium was measured in the afterglow; roughly 100 times lower than the theoretical prediction of 10{sup 17} m{sup −3}. The neutral temperatures measured were on the order of 1 eV. Temporally and spatially resolved neutral density and temperature data are presented.

  7. Two-photon activation of endogenous store-operated calcium channels without optogenetics

    Science.gov (United States)

    Cheng, Pan; Tang, Wanyi; He, Hao

    2018-02-01

    Store-operated calcium (SOC) channels, regulated by intracellular Ca2+ store, are the essential pathway of calcium signaling and participate in a wide variety of cellular activities such as gene expression, secretion and immune response1. However, our understanding and regulation of SOC channels are mainly based on pharmacological methods. Considering the unique advantages of optical control, optogenetic control of SOC channels has been developed2. However, the process of genetic engineering to express exogenous light-sensitive protein is complicated, which arouses concerns about ethic difficulties in some research of animal and applications in human. In this report, we demonstrate rapid, robust and reproducible two-photon activation of endogenous SOC channels by femtosecond laser without optogenetics. We present that the short-duration two-photon scanning on subcellular microregion induces slow Ca2+ influx from extracellular medium, which can be eliminated by removing extracellular Ca2+. Block of SOC channels using various pharmacological inhibitors or knockdown of SOC channels by RNA interference reduce the probability of two-photon activated Ca2+ influx. On the contrary, overexpression of SOC channels can increase the probability of Ca2+ influx by two-photon scanning. These results collectively indicate Ca2+ influx through two-photon activated SOC channels. Different from classical pathway of SOC entry activated by Ca2+ store depletion, STIM1, the sensor protein of Ca2+ level in endoplasmic reticulum, does not show any aggregation or migration in this two-photon activated Ca2+ influx, which rules out the possibility of intracellular Ca2+ store depletion. Thereby, we propose this all-optical method of two-photon activation of SOC channels is of great potential to be widely applied in the research of cell calcium signaling and related biological research.

  8. The development of efficient two-photon singlet oxygen sensitizers

    DEFF Research Database (Denmark)

    Nielsen, Christian Benedikt

    The development of efficient two-photon singlet oxygen sensitizers is addressed focusing on organic synthesis. Photophysical measurements were carried out on new lipophilic molecules, where two-photon absorption cross sections and singlet oxygen quantumyields were measured. Design principles...... for making efficient two-photon singlet oxygen sensitizers were then constructed from these results. Charge-transfer in the excited state of the prepared molecules was shown to play a pivotal role in the generationof singlet oxygen. This was established through studies of substituent effects on both...... the singlet oxygen yield and the two-photon absorption cross section, where it was revealed that a careful balancing of the amount of charge transfer present in theexcited state of the sensitizer is necessary to obtain both a high singlet oxygen quantum yield and a high two-photon cross section. An increasing...

  9. Remote sensing vegetation status by laser-induced fluorescence

    International Nuclear Information System (INIS)

    Günther, K.P.; Dahn, H.G.; Lüdeker, W.

    1994-01-01

    In November 1989 the EUREKA project LASFLEUR (EU 380) started as an European research effort to investigate the future application of far-field laser-induced plant fluorescence for synoptic, airborne environmental monitoring of vegetation. This report includes a brief introduction in a theoretically approach for the laser-induced fluorescence signals of leaves and their spectral and radiometric behaviour. In addition, a detailed description of the design and realization of the second generation of the far-field fluorescence lidar (DLidaR-2) is given with special regard to the optical and electronical setup, followed by a short explanation of the data processing. The main objectives of the far field measurements are to demonstrate the link between laser-induced fluorescence data and plant physiology and to show the reliability of remote single shot lidar measurements. The data sets include the typical daily cycles of the fluorescence for different global irradiation. As expected from biophysical models, the remotely sensed chlorophyll fluorescence is highly correlated with the carbon fixation rate, while the fluorescence ratio F685 / F730 is only dependent on the chlorophyll concentration. Drought stress measurement of evergreen oaks Quercus pubescens confirm the findings of healthy plants with regard to the fluorescence ratio F685 / F730 while the fluorescence signals of stressed plants show a different behavior than nonstressed plants. Additionally, the corresponding physiological data (porometer and PAM data) are presented. (author)

  10. Binding-Induced Fluorescence of Serotonin Transporter Ligands

    DEFF Research Database (Denmark)

    Wilson, James; Ladefoged, Lucy Kate; Babinchak, Michael

    2014-01-01

    The binding-induced fluorescence of 4-(4-(dimethylamino)-phenyl)-1-methylpyridinium (APP(+)) and two new serotonin transporter (SERT)-binding fluorescent analogues, 1-butyl-4-[4-(1-dimethylamino)phenyl]-pyridinium bromide (BPP(+)) and 1-methyl-4-[4-(1-piperidinyl)phenyl]-pyridinium (PPP(+)), has...

  11. Two-photon collisions and short-distance tests of quantum chromodynamics

    International Nuclear Information System (INIS)

    Brodsky, S.J.

    1978-12-01

    The physics of two-photon collisions in e +- storage rings is reviewed with emphasis on the predictions of perturbative quantum chromodynamics for high transverse momentum reactions. It is noted that because of the remarkable scaling properties predicted by the theory, two-photon collisions may be proved one of the cleanest tests of the quantum chromodynamics picture of short distance hadron dynamics. In order to contrast these predictions for photon-induced reactions with those for incident hadrons, predictions from quantum chromodynamics for hadron structure functions and form factors at large momentum transfer are also discussed. 55 references

  12. Two-photon calcium imaging in mice navigating a virtual reality environment.

    Science.gov (United States)

    Leinweber, Marcus; Zmarz, Pawel; Buchmann, Peter; Argast, Paul; Hübener, Mark; Bonhoeffer, Tobias; Keller, Georg B

    2014-02-20

    In recent years, two-photon imaging has become an invaluable tool in neuroscience, as it allows for chronic measurement of the activity of genetically identified cells during behavior(1-6). Here we describe methods to perform two-photon imaging in mouse cortex while the animal navigates a virtual reality environment. We focus on the aspects of the experimental procedures that are key to imaging in a behaving animal in a brightly lit virtual environment. The key problems that arise in this experimental setup that we here address are: minimizing brain motion related artifacts, minimizing light leak from the virtual reality projection system, and minimizing laser induced tissue damage. We also provide sample software to control the virtual reality environment and to do pupil tracking. With these procedures and resources it should be possible to convert a conventional two-photon microscope for use in behaving mice.

  13. Two-photon processes in highly charged ions

    International Nuclear Information System (INIS)

    Jahrsetz, Thorsten

    2015-01-01

    Two-photon processes are atomic processes in which an atom interacts simultaneously with two photons. Such processes describe a wide range of phenomena, such as two-photon decay and elastic or inelastic scattering of photons. In recent years two-photon processes involving highly charged heavy ions have become an active area of research. Such studies do not only consider the total transition or scattering rates but also their angular and polarization dependence. To support such examinations in this thesis I present a theoretical framework to describe these properties in all two-photon processes with bound initial and final states and involving heavy H-like or He-like ions. I demonstrate how this framework can be used in some detailed studies of different two-photon processes. Specifically a detailed analysis of two-photon decay of H-like and He-like ions in strong external electromagnetic fields shows the importance of considering the effect of such fields for the physics of such systems. Furthermore I studied the elastic Rayleigh as well as inelastic Raman scattering by heavy H-like ions. I found a number of previously unobserved phenomena in the angular and polarization dependence of the scattering cross-sections that do not only allow to study interesting details of the electronic structure of the ion but might also be useful for the measurement of weak physical effects in such systems.

  14. Two-photon processes in highly charged ions

    Energy Technology Data Exchange (ETDEWEB)

    Jahrsetz, Thorsten

    2015-03-05

    Two-photon processes are atomic processes in which an atom interacts simultaneously with two photons. Such processes describe a wide range of phenomena, such as two-photon decay and elastic or inelastic scattering of photons. In recent years two-photon processes involving highly charged heavy ions have become an active area of research. Such studies do not only consider the total transition or scattering rates but also their angular and polarization dependence. To support such examinations in this thesis I present a theoretical framework to describe these properties in all two-photon processes with bound initial and final states and involving heavy H-like or He-like ions. I demonstrate how this framework can be used in some detailed studies of different two-photon processes. Specifically a detailed analysis of two-photon decay of H-like and He-like ions in strong external electromagnetic fields shows the importance of considering the effect of such fields for the physics of such systems. Furthermore I studied the elastic Rayleigh as well as inelastic Raman scattering by heavy H-like ions. I found a number of previously unobserved phenomena in the angular and polarization dependence of the scattering cross-sections that do not only allow to study interesting details of the electronic structure of the ion but might also be useful for the measurement of weak physical effects in such systems.

  15. Exciton molecule in semiconductors by two-photon absorption

    International Nuclear Information System (INIS)

    Arya, K.; Hassan, A.R.

    1976-07-01

    Direct creation of bi-exciton states by two-photon absorption in direct gap semiconductors is investigated theoretically. A numerical application to the case of CuCl shows that the two-photon absorption coefficient for bi-excitonic transitions is larger than that for two-photon interband transitions by three orders of magnitude. It becomes comparable to that for one-photon excitonic transitions for available laser intensities. The main contribution to this enhancement of the absorption coefficient for the transitions to the bi-exciton states is found to be from the resonance effect

  16. Two-photon microscopy for non-invasive, quantitative monitoring of stem cell differentiation.

    Directory of Open Access Journals (Sweden)

    William L Rice

    Full Text Available BACKGROUND: The engineering of functional tissues is a complex multi-stage process, the success of which depends on the careful control of culture conditions and ultimately tissue maturation. To enable the efficient optimization of tissue development protocols, techniques suitable for monitoring the effects of added stimuli and induced tissue changes are needed. METHODOLOGY/PRINCIPAL FINDINGS: Here, we present the quantitative use of two-photon excited fluorescence (TPEF and second harmonic generation (SHG as a noninvasive means to monitor the differentiation of human mesenchymal stem cells (hMSCs using entirely endogenous sources of contrast. We demonstrate that the individual fluorescence contribution from the intrinsic cellular fluorophores NAD(PH, flavoproteins and lipofuscin can be extracted from TPEF images and monitored dynamically from the same cell population over time. Using the redox ratio, calculated from the contributions of NAD(PH and flavoproteins, we identify distinct patterns in the evolution of the metabolic activity of hMSCs maintained in either propagation, osteogenic or adipogenic differentiation media. The differentiation of these cells is mirrored by changes in cell morphology apparent in high resolution TPEF images and by the detection of collagen production via SHG imaging. Finally, we find dramatic increases in lipofuscin levels in hMSCs maintained at 20% oxygen vs. those in 5% oxygen, establishing the use of this chromophore as a potential biomarker for oxidative stress. CONCLUSIONS/SIGNIFICANCE: In this study we demonstrate that it is possible to monitor the metabolic activity, morphology, ECM production and oxidative stress of hMSCs in a non-invasive manner. This is accomplished using generally available multiphoton microscopy equipment and simple data analysis techniques, such that the method can widely adopted by laboratories with a diversity of comparable equipment. This method therefore represents a powerful tool

  17. Standard Model Higgs decay for two Photons in CMS

    CERN Multimedia

    Daniel Denegri

    2000-01-01

    Simulated two-photon mass distribution for SM Higgs and expected background in the CMS PbW04 crystal calorimeter for an integrated luminosity of 10 . 5 pb-1, with detailed simulation of calorimeter response.

  18. All-optical femtosecond switch using two-photon absorption

    International Nuclear Information System (INIS)

    Yavuz, D. D.

    2006-01-01

    Utilizing a two-photon absorption scheme in an alkali-metal vapor cell, we suggest a technique where a strong laser beam switches off another laser beam of different wavelength in femtosecond time scales

  19. Mass distribution for the two-photon channel

    CERN Multimedia

    ATLAS, collaboration

    2012-01-01

    Mass distribution for the two-photon channel. The strongest evidence for this new particle comes from analysis of events containing two photons. The smooth dotted line traces the measured background from known processes. The solid line traces a statistical fit to the signal plus background. The new particle appears as the excess around 126.5 GeV. The full analysis concludes that the probability of such a peak is three chances in a million.

  20. Two-photon quantum interference in a Michelson interferometer

    International Nuclear Information System (INIS)

    Odate, Satoru; Wang Haibo; Kobayashi, Takayoshi

    2005-01-01

    We have observed two-photon quantum interference in a Michelson interferometer. For the first time, we experimentally demonstrated two-photon quantum interference patterns, which show the transition from nonsubwavelength interference fringes to the general subwavelength interference. At the same time, a photon bunching effect was also shown by a postselection. The |1, 1> state with a single photon in a mode corresponding to each arm of the interferometer was exclusively postselected by using path difference between two arms

  1. Coherence revivals in two-photon frequency combs

    International Nuclear Information System (INIS)

    Torres-Company, Victor; Lancis, Jesus; Lajunen, Hanna; Friberg, Ari T.

    2011-01-01

    We describe and theoretically analyze the self-imaging Talbot effect of entangled photon pairs in the time domain. Rich phenomena are observed in coherence propagation along dispersive media of mode-locked two-photon states with frequency entanglement exhibiting a comblike correlation function. Our results can be used to remotely transfer frequency standards through optical fiber networks with two-photon light, avoiding the requirement of dispersion compensation.

  2. Volumetric Two-photon Imaging of Neurons Using Stereoscopy (vTwINS)

    Science.gov (United States)

    Song, Alexander; Charles, Adam S.; Koay, Sue Ann; Gauthier, Jeff L.; Thiberge, Stephan Y.; Pillow, Jonathan W.; Tank, David W.

    2017-01-01

    Two-photon laser scanning microscopy of calcium dynamics using fluorescent indicators is a widely used imaging method for large scale recording of neural activity in vivo. Here we introduce volumetric Two-photon Imaging of Neurons using Stereoscopy (vTwINS), a volumetric calcium imaging method that employs an elongated, V-shaped point spread function to image a 3D brain volume. Single neurons project to spatially displaced “image pairs” in the resulting 2D image, and the separation distance between images is proportional to depth in the volume. To demix the fluorescence time series of individual neurons, we introduce a novel orthogonal matching pursuit algorithm that also infers source locations within the 3D volume. We illustrate vTwINS by imaging neural population activity in mouse primary visual cortex and hippocampus. Our results demonstrate that vTwINS provides an effective method for volumetric two-photon calcium imaging that increases the number of neurons recorded while maintaining a high frame-rate. PMID:28319111

  3. Photo-induced processes in collagen-hypericin system revealed by fluorescence spectroscopy and multiphoton microscopy

    OpenAIRE

    Hovhannisyan, V.; Guo, H. W.; Hovhannisyan, A.; Ghukasyan, V.; Buryakina, T.; Chen, Y. F.; Dong, C. Y.

    2014-01-01

    Collagen is the main structural protein and the key determinant of mechanical and functional properties of tissues and organs. Proper balance between synthesis and degradation of collagen molecules is critical for maintaining normal physiological functions. In addition, collagen influences tumor development and drug delivery, which makes it a potential cancer therapy target. Using second harmonic generation, two-photon excited fluorescence microscopy, and spectrofluorimetry, we show that the ...

  4. Theory of two-photon interactions with broadband down-converted light and entangled photons

    International Nuclear Information System (INIS)

    Dayan, Barak

    2007-01-01

    When two-photon interactions are induced by down-converted light with a bandwidth that exceeds the pump bandwidth, they can obtain a behavior that is pulselike temporally, yet spectrally narrow. At low photon fluxes this behavior reflects the time and energy entanglement between the down-converted photons. However, two-photon interactions such as two-photon absorption (TPA) and sum-frequency generation (SFG) can exhibit such a behavior even at high power levels, as long as the final state (i.e., the atomic level in TPA, or the generated light in SFG) is narrow-band enough. This behavior does not depend on the squeezing properties of the light, is insensitive to linear losses, and has potential applications. In this paper we describe analytically this behavior for traveling-wave down conversion with continuous or pulsed pumping, both for high- and low-power regimes. For this we derive a quantum-mechanical expression for the down-converted amplitude generated by an arbitrary pump, and formulate operators that represent various two-photon interactions induced by broadband light. This model is in excellent agreement with experimental results of TPA and SFG with high-power down-converted light and with entangled photons [Dayan et al., Phys. Rev. Lett. 93, 023005 (2004); 94, 043602 (2005); Pe'er et al., ibid. 94, 073601 (2005)

  5. Efficient multi-site two-photon functional imaging of neuronal circuits.

    Science.gov (United States)

    Castanares, Michael Lawrence; Gautam, Vini; Drury, Jack; Bachor, Hans; Daria, Vincent R

    2016-12-01

    Two-photon imaging using high-speed multi-channel detectors is a promising approach for optical recording of cellular membrane dynamics at multiple sites. A main bottleneck of this technique is the limited number of photons captured within a short exposure time (~1ms). Here, we implement temporal gating to improve the two-photon fluorescence yield from holographically projected multiple foci whilst maintaining a biologically safe incident average power. We observed up to 6x improvement in the signal-to-noise ratio (SNR) in Fluorescein and cultured hippocampal neurons showing evoked calcium transients. With improved SNR, we could pave the way to achieving multi-site optical recording of fluorogenic probes with response times in the order of ~1ms.

  6. 5-ALA induced fluorescent image analysis of actinic keratosis

    Science.gov (United States)

    Cho, Yong-Jin; Bae, Youngwoo; Choi, Eung-Ho; Jung, Byungjo

    2010-02-01

    In this study, we quantitatively analyzed 5-ALA induced fluorescent images of actinic keratosis using digital fluorescent color and hyperspectral imaging modalities. UV-A was utilized to induce fluorescent images and actinic keratosis (AK) lesions were demarcated from surrounding the normal region with different methods. Eight subjects with AK lesion were participated in this study. In the hyperspectral imaging modality, spectral analysis method was utilized for hyperspectral cube image and AK lesions were demarcated from the normal region. Before image acquisition, we designated biopsy position for histopathology of AK lesion and surrounding normal region. Erythema index (E.I.) values on both regions were calculated from the spectral cube data. Image analysis of subjects resulted in two different groups: the first group with the higher fluorescence signal and E.I. on AK lesion than the normal region; the second group with lower fluorescence signal and without big difference in E.I. between two regions. In fluorescent color image analysis of facial AK, E.I. images were calculated on both normal and AK lesions and compared with the results of hyperspectral imaging modality. The results might indicate that the different intensity of fluorescence and E.I. among the subjects with AK might be interpreted as different phases of morphological and metabolic changes of AK lesions.

  7. Laser induced fluorescence of biochemical for UV LIDAR application.

    Science.gov (United States)

    Gupta, L; Sharma, R C; Razdan, A K; Maini, A K

    2014-05-01

    Laser induced fluorescence spectroscopy in the ultraviolet regime has been used for the detection of biochemical through a fiber coupled CCD detector from a distance of 2 m. The effect of concentration and laser excitation energy on the fluorescence spectra of nicotinamide adenine dinucleotide (NADH) has been investigated. The signature fluorescence peak of NADH was centred about 460 nm. At lower concentration Raman peak centred at 405 nm was also observed. The origin of this peak has been discussed. Detection limit with the proposed set up is found to be 1 ppm.

  8. Consolidated Laser-Induced Fluorescence Diagnostic Systems for the NASA Ames Arc Jet Facilities

    Science.gov (United States)

    Grinstead, Jay; Wilder, Michael C.; Porter, Barry; Brown, Jeff; Yeung, Dickson; Battazzo, Steve; Brubaker, Tim

    2016-01-01

    The spectroscopic diagnostic technique of two photon absorption laser-induced fluorescence (TALIF) of atomic species for non-intrusive arc jet flow property measurement was first implemented at NASA Ames in the mid-1990s. Use of TALIF expanded at NASA Ames and to NASA Johnsons arc jet facility in the late 2000s. In 2013-2014, NASA combined the agency's large-scale arc jet test capabilities at NASA Ames. Concurrent with that effort, the agency also sponsored a project to establish two comprehensive LIF diagnostic systems for the Aerodynamic Heating Facility (AHF) and Interaction Heating Facility (IHF) arc jets. The scope of the project enabled further engineering development of the existing IHF LIF system as well as the complete reconstruction of the original AHF LIF system. The updated LIF systems are identical in design and capability. They represent the culmination of over 20 years of development experience in transitioning a specialized laboratory research tool into a measurement system for large-scale, high-demand test facilities. This paper documents the overall system design from measurement requirements to implementation. Representative data from the redeveloped AHF and IHF LIF systems are also presented.

  9. Surface plasmon-enhanced molecular fluorescence induced by gold nanostructures

    International Nuclear Information System (INIS)

    Teng, Y.; Ueno, K.; Shi, X.; Aoyo, D.; Misawa, H.; Qiu, J.

    2012-01-01

    The authors report on surface plasmon-enhanced fluorescence of Eosin Y molecules induced by gold nanostructures. Al 2 O 3 films deposited by atomic layer deposition with sub-nanometer resolution were used as the spacer layer to control the distance between molecules and the gold surface. As the thickness of the Al 2 O 3 film increased, the fluorescence intensity first increased and then decreased. The highest enhancement factor is achieved with a 1 nm Al 2 O 3 film. However, the trend for the fluorescence lifetime is the opposite. It first decreased and then increased. The changes in the fluorescence quantum yield were also calculated. The yield shows a similar trend to the fluorescence intensity. The competition between the surface plasmon-induced increase in the radiative decay rate and the gold-induced fluorescence quenching is responsible for the observed phenomenon. In addition, this competition strongly depends on the thickness of the spacer layer between Eosin Y molecules and the gold surface. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  10. Ion beam induced fluorescence imaging in biological systems

    International Nuclear Information System (INIS)

    Bettiol, Andrew A.; Mi, Zhaohong; Vanga, Sudheer Kumar; Chen, Ce-belle; Tao, Ye; Watt, Frank

    2015-01-01

    Imaging fluorescence generated by MeV ions in biological systems such as cells and tissue sections requires a high resolution beam (<100 nm), a sensitive detection system and a fluorescent probe that has a high quantum efficiency and low bleaching rate. For cutting edge applications in bioimaging, the fluorescence imaging technique needs to break the optical diffraction limit allowing for sub-cellular structure to be visualized, leading to a better understanding of cellular function. In a nuclear microprobe this resolution requirement can be readily achieved utilizing low beam current techniques such as Scanning Transmission Ion Microscopy (STIM). In recent times, we have been able to extend this capability to fluorescence imaging through the development of a new high efficiency fluorescence detection system, and through the use of new novel fluorescent probes that are resistant to ion beam damage (bleaching). In this paper we demonstrate ion beam induced fluorescence imaging in several biological samples, highlighting the advantages and challenges associated with using this technique

  11. Interference of two photons in parametric down conversion

    International Nuclear Information System (INIS)

    Ghosh, R.; Hong, C.K.; Ou, Z.Y.; Mandel, L.

    1986-01-01

    A theoretical treatment is given of the process in which the two photons produced simultaneously in the parametric frequency splitting of light are allowed to interfere. It is shown that, while there is no interference in the usual sense involving quantities that are of the second order in the field, fourth-order interference effects are present. These may be revealed by measuring the joint probability of detecting two photons at two points x,x' in the interference plane with photoelectric detectors as a function of the separation x-x'. The probability exhibits a cosine modulation with x-x', with visibility that can approach 100%, even though the integration time in the experiment may greatly exceed the reciprocal bandwidth of the photons. The interference effect has a nonclassical origin and implies a violation of local realism in the highly correlated two-photon state

  12. Characterisation of estuarine intertidal macroalgae by laser-induced fluorescence

    DEFF Research Database (Denmark)

    Gameiro, Carla; Utkin, Andrei B.; Sousa Dias Cartaxana, Paulo Jorge

    2015-01-01

    The article reports the application of laser-induced fluorescence (LIF) for the assessment of macroalgae communities of estuarine intertidal areas. The method was applied for the characterisation of fifteen intertidal macroalgae species of the Tagus estuary, Portugal, and adjacent coastal area...... spectra were determined by differences in the main fluorescing pigments: phycoerythrin, phycocyanin and chlorophyll a (Chl a). In the green and brown macroalgae groups, the relative significance of the two emission maxima seems to be related to the thickness of the photosynthetic layer. In thick...... macroalgae, like Codium tomentosum or Fucus vesiculosus, the contribution of the far-red emission fluorescence peak was more significant, most probably due to re-absorption of the emitted red Chl a fluorescence within the dense photosynthetic layer. Similarly, an increase in the number of layers of the thin...

  13. An integrated single- and two-photon non-diffracting light-sheet microscope

    Science.gov (United States)

    Lau, Sze Cheung; Chiu, Hoi Chun; Zhao, Luwei; Zhao, Teng; Loy, M. M. T.; Du, Shengwang

    2018-04-01

    We describe a fluorescence optical microscope with both single-photon and two-photon non-diffracting light-sheet excitations for large volume imaging. With a special design to accommodate two different wavelength ranges (visible: 400-700 nm and near infrared: 800-1200 nm), we combine the line-Bessel sheet (LBS, for single-photon excitation) and the scanning Bessel beam (SBB, for two-photon excitation) light sheet together in a single microscope setup. For a transparent thin sample where the scattering can be ignored, the LBS single-photon excitation is the optimal imaging solution. When the light scattering becomes significant for a deep-cell or deep-tissue imaging, we use SBB light-sheet two-photon excitation with a longer wavelength. We achieved nearly identical lateral/axial resolution of about 350/270 nm for both imagings. This integrated light-sheet microscope may have a wide application for live-cell and live-tissue three-dimensional high-speed imaging.

  14. Two-photon NADH imaging exposes boundaries of oxygen diffusion in cortical vascular supply regions.

    Science.gov (United States)

    Kasischke, Karl A; Lambert, Elton M; Panepento, Ben; Sun, Anita; Gelbard, Harris A; Burgess, Robert W; Foster, Thomas H; Nedergaard, Maiken

    2011-01-01

    Oxygen transport imposes a possible constraint on the brain's ability to sustain variable metabolic demands, but oxygen diffusion in the cerebral cortex has not yet been observed directly. We show that concurrent two-photon fluorescence imaging of endogenous nicotinamide adenine dinucleotide (NADH) and the cortical microcirculation exposes well-defined boundaries of tissue oxygen diffusion in the mouse cortex. The NADH fluorescence increases rapidly over a narrow, very low pO(2) range with a p(50) of 3.4 ± 0.6 mm Hg, thereby establishing a nearly binary reporter of significant, metabolically limiting hypoxia. The transient cortical tissue boundaries of NADH fluorescence exhibit remarkably delineated geometrical patterns, which define the limits of tissue oxygen diffusion from the cortical microcirculation and bear a striking resemblance to the ideal Krogh tissue cylinder. The visualization of microvessels and their regional contribution to oxygen delivery establishes penetrating arterioles as major oxygen sources in addition to the capillary network and confirms the existence of cortical oxygen fields with steep microregional oxygen gradients. Thus, two-photon NADH imaging can be applied to expose vascular supply regions and to localize functionally relevant microregional cortical hypoxia with micrometer spatial resolution.

  15. Generation of laser-induced periodic surface structures in indium-tin-oxide thin films and two-photon lithography of ma-N photoresist by sub-15 femtosecond laser microscopy for liquid crystal cell application

    Science.gov (United States)

    Klötzer, Madlen; Afshar, Maziar; Feili, Dara; Seidel, Helmut; König, Karsten; Straub, Martin

    2015-03-01

    Indium-tin-oxide (ITO) is a widely used electrode material for liquid crystal cell applications because of its transparency in the visible spectral range and its high electrical conductivity. Important examples of applications are displays and optical phase modulators. We report on subwavelength periodic structuring and precise laser cutting of 150 nm thick indium-tin-oxide films on glass substrates, which were deposited by magnetron reactive DC-sputtering from an indiumtin target in a low-pressure oxygen atmosphere. In order to obtain nanostructured electrodes laser-induced periodic surface structures with a period of approximately 100 nm were generated using tightly focused high-repetition rate sub-15 femtosecond pulsed Ti:sapphire laser light, which was scanned across the sample by galvanometric mirrors. Three-dimensional spacers were produced by multiphoton photopolymerization in ma-N 2410 negative-tone photoresist spin-coated on top of the ITO layers. The nanostructured electrodes were aligned in parallel to set up an electrically switchable nematic liquid crystal cell.

  16. Nonlinear behavior of photoluminescence from silicon particles under two-photon excitation

    International Nuclear Information System (INIS)

    Xu Xingsheng; Yokoyama, Shiyoshi

    2011-01-01

    Two-photon excited fluorescence (TPEF) under continuous-wave excitation from silicon particles produced by a pulsed laser is investigated. Spectra and images of TPEF from silicon particles are studied under different excitation intensities and operation modes (continuous wave or pulse). It is found that the photoluminescence depends superlinearly on the excitation intensity and that the spectral shape and peaks vary with different silicon particles. The above phenomena show the nonlinear behavior of TPEF from silicon particles, and stimulated emission is a possible process.

  17. Beta-induced fluorescence detection in liquid chromatography

    International Nuclear Information System (INIS)

    Malcolme-Lawes, D.J.; Massey, S.; Warwick, P.

    1981-01-01

    A theoretical analysis of beta-induced fluorescence is used to determine the factors which influence the sensitivity of the technique as applied to liquid chromatography. Equations are presented for detector response and for signal-to-noise ratios and the theoretical response for a typical detector is compared with experimentally determined values. (author)

  18. Laser-induced fluorescence of oral mucosa cancer

    Science.gov (United States)

    Jaliashvili, Z. V.; Medoidze, T. D.; Melikishvili, Z. G.; Gogilashvili, K. T.

    2017-10-01

    The laser-induced fluorescence (LIF) spectra have been measured for cancer-infused and control mice mucosa tissues. It was established that there is quite a difference between their LIF spectral shapes. These spectral shapes are used to express the diagnostic of different states of tissues: from normal to cancer.

  19. NIR-emitting molecular-based nanoparticles as new two-photon absorbing nanotools for single particle tracking

    Science.gov (United States)

    Daniel, J.; Godin, A. G.; Clermont, G.; Lounis, B.; Cognet, L.; Blanchard-Desce, M.

    2015-07-01

    In order to provide a green alternative to QDs for bioimaging purposes and aiming at designing bright nanoparticles combining both large one- and two-photon brightness, a bottom-up route based on the molecular engineering of dedicated red to NIR emitting dyes that spontaneously form fluorescent organic nanoparticles (FONs) has been implemented. These fully organic nanoparticles built from original quadrupolar dyes are prepared using a simple, expeditious and green protocol that yield very small molecular-based nanoparticles (radius ~ 7 nm) suspension in water showing a nice NIR emission (λem=710 nm). These FONs typically have absorption coefficient more than two orders larger than popular NIR-emitting dyes (such as Alexa Fluor 700, Cy5.5 ….) and much larger Stokes shift values (i.e. up to over 5500 cm-1). They also show very large two-photon absorption response in the 800-1050 nm region (up to about 106 GM) of major promise for two-photon excited fluorescence microscopy. Thanks to their brightness and enhanced photostability, these FONs could be imaged as isolated nanoparticles and tracked using wide-field imaging. As such, thanks to their size and composition (absence of heavy metals), they represent highly promising alternatives to NIR-emitting QDs for use in bioimaging and single particle tracking applications. Moreover, efficient FONs coating was achieved by using a polymeric additive built from a long hydrophobic (PPO) and a short hydrophilic (PEO) segment and having a cationic head group able to interact with the highly negative surface of FONs. This electrostatically-driven interaction promotes both photoluminescence and two-photon absorption enhancement leading to an increase of two-photon brightness of about one order of magnitude. This opens the way to wide-field single particle tracking under two-photon excitation

  20. Time-resolved laser-induced fluorescence system

    Science.gov (United States)

    Bautista, F. J.; De la Rosa, J.; Gallegos, F. J.

    2006-02-01

    Fluorescence methods are being used increasingly in the measurement of species concentrations in gases, liquids and solids. Laser induced fluorescence is spontaneous emission from atoms or molecules that have been excited by laser radiation. Here we present a time resolved fluorescence instrument that consists of a 5 μJ Nitrogen laser (337.1 nm), a sample holder, a quartz optical fiber, a spectrometer, a PMT and a PC that allows the measurement of visible fluorescence spectra (350-750 nm). Time response of the system is approximately 5 ns. The instrument has been used in the measurement of colored bond paper, antifreeze, diesel, cochineal pigment and malignant tissues. The data acquisition was achieved through computer control of a digital oscilloscope (using General Purpose Interface Bus GPIB) and the spectrometer via serial (RS232). The instrument software provides a graphic interface that lets make some data acquisition tasks like finding fluorescence spectra, and fluorescence lifetimes. The software was developed using the Lab-View 6i graphic programming package and can be easily managed in order to add more functions to it.

  1. Laser-induced fluorescence in the detection of esophageal carcinoma

    Science.gov (United States)

    Wang, Kenneth K.; Gutta, Kumar; Laukka, Mark A.; Densmore, John

    1995-01-01

    Laser induced fluorescence (LIF) is a technique which can perform an 'optical biopsy' of gastrointestinal mucosa. LIF was performed in resected specimens using a pulsed N2-laser coupled fiberoptically to a probe. Fluorescence was measured using a 0.2 meter spectroscope with an intensified photodiode array. Measurements were made on fresh (esophagus, and adenocarcinoma. Each tissue section was examined using an optical probe consisting of a central fiber for delivering the excitation energy and a 6 fiber bundle surrounding the central fiber for detection of the fluorescence. An excitation wavelength of 337 nm was used which generated 3-ns pulses while fluorescence intensities were acquired from 300-800 nm. Spectra were obtained from each section in a standardized fashion and background spectra subtracted. Fluorescence readings were taken from 54 normal esophageal sections and 32 sections of adenocarcinoma. A fluorescence index obtained from the tumor sections was 0.68+/- 0.01 compared with 0.51+/- 0.01 for the normal sections (pesophagus with good accuracy.

  2. Precision two-photon spectroscopy of alkali elements

    Indian Academy of Sciences (India)

    effect is eliminated if the wave vector of the photons is ka = −kb, i.e., the two beams .... atomic cesium, and the metre is (indirectly) defined from the wavelength of .... plasma absorb radiation, the electrical circuit parameters may vary and this .... two-photon excitation followed by an ionization step in Ca [71], Sr [72] and Gd [73] ...

  3. Two-photon couplings of quarkonia with arbitrary JPC

    International Nuclear Information System (INIS)

    Barnes, T.; Tennessee Univ., Knoxville, TN

    1992-01-01

    We present theoretical results for the two-photon widths of relativistic quarkonium states with arbitrary angular momenta. These relativistic formulas are required to obtain reasonable agreement with the absolute scale of quarkonium decay rates to two photons, and have previously only been derived for spin-singlet q bar q states. We also evaluate these formulas numerically for ell ≤3 q = u, d states in a Coulomb-plus-linear q bar q potential model. Light-quark higher-ell and radially-excited q bar q states should be observable experimentally, as their two-photon widths are typically found to be ∼1 KeV. The radially-excited 1 S 0 higher-mass quarkonium states such as c bar c and b bar b should also be observable in γγ, but orbitally-excited c bar c states with ell>1 and b bar b states with ell>0 are expected to have very small two-photon widths. The helicity structure of the higher-ell q bar q couplings is predicted to be nontrivial, with both λ=0 and λ=2γγ final states contributing significantly; these results may be useful as signatures for q bar q states

  4. Two-photon optics of Bessel-Gaussian modes

    CSIR Research Space (South Africa)

    McLaren, M

    2013-09-01

    Full Text Available In this paper we consider geometrical two-photon optics of Bessel-Gaussian modes generated in spontaneous parametric down-conversion of a Gaussian pump beam. We provide a general theoretical expression for the orbital angular momentum (OAM) spectrum...

  5. Space-time description of the two-photon decay

    International Nuclear Information System (INIS)

    Hrasko, P.

    1981-09-01

    The time correlation of photons in a two-photon decay is shown to depend on the instantaneous nature of the wave-function collapse in an essential way so the latter hypothesis can be verified by the experimental study of these correlations. (author)

  6. Positron two-photon annihilation coincidence technique: difference mode

    Energy Technology Data Exchange (ETDEWEB)

    Karol, P J; Klobuchar, R L [Carnegie-Mellon Univ., Pittsburgh, Pa. (USA). Dept. of Chemistry

    1978-05-01

    A difference (or comparative) mode in the measurement of two-photon coincidences from positron or positronium annihilation has been developed. The method can be used to advantage, particularly in gases, in determining annihilation parameters such as quenching cross sections for low concentration strongly quenching chemical species in a reference medium which is relatively non-quenching.

  7. Impurity monitoring by laser-induced fluorescence techniques

    International Nuclear Information System (INIS)

    Gelbwachs, J.A.

    1984-01-01

    Laser-induced fluorescence spectroscopy can provide a highly sensitive and selective means of detecting atomic and ionic impurities. Because the photodetector can be physically isolated from the laser-excited region, these techniques can be applied to monitoring in hostile environments. The basic concepts behind fluorescence detection are reviewed. Saturated optical excitation is shown to maximize impurity atom emission yield while mitigating effects of laser intensity fluctuations upon absolute density calibration. Monitoring in high- and low-pressure monitoring environments is compared. Methods to improve detection sensitivity by luminescence background suppression are presented

  8. Instantaneous temperature field measurements using planar laser-induced fluorescence.

    Science.gov (United States)

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

    1985-09-01

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

  9. Tuning Ag29 nanocluster light emission from red to blue with one and two-photon excitation.

    Science.gov (United States)

    Russier-Antoine, Isabelle; Bertorelle, Franck; Hamouda, Ramzi; Rayane, Driss; Dugourd, Philippe; Sanader, Željka; Bonačić-Koutecký, Vlasta; Brevet, Pierre-François; Antoine, Rodolphe

    2016-02-07

    We demonstrate that the tuning of the light emission from red to blue in dihydrolipoic acid (DHLA) capped Ag29 nanoclusters can be trigged with one and two photon excitations. The cluster stoichiometry was determined with mass spectrometry and found to be Ag29(DHLA)12. In a detailed optical investigation, we show that these silver nanoclusters exhibit a strong red photoluminescence visible to the naked eye and characterized by a quantum yield of nearly ∼2% upon one-photon excitation. In the nonlinear optical (NLO) study of the properties of the clusters, the two-photon excited fluorescence spectra were recorded and their first hyperpolarizability obtained. The two-photon absorption cross-section at ∼800 nm for Ag29(DHLA)12 is higher than 10(4) GM and the hyperpolarizability is 106 × 10(-30) esu at the same excitation wavelength. The two-photon excited fluorescence spectrum appears strongly blue-shifted as compared to the one-photon excited spectrum, displaying a broad band between 400 and 700 nm. The density functional theory (DFT) provides insight into the structural and electronic properties of Ag29(DHLA)12 as well as into interplay between metallic subunit or core and ligands which is responsible for unique optical properties.

  10. Two-photon absorption and two-photon circular dichroism of hexahelicene derivatives: a study of the effect of the nature of intramolecular charge transfer

    Czech Academy of Sciences Publication Activity Database

    Díaz, C.; Vesga, Y.; Echevarria, L.; Stará, Irena G.; Starý, Ivo; Anger, E.; Shen, C.; Moussa, M. E. S.; Vanthuyne, N.; Crassous, J.; Rizzo, A.; Hernández, F. E.

    2015-01-01

    Roč. 5, č. 23 (2015), s. 17429-17437 ISSN 2046-2069 Institutional support: RVO:61388963 Keywords : [6]helicene * two photon absorption * two photon CD Subject RIV: CC - Organic Chemistry Impact factor: 3.289, year: 2015

  11. Inhibition of two-photon absorption due to dipole-dipole interaction in nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Mahi R. [Department of Physics and Astronomy, University of Western Ontario, London, N6A 3K7 (Canada)], E-mail: msingh@uwo.ca

    2008-07-21

    We have investigated the inhibition of two-photon absorption in photonic crystals doped with an ensemble of four-level nanoparticles. The particles are interacting with one another by the dipole-dipole interaction. Dipoles in nanoparticles are induced by a selected transition. Numerical simulations have been performed for an isotropic photonic crystal. Interesting phenomena have been predicted such as the inhibition of the two-photon absorption due to the dipole-dipole interaction. It has also been found that the inhibition effect can be switched on and off by tuning a decay resonance energy within the energy band of the crystal. A theory of dressed states has been used to explain the results.

  12. Two-photon polymerization of an epoxy-acrylate resin material system

    International Nuclear Information System (INIS)

    Winfield, R.J.; O'Brien, S.

    2011-01-01

    Improved material systems are of great interest in the development of two-photon polymerization techniques for the fabrication of three dimensional micro- and nano-structures. The properties of the photosensitive resin are important in the realisation of structures with submicron dimensions. In this study investigation of a custom organic resin, cross-linked by a two-photon induced process, using a femtosecond Ti:sapphire laser, is described. A structural, optical and mechanical analysis of the optimised material is presented. The influence of both material system and laser processing parameters on achievable micro-structure and size is presented as are representative structures. Parameters include: laser power, photo-initiator concentration and material composition.

  13. Vibrational excitation of hydrogen molecules by two-photon absorption and third-harmonic generation

    Science.gov (United States)

    Miyamoto, Yuki; Hara, Hideaki; Hiraki, Takahiro; Masuda, Takahiko; Sasao, Noboru; Uetake, Satoshi; Yoshimi, Akihiro; Yoshimura, Koji; Yoshimura, Motohiko

    2018-01-01

    We report the coherent excitation of the vibrational state of hydrogen molecules by two-photon absorption and the resultant third-harmonic generation (THG). Parahydrogen molecules cooled by liquid nitrogen are irradiated by mid-infrared nanosecond pulses at 4.8 μm with a nearly Fourier-transform-limited linewidth. The first excited vibrational state of parahydrogen is populated by two-photon absorption of the mid-infrared photons. Because of the narrow linewidth of the mid-infrared pulses, coherence between the ground and excited states is sufficient to induce higher-order processes. Near-infrared photons from the THG are observed at 1.6 μm. The dependence of the intensity of the near-infrared radiation on mid-infrared pulse energy, target pressure, and cell length is determined. We used a simple formula for THG with consideration of realistic experimental conditions to explain the observed results.

  14. Neural plasticity explored by correlative two-photon and electron/SPIM microscopy

    Science.gov (United States)

    Allegra Mascaro, A. L.; Silvestri, L.; Costantini, I.; Sacconi, L.; Maco, B.; Knott, G. W.; Pavone, F. S.

    2013-06-01

    Plasticity of the central nervous system is a complex process which involves the remodeling of neuronal processes and synaptic contacts. However, a single imaging technique can reveal only a small part of this complex machinery. To obtain a more complete view, complementary approaches should be combined. Two-photon fluorescence microscopy, combined with multi-photon laser nanosurgery, allow following the real-time dynamics of single neuronal processes in the cerebral cortex of living mice. The structural rearrangement elicited by this highly confined paradigm of injury can be imaged in vivo first, and then the same neuron could be retrieved ex-vivo and characterized in terms of ultrastructural features of the damaged neuronal branch by means of electron microscopy. Afterwards, we describe a method to integrate data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy, based on the use of major blood vessels as reference chart. We show how the apical dendritic arbor of a single cortical pyramidal neuron imaged in living mice can be found in the large-scale brain reconstruction obtained with light sheet microscopy. Starting from its apical portion, the whole pyramidal neuron can then be segmented and located in the correct cortical layer. With the correlative approach presented here, researchers will be able to place in a three-dimensional anatomic context the neurons whose dynamics have been observed with high detail in vivo.

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

    Science.gov (United States)

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

    2012-10-01

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

  16. Time-resolved stimulated emission depletion and energy transfer dynamics in two-photon excited EGFP

    Science.gov (United States)

    Masters, T. A.; Robinson, N. A.; Marsh, R. J.; Blacker, T. S.; Armoogum, D. A.; Larijani, B.; Bain, A. J.

    2018-04-01

    Time and polarization-resolved stimulated emission depletion (STED) measurements are used to investigate excited state evolution following the two-photon excitation of enhanced green fluorescent protein (EGFP). We employ a new approach for the accurate STED measurement of the hitherto unmeasured degree of hexadecapolar transition dipole moment alignment ⟨α40 ⟩ present at a given excitation-depletion (pump-dump) pulse separation. Time-resolved polarized fluorescence measurements as a function of pump-dump delay reveal the time evolution of ⟨α40 ⟩ to be considerably more rapid than predicted for isotropic rotational diffusion in EGFP. Additional depolarization by homo-Förster resonance energy transfer is investigated for both ⟨α20 ⟩ (quadrupolar) and ⟨α40 ⟩ transition dipole alignments. These results point to the utility of higher order dipole correlation measurements in the investigation of resonance energy transfer processes.

  17. Deformed two-photon squeezed states in noncommutative space

    International Nuclear Information System (INIS)

    Zhang Jianzu

    2004-01-01

    Recent studies on nonperturbation aspects of noncommutative quantum mechanics explored a new type of boson commutation relations at the deformed level, described by deformed annihilation-creation operators in noncommutative space. This correlated boson commutator correlates different degrees of freedom, and shows an essential influence on dynamics. This Letter devotes to the development of formalism of deformed two-photon squeezed states in noncommutative space. General representations of deformed annihilation-creation operators and the consistency condition for the electromagnetic wave with a single mode of frequency in noncommunicative space are obtained. Two-photon squeezed states are studied. One finds that variances of the dimensionless Hermitian quadratures of the annihilation operator in one degree of freedom include variances in the other degree of freedom. Such correlations show the new feature of spatial noncommutativity and allow a deeper understanding of the correlated boson commutator

  18. Meson production in two-photon interactions at LHC energies

    Energy Technology Data Exchange (ETDEWEB)

    Da Silva, D. T.; Goncalves, V. P.; Sauter, W. K. [Instituto de Fisica e Matematica, Universidade Federal de Pelotas, Campus Universitario UFPel, CP 354, 96010-900, Capao do Leao-RS (Brazil)

    2013-03-25

    The LHC opens a new kinematical regime at high energy, where several questions related to the description of the high-energy regime of the Quantum Chromodynamics (QCD) remain without satisfactory answers. Some open questions are the search for non-q-bar q resonances, the determination of the spectrum of q-bar q states and the identification of states with anomalous {gamma}{gamma} couplings. A possible way to study these problems is the study of meson production in two-photon interactions. In this contribution we calculate the meson production in two-photon interactions at LHC energies considering proton - proton collisions and estimate the total cross section for the production of the mesons {pi}, a, f, {eta} and {chi}.

  19. Two-photon polarization Fourier spectroscopy of metastable atomic hydrogen

    International Nuclear Information System (INIS)

    Duncan, A.J.; Beyer, H.-J.; Kleinpoppen, H.; Sheikh, Z.A,; B-Z Univ., Multan

    1997-01-01

    A novel Fourier-transform spectroscopic method using two-photon polarization to determine the spectral distribution of the two photons emitted in the spontaneous decay of metastable atomic hydrogen is described. The method uses birefringent retardation plates and takes advantage of the subtle interplay between the spectral properties and the entangled polarization properties of the radiation emitted in the decay. Assuming the validity of the theoretical spectral distribution, it is shown that the experimental results agree well with theory. On the other hand, success in solving the inverse problem of determining the spectral distribution from the experimental results is limited by the small number of experimental points. However, making reasonable assumptions it is deduced that the observed spectrum is characterized by a broadband signal of width (0.43 ± 0.06) x 10 16 rad s -1 and centre angular frequency (0.77 ± 0.03) x 10 16 rad s -1 in good agreement with the predictions of 0.489 x 10 16 rad s -1 and 0.775 x 10 16 rad s -1 , respectively, obtained from the theoretical spectral distribution modified to take account of the absorption of the two-photon radiation in air. The values of 1.5 fs for the coherence time and 440 nm for the coherence length for single photons of the two-photon pair which are obtained from the measured bandwidth imply that, in the ideal case, these values are determined by the essentially zero lifetime of the virtual intermediate state of the decay process rather than the long lifetime of the metastable state which, it is suggested, determines the coherence time and coherence length appropriate to certain types of fourth-order interference experiments. (Author)

  20. Two-Photon Absorption in Organometallic Bromide Perovskites

    KAUST Repository

    Walters, Grant

    2015-07-21

    Organometallic trihalide perovskites are solution processed semiconductors that have made great strides in third generation thin film light harvesting and light emitting optoelectronic devices. Recently it has been demonstrated that large, high purity single crystals of these perovskites can be synthesized from the solution phase. These crystals’ large dimensions, clean bandgap, and solid-state order, have provided us with a suitable medium to observe and quantify two-photon absorption in perovskites. When CH3NH3PbBr3 single crystals are pumped with intense 800 nm light, we observe band-to-band photoluminescence at 572 nm, indicative of two-photon absorption. We report the nonlinear absorption coefficient of CH3NH3PbBr3 perovskites to be 8.6 cm GW-1 at 800 nm, comparable to epitaxial single crystal semiconductors of similar bandgap. We have leveraged this nonlinear process to electrically autocorrelate a 100 fs pulsed laser using a two-photon perovskite photodetector. This work demonstrates the viability of organometallic trihalide perovskites as a convenient and low-cost nonlinear absorber for applications in ultrafast photonics.

  1. Particle Production in Two-Photon Collisions at Belle

    International Nuclear Information System (INIS)

    Nakazawa, Hideyuki

    2010-01-01

    Experimental study of η η production in two-photon collisions: The differential cross section for the process γ γ → η η has been measured in the kinematic range above the η η threshold, 1.096 GeV -1 data sample collected with the Belle detector at the KEKB e + e - collider. In the W range 1.1-2.0 GeV/c 2 we perform an analysis of resonance amplitudes for various partial waves; at higher energy we extract the contributions of χ cJ charmonia and compare the energy and angular dependence of the cross section with the predictions of theoretical models. Observation of η c (2S) in six-prong final states produced in two-photon collisions: We report the observation of η c (2S), produced in two-photon collisions, and decaying to the six-prong final states 3(π + π - ), K + K - 2(π + π - ), and K 0 S K + ππ + π - (including the charge-conjugate state). This analysis is based on a large data sample accumulated by the Belle experiment at the KEKB asymmetric-energy electron-positron collider. This is the first observation of decay modes of the η c (2S) other than K 0 S K + π - . (author)

  2. Two-photon exchange in elastic electron-nucleon scattering

    International Nuclear Information System (INIS)

    Blunden, P.G.; Melnitchouk, W.; Tjon, J.A.

    2005-01-01

    A detailed study of two-photon exchange in unpolarized and polarized elastic electron-nucleon scattering is presented, taking particular account of nucleon finite size effects. Contributions from nucleon elastic intermediate states are found to have a strong angular dependence, which leads to a partial resolution of the discrepancy between the Rosenbluth and polarization transfer measurements of the proton electric to magnetic form factor ratio, G E /G M . The two-photon exchange contribution to the longitudinal polarization transfer P L is small, whereas the contribution to the transverse polarization transfer P T is enhanced at backward angles by several percent, increasing with Q 2 . This gives rise to a small, E /G M obtained from the polarization transfer ratio P T /P L at large Q 2 . We also compare the two-photon exchange effects with data on the ratio of e + p to e - p cross sections, which is predicted to be enhanced at backward angles. Finally, we evaluate the corrections to the form factors of the neutron and estimate the elastic intermediate state contribution to the 3 He form factors

  3. Two-Photon Absorption in Organometallic Bromide Perovskites

    KAUST Repository

    Walters, Grant; Sutherland, Brandon R; Hoogland, Sjoerd; Shi, Dong; Comin, Riccardo; Sellan, Daniel P.; Bakr, Osman; Sargent, Edward H.

    2015-01-01

    Organometallic trihalide perovskites are solution processed semiconductors that have made great strides in third generation thin film light harvesting and light emitting optoelectronic devices. Recently it has been demonstrated that large, high purity single crystals of these perovskites can be synthesized from the solution phase. These crystals’ large dimensions, clean bandgap, and solid-state order, have provided us with a suitable medium to observe and quantify two-photon absorption in perovskites. When CH3NH3PbBr3 single crystals are pumped with intense 800 nm light, we observe band-to-band photoluminescence at 572 nm, indicative of two-photon absorption. We report the nonlinear absorption coefficient of CH3NH3PbBr3 perovskites to be 8.6 cm GW-1 at 800 nm, comparable to epitaxial single crystal semiconductors of similar bandgap. We have leveraged this nonlinear process to electrically autocorrelate a 100 fs pulsed laser using a two-photon perovskite photodetector. This work demonstrates the viability of organometallic trihalide perovskites as a convenient and low-cost nonlinear absorber for applications in ultrafast photonics.

  4. Doppler-free two-photon spectroscopy of Yb atoms and efficient generation of a cascade of two photons at 611.3 nm and 555.8 nm

    International Nuclear Information System (INIS)

    Song, Minsoo; Yoon, Duseong; Yoon, Taihyun

    2011-01-01

    We performed high-resolution Doppler-free two-photon spectroscopy of Yb atoms in an effusive atomic beam and generated a cascade of two photons at 611.3 nm (idler) and 555.8 nm (signal) with a narrow bandwidth of 37 MHz. Efficient population transfer from the ground state (6s 2 1 S 0 ) to the upper state (6s7s 1 S 0 ), where direct transition at 291.1 nm is dipole forbidden, was achieved through a resonant two-photon excitation enhanced by the electromagnetically-induced transparency mediated by the intermediate state (6s6p 1 P 1 ). From the upper state, a cascade of two photons in sequence was emitted via the spin triplet state (6s 2 3 P 1 ). Numerical calculations of the density matrix equations taking into account the residual Doppler effect and strong driving fields explain quantitatively the experimental results for the dependences of the idler and the signal beam intensities on the various parameters of the driving fields. We report on the generation of a cascade of two photons with fluxes at the level of a few times 10 6 photons/s detected at a solid angle of 0.01 sr.

  5. Doppler-free two-photon spectroscopy of Yb atoms and efficient generation of a cascade of two photons at 611.3 nm and 555.8 nm

    Energy Technology Data Exchange (ETDEWEB)

    Song, Minsoo; Yoon, Duseong; Yoon, Taihyun [Korea University, Seoul (Korea, Republic of)

    2011-10-15

    We performed high-resolution Doppler-free two-photon spectroscopy of Yb atoms in an effusive atomic beam and generated a cascade of two photons at 611.3 nm (idler) and 555.8 nm (signal) with a narrow bandwidth of 37 MHz. Efficient population transfer from the ground state (6s{sup 2} {sup 1}S{sub 0}) to the upper state (6s7s {sup 1}S{sub 0}), where direct transition at 291.1 nm is dipole forbidden, was achieved through a resonant two-photon excitation enhanced by the electromagnetically-induced transparency mediated by the intermediate state (6s6p {sup 1}P{sub 1}). From the upper state, a cascade of two photons in sequence was emitted via the spin triplet state (6s{sup 2} {sup 3}P{sub 1}). Numerical calculations of the density matrix equations taking into account the residual Doppler effect and strong driving fields explain quantitatively the experimental results for the dependences of the idler and the signal beam intensities on the various parameters of the driving fields. We report on the generation of a cascade of two photons with fluxes at the level of a few times 10{sup 6} photons/s detected at a solid angle of 0.01 sr.

  6. Led induced chlorophyll fluorescence transient imager for measurements of health and stress status of whole plants

    NARCIS (Netherlands)

    Jalink, H.; Schoor, van der R.

    2011-01-01

    We have developed LED (light emitting diode) induced fluorescence transient imaging instrumentation to image the plant health/stress status by calculation of two images: Fv/Fm (variable fluorescence over saturation level of fluorescence) and the time response, tTR, of the fluorescence time curve.

  7. Detection of vegetation stress from laser-induced fluorescence signatures

    International Nuclear Information System (INIS)

    Subhash, N.

    1995-01-01

    The in vivo laser-induced fluorescence (LIF) signatures of UV irradiated Salvia splendens plants were measured using an Optical Multichannel Analyser (OMA) system with Nitrogen laser excitation. The LIF spectra which consisted of the blue-green and the red chlorophyll bands were analysed with a non-linear interactive procedure using Gaussian spectral functions. The fluorescence intensity ratios of the various bands obtained from curve fitted parameters were found to be more sensitive to changes in the photosynthetic activity of the plant. The variation in the intensity ratio for the chlorophyll bands for nutrient stressed sunflower, cotton and groundnut plants as well as the nutrient and water stressed rice plants are also presented. It is observed that vegetation stress not only changes the fluorescence intensity ratios and the vitality index of the plant but also changes the peak position of the emission bands, in some cases. It is also seen that analysis of the fluorescence spectra in vegetation remote sensing applications would require a deconvolution procedure to evaluate the exact contribution of each band in the total spectra. (author). 23 refs, 8 figs, 5 tabs

  8. Novel biocompatible materials for in vivo two-photon polymerisation

    Energy Technology Data Exchange (ETDEWEB)

    Torgersen, J.

    2013-07-01

    Two-photon polymerisation (2PP) is a versatile laser fabrication technique that allows the creation of 3D structures at micro- and nanometre precision. The structures are created additively in direct accordance to a computer-aided design (CAD). It requires tightly focused fs-pulsed light sources usually operating in the near infrared wavelength range. In this region, biological tissues exhibit a window of transparency and only absorb light minimally. When operating below a certain pulse energy threshold, the laser light does not cause any cellular damage. This theoretically allows inducing 2PP in the presence of living biological tissues and cells. Suitable biocompatible formulations that can render bioactive constructs would potentially allow building a dynamic environment with topographical, chemical and mechanical cues similar to that of the natural extracellular matrix. In that way, 2PP would allow to alter key elements of this environment without changing any other influencing factors. To explore these possibilities, 2PP has to overcome two main limitations, the slow process speeds and the lack of available optimised formulations. In this thesis, we report the design and realisation of a 2PP experimental setup, which allows fabricating hydrogel structures from novel water-based formulations. Writing speeds of above 100 mm/s are feasible, which is the highest speed reported in 2PP. Moreover, the presented components have the potential to be formed in vivo, in the presence of living cells and tissues. Using water-soluble two-photon optimised photoinitiators, we could effectively cross-link acrylates in formulations of up to 80% water content. As acrylates show a tendency towards Michael addition to proteins, we explored the use of vinyl ester and vinyl carbonate monomers for 2PP. In contrast to acrylic polymers, which form potentially toxic poly (acrylic acid), vinyl ester and carbonate polymers form biocompatible poly (vinyl alcohol) during degradation

  9. Comparison of in Situ and ex Situ Methods for Synthesis of Two-Photon Polymerization Polymer Nanocomposites

    Directory of Open Access Journals (Sweden)

    Qingchuan Guo

    2014-07-01

    Full Text Available This article reports about nanocomposites, which refractive index is tuned by adding TiO2 nanoparticles. We compare in situ/ex situ preparation of nanocomposites. Preparation procedure is described, properties of nanocomposites are compared, and especially we examine the applicability of two-photon polymerization (2PP of synthesized nanocomposites. All prepared samples exhibit suitable optical transparency at specific laser wavelengths. Three-dimensional structures were generated by means of two-photon polymerization effect induced by a femtosecond laser.

  10. Studying electron-PAG interactions using electron-induced fluorescence

    Science.gov (United States)

    Narasimhan, Amrit; Grzeskowiak, Steven; Ostrander, Jonathan; Schad, Jonathon; Rebeyev, Eliran; Neisser, Mark; Ocola, Leonidas E.; Denbeaux, Gregory; Brainard, Robert L.

    2016-03-01

    In extreme ultraviolet (EUV) lithography, 92 eV photons are used to expose photoresists. Typical EUV resists are organic-based and chemically amplified using photoacid generators (PAGs). Upon exposure, PAGs produce acids which catalyze reactions that result in changes in solubility. In EUV lithography, photo- and secondary electrons (energies of 10- 80 eV) play a large role in PAG acid-production. Several mechanisms for electron-PAG interactions (e.g. electron trapping, and hole-initiated chemistry) have been proposed. The aim of this study is to explore another mechanism - internal excitation - in which a bound PAG electron can be excited by receiving energy from another energetic electron, causing a reaction that produces acid. This paper explores the mechanism of internal excitation through the analogous process of electron-induced fluorescence, in which an electron loses energy by transferring that energy to a molecule and that molecule emits a photon rather than decomposing. We will show and quantify electron-induced fluorescence of several fluorophores in polymer films to mimic resist materials, and use this information to refine our proposed mechanism. Relationships between the molecular structure of fluorophores and fluorescent quantum yield may aid in the development of novel PAGs for EUV lithography.

  11. Porous Porphyrin-Based Organosilica Nanoparticles for NIR Two-Photon Photodynamic Therapy and Gene Delivery in Zebrafish

    KAUST Repository

    Mauriello Jimenez, Chiara

    2018-03-30

    Periodic mesoporous organosilica nanoparticles emerge as promising vectors for nanomedicine applications. Their properties are very different from those of well-known mesoporous silica nanoparticles as there is no silica source for their synthesis. So far, they have only been synthesized from small bis-silylated organic precursors. However, no studies employing large stimuli-responsive precursors have been reported on such hybrid systems yet. Here, the synthesis of porphyrin-based organosilica nanoparticles from a large octasilylated metalated porphyrin precursor is described for applications in near-infrared two-photon-triggered spatiotemporal theranostics. The nanoparticles display unique interconnected large cavities of 10-80 nm. The framework of the nanoparticles is constituted with J-aggregates of porphyrins, which endows them with two-photon sensitivity. The nanoparticle efficiency for intracellular tracking is first demonstrated by the in vitro near-infrared imaging of breast cancer cells. After functionalization of the nanoparticles with aminopropyltriethoxysilane, two-photon-excited photodynamic therapy in zebrafish is successfully achieved. Two-photon photochemical internalization in cancer cells of the nanoparticles loaded with siRNA is also performed for the first time. Furthermore, siRNA targeting green fluorescent protein complexed with the nanoparticles is delivered in vivo in zebrafish embryos, which demonstrates the versatility of the nanovectors for biomedical applications.

  12. Towards Precision Measurement of the 21S0-31D2 Two-Photon Transition in Atomic Helium

    Science.gov (United States)

    Huang, Yi-Jan; Guan, Yu-Chan; Suen, Te-Hwei; Wang, Li-Bang; Shy, Jow-Tsong

    2017-04-01

    We intend to accurately measure the frequency for 2S-3D two-photon transition and to deduce the 2S ionization energy to an accuracy below 100 kHz from the theoretical calculation of the 3D state. In this talk, we present a precision measurement of the 21S0 -31D2 two-photon transition in atomic helium at 1009 nm. A master oscillator power amplifier (MOPA) is seeded by an external cavity diode laser (ECDL) is constructed to generate more than 700 mW laser power with TEM00 beam profile at 1009 nm. To observe the two-photon transition, a helium cell is placed inside a power enhancement optical cavity and the helium atoms at 21S metastable level are prepared by a pulsed RF discharge and monitor the 668 nm 31D2 to 21P1 fluorescence after RF discharge is turned off . The absolute frequency metrology of the ECDL is carried out by an Er-fiber optical frequency comb (OFC). The two-photon spectrum is obtained by tuning the repetition frequency of the OFC. The 21S0-31D2 frequency is determined to be 594414291.967 (80) MHz in He-4. More results will be presented at the annual meeting.

  13. Design of remote laser-induced fluorescence system's acquisition circuit

    Science.gov (United States)

    Wang, Guoqing; Lou, Yue; Wang, Ran; Yan, Debao; Li, Xin; Zhao, Xin; Chen, Dong; Zhao, Qi

    2017-10-01

    Laser-induced fluorescence system(LIfS) has been found its significant application in identifying one kind of substance from another by its properties even it's thimbleful, and becomes useful in plenty of fields. Many superior works have reported LIfS' theoretical analysis , designs and uses. However, the usual LIPS is always constructed in labs to detect matter quite closely, for the system using low-power laser as excitation source and charge coupled device (CCD) as detector. Promoting the detectivity of LIfS is of much concern to spread its application. Here, we take a high-energy narrow-pulse laser instead of commonly used continuous wave laser to operate sample, thus we can get strong fluorescent. Besides, photomultiplier (PMT) with high sensitivity is adopted in our system to detect extremely weak fluorescence after a long flight time from the sample to the detector. Another advantage in our system, as the fluorescence collected into spectroscopy, multiple wavelengths of light can be converted to the corresponding electrical signals with the linear array multichannel PMT. Therefore, at the cost of high-powered incentive and high-sensitive detector, a remote LIFS is get. In order to run this system, it is of importance to turn light signal to digital signal which can be processed by computer. The pulse width of fluorescence is deeply associated with excitation laser, at the nanosecond(ns) level, which has a high demand for acquisition circuit. We design an acquisition circuit including, I/V conversion circuit, amplifying circuit and peak-holding circuit. The simulation of circuit shows that peak-holding circuit can be one effective approach to reducing difficulty of acquisition circuit.

  14. Laser induced uranium fluorescence as an analytical method

    International Nuclear Information System (INIS)

    Krutman, I.

    1985-01-01

    A laser induced fluorescence system was developed to measure uranium trace level amounts in aqueous solution with reliable and simple materials and electronics. A nitrogen pulsed laser was built with the storage energy capacitor directly coupled to laser tube electrodes as a transmission line device. This laser operated at 3Hz repetition rate with peak intensity around 21 Kw and temporal width of 4.5 x 10 -9 s. A sample compartment made of rigid PVC and a photomultiplier housing of aluminium were constructed and assembled forming a single integrated device. As a result of this prototype system we made several analytical measurements with U dissolved in nitric acid to obtain a calibration curve. We obtained a straight line from a plot of U concentration versus fluorescence intensity fitted by a least square method that produced a regression coefficient of 0.994. The lower limit of U determination was 30 ppb -+ 3.5%. (Author) [pt

  15. Inclusive $D*^{+-}$ Production in Two-Photon Collisions at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery P.; Anselmo, F.; Arefev, A.; Azemoon, T.; Aziz, T.; Bagnaia, P.; Bajo, A.; Baksay, G.; Baksay, L.; Baldew, S.V.; Banerjee, S.; Banerjee, Sw.; Barczyk, A.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Echenard, B.; Eline, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Ewers, A.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hakobyan, R.S.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Krenz, W.; Kruger, A.; Kunin, A.; Ladron de Guevara, P.; Laktineh, I.; Landi, G.; Lebeau, M.; Lebedev, A.; Lebrun, P.; Lecomte, P.; Lecoq, P.; Le Coultre, P.; Le Goff, J.M.; Leiste, R.; Levtchenko, M.; Levtchenko, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Lubelsmeyer, K.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mangeol, D.; Mans, J.; Martin, J.P.; Marzano, F.; Mazumdar, K.; McNeil, R.R.; Mele, S.; Merola, L.; Meschini, M.; Metzger, W.J.; Mihul, A.; Milcent, H.; Mirabelli, G.; Mnich, J.; Mohanty, G.B.; Muanza, G.S.; Muijs, A.J.M.; Musicar, B.; Musy, M.; Nagy, S.; Natale, S.; Napolitano, M.; Nessi-Tedaldi, F.; Newman, H.; Niessen, T.; Nisati, A.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Palomares, C.; Pandoulas, D.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.O.; Prokofiev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, M.A.; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Roux, B.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Sanders, M.P.; Schafer, C.; Schegelsky, V.; Schmidt-Kaerst, S.; Schmitz, D.; Schopper, H.; Schotanus, D.J.; Schwering, G.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Siedenburg, T.; Son, D.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, I.; Vogel, H.; Vogt, H.; Vorobev, I.; Vorobyov, A.A.; Wadhwa, M.; Wallraff, W.; Wang, X.L.; Wang, Z.M.; Weber, M.; Wienemann, P.; Wilkens, H.; Wynhoff, S.; Xia, L.; Xu, Z.Z.; Yamamoto, J.; Yang, B.Z.; Yang, C.G.; Yang, H.J.; Yang, M.; Yeh, S.C.; Zalite, A.; Zalite, Yu.; Zhang, Z.P.; Zhao, J.; Zhu, G.Y.; Zhu, R.Y.; Zhuang, H.L.; Zichichi, A.; Zilizi, G.; Zimmermann, B.; Zoller, M.

    2002-01-01

    Inclusive D^{*+-} production in two-photon collisions is studied with the L3 detector at LEP, using 683 pb^{-1} of data collected at centre-of-mass energies from 183 to 208 GeV. Differential cross sections are determined as functions of the transverse momentum and pseudorapidity of the D^{*+-} mesons in the kinematic region 1 GeV e^+e^-D^{*+-}X)$ in this kinematical region is measured and the sigma(e^+e^- ---> e^+e^- cc{bar}X) cross section is derived. The measurements are compared with next-to-leading order perturbative QCD calculations.

  16. New two-photon based nanoscopic modalities and optogenetics

    DEFF Research Database (Denmark)

    Glückstad, Jesper

    that can be equipped with optical trapping handles for convenient mechanical control using only optical forces [6]. These microstructures illustrated above can be effectively handled with simultaneous top- and side-view on our BioPhotonics Workstation to undertake six-degree-of-freedom optical actuation...... of two-photon polymerised microstructures equipped with features easily entering the submicron-regime. Aided by European collaborators who fabricated test structures with built-in waveguides for us, we were able to put the idea of optically steerable freestanding waveguides – coined: wave-guided optical...

  17. Two photon production of charm states - a charmonium calculation

    International Nuclear Information System (INIS)

    Suaya, R.

    1980-03-01

    The comparison of the nonrelativistic multichannel calculation with the double Rutherford process shows that duality is satisfied, in the sense that if one averages the actual cross sections for a wide enough range of W one gets the same result as for nonstrongly interacting point like quarks. It has been shown for the case of e + e - annihilation that duality in this sense follows from nonrelativistic potential models; and it has been verified that this proof can be extended to two-photon processes

  18. Two-photon exclusive processes in quantum chromodynamics

    International Nuclear Information System (INIS)

    Brodsky, S.J.

    1986-07-01

    QCD predictions for γγ annihilation into single mesons, meson pairs, and baryon pairs are reviewed. Two-photon exclusive processes provide the most sensitive and practical measure of the distribution amplitudes, and thus a critical confrontation between QCD and experiment. Both the angular distribution and virtual photon mass dependence of these amplitudes are sensitive to the shapes of the phi (chi, Q). Novel effects involving the production of qq anti q anti q states at threshold are also discussed, and a new method is presented for systematically incorporating higher-order QCD corrections in γγ reactions

  19. Two-photon exclusive processes in quantum chromodynamics

    Energy Technology Data Exchange (ETDEWEB)

    Brodsky, S.J.

    1986-07-01

    QCD predictions for ..gamma gamma.. annihilation into single mesons, meson pairs, and baryon pairs are reviewed. Two-photon exclusive processes provide the most sensitive and practical measure of the distribution amplitudes, and thus a critical confrontation between QCD and experiment. Both the angular distribution and virtual photon mass dependence of these amplitudes are sensitive to the shapes of the phi (chi, Q). Novel effects involving the production of qq anti q anti q states at threshold are also discussed, and a new method is presented for systematically incorporating higher-order QCD corrections in ..gamma gamma.. reactions.

  20. Direct and indirect two-photon processes in semiconductors

    International Nuclear Information System (INIS)

    Hassan, A.R.

    1986-07-01

    The expressions describing direct and indirect two-photon absorption in crystals are given. They are valid both near and far from the energy gap. A perturbative approach through two different band models is adopted. The effects of the non-parabolicity and the degeneracy of the energy bands are considered. The numerical results are compared with the other theories and with a recent experimental data in Zn and AgCl. It is shown that the dominant transition mechanisms are of the allowed-allowed type near and far from the gap for both direct and indirect processes. (author)

  1. {Lambda}{bar {Lambda}} production in two-photon interactions

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, S.; Kubota, Y.; Lattery, M.; ONeill, J.J.; Patton, S.; Poling, R.; Riehle, T.; Savinov, V.; Smith, A. [University of Minnesota, Minneapolis, Minnesota 55455 (United States); Alam, M.S.; Athar, S.B.; Ling, Z.; Mahmood, A.H.; Severini, H.; Timm, S.; Wappler, F. [State University of New York at Albany, Albany, New York 12222 (United States); Anastassov, A.; Blinov, S.; Duboscq, J.E.; Fujino, D.; Fulton, R.; Gan, K.K.; Hart, T.; Honscheid, K.; Kagan, H.; Kass, R.; Lee, J.; Spencer, M.B.; Sung, M.; Undrus, A.; Wanke, R.; Wolf, A.; Zoeller, M.M. [Ohio State University, Columbus, Ohio 43210 (United States); Nemati, B.; Richichi, S.J.; Ross, W.R.; Skubic, P.; Wood, M. [University of Oklahoma, Norman, Oklahoma 73019 (United States); Bishai, M.; Fast, J.; Gerndt, E.; Hinson, J.W.; Menon, N.; Miller, D.H.; Shibata, E.I.; Shipsey, I.P.; Yurko, M. [Purdue University, West Lafayette, Indiana 47907 (United States); Gibbons, L.; Johnson, S.D.; Kwon, Y.; Roberts, S.; Thorndike, E.H. [University of Rochester, Rochester, New York 14627 (United States); Jessop, C.P.; Lingel, K.; Marsiske, H.; Perl, M.L.; Schaffner, S.F.; Ugolini, D.; Wang, R.; Zhou, X. [Stanford Linear Accelerator Center, Stanford University, Stanford, California 94309 (United States); Coan, T.E.; Fadeyev, V.; Korolkov, I.; Maravin, Y.; Narsky, I.; Shelkov, V.; Staeck, J.; Stroynowski, R.; Volobouev, I.; Ye, J. [Southern Methodist University, Dallas, Texas 75275 (United States); Artuso, M.; Efimov, A.; Frasconi, F.; Gao, M.; Goldberg, M.; He, D.; Kopp, S.; Moneti, G.C.; Mountain, R.; Mukhin, Y.; Schuh, S.; Skwarnicki, T.; Stone, S.; Viehhauser, G.; Xing, X. [Syracuse University, Syracuse, New York 13244 (United States); Bartelt, J.; Csorna, S.E.; Jain, V.; Marka, S. [Vanderbilt University, Nashville, Tennessee 37235 (United States); Freyberger, A.; Gibaut, D.; Godang, R.; Kinoshita, K.; Lai, I.C.; Pomianowski, P.; Schrenk, S.; and others

    1997-09-01

    Using the CLEO detector at the Cornell e{sup +}e{sup {minus}} storage ring CESR we study the two-photon production of {Lambda}{bar {Lambda}}, making the first observation of {gamma}{gamma}{r_arrow}{Lambda}{bar {Lambda}}. We present the cross section for {gamma}{gamma}{r_arrow}{Lambda}{bar {Lambda}} as a function of the {gamma}{gamma} center of mass energy and compare it to that predicted by the quark-diquark model. {copyright} {ital 1997} {ital The American Physical Society}

  2. Two-photon excited autofluorescence imaging of freshly isolated frog retinas.

    Science.gov (United States)

    Lu, Rong-Wen; Li, Yi-Chao; Ye, Tong; Strang, Christianne; Keyser, Kent; Curcio, Christine A; Yao, Xin-Cheng

    2011-06-01

    The purpose of this study was to investigate cellular sources of autofluorescence signals in freshly isolated frog (Rana pipiens) retinas. Equipped with an ultrafast laser, a laser scanning two-photon excitation fluorescence microscope was employed for sub-cellular resolution examination of both sliced and flat-mounted retinas. Two-photon imaging of retinal slices revealed autofluorescence signals over multiple functional layers, including the photoreceptor layer (PRL), outer nuclear layer (ONL), outer plexiform layer (OPL), inner nuclear layer (INL), inner plexiform layer (IPL), and ganglion cell layer (GCL). Using flat-mounted retinas, depth-resolved imaging of individual retinal layers further confirmed multiple sources of autofluorescence signals. Cellular structures were clearly observed at the PRL, ONL, INL, and GCL. At the PRL, the autofluorescence was dominantly recorded from the intracellular compartment of the photoreceptors; while mixed intracellular and extracellular autofluorescence signals were observed at the ONL, INL, and GCL. High resolution autofluorescence imaging clearly revealed mosaic organization of rod and cone photoreceptors; and sub-cellular bright autofluorescence spots, which might relate to connecting cilium, was observed in the cone photoreceptors only. Moreover, single-cone and double-cone outer segments could be directly differentiated.

  3. Simultaneous morphological and functional imaging of the honeybee's brain by two-photon microscopy

    International Nuclear Information System (INIS)

    Haase, A.

    2011-01-01

    Thanks to its rather simply structured but highly performing brain, the honeybee (Apis mellifera) is an important model for neurobiological studies. Therefore there is a great need for new functional imaging modalities adapted to this species. Herein we give a detailed report on the development and performance of a platform for in vivo functional and morphological imaging of the honeybee's brain, focusing on its primary olfactory centres, the antennal lobes (ALs). The experimental setup consists of a two-photon microscope combined with a synchronized odour stimulus generator. Our imaging platform allows to simultaneously obtain both morphological measurements of the ALs functional units, the glomeruli, and in vivo calcium recording of their neural activity. We were able to record the characteristic glomerular response maps to odour stimuli applied to the bee's antennae. Our approach offers several advantages over the commonly used conventional fluorescence microscopy. Two-photon microscopy provides substantial enhancement in both spatial and temporal resolutions, while minimizing photo damage. Calcium recordings show a more than fourfold improvement in the functional signal with respect to the techniques available up to now. Finally, the extended penetration depth, thanks to the infrared excitation, allows the functional imaging of profound glomeruli which have not been optically accessible up to now.

  4. Multicolor Fluorescence Writing Based on Host-Guest Interactions and Force-Induced Fluorescence-Color Memory.

    Science.gov (United States)

    Matsunaga, Yuki; Yang, Jye-Shane

    2015-06-26

    A new strategy is reported for multicolor fluorescence writing on thin solid films with mechanical forces. This concept is illustrated by the use of a green-fluorescent pentiptycene derivative 1, which forms variably colored fluorescent exciplexes: a change from yellow to red was observed with anilines, and fluorescence quenching (a change to black) occurred in the presence of benzoquinone. Mechanical forces, such as grinding and shearing, induced a crystalline-to-amorphous phase transition in both the pristine and guest-adsorbed solids that led to a change in the fluorescence color (mechanofluorochromism) and a memory of the resulting color. Fluorescence drawings of five or more colors were created on glass or paper and could be readily erased by exposure to air and dichloromethane fumes. The structural and mechanistic aspects of the observations are also discussed. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Two-photon polarization data storage in bacteriorhodopsin films and its potential use in security applications

    Energy Technology Data Exchange (ETDEWEB)

    Imhof, Martin; Hampp, Norbert, E-mail: hampp@staff.uni-marburg.de [Department of Chemistry, Material Sciences Center, University of Marburg, Hans-Meerwein-Str., D-35032 Marburg (Germany); Rhinow, Daniel [Max-Planck-Institute of Biophysics, Max-von-Laue-Straße 3, D-60438 Frankfurt (Germany)

    2014-02-24

    Bacteriorhodopsin (BR) films allow write-once-read-many recording of polarization data by a two-photon-absorption (TPA) process. The optical changes in BR films induced by the TPA recording were measured and the Müller matrix of a BR film was determined. A potential application of BR films in security technology is shown. Polarization data can be angle-selective retrieved with high signal-to-noise ratio. The BR film does not only carry optical information but serves also as a linear polarizer. This enables that polarization features recorded in BR films may be retrieved by merely using polarized light from a mobile phone display.

  6. Transient behaviour of EIT and EIA in an optical-radio two-photon coupling configuration

    Science.gov (United States)

    Li, Xiaoli; Yang, Zicai; Shang, Yaxuan

    2012-11-01

    Both electromagnetically induced absorption (EIA) and transparency (EIT) can be obtained in a modified quasi-lambda four level system consisting of an optical-radio two-photon coupling field and a probing field. A physical account of EIA and EIT is given in terms of a transient state picture in this paper. It can be seen that the optical coupling field in this quasi-lambda four level system has a crucial effect on the forming of EIA and EIT. An EIA is observed under a resonant optical coupling and it evolves into an EIT when there is a detuning.

  7. Three-dimensional cell organization leads to almost immediate HRE activity as demonstrated by molecular imaging of MG-63 spheroids using two-photon excitation microscopy.

    Science.gov (United States)

    Indovina, Paola; Collini, Maddalena; Chirico, Giuseppe; Santini, Maria Teresa

    2007-02-20

    Hypoxia through HRE (hypoxia-responsive element) activity in MG-63 human osteosarcoma cells grown in monolayer and as very small, three-dimensional tumor spheroids was investigated using molecular imaging techniques. MG-63 cells were stably transfected with a vector constructed with multiple copies of the HRE sequence of the human vascular endothelial growth factor (VEGF) gene and with the enhanced green fluorescent protein (EGFP) coding sequence. During hypoxia when HIF-1alpha (hypoxia-inducible factor-1alpha) is stabilized, the binding of HIF-1 to the HRE sequences of the vector allows the transcription of EGFP and the appearance of fluorescence. Transfected monolayer cells were characterized by flow cytometric analysis in response to various hypoxic conditions and HIF-1alpha expression in these cells was assessed by Western blotting. Two-photon excitation (TPE) microscopy was then used to examine both MG-63-transfected monolayer cells and spheroids at 2 and 5 days of growth in normoxic conditions. Monolayer cells reveal almost no fluorescence, whereas even very small spheroids (HRE activation. This activation of the HRE sequences, which control a wide variety of genes, suggests that monolayer cells and spheroids of the MG-63 cell line have different genes activated and thus diverse functional activities.

  8. THE REGIONAL DIFFERENCES OF GPP ESTIMATION BY SOLAR INDUCED FLUORESCENCE

    Directory of Open Access Journals (Sweden)

    X. Wang

    2018-04-01

    Full Text Available Estimating gross primary productivity (GPP at large spatial scales is important for studying the global carbon cycle and global climate change. In this study, the relationship between solar-induced chlorophyll fluorescence (SIF and GPP is analysed in different levels of annual average temperature and annual total precipitation respectively using simple linear regression analysis. The results showed high correlation between SIF and GPP, when the area satisfied annual average temperature in the range of −5 °C to 15 °C and the annual total precipitation is higher than 200 mm. These results can provide a basis for future estimation of GPP research.

  9. The Regional Differences of Gpp Estimation by Solar Induced Fluorescence

    Science.gov (United States)

    Wang, X.; Lu, S.

    2018-04-01

    Estimating gross primary productivity (GPP) at large spatial scales is important for studying the global carbon cycle and global climate change. In this study, the relationship between solar-induced chlorophyll fluorescence (SIF) and GPP is analysed in different levels of annual average temperature and annual total precipitation respectively using simple linear regression analysis. The results showed high correlation between SIF and GPP, when the area satisfied annual average temperature in the range of -5 °C to 15 °C and the annual total precipitation is higher than 200 mm. These results can provide a basis for future estimation of GPP research.

  10. Inclusive D*(+/-) production in two photon collisions at LEP

    CERN Document Server

    Prokofiev, Denis Olegovich

    2001-01-01

    In this thesis I present my results on the measurement of the open charm production in two-photon collision events done with the L3 detector at Large Electron Positron machine (LEP). The data sample was collected from 1997 through 2000 at center-of-mass energies ranging from 183 GeV to 209 GeV, corresponding to a total integrated luminosity of 683.4pb −1. The open charm production in two-photon collision events extrapolated to the full phase space is estimated to be: s&parl0;e+e-&rarrr;e +e-cc&d1;X&parr0;=9 23±69±109±222pb. The differential cross sections d s /dpT(D*±) and d s /d:η(D*±): are also measured as functions of transverse momentum pT(D*±) and the absolute value of pseudorapidity :η(D*±):, respectively. A fit to the data estimating the relative contributions of Direct and Resolved open charm production mechanisms is performed, giving (28.7 ± 5.6)% and (71.3 ± 8.8)%, respectively. Using those relative fractions, the Direct and Resolved process cross sections yield: s&p...

  11. Temperature dependence of the two photon absorption in indium arsenide

    International Nuclear Information System (INIS)

    Berryman, K.W.; Rella, C.W.

    1995-01-01

    Nonlinear optical processes in semiconductors have long been a source of interesting physics. Two photon absorption (TPA) is one such process, in which two photons provide the energy for the creation of an electron-hole pair. Researchers at other FEL centers have studied room temperature TPA in InSb, InAs, and HgCdTe. Working at the Stanford Picosecond FEL Center, we have extended and refined this work by measuring the temperature dependence of the TPA coefficient in InAs over the range from 80 to 350 K at four wavelengths: 4.5, 5.06, 6.01, and 6.3 microns. The measurements validate the functional dependence of recent band structure calculations with enough precision to discriminate parabolic from non-parabolic models, and to begin to observe smaller effects, such as contributions due to the split-off band. These experiments therefore serve as a strong independent test of the Kane band theory, as well as providing a starting point for detailed observations of other nonlinear absorption mechanisms

  12. Two-photon emission and multiphoton absorption by atoms

    International Nuclear Information System (INIS)

    Mu, X.

    1988-01-01

    This thesis consists of investigations of two problems concerning photon-atom interactions. The first topic deals with two-photon transitions in atomic inner shells. An independent-particle model has been used to describe the two-photon transitions between different inner-shell electron states. The first relativistic self-consistent-field calculation of these transition rates in Ag, Mo, and Xe has been carried out. The theoretical results are compared with recent measurements. Good agreement with measured rates is found except in some cases where more reliable experiments still need to be done. The second topic is multiphoton multiionization of atoms. The maximum entropy principle has been employed in this theoretical investigation. A detailed statistical analysis of measured ionic charge distributions produced in strong laser pulses has been carried out. The results of this analysis indicates that the charge-state distribution is a Poissonian, rather than the binomial which prevails under infrared radiation, and hence that ionization occurs stepwise during the pulse. This result is shown to be consistent with experimental data

  13. Monte Carlo simulation of two-photon processes

    International Nuclear Information System (INIS)

    Daverveldt, P.H.W.M.

    1985-01-01

    During the last two decades e + e - collider experiments provided physicists with a wealth of important discoveries concerning elementary particle physics. This thesis explains in detail how the Monte Carlo approach can be applied to establish the comparison between two-photon experiments and theory. The author describes the main motives for and objectives of two-photon research. He defines the kinematics and pays attention to some special kinematical regions. Also a popular approximation for the exact differential cross section is reviewed. Next he discusses the calculation of the complete lowest order cross section for processes with four leptons in the final state and for reactions such as e + e - →e + e - qanti q, e + e - →μ + μ - qanti q. Radiative corrections to the multiperipheral diagrams are considered. The author explains in detail the distinction between soft and hard photon corrections which turns out to be somewhat more tricky than in the case of radiative corrections to one-photon processes. Finally, he presents some results which were obtained by using the event generators. (Auth.)

  14. Introducing inducible fluorescent split cholesterol oxidase to mammalian cells.

    Science.gov (United States)

    Chernov, Konstantin G; Neuvonen, Maarit; Brock, Ivonne; Ikonen, Elina; Verkhusha, Vladislav V

    2017-05-26

    Cholesterol oxidase (COase) is a bacterial enzyme catalyzing the first step in the biodegradation of cholesterol. COase is an important biotechnological tool for clinical diagnostics and production of steroid drugs and insecticides. It is also used for tracking intracellular cholesterol; however, its utility is limited by the lack of an efficient temporal control of its activity. To overcome this we have developed a regulatable fragment complementation system for COase cloned from Chromobacterium sp. The enzyme was split into two moieties that were fused to FKBP (FK506-binding protein) and FRB (rapamycin-binding domain) pair and split GFP fragments. The addition of rapamycin reconstituted a fluorescent enzyme, termed split GFP-COase, the fluorescence level of which correlated with its oxidation activity. A rapid decrease of cellular cholesterol induced by intracellular expression of the split GFP-COase promoted the dissociation of a cholesterol biosensor D4H from the plasma membrane. The process was reversible as upon rapamycin removal, the split GFP-COase fluorescence was lost, and cellular cholesterol levels returned to normal. These data demonstrate that the split GFP-COase provides a novel tool to manipulate cholesterol in mammalian cells. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  15. Simple approach to three-color two-photon microscopy by a fiber-optic wavelength convertor.

    Science.gov (United States)

    Li, Kuen-Che; Huang, Lynn L H; Liang, Jhih-Hao; Chan, Ming-Che

    2016-11-01

    A simple approach to multi-color two-photon microscopy of the red, green, and blue fluorescent indicators was reported based on an ultra-compact 1.03-μm femtosecond laser and a nonlinear fiber. Inside the nonlinear fiber, the 1.03-μm laser pulses were simultaneously blue-shifted to 0.6~0.8 μm and red-shifted to 1.2~1.4 μm region by the Cherenkov radiation and fiber Raman gain effects. The wavelength-shifted 0.6~0.8 μm and 1.2~1.4 μm radiations were co-propagated with the residual non-converted 1.03-μm pulses inside the same nonlinear fiber to form a fiber-output three-color femtosecond source. The application of the multi-wavelength sources on multi-color two-photon fluorescence microscopy were also demonstrated. Overall, due to simple system configuration, convenient wavelength conversion, easy wavelength tunability within the entire 0.7~1.35 μm bio-penetration window and less requirement for high power and bulky light sources, the simple approach to multi-color two-photon microscopy could be widely applicable as an easily implemented and excellent research tool for future biomedical and possibly even clinical applications.

  16. Two-photon transitions driven by a combination of diode and femtosecond lasers.

    Science.gov (United States)

    Moreno, Marco P; Nogueira, Giovana T; Felinto, Daniel; Vianna, Sandra S

    2012-10-15

    We report on the combined action of a cw diode laser and a train of ultrashort pulses when each of them drives one step of the 5S-5P-5D two-photon transition in rubidium vapor. The fluorescence from the 6P(3/2) state is detected for a fixed repetition rate of the femtosecond laser while the cw-laser frequency is scanned over the rubidium D(2) lines. This scheme allows for a velocity selective spectroscopy in a large spectral range including the 5D(3/2) and 5D(5/2) states. The results are well described in a simplified frequency domain picture, considering the interaction of each velocity group with the cw laser and a single mode of the frequency comb.

  17. Quantum theory of two-photon wavepacket interference in a beamsplitter

    International Nuclear Information System (INIS)

    Wang, Kaige

    2006-01-01

    A general theory is derived for the interference of a two-photon wavepacket in a beamsplitter. The theory is presented in the Schroedinger picture so that the quantum nature of the two-photon interference is explicitly revealed. We find that the topological symmetry of the probability-amplitude spectrum of the two-photon wavepacket dominates the nature of the two-photon interference, which may be distinguished by the increase or decrease of the coincidence probability in the absence of interference. However, two-photon entanglement can be identified by the nature of the interference. We demonstrate the necessary and sufficient conditions for perfect two-photon interference. It is shown that a two-photon entangled state with an anti-symmetric spectrum passes through a 50/50 beamsplitter with perfect transparency. The theory provides us with a unified understanding of the various two-photon interference effects. (topical review)

  18. Reactive quenching of two-photon excited xenon atoms by Cl2

    International Nuclear Information System (INIS)

    Bruce, M.R.; Layne, W.B.; Meyer, E.; Keto, J.W.

    1987-01-01

    Total binary and tertiary quench rates have been measured for the reaction Xe (5p 5 6p) + Cl 2 at thermal temperatures. Xenon atoms are excited by state-selective, two-photon absorption with a uv laser. The time dependent fluorescence from the excited atom in the IR and from XeCl* (B) product near 308 nm have been measured with subnanosecond time resolution. The decay rates are measured as a function of Cl 2 pressure to 20 Torr and Xe pressure to 400 Torr. The measured reaction rates (k 2 ∼ 10 -9 cm 3 sec -1 ) are consistent with a harpoon model described in a separate paper. We also measure large termolecular reaction rates for collisions with xenon atoms (k 3 ∼ 10 -28 cm 6 sec -1 ). Total product fluorescence has been examined using a gated optical multichannel analyzer. We measure unit branching fractions for high vibrational levels of XeCl* (B) with very little C state fluorescence observed. The measured termolecular rates suggest similar processes will dominate at the high buffer-gas pressures used in XeCl lasers. The effect of these large reactive cross sections for neutral xenon atoms on models of the XeCl laser will be discussed

  19. Development of a two photon microscope for tracking Drosophila larvae

    Science.gov (United States)

    Karagyozov, Doycho; Mihovilovic Skanata, Mirna; Gershow, Marc

    Current in vivo methods for measuring neural activity in Drosophila larva require immobilization of the animal. Although we can record neural signals while stimulating the sensory organs, we cannot read the behavioral output because we have prevented the animal from moving. Many research questions cannot be answered without observation of neural activity in behaving (freely-moving) animals. We incorporated a Tunable Acoustic Gradient (TAG) lens into a two-photon microscope to achieve a 70kHz axial scan rate, enabling volumetric imaging at tens of hertz. We then implemented a tracking algorithm based on a Kalman filter to maintain the neurons of interest in the field of view and in focus during the rapid three dimensional motion of a free larva. Preliminary results show successful tracking of a neuron moving at speeds reaching 500 μm/s. NIH Grant 1DP2EB022359 and NSF Grant PHY-1455015.

  20. Two-photon resonant, stimulated processes in krypton and xenon

    International Nuclear Information System (INIS)

    Miller, J.C.

    1988-11-01

    Both on-axis and conical emissions have been observed following two-photon pumping of the 5p states of krypton and the 6p', 7p, 8p, and 4f states of xenon. In the former case, coherent emissions from the 5p states to the 5s are observed, and in the latter case, many p→s, d→p, and f→d cascade emissions are observed. By analogy to the well-studied alkali and alkaline earth examples, the emissions are discussed in terms of amplified spontaneous emission (ASE), stimulated hyper-Raman scattering, and parametric four-wave mixing. The physical processes responsible for the conical emission and for intensity anomalies in the xenon p→s emissions are not understood at present. Interference effects due to coherent cancellation between competing excitation pathways may be occurring. 4 refs., 3 figs

  1. Whole brain imaging with Serial Two-Photon Tomography

    Directory of Open Access Journals (Sweden)

    Stephen P Amato

    2016-03-01

    Full Text Available Imaging entire mouse brains at submicron resolution has historically been a challenging undertaking and largely confined to the province of dedicated atlasing initiatives. The has limited systematic investigations into important areas of neuroscience, such as neural circuits, brain mapping and neurodegeneration. In this paper, we describe in detail Serial Two-Photon (STP tomography, a robust, reliable method for imaging entire brains with histological detail. We provide examples of how the basic methodology can be extended to other imaging modalities, such as optical coherence tomography, in order to provide unique contrast mechanisms. Furthermore we provide a survey of the research that STP tomography has enabled in the field of neuroscience, provide examples of how this technology enables quantitative whole brain studies, and discuss the current limitations of STP tomography-based approaches

  2. N-dimensional integrability from two-photon coalgebra symmetry

    International Nuclear Information System (INIS)

    Ballesteros, Angel; Blasco, Alfonso; Herranz, Francisco J

    2009-01-01

    A wide class of Hamiltonian systems with N degrees of freedom and endowed with, at least, (N - 2) functionally independent integrals of motion in involution is constructed by making use of the two-photon Lie-Poisson coalgebra (h 6 , Δ). The set of (N - 2) constants of the motion is shown to be a universal one for all these Hamiltonians, irrespective of the dependence of the latter on several arbitrary functions and N free parameters. Within this large class of quasi-integrable N-dimensional Hamiltonians, new families of completely integrable systems are identified by finding explicitly a new independent integral I through the analysis of the sub-coalgebra structure of h 6 . In particular, new completely integrable N-dimensional Hamiltonians describing natural systems, geodesic flows and static electromagnetic Hamiltonians are presented

  3. Phonon-assisted two-photon exciton transitions in semiconductors

    International Nuclear Information System (INIS)

    Hassan, A.R.

    1987-08-01

    The theory of phonon-assisted two-photon transitions to excitonic states in semiconductors has been theoretically investigated. The effects of both the nonparabolicity of the band and the degeneracy of the valence band have been taken into account. Expressions for the absorption coefficient through different band models are calculated. The numerical applications to CdI 2 and GaP show that the 4-band model gives the dominant contribution which leads to a final s-exciton state. An exciton peak appears at an energy which is close to that recently observed in CdI 2 . The non-parabolic effect enhances the absorption coefficient by a two-order of magnitude. (author). 6 refs, 1 fig., 1 tab

  4. Clinical multiphoton tomography and clinical two-photon microendoscopy

    Science.gov (United States)

    König, Karsten; Bückle, Rainer; Weinigel, Martin; Elsner, Peter; Kaatz, Martin

    2009-02-01

    We report on applications of high-resolution clinical multiphoton tomography based on the femtosecond laser system DermaInspectTM with its flexible mirror arm in Australia, Asia, and Europe. Applications include early detection of melanoma, in situ tracing of pharmacological and cosmetical compounds including ZnO nanoparticles in the epidermis and upper dermis, the determination of the skin aging index SAAID as well as the study of the effects of anti-aging products. In addition, first clinical studies with novel rigid high-NA two-photon 1.6 mm GRIN microendoscopes have been conducted to study the effect of wound healing in chronic wounds (ulcus ulcera) as well as to perform intrabody imaging with subcellular resolution in small animals.

  5. Optimizing pulse compressibility in completely all-fibered Ytterbium chirped pulse amplifiers for in vivo two photon laser scanning microscopy.

    Science.gov (United States)

    Fernández, A; Grüner-Nielsen, L; Andreana, M; Stadler, M; Kirchberger, S; Sturtzel, C; Distel, M; Zhu, L; Kautek, W; Leitgeb, R; Baltuska, A; Jespersen, K; Verhoef, A

    2017-08-01

    A simple and completely all-fiber Yb chirped pulse amplifier that uses a dispersion matched fiber stretcher and a spliced-on hollow core photonic bandgap fiber compressor is applied in nonlinear optical microscopy. This stretching-compression approach improves compressibility and helps to maximize the fluorescence signal in two-photon laser scanning microscopy as compared with approaches that use standard single mode fibers as stretcher. We also show that in femtosecond all-fiber systems, compensation of higher order dispersion terms is relevant even for pulses with relatively narrow bandwidths for applications relying on nonlinear optical effects. The completely all-fiber system was applied to image green fluorescent beads, a stained lily-of-the-valley root and rat-tail tendon. We also demonstrated in vivo imaging in zebrafish larvae, where we simultaneously measure second harmonic and fluorescence from two-photon excited red-fluorescent protein. Since the pulses are compressed in a fiber, this source is especially suited for upgrading existing laser scanning (confocal) microscopes with multiphoton imaging capabilities in space restricted settings or for incorporation in endoscope-based microscopy.

  6. Quantitative laser-induced fluorescence measurements of nitric oxide in a heavy-duty Diesel engine

    NARCIS (Netherlands)

    Verbiezen, K.; Klein-Douwel, R. J. H.; van Viet, A. P.; Donkerbroek, A. J.; Meerts, W. L.; Dam, N. J.; ter Meulen, J. J.

    2007-01-01

    We present quantitative, in-cylinder, UV-laser-induced fluorescence measurements of nitric oxide in a heavy-duty Diesel engine. Processing of the raw fluorescence signals includes a detailed correction, based on additional measurements, for the effect of laser beam and fluorescence attenuation, and

  7. Laser Induced Fluorescence of Helium Ions in a Helicon Plasma

    Science.gov (United States)

    Compton, C. S.; Biloui, C.; Hardin, R. A.; Keesee, A. M.; Scime, E. E.; Boivin, R.

    2003-10-01

    The lack of a suitable Laser Induced Fluorescence (LIF) scheme for helium ions at visible wavelengths has prevented LIF from being employed in helium plasmas for measurements of ion temperature and bulk ion flow speeds. In this work, we will discuss our attempts to perform LIF of helium ions in a helicon source plasma using an infrared, tunable diode laser operating at 1012.36 nm. The infrared transition corresponds to excitation from the n = 4 level (4f ^2F) to the n = 5 (5g ^2G) level of singly ionized helium and therefore requires substantial electron temperatures (> 10 eV) to maintain an adequate ion population in the n = 4 state. Calculations using a steady state coronal model predict that the n = 4 state population will be 25% larger than the n = 5 population for our experimental conditions. The fluorescence decay from the n = 5 (5f ^2F) level of singly ionized helium level to the n = 3 (3d ^2D) level at 320.31 nm is monitored as the diode laser is swept through 10 GHz around the 1012.36 nm line. Note that the fluorescence emission requires a collisionally coupled transition between two different n = 5 quantum states. We will also present measurements of the emission intensities of both the 1012.36 nm and the 320.31 nm lines as a function of source neutral pressure, rf power, and plasma density. This work supported by the U.S. DoE EPSCoR Lab Partnership Program.

  8. Study on two-color planar laser induced fluorescence thermometry

    International Nuclear Information System (INIS)

    Li Shaodan; Tan Sichao; Gao Puzhen; Lin Yuansheng

    2014-01-01

    Many of the convection heat transfer process are involved in the research of nuclear reactor thermal hydraulics. To experimentally determine the variation of the temperature field in those processes is important for the design and safety operation of the nuclear reactor. The application of the two-color planar laser induced fluorescence (PLIF) in the measurements of fluid temperature distribution is discussed in the paper. The laser dyes used here is rhodamine B (RhB) with negative temperature coefficient and fluorescein 27 (F127) with positive temperature coefficient. The beam of the laser light is adjusted to laser sheet by using the lens group. The fluid with dyes is excited by this laser sheet in a specific plane and temperature dependent fluorescence is released. The temperature field of the plane can be determined through the intensity information. Some technical aspects encountered in the application of the two-laser PLIF are discussed in the paper, such as the spectra characteristic of the dyes and the separation of the spectra. The calibration temperature is higher than the water saturation temperature (at atmosphere pressure). (authors)

  9. Initial operation of the LEDA beam-induced fluorescence diagnostic

    International Nuclear Information System (INIS)

    Kamperschroer, James H.; Gurd, Pamela A.; Martinez, Derwin G.; Gilpatrick, J. Douglas; Shurter, R. Bradford; Stettler, Matthew W.; Madsen, David W.; O'Hara, James F.; Sage, Joan; Schaefer, Timothy L.

    2000-01-01

    A diagnostic based on beam-induced fluorescence has been developed and used to examine the expanded beam in the High-Energy Beam Transport (HEBT) section of the Low Energy Demonstration Accelerator (LEDA). The system consists of a camera, a gas injector, a spectrometer, and a control system. Gas is injected to provide a medium for the beam to excite, the camera captures the resulting image of the fluorescing gas, and the spectrometer measures the spectrum of the emitted light. EPICS was used to control the camera and acquire and store images. Data analysis is presently being performed offline. A Kodak DCS420m professional CCD camera is the primary component of the optical system. InterScience, Inc. modified the camera with the addition of a gain of 4000 image intensifier, thereby producing an intensified camera with a sensitivity of ∼0.5 milli-lux. Light is gathered with a 1 '' format, 16-160 mm, Computar zoom lens. This lens is attached to the camera via a Century Precision Optics relay lens. Images obtained using only hydrogen from the beam stop exhibited features not yet understood. Images with good signal-to-noise ratio were obtained with the injection of sufficient nitrogen to raise the HEBT pressure to 2-8x10 -6 torr. Two strong nitrogen lines, believed to be of the first negative group of N 2 + , were identified at 391 and 428 nm

  10. Two-Photon Absorbing Molecules as Potential Materials for 3D Optical Memory

    Directory of Open Access Journals (Sweden)

    Kazuya Ogawa

    2014-01-01

    Full Text Available In this review, recent advances in two-photon absorbing photochromic molecules, as potential materials for 3D optical memory, are presented. The investigations introduced in this review indicate that 3D data storage processing at the molecular level is possible. As 3D memory using two-photon absorption allows advantages over existing systems, the use of two-photon absorbing photochromic molecules is preferable. Although there are some photochromic molecules with good properties for memory, in most cases, the two-photon absorption efficiency is not high. Photochromic molecules with high two-photon absorption efficiency are desired. Recently, molecules having much larger two-photon absorption cross sections over 10,000 GM (GM= 10−50 cm4 s molecule−1 photon−1 have been discovered and are expected to open the way to realize two-photon absorption 3D data storage.

  11. Sputtering of amorphous carbon layers studied by laser induced fluorescence

    International Nuclear Information System (INIS)

    Pasch, E.

    1992-07-01

    In order to minimize the radiation losses, it is desirable to keep the plasmas in nuclear fusion devices free of high-Z-impurities. Therefore, the walls of TEXTOR and other tokamaks are covered with thin layers of amorphous carbon layers (a-C:H) or amorphous carbon/boron layers (a-C/B:H). The sputtering behaviour of these layers has been studied under bombardment by Ar + ions with energies of 1.5 keV and current densities of a few mA/cm 2 . Investigations of these coatings were carried out with the object to measure the velocity distribution of the sputtered atoms and the sputtered yields by laser induced fluorescence in the vacuum ultraviolet. (orig.)

  12. Two-Photon Ghost Image and Interference-Diffraction

    Science.gov (United States)

    Shih, Y. H.; Sergienko, A. V.; Pittman, T. B.; Strekalov, D. V.; Klyshko, D. N.

    1996-01-01

    One of the most surprising consequences of quantum mechanics is entanglement of two or more distance particles. The two-particle entangled state was mathematically formulated by Schrodinger. Based on this unusual quantum behavior, EPR defined their 'physical reality' and then asked the question: 'Can Quantum-Mechanical Description of Physical Reality Be Considered Complete?' One may not appreciate EPR's criterion of physical reality and insist that 'no elementary quantum phenomenon is a phenomenon until it is a recorded phenomenon'. Optical spontaneous parametric down conversion (SPDC) is the most effective mechanism to generate an EPR type entangled two-photon state. In SPDC, an optical beam, called the pump, is incident on a birefringent crystal. The pump is intense enough so that nonlinear effects lead to the conversion of pump photons into pairs of photons, historically called signal and idler. Technically, the SPDC is said to be type-1 or type-2, depending on whether the signal and idler beams have parallel or orthogonal polarization. The SPDC conversion efficiency is typically on the order of 10(exp -9) to 10(exp -11), depending on the SPDC nonlinear material. The signal and idler intensities are extremely low, only single photon detection devices can register them. The quantum entanglement nature of SPDC has been demonstrated in EPR-Bohm experiments and Bell's inequality measurements. The following two experiments were recently performed in our laboratory, which are more closely related to the original 1935 EPR gedankenezperiment. The first experiment is a two-photon optical imaging type experiment, which has been named 'ghost image' by the physics community. The signal and idler beams of SPDC are sent in different directions, so that the detection of the signal and idler photons can be performed by two distant photon counting detectors. An aperture object (mask) is placed in front of the signal photon detector and illuminated by the signal beam through a

  13. Laser-induced fluorescence detection strategies for sodium atoms and compounds in high-pressure combustors

    Science.gov (United States)

    Weiland, Karen J. R.; Wise, Michael L.; Smith, Gregory P.

    1993-01-01

    A variety of laser-induced fluorescence schemes were examined experimentally in atmospheric pressure flames to determine their use for sodium atom and salt detection in high-pressure, optically thick environments. Collisional energy transfer plays a large role in fluorescence detection. Optimum sensitivity, at the parts in 10 exp 9 level for a single laser pulse, was obtained with the excitation of the 4p-3s transition at 330 nm and the detection of the 3d-3p fluorescence at 818 nm. Fluorescence loss processes, such as ionization and amplified spontaneous emission, were examined. A new laser-induced atomization/laser-induced fluorescence detection technique was demonstrated for NaOH and NaCl. A 248-nm excimer laser photodissociates the salt molecules present in the seeded flames prior to atom detection by laser-induced fluorescence.

  14. In situ electrical and thermal monitoring of printed electronics by two-photon mapping.

    Science.gov (United States)

    Pastorelli, Francesco; Accanto, Nicolò; Jørgensen, Mikkel; van Hulst, Niek F; Krebs, Frederik C

    2017-06-19

    Printed electronics is emerging as a new, large scale and cost effective technology that will be disruptive in fields such as energy harvesting, consumer electronics and medical sensors. The performance of printed electronic devices relies principally on the carrier mobility and molecular packing of the polymer semiconductor material. Unfortunately, the analysis of such materials is generally performed with destructive techniques, which are hard to make compatible with in situ measurements, and pose a great obstacle for the mass production of printed electronics devices. A rapid, in situ, non-destructive and low-cost testing method is needed. In this study, we demonstrate that nonlinear optical microscopy is a promising technique to achieve this goal. Using ultrashort laser pulses we stimulate two-photon absorption in a roll coated polymer semiconductor and map the resulting two-photon induced photoluminescence and second harmonic response. We show that, in our experimental conditions, it is possible to relate the total amount of photoluminescence detected to important material properties such as the charge carrier density and the molecular packing of the printed polymer material, all with a spatial resolution of 400 nm. Importantly, this technique can be extended to the real time mapping of the polymer semiconductor film, even during the printing process, in which the high printing speed poses the need for equally high acquisition rates.

  15. Determining the quark charges by one and two photon processes

    International Nuclear Information System (INIS)

    Janah, A.

    1982-01-01

    Testable predictions are presented, which may be used to decide between the gauge theories of integer and fractionally charged quarks (icq and fcq). Two distinctive features of icq are exploited, namely (a) presence of color non-singlet components in weak and electromagnetic currents and (b) possible liberation of color non-singlet states above a threshold energy. Consequences are sought in lepton-hadron interaction processes, taking into account the known color-suppression effect. Single photon/weak-boson processes such as nuN → nuX distinguish between icq and fcq only above color-threshold. Experimental consequences of color-liberation in the above process are obtained. It is found that the gluon-parton contribution survives color-suppression to produce a significant rise in the structure functions when color-threshold is exceeded. Two-photon processes such as e + e - → e + e - + 2 jets distinguish between the two theories even below color threshold. To obtain the icq predictions for this process, one must take into account (a) the (momentum-dependent) color suppression and (b) the added contribution from pair production of charged gluons

  16. Two-Photon-Absorption Scheme for Optical Beam Tracking

    Science.gov (United States)

    Ortiz, Gerardo G.; Farr, William H.

    2011-01-01

    A new optical beam tracking approach for free-space optical communication links using two-photon absorption (TPA) in a high-bandgap detector material was demonstrated. This tracking scheme is part of the canonical architecture described in the preceding article. TPA is used to track a long-wavelength transmit laser while direct absorption on the same sensor simultaneously tracks a shorter-wavelength beacon. The TPA responsivity was measured for silicon using a PIN photodiode at a laser beacon wavelength of 1,550 nm. As expected, the responsivity shows a linear dependence with incident power level. The responsivity slope is 4.5 x 10(exp -7) A/W2. Also, optical beam spots from the 1,550-nm laser beacon were characterized on commercial charge coupled device (CCD) and complementary metal-oxide semiconductor (CMOS) imagers with as little as 13.7 microWatts of optical power (see figure). This new tracker technology offers an innovative solution to reduce system complexity, improve transmit/receive isolation, improve optical efficiency, improve signal-to-noise ratio (SNR), and reduce cost for free-space optical communications transceivers.

  17. Higgs decay into two photons in a warped extra dimension

    Science.gov (United States)

    Hahn, Juliane; Hörner, Clara; Malm, Raoul; Neubert, Matthias; Novotny, Kristiane; Schmell, Christoph

    2014-05-01

    A detailed five-dimensional calculation of the Higgs-boson decay into two photons is performed in both the minimal and the custodially protected Randall-Sundrum (RS) model, where the Standard Model (SM) fields propagate in the bulk and the scalar sector lives on or near the IR brane. It is explicitly shown that the gauge invariance of the sum of diagrams involving bosonic fields in the SM also applies to the case of these RS scenarios. An exact expression for the amplitude in terms of the five-dimensional (5D) gauge-boson and fermion propagators is presented, which includes the full dependence on the Higgs-boson mass. Closed expressions for the 5D -boson propagators in the minimal and the custodial RS model are derived, which are valid to all orders in . In contrast to the fermion case, the result for the bosonic contributions to the amplitude is insensitive to the details of the localization of the Higgs profile on or near the IR brane. The various RS predictions for the rate of the process are compared with the latest LHC data, and exclusion regions for the RS model parameters are derived.

  18. Review of two-photon exchange in electron scattering

    Energy Technology Data Exchange (ETDEWEB)

    J. Arrington, P. G. Blunden, W. Melnitchouk

    2011-10-01

    We review the role of two-photon exchange (TPE) in electron-hadron scattering, focusing in particular on hadronic frameworks suitable for describing the low and moderate Q^2 region relevant to most experimental studies. We discuss the effects of TPE on the extraction of nucleon form factors and their role in the resolution of the proton electric to magnetic form factor ratio puzzle. The implications of TPE on various other observables, including neutron form factors, electroproduction of resonances and pions, and nuclear form factors, are summarized. Measurements seeking to directly identify TPE effects, such as through the angular dependence of polarization measurements, nonlinear epsilon contributions to the cross sections, and via e+p to e-p cross section ratios, are also outlined. In the weak sector, we describe the role of TPE and gamma-Z interference in parity-violating electron scattering, and assess their impact on the extraction of the strange form factors of the nucleon and the weak charge of the proton.

  19. Method for universal detection of two-photon polarization entanglement

    Science.gov (United States)

    Bartkiewicz, Karol; Horodecki, Paweł; Lemr, Karel; Miranowicz, Adam; Życzkowski, Karol

    2015-03-01

    Detecting and quantifying quantum entanglement of a given unknown state poses problems that are fundamentally important for quantum information processing. Surprisingly, no direct (i.e., without quantum tomography) universal experimental implementation of a necessary and sufficient test of entanglement has been designed even for a general two-qubit state. Here we propose an experimental method for detecting a collective universal witness, which is a necessary and sufficient test of two-photon polarization entanglement. It allows us to detect entanglement for any two-qubit mixed state and to establish tight upper and lower bounds on its amount. A different element of this method is the sequential character of its main components, which allows us to obtain relatively complicated information about quantum correlations with the help of simple linear-optical elements. As such, this proposal realizes a universal two-qubit entanglement test within the present state of the art of quantum optics. We show the optimality of our setup with respect to the minimal number of measured quantities.

  20. Evaluation of mitochondrial activity by two-photon absorption with near-field multioptical fiber probes

    Science.gov (United States)

    Kanazashi, Yasuaki; Takara, Naoshi; Iwami, Kentaro; Ohta, Yoshihiro; Umeda, Norihiro

    2018-04-01

    pH measurements enable the direct monitoring and evaluation of mitochondrial activity. We constructed a scanning near-field optical microscopy system with multioptical fiber probes using the two-photon absorption of a pH-sensitive fluorescent dye, SNARF-4F, to measure the activity difference of mitochondrial aggregates. pH can be monitored through the fluorescence intensity ratio (FIR) of SNARF-4F. We derived a calibration curve of the FIR as a function of pH. The FIR dynamic responses were measured by adding hydrochloric acid to the buffer solution. Using the developed system, we simultaneously measured the pH changes at two different locations in the SNARF-4F solution. Mitochondrial samples were prepared using optical tweezers to control the number and position of mitochondria. Mitochondrial pH changes (ΔpH) between 0.05 and 0.57 were observed after adding a nutritional supplement (malate and glutamate). In addition, in the comparative experiment on the activities of two mitochondrial populations, the obtained result suggested that the activity differs depending on the difference in the number of mitochondria.

  1. Intravital Confocal and Two-photon Imaging of Dual-color Cells and Extracellular Matrix Mimics

    Science.gov (United States)

    Bal, Ufuk; Andresen, Volker; Baggett, Brenda; Utzinger, Urs

    2013-01-01

    To optimize imaging of cells in three dimensional culture we studied confocal backscattering, Second Harmonic Generation (SHG) and autofluorescence as source of contrast in extracellular matrix (ECM) mimics and evaluated the attenuation as well as bleaching of endogenous cellular fluorescence signals. All common ECM mimics exhibit contrast observable with confocal reflectance microscopy. SHG imaging on collagen I based hydrogels provides high contrast and good optical penetration depth. Agarose is a useful embedding medium because it allows for large optical penetration and exhibits minimal autofluorescence while still providing good reflectance to detect voids in the embedding medium. We labeled breast cancer cells’ outline with DsRed2 and nucleus with eGFP. DsRed2 can be excited with confocal imaging at 568nm, and with two photon excitation (TPE) in the red and longer NIR. eGFP was excited at 488nm for confocal and in the NIR for TPE. While there is small difference in the bleaching rate for eGFP between confocal and TPE we observed significant difference for DsRed2 where bleaching is strongest during TPE in the red wavelengths and smallest during confocal imaging. After a few hundred microns depth in a collagen I hydrogel, TPE fluorescence becomes twice as strong compared to confocal imaging. PMID:23380006

  2. Ratiometric two-photon excited photoluminescence of quantum dots triggered by near-infrared-light for real-time detection of nitric oxide release in situ

    International Nuclear Information System (INIS)

    Jin, Hui; Gui, Rijun; Sun, Jie; Wang, Yanfeng

    2016-01-01

    Probe-donor integrated nanocomposites were developed from conjugating silica-coated Mn"2"+:ZnS quantum dots (QDs) with MoS_2 QDs and photosensitive nitric oxide (NO) donors (Fe_4S_3(NO)_7"−, RBS). Under excitation with near-infrared (NIR) light at 808 nm, the Mn"2"+:ZnS@SiO_2/MoS_2-RBS nanocomposites showed the dual-emissive two-photon excited photoluminescence (TPEPL) that induced RBS photolysis to release NO in situ. NO caused TPEPL quenching of Mn"2"+:ZnS QDs, but it produced almost no impact on the TPEPL of MoS_2 QDs. Hence, the nanocomposites were developed as a novel QDs-based ratiometric TPEPL probe for real-time detection of NO release in situ. The ratiometric TPEPL intensity is nearly linear (R"2 = 0.9901) with NO concentration in the range of 0.01∼0.8 μM, which corresponds to the range of NO release time (0∼15 min). The detection limit was calculated to be approximately 4 nM of NO. Experimental results confirmed that this novel ratiometric TPEPL probe possessed high selectivity and sensitivity for the detection of NO against potential competitors, and especially showed high detection performance for NIR-light triggered NO release in tumor intracellular microenvironments. These results would promote the development of versatile probe-donor integrated systems, also providing a facile and efficient strategy to real-time detect the highly controllable drug release in situ, especially in physiological microenvironments. - Highlights: • Mn"2"+:ZnS@SiO_2/MoS_2-RBS nanocomposites were developed as a novel ratiometric two-photon excited fluorescence probe. • This probe could conduct real-time detection of nitric oxide release in situ. • High feasibility of this probe was confirmed in tumor intracellular microenvironments.

  3. Large scale serial two-photon microscopy to investigate local vascular changes in whole rodent brain models of Alzheimer's disease

    Science.gov (United States)

    Delafontaine-Martel, P.; Lefebvre, J.; Damseh, R.; Castonguay, A.; Tardif, P.; Lesage, F.

    2018-02-01

    In this study, an automated serial two-photon microscope was used to image a fluorescent gelatin filled rodent's brain in 3D. A method to compute vascular density using automatic segmentation was combined with coregistration techniques to build group-level vasculature metrics. By studying the medial prefrontal cortex and the hippocampal formation of 3 age groups (2, 4.5 and 8 months old), we compared vascular density for both WT and an Alzheimer model transgenic brain (APP/PS1). We observe a loss of vascular density caused by the ageing process and we propose further analysis to confirm our results.

  4. Opposite reactivity of meningeal versus cortical microvessels to the nitric oxide donor glyceryl trinitrate evaluated in vivo with two-photon imaging.

    Directory of Open Access Journals (Sweden)

    Evgeny Pryazhnikov

    Full Text Available Vascular changes underlying headache in migraine patients induced by Glyceryl trinitrate (GTN were previously studied with various imaging techniques. Despite the long history of medical and experimental use of GTN, its effects on the brain vasculature are still poorly understood presumably due to low spatial resolution of the imaging modalities used so far. We took advantage of the micrometer-scale vertical resolution of two-photon microscopy to differentiate between the vasodynamic effects of GTN on meningeal versus cortical vessels imaged simultaneously in anesthetized rats through either thinned skull or glass-sealed cranial window. Intermediate and small calibre vessels were visualized in vivo by imaging intravascular fluorescent dextran, and detection of blood flow direction allowed identification of individual arterioles and venules. We found that i.p.-injected GTN induced a transient constriction of meningeal arterioles, while their cortical counterparts were, in contrast, dilated. These opposing effects of GTN were restricted to arterioles, whereas the effects on venules were insignificant. Interestingly, the NO synthase inhibitor L-NAME did not affect the diameter of meningeal vessels but induced a constriction of cortical vessels. The different cellular environment in cortex versus meninges as well as distinct vessel wall anatomical features probably play crucial role in the observed phenomena. These findings highlight differential region- and vessel-type-specific effects of GTN on cranial vessels, and may implicate new vascular mechanisms of NO-mediated primary headaches.

  5. Fluorescent tagged episomals for stoichiometric induced pluripotent stem cell reprogramming.

    Science.gov (United States)

    Schmitt, Christopher E; Morales, Blanca M; Schmitz, Ellen M H; Hawkins, John S; Lizama, Carlos O; Zape, Joan P; Hsiao, Edward C; Zovein, Ann C

    2017-06-05

    Non-integrating episomal vectors have become an important tool for induced pluripotent stem cell reprogramming. The episomal vectors carrying the "Yamanaka reprogramming factors" (Oct4, Klf, Sox2, and L-Myc + Lin28) are critical tools for non-integrating reprogramming of cells to a pluripotent state. However, the reprogramming process remains highly stochastic, and is hampered by an inability to easily identify clones that carry the episomal vectors. We modified the original set of vectors to express spectrally separable fluorescent proteins to allow for enrichment of transfected cells. The vectors were then tested against the standard original vectors for reprogramming efficiency and for the ability to enrich for stoichiometric ratios of factors. The reengineered vectors allow for cell sorting based on reprogramming factor expression. We show that these vectors can assist in tracking episomal expression in individual cells and can select the reprogramming factor dosage. Together, these modified vectors are a useful tool for understanding the reprogramming process and improving induced pluripotent stem cell isolation efficiency.

  6. Grey-grey separate spatial soliton pairs in a biased series two-photon centrosymmetric photorefractive crystals circuit

    International Nuclear Information System (INIS)

    Ji, Xuanmang; Wang, Jinlai; Jiang, Qichang; Liu, Jinsong

    2012-01-01

    Grey-grey separate spatial soliton pairs are predicted in a biased series circuit consisting of two centrosymmetric photorefractive (PR) crystals with the two-photon PR effect. The numerical results show that two grey solitons in a soliton pair can affect each other by the light-induced current. The effects of the intensity of solitary waves and gating lights on the normalized profiles and the dynamical evolutions of solitons are discussed.

  7. Effect of the coherent cancellation of the two-photon resonance on the generation of vacuum ultraviolet light by two-photon reasonantly enhanced four-wave mixing

    International Nuclear Information System (INIS)

    Payne, M.G.; Garrett, W.R.; Judish, J.P.; Wunderlich, R.

    1988-11-01

    Many of the most impressive demonstrations of the efficient generation of vacuum ultraviolet (VUV) light have made use of two- photon resonantly enhanced four-wave mixing to generate light at ω/sub VUV/ = 2ω/sub L1/ +- ω/sub L2/. The two-photon resonance state is coupled to the ground state both by two photons from the first laser, or by a photon from the second laser and one from the generated VUV beam. We show here that these two coherent pathways destructively interfere once the second laser is made sufficiently intense, thereby leading to an important limiting effect on the achievable conversion efficiency. 4 refs

  8. Recent results on two-photon physics from Tasso and a review of measurements of the two-photon total cross section

    International Nuclear Information System (INIS)

    Kolanoski, H.

    1983-03-01

    Recent results on two-photon physics from the Tasso experiment are presented: the measurement of the two-photon production of Kanti K with the determination of the #betta##betta#-width of the f'(1515), an analysis of the angular correlations in the reaction #betta##betta#->rho 0 rho 0 ->π + π - π + π - and the observation of a narrow structure in the four pion mass spectrum around 2.1 GeV. In a separate part the experimental results on the total cross section for hadron production by two photons are reviewed. (orig.)

  9. A high performance, cost-effective, open-source microscope for scanning two-photon microscopy that is modular and readily adaptable.

    Directory of Open Access Journals (Sweden)

    David G Rosenegger

    Full Text Available Two-photon laser scanning microscopy has revolutionized the ability to delineate cellular and physiological function in acutely isolated tissue and in vivo. However, there exist barriers for many laboratories to acquire two-photon microscopes. Additionally, if owned, typical systems are difficult to modify to rapidly evolving methodologies. A potential solution to these problems is to enable scientists to build their own high-performance and adaptable system by overcoming a resource insufficiency. Here we present a detailed hardware resource and protocol for building an upright, highly modular and adaptable two-photon laser scanning fluorescence microscope that can be used for in vitro or in vivo applications. The microscope is comprised of high-end componentry on a skeleton of off-the-shelf compatible opto-mechanical parts. The dedicated design enabled imaging depths close to 1 mm into mouse brain tissue and a signal-to-noise ratio that exceeded all commercial two-photon systems tested. In addition to a detailed parts list, instructions for assembly, testing and troubleshooting, our plan includes complete three dimensional computer models that greatly reduce the knowledge base required for the non-expert user. This open-source resource lowers barriers in order to equip more laboratories with high-performance two-photon imaging and to help progress our understanding of the cellular and physiological function of living systems.

  10. A High Performance, Cost-Effective, Open-Source Microscope for Scanning Two-Photon Microscopy that Is Modular and Readily Adaptable

    Science.gov (United States)

    Rosenegger, David G.; Tran, Cam Ha T.; LeDue, Jeffery; Zhou, Ning; Gordon, Grant R.

    2014-01-01

    Two-photon laser scanning microscopy has revolutionized the ability to delineate cellular and physiological function in acutely isolated tissue and in vivo. However, there exist barriers for many laboratories to acquire two-photon microscopes. Additionally, if owned, typical systems are difficult to modify to rapidly evolving methodologies. A potential solution to these problems is to enable scientists to build their own high-performance and adaptable system by overcoming a resource insufficiency. Here we present a detailed hardware resource and protocol for building an upright, highly modular and adaptable two-photon laser scanning fluorescence microscope that can be used for in vitro or in vivo applications. The microscope is comprised of high-end componentry on a skeleton of off-the-shelf compatible opto-mechanical parts. The dedicated design enabled imaging depths close to 1 mm into mouse brain tissue and a signal-to-noise ratio that exceeded all commercial two-photon systems tested. In addition to a detailed parts list, instructions for assembly, testing and troubleshooting, our plan includes complete three dimensional computer models that greatly reduce the knowledge base required for the non-expert user. This open-source resource lowers barriers in order to equip more laboratories with high-performance two-photon imaging and to help progress our understanding of the cellular and physiological function of living systems. PMID:25333934

  11. Distribution of quantum information between an atom and two photons

    International Nuclear Information System (INIS)

    Weber, Bernhard

    2008-01-01

    The construction of networks consisting of optically interconnected processing units is a promising way to scale up quantum information processing systems. To store quantum information, single trapped atoms are among the most proven candidates. By placing them in high finesse optical resonators, a bidirectional information exchange between the atoms and photons becomes possible with, in principle, unit efficiency. Such an interface between stationary and ying qubits constitutes a possible node of a future quantum network. The results presented in this thesis demonstrate the prospects of a quantum interface consisting of a single atom trapped within the mode of a high-finesse optical cavity. In a two-step process, we distribute entanglement between the stored atom and two subsequently emitted single photons. The long atom trapping times achieved in the system together with the high photon collection efficiency of the cavity make the applied protocol in principle deterministic, allowing for the creation of an entangled state at the push of a button. Running the protocol on this quasi-stationary quantum interface, the internal state of the atom is entangled with the polarization state of a single emitted photon. The entanglement is generated by driving a vacuum-stimulated Raman adiabatic passage between states of the coupled atom-cavity system. In a second process, the atomic part of the entangled state is mapped onto a second emitted photon using a similar technique and resulting in a polarization-entangled two-photon state. To verify and characterize the photon-photon entanglement, we measured a violation of a Bell inequality and performed a full quantum state tomography. The results prove the prior atom-photon entanglement and demonstrate a quantum information transfer between the atom and the two emitted photons. This reflects the advantages of a high-finesse cavity as a quantum interface in future quantum networks. (orig.)

  12. Dynamical modeling of pulsed two-photon interference

    International Nuclear Information System (INIS)

    Fischer, Kevin A; Lagoudakis, Konstantinos G; Vučković, Jelena; Müller, Kai

    2016-01-01

    Single-photon sources are at the heart of quantum-optical networks, with their uniquely quantum emission and phenomenon of two-photon interference allowing for the generation and transfer of nonclassical states. Although a few analytical methods have been briefly investigated for describing pulsed single-photon sources, these methods apply only to either perfectly ideal or at least extremely idealized sources. Here, we present the first complete picture of pulsed single-photon sources by elaborating how to numerically and fully characterize non-ideal single-photon sources operating in a pulsed regime. In order to achieve this result, we make the connection between quantum Monte-Carlo simulations, experimental characterizations, and an extended form of the quantum regression theorem. We elaborate on how an ideal pulsed single-photon source is connected to its photocount distribution and its measured degree of second- and first-order optical coherence. By doing so, we provide a description of the relationship between instantaneous source correlations and the typical experimental interferometers (Hanbury-Brown and Twiss, Hong–Ou–Mandel, and Mach–Zehnder) used to characterize such sources. Then, we use these techniques to explore several prototypical quantum systems and their non-ideal behaviors. As an example numerical result, we show that for the most popular single-photon source—a resonantly excited two-level system—its error probability is directly related to its excitation pulse length. We believe that the intuition gained from these representative systems and characters can be used to interpret future results with more complicated source Hamiltonians and behaviors. Finally, we have thoroughly documented our simulation methods with contributions to the Quantum Optics Toolbox in Python in order to make our work easily accessible to other scientists and engineers. (paper)

  13. Distribution of quantum information between an atom and two photons

    Energy Technology Data Exchange (ETDEWEB)

    Weber, Bernhard

    2008-11-03

    The construction of networks consisting of optically interconnected processing units is a promising way to scale up quantum information processing systems. To store quantum information, single trapped atoms are among the most proven candidates. By placing them in high finesse optical resonators, a bidirectional information exchange between the atoms and photons becomes possible with, in principle, unit efficiency. Such an interface between stationary and ying qubits constitutes a possible node of a future quantum network. The results presented in this thesis demonstrate the prospects of a quantum interface consisting of a single atom trapped within the mode of a high-finesse optical cavity. In a two-step process, we distribute entanglement between the stored atom and two subsequently emitted single photons. The long atom trapping times achieved in the system together with the high photon collection efficiency of the cavity make the applied protocol in principle deterministic, allowing for the creation of an entangled state at the push of a button. Running the protocol on this quasi-stationary quantum interface, the internal state of the atom is entangled with the polarization state of a single emitted photon. The entanglement is generated by driving a vacuum-stimulated Raman adiabatic passage between states of the coupled atom-cavity system. In a second process, the atomic part of the entangled state is mapped onto a second emitted photon using a similar technique and resulting in a polarization-entangled two-photon state. To verify and characterize the photon-photon entanglement, we measured a violation of a Bell inequality and performed a full quantum state tomography. The results prove the prior atom-photon entanglement and demonstrate a quantum information transfer between the atom and the two emitted photons. This reflects the advantages of a high-finesse cavity as a quantum interface in future quantum networks. (orig.)

  14. Two-photon interference of polarization-entangled photons in a Franson interferometer.

    Science.gov (United States)

    Kim, Heonoh; Lee, Sang Min; Kwon, Osung; Moon, Han Seb

    2017-07-18

    We present two-photon interference experiments with polarization-entangled photon pairs in a polarization-based Franson-type interferometer. Although the two photons do not meet at a common beamsplitter, a phase-insensitive Hong-Ou-Mandel type two-photon interference peak and dip fringes are observed, resulting from the two-photon interference effect between two indistinguishable two-photon probability amplitudes leading to a coincidence detection. A spatial quantum beating fringe is also measured for nondegenerate photon pairs in the same interferometer, although the two-photon states have no frequency entanglement. When unentangled polarization-correlated photons are used as an input state, the polarization entanglement is successfully recovered through the interferometer via delayed compensation.

  15. Development of a new light collection and detection system optimized for ion beam induced fluorescence microscopy

    International Nuclear Information System (INIS)

    Vanga, Sudheer Kumar; Mi, Zhaohong; Koh, Long Cheng; Tao, Ye; Bettiol, Andrew A.; Watt, Frank

    2015-01-01

    Ion beam induced fluorescence microscopy is a new imaging technique which has the potential to achieve sub-50 nm spatial resolution fluorescence images. Currently the resolution of the technique has been limited to around 150 nm mainly because of inefficient collection and detection of emitted photons from the sample. To overcome this limitation, a new light collection system based on a custom made parabolic mirror is employed to enhance the fluorescence collection. The custom made mirror is designed so as to obtain both structural (scanning transmission ion microscopy) and ion beam induced fluorescence imaging simultaneously. The design and characterization of the parabolic mirror is discussed in detail

  16. Two-photon polarization microscopy reveals protein structure and function

    Czech Academy of Sciences Publication Activity Database

    Lazar, Josef; Bondar, Alexey; Timr, S.; Firestein, S. J.

    2011-01-01

    Roč. 8, č. 8 (2011), s. 684-U120 ISSN 1548-7091 Institutional research plan: CEZ:AV0Z60870520 Keywords : green fluorescent proteins * living cells * in-vivo * indicators * anisotropy * activacion * dissociation * orientation * calmodulin * membranes Subject RIV: CE - Biochemistry Impact factor: 19.276, year: 2011

  17. One- and two-photon ionization of hydrogen atom embedded in Debye plasmas

    International Nuclear Information System (INIS)

    Chang, T. N.; Fang, T. K.; Ho, Y. K.

    2013-01-01

    We present a detailed analysis of the plasma-induced resonance-like atomic structures near the ionization threshold in one- and two-photon ionization of hydrogen atom. Such resonance-like structures result from the migration of the upper bound excited states of bound-bound atomic transitions into the continuum due to the less attractive screened Coulomb potential which simulates the external environmental effect for an atom embedded in Debye plasma. The change from the resonance-like narrow structures into broad continuous spectra as the plasma effect increases could be accounted for by the overlap between the respective wavefunctions of the atomic electron in the initial state and its corresponding outgoing ionized state in the continuum

  18. Assisted Interpretation of Laser-Induced Fluorescence Spectra of Egg-Based Binding Media Using Total Emission Fluorescence Spectroscopy

    International Nuclear Information System (INIS)

    Anglos, D.; Nevin, A.

    2006-01-01

    Laser-induced fluorescence (LIF) spectroscopy can provide nondestructive, qualitative analysis of protein-based binding media found in artworks. Fluorescence emissions from proteins in egg yolk and egg white are due to auto fluorescent aromatic amino acids as well as other native and age-related fluorophores, but the potential of fluorescence spectroscopy for the differentiation between binding media is dependent on the choice of a suitable excitation wavelength and limited by problems in interpretation. However, a better understanding of emission spectra associated with LIF can be achieved following comparisons with total emission fluorescence spectra where a series of consecutive emission spectra are recorded over a specific range. Results using nanosecond UV laser sources for LIF of egg-based binding media are presented which are rationalised following comparisons with total emission spectra. Specifically, fluorescence is assigned to tryptophan and oxidation products of amino acids; in the case of egg yolk, fatty-acid polymerisation and age-related degradation products account for the formation of fluorophores.

  19. Solution-dispersible Au nanocube dimers with greatly enhanced two-photon luminescence and SERS

    Science.gov (United States)

    Liu, Xiao-Li; Liang, Shan; Nan, Fan; Yang, Zhong-Jian; Yu, Xue-Feng; Zhou, Li; Hao, Zhong-Hua; Wang, Qu-Quan

    2013-05-01

    We report the synthesis of 43-nm diameter Au nanocube dimers by using Ag+ ions as competitive ligands to freeze l-cysteine-induced assembly process of the nanocubes to a desirable stage. Ascribed to the resonant interparticle coupling with an newly arising plasmon band at 710 nm and local field enhancement, the two-photon luminescence intensity of the Au nanocube dimers in solution was over 20 times stronger than that of the monomers in the wavelength range 555-620 nm. Furthermore, by coupling Raman tags onto the nanocube surface, a solution-based surface-enhanced Raman scattering (SERS) of the nanocube dimers had an enhancement factor of over 10 times compared to the isolated nanocubes. To sum up, with high stability in solution and attractive optical properties, the Au nanocube dimers have potential applications in in vivo bio-imaging and solution-based SERS.We report the synthesis of 43-nm diameter Au nanocube dimers by using Ag+ ions as competitive ligands to freeze l-cysteine-induced assembly process of the nanocubes to a desirable stage. Ascribed to the resonant interparticle coupling with an newly arising plasmon band at 710 nm and local field enhancement, the two-photon luminescence intensity of the Au nanocube dimers in solution was over 20 times stronger than that of the monomers in the wavelength range 555-620 nm. Furthermore, by coupling Raman tags onto the nanocube surface, a solution-based surface-enhanced Raman scattering (SERS) of the nanocube dimers had an enhancement factor of over 10 times compared to the isolated nanocubes. To sum up, with high stability in solution and attractive optical properties, the Au nanocube dimers have potential applications in in vivo bio-imaging and solution-based SERS. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr01170d

  20. The motional stark effect with laser-induced fluorescence diagnostic

    Science.gov (United States)

    Foley, E. L.; Levinton, F. M.

    2010-05-01

    The motional Stark effect (MSE) diagnostic is the worldwide standard technique for internal magnetic field pitch angle measurements in magnetized plasmas. Traditionally, it is based on using polarimetry to measure the polarization direction of light emitted from a hydrogenic species in a neutral beam. As the beam passes through the magnetized plasma at a high velocity, in its rest frame it perceives a Lorentz electric field. This field causes the H-alpha emission to be split and polarized. A new technique under development adds laser-induced fluorescence (LIF) to a diagnostic neutral beam (DNB) for an MSE measurement that will enable radially resolved magnetic field magnitude as well as pitch angle measurements in even low-field (experiments. An MSE-LIF system will be installed on the National Spherical Torus Experiment (NSTX) at the Princeton Plasma Physics Laboratory. It will enable reconstructions of the plasma pressure, q-profile and current as well as, in conjunction with the existing MSE system, measurements of radial electric fields.

  1. In-situ hydrocarbon delineation using laser-induced fluorescence

    International Nuclear Information System (INIS)

    Taer, A.D.; Hastings, R.W.; Brown, A.Y.; Frend, R.

    1996-01-01

    An investigation of hydrocarbons in soils was conducted at an active Shell Oil Company petroleum products terminal, located in Carson, California. An investigation approach involving Laser-Induced Fluorescence (LIF) and Cone Penetrometer Testing (CPT) technologies was implemented to provide real-time, in-situ characterization of site stratigraphy, hydrocarbon distribution and importantly, hydrocarbon product differentiation. The area of investigation is located along a property boundary, where a plume of separate phase hydrocarbons has been actively recovered for several years. CPT/LIF technology was selected for the investigation since previous delineation efforts using hydrocarbon fingerprinting methods proved inconclusive. Additionally, the CPT/LIF technology had the potential to provide a cost effective solution to accomplish project objectives. Based on the information obtained during this investigation, it was determined that the plume of separate phase hydrocarbons along the northern property boundary is from a source distinctly different than any identified hydrocarbons known to be from on-site sources. In addition, the plume was determined to not be connected with any other known on-site hydrocarbon plumes. The results of this CPT/LIF investigation were consistent with the known hydrogeologic conditions. This evaluation determined that CPT/LIF technology was very effective in addressing project objectives and resulted in a significant cost savings

  2. Airborne laser induced fluorescence imaging. Innovative technology summary report

    International Nuclear Information System (INIS)

    1999-06-01

    Laser-Induced Fluorescence (LIF) was demonstration as part of the Fernald Environmental Management Project (FEMP) Plant 1 Large Scale Demonstration and Deployment Project (LSDDP) sponsored by the US Department of Energy (DOE) Office of Science and Technology, Deactivation and Decommissioning Focus Area located at the Federal Energy Technology Center (FETC) in Morgantown, West Virginia. The demonstration took place on November 19, 1996. In order to allow the contaminated buildings undergoing deactivation and decommissioning (D and D) to be opened to the atmosphere, radiological surveys of floors, walls and ceilings must take place. After successful completion of the radiological clearance survey, demolition of the building can continue. Currently, this process is performed by collecting and analyzing swipe samples for radiological analysis. Two methods are used to analyze the swipe samples: hand-held frisker and laboratory analysis. For the purpose of this demonstration, the least expensive method, swipe samples analyzed by hand-held frisker, is the baseline technology. The objective of the technology demonstration was to determine if the baseline technology could be replaced using LIF

  3. Laser Induced Fluorescence Diagnostic for the Plasma Couette Experiment

    Science.gov (United States)

    Katz, Noam; Skiff, Fred; Collins, Cami; Weisberg, Dave; Wallace, John; Clark, Mike; Garot, Kristine; Forest, Cary

    2010-11-01

    The Plasma Couette Experiment (PCX) at U. Wisconsin-Madison consists of a rotating high-beta plasma and is well-suited to the study of flow-driven, astrophysically-relevant plasma phenomena. PCX confinement relies on alternating rings of 1kG permanent magnets and the rotation is driven by electrode rings, interspersed between the magnets, which provide an azimuthal ExB. I will discuss the development of a laser-induced fluorescence diagnostic (LIF) to characterize the ion distribution function of argon plasmas in PCX. The LIF system--which will be scanned radially--will be used to calibrate internal Mach probes, as well as to measure the time-resolved velocity profile, ion temperature and density non-perturbatively. These diagnostics will be applied to study the magneto-rotational instability in a plasma, as well as the buoyancy instability thought to be involved in producing the solar magnetic field. This work is supported by NSF and DOE.

  4. Stink Bug Feeding Induces Fluorescence in Developing Cotton Bolls

    Directory of Open Access Journals (Sweden)

    Toews Michael D

    2011-08-01

    Full Text Available Abstract Background Stink bugs (Hemiptera: Pentatomidae comprise a critically important insect pest complex affecting 12 major crops worldwide including cotton. In the US, stink bug damage to developing cotton bolls causes boll abscission, lint staining, reduced fiber quality, and reduced yields with estimated losses ranging from 10 to 60 million dollars annually. Unfortunately, scouting for stink bug damage in the field is laborious and excessively time consuming. To improve scouting accuracy and efficiency, we investigated fluorescence changes in cotton boll tissues as a result of stink bug feeding. Results Fluorescent imaging under long-wave ultraviolet light showed that stink bug-damaged lint, the inner carpal wall, and the outside of the boll emitted strong blue-green fluorescence in a circular region near the puncture wound, whereas undamaged tissue emissions occurred at different wavelengths; the much weaker emission of undamaged tissue was dominated by chlorophyll fluorescence. We further characterized the optimum emission and excitation spectra to distinguish between stink bug damaged bolls from undamaged bolls. Conclusions The observed characteristic fluorescence peaks associated with stink bug damage give rise to a fluorescence-based method to rapidly distinguish between undamaged and stink bug damaged cotton bolls. Based on the fluorescent fingerprint, we envision a fluorescence reflectance imaging or a fluorescence ratiometric device to assist pest management professionals with rapidly determining the extent of stink bug damage in a cotton field.

  5. Methotrexate-Induced Accumulation of Fluorescent Annexin V in Collagen-Induced Arthritis

    Directory of Open Access Journals (Sweden)

    Andreas Wunder

    2005-01-01

    Full Text Available We examined the accumulation of Cy5.5-labeled annexin V in the paws of mice with and without collagen-induced arthritis, with and without methotrexate (MTX treatment, by near-infrared fluorescence imaging. Fluorescence reflectance imaging (FRI of paws was performed 48 hr after MTX injection and at 10 min and 3 hr after the injection of Cy5.5-annexin V (1 nmol dye per mouse. With arthritic paws, MTX treatment caused a 7-fold increase in fluorescence intensity compared with the paws of untreated mice and a 4-fold increase compared to nonarthritic paws of MTX-treated mice (p < .001 each. Tissue samples of paws were examined histologically for Cy5.5 fluorescence and by TUNEL staining for apoptosis. Cy5.5-annexin V was seen in the hyperplastic synovia of MTX-treated mice, and TUNEL staining for apoptosis showed apoptotic cells in the hyperplastic synovia. Monitoring the uptake of Cy5.5-annexin V in arthritic paws by FRI provided a method of assessing a response to MTX, a response that was readily quantitated with simple instrumentation and that occurred before conventional measurements of treatment response.

  6. Photo-redox activated drug delivery systems operating under two photon excitation in the near-IR.

    Science.gov (United States)

    Guardado-Alvarez, Tania M; Devi, Lekshmi Sudha; Vabre, Jean-Marie; Pecorelli, Travis A; Schwartz, Benjamin J; Durand, Jean-Olivier; Mongin, Olivier; Blanchard-Desce, Mireille; Zink, Jeffrey I

    2014-05-07

    We report the design and synthesis of a nano-container consisting of mesoporous silica nanoparticles with the pore openings covered by "snap-top" caps that are opened by near-IR light. A photo transducer molecule that is a reducing agent in an excited electronic state is covalently attached to the system. Near IR two-photon excitation causes inter-molecular electron transfer that reduces a disulfide bond holding the cap in place, thus allowing the cargo molecules to escape. We describe the operation of the "snap-top" release mechanism by both one- and two-photon activation. This system presents a proof of concept of a near-IR photoredox-induced nanoparticle delivery system that may lead to a new type of photodynamic drug release therapy.

  7. Calcium rubies: a family of red-emitting functionalizable indicators suitable for two-photon Ca2+ imaging.

    Science.gov (United States)

    Collot, Mayeul; Loukou, Christina; Yakovlev, Aleksey V; Wilms, Christian D; Li, Dongdong; Evrard, Alexis; Zamaleeva, Alsu; Bourdieu, Laurent; Léger, Jean-François; Ropert, Nicole; Eilers, Jens; Oheim, Martin; Feltz, Anne; Mallet, Jean-Maurice

    2012-09-12

    We designed Calcium Rubies, a family of functionalizable BAPTA-based red-fluorescent calcium (Ca(2+)) indicators as new tools for biological Ca(2+) imaging. The specificity of this Ca(2+)-indicator family is its side arm, attached on the ethylene glycol bridge that allows coupling the indicator to various groups while leaving open the possibility of aromatic substitutions on the BAPTA core for tuning the Ca(2+)-binding affinity. Using this possibility we now synthesize and characterize three different CaRubies with affinities between 3 and 22 μM. Their long excitation and emission wavelengths (peaks at 586/604 nm) allow their use in otherwise challenging multicolor experiments, e.g., when combining Ca(2+) uncaging or optogenetic stimulation with Ca(2+) imaging in cells expressing fluorescent proteins. We illustrate this capacity by the detection of Ca(2+) transients evoked by blue light in cultured astrocytes expressing CatCh, a light-sensitive Ca(2+)-translocating channelrhodopsin linked to yellow fluorescent protein. Using time-correlated single-photon counting, we measured fluorescence lifetimes for all CaRubies and demonstrate a 10-fold increase in the average lifetime upon Ca(2+) chelation. Since only the fluorescence quantum yield but not the absorbance of the CaRubies is Ca(2+)-dependent, calibrated two-photon fluorescence excitation measurements of absolute Ca(2+) concentrations are feasible.

  8. Fluorescent-light-induced lethality and DNA repair in normal and xeroderma pigmentosum fibroblasts

    International Nuclear Information System (INIS)

    Ritter, M.A.; Williams, J.R.

    1981-01-01

    Cell survival and induction of endonuclease-sensitive sites in DNA were measured in human fibroblast cells exposed to fluorescent light or germicidal ultraviolet light. Cells from a xeroderma pigmentosum patient were hypersensitive to cell killing by fluorescent light, although less so than for germicidal ultraviolet light. Xeroderma pigmentosum cells were deficient in the removal of fluorescent light-induced endonuclease sites that are probably pyrimidine dimers, and both the xeroderma pigmentosum and normal cells removed these sites with kinetics indistinguishable from those for ultraviolet light-induced sites. A comparison of fluorescent with ultraviolet light data demonstrates that there are markedly fewer pyrimidine dimers per lethal event for fluorescent than for ultraviolet light, suggesting a major role for non-dimer damage in fluorescent lethality. (Auth.)

  9. Laser induced fluorescence technique for detecting organic matter in East China Sea

    Science.gov (United States)

    Chen, Peng; Wang, Tianyu; Pan, Delu; Huang, Haiqing

    2017-10-01

    A laser induced fluorescence (LIF) technique for fast diagnosing chromophoric dissolved organic matter (CDOM) in water is discussed. We have developed a new field-portable laser fluorometer for rapid fluorescence measurements. In addtion, the fluorescence spectral characteristics of fluorescent constituents (e.g., CDOM, chlorophyll-a) were analyzed with a spectral deconvolution method of bi-Gaussian peak function. In situ measurements by the LIF technique compared well with values measured by conventional spectrophotometer method in laboratory. A significant correlation (R2 = 0.93) was observed between fluorescence by the technique and absorption by laboratory spectrophotometer. Influence of temperature variation on LIF measurement was investigated in lab and a temperature coefficient was deduced for fluorescence correction. Distributions of CDOM fluorescence measured using this technique in the East China Sea coast were presented. The in situ result demonstrated the utility of the LIF technique for rapid detecting dissolved organic matter.

  10. Cell and brain tissue imaging of the flavonoid fisetin using label-free two-photon microscopy.

    Science.gov (United States)

    Krasieva, Tatiana B; Ehren, Jennifer; O'Sullivan, Thomas; Tromberg, Bruce J; Maher, Pamela

    2015-10-01

    Over the last few years, we have identified an orally active, novel neuroprotective and cognition-enhancing molecule, the flavonoid fisetin. Fisetin not only has direct antioxidant activity but it can also increase the intracellular levels of glutathione, the major intracellular antioxidant. Fisetin can also activate key neurotrophic factor signaling pathways. In addition, it has anti-inflammatory activity against microglia and astrocytes and inhibits the activity of lipoxygenases, thereby reducing the production of pro-inflammatory eicosanoids and their by-products. However, key questions about its targets and brain penetration remain. In this study, we used label-free two-photon microscopy of intrinsic fisetin fluorescence to examine the localization of fisetin in living nerve cells and the brains of living mice. In cells, fisetin but not structurally related flavonols with different numbers of hydroxyl groups, localized to the nucleoli suggesting that key targets of fisetin may reside in this organelle. In the mouse brain, following intraperitoneal injection and oral administration, fisetin rapidly distributed to the blood vessels of the brain followed by a slower dispersion into the brain parenchyma. Thus, these results provide further support for the effects of fisetin on brain function. In addition, they suggest that label-free two-photon microscopy may prove useful for studying the intracellular and tissue distribution of other intrinsically-fluorescent flavonoids. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Nanoantenna array-induced fluorescence enhancement and reduced lifetimes

    DEFF Research Database (Denmark)

    Bakker, R. M.; Drachev, V. P.; Liu, Z.

    2008-01-01

    Enhanced fluorescence is observed from dye molecules interacting with optical nanoantenna arrays. Elliptical gold dimers form individual nanoantennae with tunable plasmon resonances depending upon the geometry of the two particles and the size of the gap between them. A fluorescent dye, Rhodamine...... 800, is uniformly embedded in a dielectric host that coats the nanoantennae. The nanoantennae act to enhance the dye absorption. In turn, emission from the dye drives the plasmon resonance of the antennae; the nanoantennae act to enhance the fluorescence signal and change the angular distribution...... of emission. These effects depend upon the overlap of the plasmon resonance with the excitation wavelength and the fluorescence emission band. A decreased fluorescence lifetime is observed along with highly polarized emission that displays the characteristics of the nanoantenna's dipole mode. Being able...

  12. Laser-Induced Fluorescence (LIF) from plant foliage

    Science.gov (United States)

    Chappelle, Emmett W.; Williams, Darrel L.

    1987-01-01

    The fluorescence spectra and fluorescence induction kinetics of green plants excited at 337 nm by a laser were studied. They correlate with plant type, as well as with changes in the physiology of the plant as the result of stress. The plant types studied include herbaceous dicots, monocots, hardwoods, conifers, and algae. These plant types could be identified on the basis of differences in either the number of fluorescent bands or the relative intensity of the bands. Differences in fluorescent spectra which could be related to vigor status are observed in conifers located in an area of high atmospheric deposition. Changes in the fluorescence spectra and induction kinetics are also seen in plants grown under conditions of nutrient deficiency and drought stress.

  13. In vivo study of the human skin by the method of laser-induced fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Borisova, E.; Avramov, L.

    2000-01-01

    The goals of this study are to perform a preliminary evaluation of the diagnostic potential of noninvasive laser-induced auto-fluorescence spectroscopy (LIAFS) for human skin and optimize of detection and diagnosis of hollow organs and skin. In recent years, there has been growing interest in the use of laser-induced fluorescence to discriminate disease from normal surrounding tissue. The most fluorescence studies have used exogenous fluorophores of this discrimination. The laser-induced auto-fluorescence which is used for diagnosis of tissues in the human body avoids administration of any drugs. In this study a technique for optical biopsy of in vivo human skin is presented. The auto-fluorescence characterization of tissue relies on different spectral properties of tissues. It was demonstrated a differentiation between normal skin and skin with vitiligo. Two main endogenous fluorophores in the human skin account for most of the cellular auto-fluorescence for excitation wavelength 337 nm reduced from of nicotinamide adenine dinucleotide and collagen. The auto-fluorescence spectrum of human skin depend on main internal absorbers which are blood and melanin. In this study was described the effect caused by blood and melanin content on the shape of the auto-fluorescence spectrum of human skin. Human skin fluorescence spectrum might provide dermatologists with important information and such investigations are successfully used now in skin disease diagnostics, in investigation of the environmental factor impact or for evaluation of treatment efficiency. (authors)

  14. Laser induced fluorescence measurements of the mixing of fuel oil with air

    Energy Technology Data Exchange (ETDEWEB)

    Arnold, A; Bombach, R; Hubschmid, W; Kaeppeli, B [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1999-08-01

    We report on measurements of the mixing of fuel oil with air at atmospheric pressure in an industrial premixed gas turbine burner. The concentration of the vaporized fuel oil was measured with laser induced fluorescence. We reason that the fuel oil concentration can be considered with good accuracy as proportional to the fluorescence intensity. (author) 6 fig., 3 refs.

  15. Electron beam induced fluorescence measurements of the degree of hydrogen dissociation in hydrogen plasmas

    NARCIS (Netherlands)

    Smit, C.; Brussaard, G.J.H.; de Beer, E.C.M.; Schram, D.C.; Sanden, van de M.C.M.

    2004-01-01

    The degree of dissociation of hydrogen in a hydrogen plasma has been measured using electron beam induced fluorescence. A 20 kV, 1 mA electron beam excites both the ground state H atom and H2 molecule into atomic hydrogen in an excited state. From the resulting fluorescence the degree of

  16. Detection of fecal residue on poultry carcasses by laser induced fluorescence imaging techniques

    Science.gov (United States)

    The potential use of laser-induced fluorescence imaging techniques was investigated for the detection of diluted fecal matters from various parts of the digestive tract, including colon, ceca, small intestine, and duodenum, on poultry carcasses. One of the challenges for using fluorescence imaging f...

  17. Applying fluorescence correlation spectroscopy to investigate peptide-induced membrane disruption

    DEFF Research Database (Denmark)

    Kristensen, Kasper; Henriksen, Jonas Rosager; Andresen, Thomas Lars

    2017-01-01

    to quantify leakage of fluorescent molecules of different sizes from large unilamellar lipid vesicles, thereby providing a tool for estimating the size of peptide-induced membrane disruptions. If fluorescently labeled lipids are incorporated into the membranes of the vesicles, FCS can also be used to obtain...

  18. Recognition of edible oil by using BP neural network and laser induced fluorescence spectrum

    Science.gov (United States)

    Mu, Tao-tao; Chen, Si-ying; Zhang, Yin-chao; Guo, Pan; Chen, He; Zhang, Hong-yan; Liu, Xiao-hua; Wang, Yuan; Bu, Zhi-chao

    2013-09-01

    In order to accomplish recognition of the different edible oil we set up a laser induced fluorescence spectrum system in the laboratory based on Laser induced fluorescence spectrum technology, and then collect the fluorescence spectrum of different edible oil by using that system. Based on this, we set up a fluorescence spectrum database of different cooking oil. It is clear that there are three main peak position of different edible oil from fluorescence spectrum chart. Although the peak positions of all cooking oil were almost the same, the relative intensity of different edible oils was totally different. So it could easily accomplish that oil recognition could take advantage of the difference of relative intensity. Feature invariants were extracted from the spectrum data, which were chosen from the fluorescence spectrum database randomly, before distinguishing different cooking oil. Then back propagation (BP) neural network was established and trained by the chosen data from the spectrum database. On that basis real experiment data was identified by BP neural network. It was found that the overall recognition rate could reach as high as 83.2%. Experiments showed that the laser induced fluorescence spectrum of different cooking oil was very different from each other, which could be used to accomplish the oil recognition. Laser induced fluorescence spectrum technology, combined BP neural network,was fast, high sensitivity, non-contact, and high recognition rate. It could become a new technique to accomplish the edible oil recognition and quality detection.

  19. Visualization of laser tattoo removal treatment effects in a mouse model by two-photon microscopy.

    Science.gov (United States)

    Jang, Won Hyuk; Yoon, Yeoreum; Kim, Wonjoong; Kwon, Soonjae; Lee, Seunghun; Song, Duke; Choi, Jong Woon; Kim, Ki Hean

    2017-08-01

    Laser tattoo removal is an effective method of eliminating tattoo particles in the skin. However, laser treatment cannot always remove the unwanted tattoo completely, and there are risks of either temporary or permanent side effects. Studies using preclinical animal models could provide detailed information on the effects of laser treatment in the skin, and might help to minimize side effects in clinical practices. In this study, two-photon microscopy (TPM) was used to visualize the laser treatment effects on tattoo particles in both phantom specimens and in vivo mouse models. Fluorescent tattoo ink was used for particle visualization by TPM, and nanosecond (ns) and picosecond (ps) lasers at 532 nm were used for treatment. In phantom specimens, TPM characterized the fragmentation of individual tattoo particles by tracking them before and after the laser treatment. These changes were confirmed by field emission scanning electron microscopy (FE-SEM). TPM was used to measure the treatment efficiency of the two lasers at different laser fluences. In the mouse model, TPM visualized clusters of tattoo particles in the skin and detected their fragmentation after the laser treatment. Longitudinal TPM imaging observed the migration of cells containing tattoo particles after the laser treatment. These results show that TPM may be useful for the assessment of laser tattoo removal treatment in preclinical studies.

  20. Automatic quantification of mitochondrial fragmentation from two-photon microscope images of mouse brain tissue.

    Science.gov (United States)

    Lihavainen, E; Kislin, M; Toptunov, D; Khiroug, L; Ribeiro, A S

    2015-12-01

    The morphology of mitochondria can inform about their functional state and, thus, about cell vitality. For example, fragmentation of the mitochondrial network is associated with many diseases. Recent advances in neuronal imaging have enabled the observation of mitochondria in live brains for long periods of time, enabling the study of their dynamics in animal models of diseases. To aid these studies, we developed an automatic method, based on supervised learning, for quantifying the degree of mitochondrial fragmentation in tissue images acquired via two-photon microscopy from transgenic mice, which exclusively express Enhanced cyan fluorescent protein (ECFP) under Thy1 promoter, targeted to the mitochondrial matrix in subpopulations of neurons. We tested the method on images prior to and after cardiac arrest, and found it to be sensitive to significant changes in mitochondrial morphology because of the arrest. We conclude that the method is useful in detecting morphological abnormalities in mitochondria and, likely, in other subcellular structures as well. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  1. Exploring the brain on multiple scales with correlative two-photon and light sheet microscopy

    Science.gov (United States)

    Silvestri, Ludovico; Allegra Mascaro, Anna Letizia; Costantini, Irene; Sacconi, Leonardo; Pavone, Francesco S.

    2014-02-01

    One of the unique features of the brain is that its activity cannot be framed in a single spatio-temporal scale, but rather spans many orders of magnitude both in space and time. A single imaging technique can reveal only a small part of this complex machinery. To obtain a more comprehensive view of brain functionality, complementary approaches should be combined into a correlative framework. Here, we describe a method to integrate data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy, taking advantage of blood vessels as reference chart. We show how the apical dendritic arbor of a single cortical pyramidal neuron imaged in living thy1-GFP-M mice can be found in the large-scale brain reconstruction obtained with light sheet microscopy. Starting from the apical portion, the whole pyramidal neuron can then be segmented. The correlative approach presented here allows contextualizing within a three-dimensional anatomic framework the neurons whose dynamics have been observed with high detail in vivo.

  2. Imaging-guided two-photon excitation-emission-matrix measurements of human skin tissues

    Science.gov (United States)

    Yu, Yingqiu; Lee, Anthony M. D.; Wang, Hequn; Tang, Shuo; Zhao, Jianhua; Lui, Harvey; Zeng, Haishan

    2012-07-01

    There are increased interests on using multiphoton imaging and spectroscopy for skin tissue characterization and diagnosis. However, most studies have been done with just a few excitation wavelengths. Our objective is to perform a systematic study of the two-photon fluorescence (TPF) properties of skin fluorophores, normal skin, and diseased skin tissues. A nonlinear excitation-emission-matrix (EEM) spectroscopy system with multiphoton imaging guidance was constructed. A tunable femtosecond laser was used to vary excitation wavelengths from 730 to 920 nm for EEM data acquisition. EEM measurements were performed on excised fresh normal skin tissues, seborrheic keratosis tissue samples, and skin fluorophores including: NADH, FAD, keratin, melanin, collagen, and elastin. We found that in the stratum corneum and upper epidermis of normal skin, the cells have large sizes and the TPF originates from keratin. In the lower epidermis, cells are smaller and TPF is dominated by NADH contributions. In the dermis, TPF is dominated by elastin components. The depth resolved EEM measurements also demonstrated that keratin structure has intruded into the middle sublayers of the epidermal part of the seborrheic keratosis lesion. These results suggest that the imaging guided TPF EEM spectroscopy provides useful information for the development of multiphoton clinical devices for skin disease diagnosis.

  3. Spin polarization in top pair production in association with two photons at NLO+PS

    CERN Document Server

    Luisoni, Gionata

    2018-01-01

    This talk focuses on the impact of top-quark spin polarization effects in Higgs boson production in association with a top-quark pair, where the Higgs boson decays to two photons. Predictions for the signal are compared with direct top-quark pair production in association with two photons at NLO+PS.

  4. Measurement and model of the infrared two-photon emission spectrum of GaAs.

    Science.gov (United States)

    Hayat, Alex; Ginzburg, Pavel; Orenstein, Meir

    2009-07-10

    Two-photon emission from semiconductors was recently observed, but not fully interpreted. We develop a dressed-state model incorporating intraband scattering-related level broadening, yielding nondivergent emission rates. The spectrum calculations for high carrier concentrations including the time dependence of the screening buildup correspond well to our measured two-photon emission spectrum from GaAs.

  5. Effect of morphology and solvent on two-photon absorption of nano zinc oxide

    Energy Technology Data Exchange (ETDEWEB)

    Kavitha, M.K. [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram 695547, Kerala (India); Haripadmam, P.C.; Gopinath, Pramod; Krishnan, Bindu [Department of Physics, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram 695547, Kerala (India); John, Honey, E-mail: honey@iist.ac.in [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram 695547, Kerala (India)

    2013-05-15

    Highlights: ► ZnO nanospheres and triangular structures synthesis by novel precipitation technique. ► The effect of precursor concentration on the size and shape of nano ZnO. ► Open aperture Z-scan measurements of the ZnO nanoparticle dispersions. ► Nanospheres exhibit higher two photon absorption coefficient than triangular nanostructures. ► Nanospheres dispersed in water exhibit higher two photon absorption coefficient than its dispersion in 2-propanol. - Abstract: In this paper, we report the effect of morphology and solvent on the two-photon absorption of nano zinc oxide. Zinc oxide nanoparticles in two different morphologies like nanospheres and triangular nanostructures are synthesized by novel precipitation technique and their two-photon absorption coefficient is measured using open aperture Z-scan technique. Experimental results show that the zinc oxide nanospheres exhibit higher two-photon absorption coefficient than the zinc oxide triangular nanostructures. The zinc oxide nanospheres dispersed in water exhibit higher two-photon absorption coefficient than that of its dispersion in 2-propanol. The zinc oxide nanospheres dispersed in water shows a decrease in two-photon absorption coefficient with an increase in on-axis irradiance. The result confirms the dependence of shape and solvent on the two-photon absorption of nano zinc oxide.

  6. A survey of two-photon physics: What have we done, where are we going

    International Nuclear Information System (INIS)

    Kolanoski, H.

    1984-11-01

    After a brief general introduction into two-photon scattering processes the status of two-photon physics as of the beginning of this workshop is reported. Emphasis is given to those subjects which should get more attention in the future. (orig.)

  7. Event-by-event simulation of nonclassical effects in two-photon interference experiments

    NARCIS (Netherlands)

    Michielsen, K.; Jin, F.; Delina, M.; Raedt, H. De

    2012-01-01

    A corpuscular simulation model for second-order intensity interference phenomena is discussed. It is shown that both the visibility V = 1/2 predicted for two-photon interference experiments with two independent sources and the visibility V = 1 predicted for two-photon interference experiments with a

  8. Two photon emission by hydrogen-like atoms in high temperature plasmas

    International Nuclear Information System (INIS)

    Costescu, A.; Manzatu, I.; Dinu, C.; Mihailescu, I.N.

    1981-08-01

    New exact solutions and a rather simple polynomial expression of the power emitted in the two photon transition from a metastable 2s state to the ground state of a hydrogen-like atom were infered with the aid of the Coulomb Green's function method. It was shown that the two photon decay represents under certain circumstances a significant power loss mechanism. (authors)

  9. Production of e, $\\mu$ and $\\tau$ Pairs in Untagged Two-Photon Collisions at LEP

    CERN Document Server

    Acciarri, M; Aguilar-Benítez, M; Ahlen, S P; Alcaraz, J; Alemanni, G; Allaby, James V; Aloisio, A; Alverson, G; Alviggi, M G; Ambrosi, G; Anderhub, H; Andreev, V P; Angelescu, T; Anselmo, F; Arefev, A; Azemoon, T; Aziz, T; Bagnaia, P; Baksay, L; Ball, R C; Banerjee, S; Banerjee, Sw; Banicz, K; Barczyk, A; Barillère, R; Barone, L; Bartalini, P; Baschirotto, A; Basile, M; Battiston, R; Bay, A; Becattini, F; Becker, U; Behner, F; Berdugo, J; Berges, P; Bertucci, B; Betev, B L; Bhattacharya, S; Biasini, M; Biland, A; Bilei, G M; Blaising, J J; Blyth, S C; Bobbink, Gerjan J; Böck, R K; Böhm, A; Boldizsar, L; Borgia, B; Boucham, A; Bourilkov, D; Bourquin, Maurice; Boutigny, D; Braccini, S; Branson, J G; Brigljevic, V; Brock, I C; Buffini, A; Buijs, A; Burger, J D; Burger, W J; Busenitz, J K; Cai, X D; Campanelli, M; Capell, M; Cara Romeo, G; Carlino, G; Cartacci, A M; Casaus, J; Castellini, G; Cavallari, F; Cavallo, N; Cecchi, C; Cerrada-Canales, M; Cesaroni, F; Chamizo-Llatas, M; Chang, Y H; Chaturvedi, U K; Chekanov, S V; Chemarin, M; Chen, A; Chen, G; Chen, G M; Chen, H F; Chen, H S; Chen, M; Chiefari, G; Chien, C Y; Cifarelli, Luisa; Cindolo, F; Civinini, C; Clare, I; Clare, R; Cohn, H O; Coignet, G; Colijn, A P; Colino, N; Commichau, V; Costantini, S; Cotorobai, F; de la Cruz, B; Csilling, Akos; Dai, T S; D'Alessandro, R; De Asmundis, R; Degré, A; Deiters, K; Denes, P; De Notaristefani, F; DiBitonto, Daryl; Diemoz, M; Van Dierendonck, D N; Di Lodovico, F; Dionisi, C; Dittmar, Michael; Dominguez, A; Doria, A; Dorne, I; Dova, M T; Drago, E; Duchesneau, D; Duinker, P; Durán, I; Dutta, S; Easo, S; Efremenko, Yu V; El-Mamouni, H; Engler, A; Eppling, F J; Erné, F C; Ernenwein, J P; Extermann, Pierre; Fabre, M; Faccini, R; Falciano, S; Favara, A; Fay, J; Fedin, O; Felcini, Marta; Fenyi, B; Ferguson, T; Ferroni, F; Fesefeldt, H S; Fiandrini, E; Field, J H; Filthaut, Frank; Fisher, P H; Fisk, I; Forconi, G; Fredj, L; Freudenreich, Klaus; Furetta, C; Galaktionov, Yu; Ganguli, S N; García-Abia, P; Gau, S S; Gentile, S; Gerald, J; Gheordanescu, N; Giagu, S; Goldfarb, S; Goldstein, J; Gong, Z F; Gougas, Andreas; Gratta, Giorgio; Grünewald, M W; Gupta, V K; Gurtu, A; Gutay, L J; Hartmann, B; Hasan, A; Hatzifotiadou, D; Hebbeker, T; Hervé, A; Van Hoek, W C; Hofer, H; Hong, S J; Hoorani, H; Hou, S R; Hu, G; Innocente, Vincenzo; Janssen, H; Jenkes, K; Jin, B N; Jones, L W; de Jong, P; Josa-Mutuberria, I; Kasser, A; Khan, R A; Kamrad, D; Kamyshkov, Yu A; Kapustinsky, J S; Karyotakis, Yu; Kaur, M; Kienzle-Focacci, M N; Kim, D; Kim, D H; Kim, J K; Kim, S C; Kim, Y G; Kinnison, W W; Kirkby, A; Kirkby, D; Kirkby, Jasper; Kiss, D; Kittel, E W; Klimentov, A; König, A C; Kopp, A; Korolko, I; Koutsenko, V F; Krämer, R W; Krenz, W; Kunin, A; Ladrón de Guevara, P; Landi, G; Lapoint, C; Lassila-Perini, K M; Laurikainen, P; Lebeau, M; Lebedev, A; Lebrun, P; Lecomte, P; Lecoq, P; Le Coultre, P; Leggett, C; Le Goff, J M; Leiste, R; Leonardi, E; Levchenko, P M; Li Chuan; Lin, C H; Lin, W T; Linde, Frank L; Lista, L; Liu, Z A; Lohmann, W; Longo, E; Lu, W; Lü, Y S; Lübelsmeyer, K; Luci, C; Luckey, D; Luminari, L; Lustermann, W; Ma Wen Gan; Maity, M; Majumder, G; Malgeri, L; Malinin, A; Maña, C; Mangeol, D J J; Mangla, S; Marchesini, P A; Marin, A; Martin, J P; Marzano, F; Massaro, G G G; McNally, D; Mele, S; Merola, L; Meschini, M; Metzger, W J; Von der Mey, M; Mi, Y; Mihul, A; Van Mil, A J W; Mirabelli, G; Mnich, J; Molnár, P; Monteleoni, B; Moore, R; Morganti, S; Moulik, T; Mount, R; Müller, S; Muheim, F; Muijs, A J M; Nahn, S; Napolitano, M; Nessi-Tedaldi, F; Newman, H; Niessen, T; Nippe, A; Nisati, A; Nowak, H; Oh, Yu D; Opitz, H; Organtini, G; Ostonen, R; Palomares, C; Pandoulas, D; Paoletti, S; Paolucci, P; Park, H K; Park, I H; Pascale, G; Passaleva, G; Patricelli, S; Paul, T; Pauluzzi, M; Paus, C; Pauss, Felicitas; Peach, D; Pei, Y J; Pensotti, S; Perret-Gallix, D; Petersen, B; Petrak, S; Pevsner, A; Piccolo, D; Pieri, M; Pinto, J C; Piroué, P A; Pistolesi, E; Plyaskin, V; Pohl, M; Pozhidaev, V; Postema, H; Produit, N; Prokofev, D; Prokofiev, D O; Rahal-Callot, G; Raja, N; Rancoita, P G; Rattaggi, M; Raven, G; Razis, P A; Read, K; Ren, D; Rescigno, M; Reucroft, S; Van Rhee, T; Riemann, S; Riles, K; Rind, O; Robohm, A; Rodin, J; Roe, B P; Romero, L; Rosier-Lees, S; Rosselet, P; Van Rossum, W; Roth, S; Rubio, Juan Antonio; Ruschmeier, D; Rykaczewski, H; Salicio, J; Sánchez, E; Sanders, M P; Sarakinos, M E; Sarkar, S; Sassowsky, M; Sauvage, G; Schäfer, C; Shchegelskii, V; Schmidt-Kärst, S; Schmitz, D; Schmitz, P; Schneegans, M; Scholz, N; Schopper, Herwig Franz; Schotanus, D J; Schwenke, J; Schwering, G; Sciacca, C; Sciarrino, D; Servoli, L; Shevchenko, S; Shivarov, N; Shoutko, V; Shukla, J; Shumilov, E; Shvorob, A V; Siedenburg, T; Son, D; Sopczak, André; Soulimov, V; Smith, B; Spillantini, P; Steuer, M; Stickland, D P; Stone, H; Stoyanov, B; Strässner, A; Strauch, K; Sudhakar, K; Sultanov, G G; Sun, L Z; Susinno, G F; Suter, H; Swain, J D; Tang, X W; Tauscher, Ludwig; Taylor, L; Ting, Samuel C C; Ting, S M; Tonutti, M; Tonwar, S C; Tóth, J; Tully, C; Tuchscherer, H; Tung, K L; Uchida, Y; Ulbricht, J; Uwer, U; Valente, E; Van de Walle, R T; Vesztergombi, G; Vetlitskii, I; Viertel, Gert M; Vivargent, M; Völkert, R; Vogel, H; Vogt, H; Vorobev, I; Vorobyov, A A; Vorvolakos, A; Wadhwa, M; Wallraff, W; Wang, J C; Wang, X L; Wang, Z M; Weber, A; Wittgenstein, F; Wu, S X; Wynhoff, S; Xu, J; Xu, Z Z; Yang, B Z; Yang, C G; Yao, X Y; Ye, J B; Yeh, S C; You, J M; Zalite, A; Zalite, Yu; Zemp, P; Zeng, Y; Zhang, Z; Zhang, Z P; Zhou, B; Zhou, Y; Zhu, G Y; Zhu, R Y; Zichichi, Antonino; Ziegler, F

    1997-01-01

    The two-photon collision reaction e+e- --> e+e-l+l- has been studied at root(s) ~ 91 GeV using the L3 detector at LEP for l = e, muon , tau. We have analysed untagged configurations where the two photons are quasi-real. Good agreement is found between our measurements and the order alpha**4 QED expectation.

  10. Spin polarization in top pair production in association with two photons at NLO+PS

    CERN Document Server

    Luisoni, Gionata

    2017-01-01

    This talk focuses on the impact of top-quark spin polarization effects in Higgs boson production in association with a top-quark pair, where the Higgs boson decays to two photons. Predictions for the signal are compared with direct top-quark pair production in association with two photons at NLO+PS.

  11. Two-Photon Excitation Microscopy for the Study of Living Cells and Tissues

    Science.gov (United States)

    Benninger, Richard K.P.; Piston, David W.

    2013-01-01

    Two-photon excitation microscopy is an alternative to confocal microscopy that provides advantages for three-dimensional and deep tissue imaging. This unit will describe the basic physical principles behind two-photon excitation and discuss the advantages and limitations of its use in laser-scanning microscopy. The principal advantages of two-photon microscopy are reduced phototoxicity, increased imaging depth, and the ability to initiate highly localized photochemistry in thick samples. Practical considerations for the application of two-photon microscopy will then be discussed, including recent technological advances. This unit will conclude with some recent applications of two-photon microscopy that highlight the key advantages over confocal microscopy and the types of experiments which would benefit most from its application. PMID:23728746

  12. Review: two-photon scanning systems for clinical high resolution in vivo tissue imaging

    Science.gov (United States)

    König, K.; Müller, J.; Höfer, M.; Müller, C.; Weinigel, M.; Bückle, R.; Elsner, P.; Kaatz, M.; Messerschmidt, B.

    2008-02-01

    The femtosecond laser multiphoton tomograph DermaInspect as well as high NA two-photon GRIN microendoscopes for in vivo tomography of human skin have been used to detect malignant melanoma as well as to study the diffusion and intradermal accumulation of topically applied cosmetical and pharmaceutical components. So far, more than 500 patients and volunteers in Europe, Australia, and Asia have been investigated with this unique tomograph. Near infrared 80 MHz picojoule femtosecond laser pulses were employed to excite endogenous fluorophores such as NAD(P)H, flavoproteins, melanin, and elastin as well as fluorescent components of a variety of ointments via a twophoton excitation process. In addition, collagen has been imaged by second harmonic generation. Using a two-PMT detection system, the ratio of elastin to collagen was determined during optical sectioning. A high submicron spatial resolution and 50 picosecond temporal resolution was achieved using galvoscan mirrors and piezodriven focusing optics as well as a time-correlated single photon counting module with a fast microchannel plate detector and fast photomultipliers. Individual intratissue cells, mitochondria, melanosomes, and the morphology of the nuclei as well as extracellular matrix elements could be clearly visualized due to molecular imaging and the calculation of fluorescence lifetime images. Nanoparticles and intratissue drugs have been detected non-invasively, in situ and over a period of up to 3 months. In addition, hydration effects and UV effects were studied by monitoring modifications of cellular morphology and autofluorescence. The system was used to observe the diffusion through the stratum corneum and the accumulation and release of functionalized nanoparticles along hair shafts and epidermal ridges. The DermaInspect been also employed to gain information on skin age and wound healing in patients with ulcers. Novel developments include a galvo/piezo-scan driven flexible articulated arm as

  13. Label-free imaging immune cells and collagen in atherosclerosis with two-photon and second harmonic generation microscopy

    Directory of Open Access Journals (Sweden)

    Chunqiang Li

    2016-01-01

    Full Text Available Atherosclerosis has been recognized as a chronic inflammation disease, in which many types of cells participate in this process, including lymphocytes, macrophages, dendritic cells (DCs, mast cells, vascular smooth muscle cells (SMCs. Developments in imaging technology provide the capability to observe cellular and tissue components and their interactions. The knowledge of the functions of immune cells and their interactions with other cell and tissue components will facilitate our discovery of biomarkers in atherosclerosis and prediction of the risk factor of rupture-prone plaques. Nonlinear optical microscopy based on two-photon excited autofluorescence and second harmonic generation (SHG were developed to image mast cells, SMCs and collagen in plaque ex vivo using endogenous optical signals. Mast cells were imaged with two-photon tryptophan autofluorescence, SMCs were imaged with two-photon NADH autofluorescence, and collagen were imaged with SHG. This development paves the way for further study of mast cell degranulation, and the effects of mast cell derived mediators such as induced synthesis and activation of matrix metalloproteinases (MMPs which participate in the degradation of collagen.

  14. Laser-Induced Fluorescence diagnostic of barium ion plasmas in the Paul Trap Simulator Experiment

    International Nuclear Information System (INIS)

    Chung, Moses; Gilson, Erik P.; Davidson, Ronald C.; Efthimion, Philip C.; Majeski, Richard; Startsev, Edward A.

    2005-01-01

    The Paul Trap Simulator Experiment (PTSX) is a cylindrical Paul trap whose purpose is to simulate the nonlinear dynamics of intense charged particle beam propagation in alternating-gradient magnetic transport systems. To investigate the ion plasma microstate in PTSX, including the ion density profile and the ion velocity distribution function, a laser-induced fluorescence diagnostic system is being developed as a nondestructive diagnostic. Instead of cesium, which has been used in the initial phase of the PTSX experiment, barium has been selected as the preferred ion for the laser-induced fluorescence diagnostic. A feasibility study of the laser-induced fluorescence diagnostic using barium ions is presented with the characterization of a tunable dye laser. The installation of the barium ion source and the development of the laser-induced fluorescence diagnostic system are also discussed

  15. Laser-Induced Fluorescence Measurements within a Laboratory Hall Thruster (Postprint)

    National Research Council Canada - National Science Library

    Hargus, Jr., W. A; Cappelli, M. A

    1999-01-01

    In this paper, we describe the results of a study of laser induced fluorescence velocimetry of ionic xenon in the plume and interior acceleration channel of a laboratory Hall type thruster operating...

  16. Fabrication of fluorescent silica nanoparticles with aggregation-induced emission luminogens for cell imaging.

    Science.gov (United States)

    Chen, Sijie; Lam, Jacky W Y; Tang, Ben Zhong

    2013-01-01

    Fluorescence-based techniques have found wide applications in life science. Among various luminogenic materials, fluorescent nanoparticles have attracted much attention due to their fabulous emission properties and potential applications as sensors. Here, we describe the fabrication of fluorescent silica nanoparticles (FSNPs) containing aggregation-induced emission (AIE) luminogens. By employing surfactant-free sol-gel reaction, FSNPs with uniform size and high surface charge and colloidal stability are generated. The FSNPs emit strong light upon photoexcitation, due to the AIE characteristic of the silole -aggregates in the hybrid nanoparticles. The FSNPs are cytocompatible and can be utilized as fluorescent visualizer for intracellular imaging for HeLa cells.

  17. Experimental Research of Reliability of Plant Stress State Detection by Laser-Induced Fluorescence Method

    Directory of Open Access Journals (Sweden)

    Yury Fedotov

    2016-01-01

    Full Text Available Experimental laboratory investigations of the laser-induced fluorescence spectra of watercress and lawn grass were conducted. The fluorescence spectra were excited by YAG:Nd laser emitting at 532 nm. It was established that the influence of stress caused by mechanical damage, overwatering, and soil pollution is manifested in changes of the spectra shapes. The mean values and confidence intervals for the ratio of two fluorescence maxima near 685 and 740 nm were estimated. It is presented that the fluorescence ratio could be considered a reliable characteristic of plant stress state.

  18. Fluorescence microscopy.

    Science.gov (United States)

    Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D

    2014-10-01

    Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

  19. Atomic frequency reference at 1033 nm for ytterbium (Yb)-doped fiber lasers and applications exploiting a rubidium (Rb) 5S_1/2 to 4D_5/2 one-colour two-photon transition

    Science.gov (United States)

    Roy, Ritayan; Condylis, Paul C.; Johnathan, Yik Jinen; Hessmo, Björn

    2017-04-01

    We demonstrate a two-photon transition of rubidium (Rb) atoms from the ground state (5$S_{1/2}$) to the excited state (4$D_{5/2}$), using a home-built ytterbium (Yb)-doped fiber amplifier at 1033 nm. This is the first demonstration of an atomic frequency reference at 1033 nm as well as of a one-colour two-photon transition for the above energy levels. A simple optical setup is presented for the two-photon transition fluorescence spectroscopy, which is useful for frequency stabilization for a broad class of lasers. This spectroscopy has potential applications in the fiber laser industry as a frequency reference, particularly for the Yb-doped fiber lasers. This two-photon transition also has applications in atomic physics as a background- free high- resolution atom detection and for quantum communication, which is outlined in this article.

  20. [Laser induced fluorescence spectrum characteristics of common edible oil and fried cooking oil].

    Science.gov (United States)

    Mu, Tao-tao; Chen, Si-ying; Zhang, Yin-chao; Chen, He; Guo, Pan; Ge, Xian-ying; Gao, Li-lei

    2013-09-01

    In order to detect the trench oil the authors built a trench oil rapid detection system based on laser induced fluorescence detection technology. This system used 355 nm laser as excitation light source. The authors collected the fluorescence spectrum of a variety of edible oil and fried cooking oil (a kind of trench oil) and then set up a fluorescence spectrum database by taking advantage of the trench oil detection system It was found that the fluorescence characteristics of fried cooking oil and common edible oil were obviously different. Then it could easily realize the oil recognition and trench oil rapid detection by using principal component analysis and BP neural network, and the overall recognition rate could reach as high as 97.5%. Experiments showed that laser induced fluorescence spectrum technology was fast, non-contact, and highly sensitive. Combined with BP neural network, it would become a new technique to detect the trench oil.

  1. Concentration of atomic hydrogen in a dielectric barrier discharge measured by two-photon absorption fluorescence

    Czech Academy of Sciences Publication Activity Database

    Dvořák, P.; Talába, M.; Obrusník, A.; Kratzer, Jan; Dědina, Jiří

    2017-01-01

    Roč. 26, č. 8 (2017), č. článku 085002. ISSN 0963-0252 R&D Projects: GA ČR(CZ) GA17-04329S; GA ČR GA14-23532S Institutional support: RVO:68081715 Keywords : TALIF * dielectric barrier discharge * H radicals Subject RIV: CB - Analytical Chemistry, Separation OBOR OECD: Analytical chemistry Impact factor: 3.302, year: 2016

  2. Brain refractive index measured in vivo with high-NA defocus-corrected full-field OCT and consequences for two-photon microscopy.

    Science.gov (United States)

    Binding, Jonas; Ben Arous, Juliette; Léger, Jean-François; Gigan, Sylvain; Boccara, Claude; Bourdieu, Laurent

    2011-03-14

    Two-photon laser scanning microscopy (2PLSM) is an important tool for in vivo tissue imaging with sub-cellular resolution, but the penetration depth of current systems is potentially limited by sample-induced optical aberrations. To quantify these, we measured the refractive index n' in the somatosensory cortex of 7 rats in vivo using defocus optimization in full-field optical coherence tomography (ff-OCT). We found n' to be independent of imaging depth or rat age. From these measurements, we calculated that two-photon imaging beyond 200 µm into the cortex is limited by spherical aberration, indicating that adaptive optics will improve imaging depth.

  3. Measuring solar induced chlorophyll fluorescence (SIF) in the Amazon rainforest

    Science.gov (United States)

    Kornfeld, A.; Stutz, J.; Berry, J. A.

    2016-12-01

    Measurement of solar induced chlorophyll fluorescence (SIF) has, in our hands, been fraught with missteps and puzzling problems. Here we describe lessons we have learned and the resulting novel system recently installed in the Amazon rainforest near Manaus, Brazil. The system is designed to measure light from 740 - 780 nm, enabling us to compare SIF computed from Fraunhofer lines in an optically transparent band of the atmosphere (745 - 759 nm) with SIF computed using the telluric O2A band (760 - 770 nm). Fraunhofer line analysis requires high optical resolution (better than 0.2 nm) to detect the relatively narrow lines, but we discovered that fiber-optic diffraction-grating spectrometers are sensitive to very small inhomogeneities in the lighting. Errors resulting from this autocorrelated but random noise were similar in magnitude to the SIF signal itself. Optical diffusers reduce this problem, leading to our final design: a sealed cylinder, dubbed Rotaprism, in which a rotatable prism selects whether light from upward- or downward-looking windows enters an axially-placed optical fiber. Cosine-correcting opal glass covering the windows not only solves the noise issue but also makes the measurements correspond to photon flux. Rotaprism also maximizes the amount of light reaching the spectrometer - maximizing the signal:noise ratio - by avoiding the need for lossy optical switches and fiber splitters. Rotaprism is driven by a pneumatic actuator that is controlled by electronic valves attached to a pressurized N2 source. The gas exhausts into the temperature-controlled spectrometer enclosure to help purge the optics. Finally, custom software provides fault-tolerant control and data acquisition, ensuring that measurements continue with little or no intervention at the remote field site despite unreliable power. Analysis of initial data demonstrates the advantage of Fraunhofer line SIF analysis: due to the atmosphere transparency in this band, the results are more

  4. Teaching laser-induced fluorescence of plant leaves

    Science.gov (United States)

    Lenk, Sándor; Gádoros, Patrik; Kocsányi, László; Barócsi, Attila

    2016-11-01

    Plants convert carbon dioxide into sugars using the energy of sunlight. Absorbed light unused for conversion is dissipated primarily as heat with a small fraction re-emitted as fluorescence at longer wavelengths. One can use the latter to estimate photosynthetic activity. The illumination of intact leaves with strong light after keeping them in dark for tens of minutes results in a rapid increase followed by a slow decay of fluorescence emission from the fluorophore chlorophyll-a, called the Kautsky effect. This paper describes a laboratory practice that introduces students of physics or engineering into this research field. It begins with the spectral measurement of the fluorescence emitted by a plant leaf upon UV excitation. Then it focuses on the red and far-red components of the fluorescence emission spectrum characteristic to the chlorophyll-a molecule and presents an inexpensive demonstration of the Kautsky effect. As researchers use more complex measurement techniques and tools, the practice ends up with the demonstration of an intelligent fluorosensor, a compact tool developed for plant physiological research and horticulture applications together with a brief interpretation of some important fluorescence parameters.

  5. Teaching laser-induced fluorescence of plant leaves

    International Nuclear Information System (INIS)

    Lenk, Sándor; Gádoros, Patrik; Kocsányi, László; Barócsi, Attila

    2016-01-01

    Plants convert carbon dioxide into sugars using the energy of sunlight. Absorbed light unused for conversion is dissipated primarily as heat with a small fraction re-emitted as fluorescence at longer wavelengths. One can use the latter to estimate photosynthetic activity. The illumination of intact leaves with strong light after keeping them in dark for tens of minutes results in a rapid increase followed by a slow decay of fluorescence emission from the fluorophore chlorophyll -a , called the Kautsky effect. This paper describes a laboratory practice that introduces students of physics or engineering into this research field. It begins with the spectral measurement of the fluorescence emitted by a plant leaf upon UV excitation. Then it focuses on the red and far-red components of the fluorescence emission spectrum characteristic to the chlorophyll -a molecule and presents an inexpensive demonstration of the Kautsky effect. As researchers use more complex measurement techniques and tools, the practice ends up with the demonstration of an intelligent fluorosensor, a compact tool developed for plant physiological research and horticulture applications together with a brief interpretation of some important fluorescence parameters. (paper)

  6. LETTER TO THE EDITOR: Green emission and bandgap narrowing due to two-photon excitation in thin film CdS formed by spray pyrolysis

    Science.gov (United States)

    Ullrich, B.; Schroeder, R.

    2001-08-01

    Thin (10 µm) film CdS on Pyrex® formed by spray pyrolysis is excited below the gap at 804 nm with 200 fs laser pulses at room temperature. Excitation intensities up to 250 GW cm-2 evoke green bandgap emission due to two-photon transitions. This two-photon photoluminescence does not show a red emission contribution in contrast to the single-photon excited emission, which is dominated by broad emission in the red spectral range. It is demonstrated that two-photon excitation causes photo-induced bandgap narrowing due to Debye screening. At 250 GW cm-2 bandgap narrowing of 47 meV is observed, which corresponds to an excited electron density of 1.6×1018 cm-3.

  7. An orthotopic glioblastoma mouse model maintaining brain parenchymal physical constraints and suitable for intravital two-photon microscopy.

    Science.gov (United States)

    Ricard, Clément; Stanchi, Fabio; Rougon, Geneviève; Debarbieux, Franck

    2014-04-21

    Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date. Murine models of this pathology rely on the injection of a suspension of glioma cells into the brain parenchyma following incision of the dura-mater. Whereas the cells have to be injected superficially to be accessible to intravital two-photon microscopy, superficial injections fail to recapitulate the physiopathological conditions. Indeed, escaping through the injection tract most tumor cells reach the extra-dural space where they expand abnormally fast in absence of mechanical constraints from the parenchyma. Our improvements consist not only in focally implanting a glioma spheroid rather than injecting a suspension of glioma cells in the superficial layers of the cerebral cortex but also in clogging the injection site by a cross-linked dextran gel hemi-bead that is glued to the surrounding parenchyma and sealed to dura-mater with cyanoacrylate. Altogether these measures enforce the physiological expansion and infiltration of the tumor cells inside the brain parenchyma. Craniotomy was finally closed with a glass window cemented to the skull to allow chronic imaging over weeks in absence of scar tissue development. Taking advantage of fluorescent transgenic animals grafted with fluorescent tumor cells we have shown that the dynamics of interactions occurring between glioma cells, neurons (e.g. Thy1-CFP mice) and vasculature (highlighted by an intravenous injection of a fluorescent dye) can be visualized by intravital two-photon microscopy during the progression of the disease. The possibility to image a tumor at microscopic resolution in a minimally compromised cerebral environment represents an improvement of current GBM animal models which should benefit the field of neuro-oncology and drug testing.

  8. Optical nonlinearities of colloidal InP@ZnS core-shell quantum dots probed by Z-scan and two-photon excited emission

    International Nuclear Information System (INIS)

    Wawrzynczyk, Dominika; Szeremeta, Janusz; Samoc, Marek; Nyk, Marcin

    2015-01-01

    Spectrally resolved nonlinear optical properties of colloidal InP@ZnS core-shell quantum dots of various sizes were investigated with the Z-scan technique and two-photon fluorescence excitation method using a femtosecond laser system tunable in the range from 750 nm to 1600 nm. In principle, both techniques should provide comparable results and can be interchangeably used for determination of the nonlinear optical absorption parameters, finding maximal values of the cross sections and optimizing them. We have observed slight differences between the two-photon absorption cross sections measured by the two techniques and attributed them to the presence of non-radiative paths of absorption or relaxation. The most significant value of two-photon absorption cross section σ 2 for 4.3 nm size InP@ZnS quantum dot was equal to 2200 GM, while the two-photon excitation action cross section σ 2 Φ was found to be 682 GM at 880 nm. The properties of these cadmium-free colloidal quantum dots can be potentially useful for nonlinear bioimaging

  9. Rotationally cooled laser induced fluorescence determination of polycyclic aromatic hydrocarbons

    International Nuclear Information System (INIS)

    Warren, J.A.; Hayes, J.M.; Small, G.J.

    1982-01-01

    In recent years the development of new highly selective and sensitive methods for the characterization and determination of polycyclic aromatic hydrocarbons (PAHs) and their derivatives in complex mixtures has received considerable attention. High selectivity is associated here with the ability to distinguish between substitutional isomers of PAHs. Attainment of this selectivity with capillary column-gas chromatography-mass spectrometry for complex mixtures is very difficult and time-consuming. Alternative approaches are, therefore, required. Given that the majority of PAHs fluoresce with reasonable quantum yields and that high sensitivities are afforded by fluorescence detection, the possibility of developing high-resolution fluorescence based techniques is attractive. This is all the more so if the technique's selectivity does not rely on physical separation, e.g., chromatography. In this paper discussion is limited to such techniques

  10. Time-resolved resonance fluorescence spectroscopy for study of chemical reactions in laser-induced plasmas.

    Science.gov (United States)

    Liu, Lei; Deng, Leimin; Fan, Lisha; Huang, Xi; Lu, Yao; Shen, Xiaokang; Jiang, Lan; Silvain, Jean-François; Lu, Yongfeng

    2017-10-30

    Identification of chemical intermediates and study of chemical reaction pathways and mechanisms in laser-induced plasmas are important for laser-ablated applications. Laser-induced breakdown spectroscopy (LIBS), as a promising spectroscopic technique, is efficient for elemental analyses but can only provide limited information about chemical products in laser-induced plasmas. In this work, time-resolved resonance fluorescence spectroscopy was studied as a promising tool for the study of chemical reactions in laser-induced plasmas. Resonance fluorescence excitation of diatomic aluminum monoxide (AlO) and triatomic dialuminum monoxide (Al 2 O) was used to identify these chemical intermediates. Time-resolved fluorescence spectra of AlO and Al 2 O were used to observe the temporal evolution in laser-induced Al plasmas and to study their formation in the Al-O 2 chemistry in air.

  11. Two-photon direct frequency comb spectroscopy of alkali atoms

    Science.gov (United States)

    Palm, Christopher; Pradhananga, Trinity; Nguyen, Khoa; Montcrieffe, Caitlin; Kimball, Derek

    2012-11-01

    We have studied transition frequencies and excited state hyperfine structure in rubidium using 2-photon transitions excited directly with the frequency-doubled output of a erbium fiber optical frequency comb. The frequency comb output is directed in two counterpropagating directions through a vapor cell containing the rubidium vapor. A pair of optical filters is used to select teeth of the comb in order to identify the transition wavelengths. A photomultiplier tube (PMT) measures fluorescence from a decay channel wavelength selected with another optical filter. Using different combinations of filters enables a wide range of transitions to be investigated. By scanning the repetition rate, a Doppler-free spectrum can be obtained enabling kHz-resolution spectral measurements. An interesting dependence of the 2-photon spectrum on the energy of the intermediate state of the 2-photon transition is discussed. Our investigations are laying the groundwork for a long-term research program to use direct frequency comb spectroscopy to understand the complex spectra of rare-earth atoms.

  12. The study of nonlinear two-photon phenomenon in photonic crystals doped with nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Mahi R [Department of Physics and Astronomy, University of Western Ontario, London, N6A 3K7 (Canada)

    2007-02-28

    A theory of the nonlinear two-photon absorption has been developed in a photonic crystal doped with an ensemble of four-level nanoparticles. We have considered that the nanoparticles are interacting with the photonic crystal. An expression of two-photon absorption has been obtained by using the density matrix method. The effect of the dipole-dipole interaction has also been included in the formulation. Interesting new phenomena have been predicted. For example, it is found that the inhibition of two-photon absorption can be turned on and off when the decay resonance energies of the four-level nanoparticles are moved within the energy band.

  13. Two-photon Doppler cooling of alkaline-earth-metal and ytterbium atoms

    International Nuclear Information System (INIS)

    Magno, Wictor C.; Cavasso Filho, Reinaldo L.; Cruz, Flavio C.

    2003-01-01

    The possibility of laser cooling of alkaline-earth-metal atoms and ytterbium atoms using a two-photon transition is analyzed. We consider a 1 S 0 - 1 S 0 transition with excitation in near resonance with the 1 P 1 level. This greatly increases the two-photon transition rate, allowing an effective transfer of momentum. The experimental implementation of this technique is discussed and we show that for calcium, for example, two-photon cooling can be used to achieve a Doppler limit of 123 μK. The efficiency of this cooling scheme and the main loss mechanisms are analyzed

  14. Dicke coherent narrowing in two-photon and Raman spectroscopy of thin vapor cells

    International Nuclear Information System (INIS)

    Dutier, Gabriel; Todorov, Petko; Hamdi, Ismahene; Maurin, Isabelle; Saltiel, Solomon; Bloch, Daniel; Ducloy, Martial

    2005-01-01

    The principle of coherent Dicke narrowing in a thin vapor cell, in which sub-Doppler spectral line shapes are observed under a normal irradiation for a λ/2 thickness, is generalized to two-photon spectroscopy. Only the sum of the two wave vectors must be normal to the cell, making the two-photon scheme highly versatile. A comparison is provided between the Dicke narrowing with copropagating fields, and the residual Doppler broadening occurring with counterpropagating geometries. The experimental feasibility is discussed on the basis of a first observation of a two-photon resonance in a 300-nm-thick Cs cell. Extension to the Raman situation is finally considered

  15. Limitations of two-level emitters as nonlinearities in two-photon controlled-PHASE gates

    DEFF Research Database (Denmark)

    Nysteen, Anders; McCutcheon, Dara P. S.; Heuck, Mikkel

    2017-01-01

    We investigate the origin of imperfections in the fidelity of a two-photon controlled-PHASE gate based on two-level-emitter nonlinearities. We focus on a passive system that operates without external modulations to enhance its performance. We demonstrate that the fidelity of the gate is limited...... by opposing requirements on the input pulse width for one-and two-photon-scattering events. For one-photon scattering, the spectral pulse width must be narrow compared with the emitter linewidth, while two-photon-scattering processes require the pulse width and emitter linewidth to be comparable. We find...

  16. Two-photon absorption of a supramolecular pseudoisocyanine J-aggregate assembly

    International Nuclear Information System (INIS)

    Belfield, Kevin D.; Bondar, Mykhailo V.; Hernandez, Florencio E.; Przhonska, Olga V.; Yao, Sheng

    2006-01-01

    Linear spectral properties, including excitation anisotropy, of pseudoisocyanine or 1,1'-diethyl-2,2'-cyanine iodide (PIC) J-aggregates in aqueous solutions with J-band position at 573 nm were investigated. Two-photon absorption of PIC J-aggregates and monomer molecules was studied using an open aperture Z-scan technique. A strong enhancement of the two-photon absorption cross-section of PIC in the supramolecular J-aggregate assembly was observed in aqueous solution. This enhancement is attributed to a strong coupling of the molecular transition dipoles. No two-photon absorption at the peak of the J-band was detected

  17. Two-photon Anderson localization in a disordered quadratic waveguide array

    International Nuclear Information System (INIS)

    Bai, Y F; Xu, P; Lu, L L; Zhong, M L; Zhu, S N

    2016-01-01

    We theoretically investigate two-photon Anderson localization in a χ (2) waveguide array with off-diagonal disorder. The nonlinear parametric down-conversion process would enhance both the single-photon and the two-photon Anderson localization. In the strong disorder regime, the two-photon position correlation exhibits a bunching distribution around the pumped waveguides, which is independent of pumping conditions and geometrical structures of waveguide arrays. Quadratic nonlinearity can be supplied as a new ingredient for Anderson localization. Also, our results pave the way for engineering quantum states through nonlinear quantum walks. (paper)

  18. Wide-field two-photon microscopy with temporal focusing and HiLo background rejection

    Science.gov (United States)

    Yew, Elijah Y. S.; Choi, Heejin; Kim, Daekeun; So, Peter T. C.

    2011-03-01

    Scanningless depth-resolved microscopy is achieved through spatial-temporal focusing and has been demonstrated previously. The advantage of this method is that a large area may be imaged without scanning resulting in higher throughput of the imaging system. Because it is a widefield technique, the optical sectioning effect is considerably poorer than with conventional spatial focusing two-photon microscopy. Here we propose wide-field two-photon microscopy based on spatio-temporal focusing and employing background rejection based on the HiLo microscope principle. We demonstrate the effects of applying HiLo microscopy to widefield temporally focused two-photon microscopy.

  19. Chlorophyll induced fluorescence retrieved from GOME2 for improving gross primary productivity estimates of vegetation

    Science.gov (United States)

    van Leth, Thomas C.; Verstraeten, Willem W.; Sanders, Abram F. J.

    2014-05-01

    Mapping terrestrial chlorophyll fluorescence is a crucial activity to obtain information on the functional status of vegetation and to improve estimates of light-use efficiency (LUE) and global primary productivity (GPP). GPP quantifies carbon fixation by plant ecosystems and is therefore an important parameter for budgeting terrestrial carbon cycles. Satellite remote sensing offers an excellent tool for investigating GPP in a spatially explicit fashion across different scales of observation. The GPP estimates, however, still remain largely uncertain due to biotic and abiotic factors that influence plant production. Sun-induced fluorescence has the ability to enhance our knowledge on how environmentally induced changes affect the LUE. This can be linked to optical derived remote sensing parameters thereby reducing the uncertainty in GPP estimates. Satellite measurements provide a relatively new perspective on global sun-induced fluorescence, enabling us to quantify spatial distributions and changes over time. Techniques have recently been developed to retrieve fluorescence emissions from hyperspectral satellite measurements. We use data from the Global Ozone Monitoring Instrument 2 (GOME2) to infer terrestrial fluorescence. The spectral signatures of three basic components atmospheric: absorption, surface reflectance, and fluorescence radiance are separated using reference measurements of non-fluorescent surfaces (desserts, deep oceans and ice) to solve for the atmospheric absorption. An empirically based principal component analysis (PCA) approach is applied similar to that of Joiner et al. (2013, ACP). Here we show our first global maps of the GOME2 retrievals of chlorophyll fluorescence. First results indicate fluorescence distributions that are similar with that obtained by GOSAT and GOME2 as reported by Joiner et al. (2013, ACP), although we find slightly higher values. In view of optimizing the fluorescence retrieval, we will show the effect of the references

  20. Hyperspectral solar-induced chlorophyll fluorescence of urban tree leaves: Analyses and applications

    Science.gov (United States)

    Van Wittenberghe, Shari

    Solar energy is the primary energy source for life on Earth which is converted into chemical energy through photosynthesis by plants, algae and cyanobacteria, releasing fuel for the organisms' activities. To dissipate excess of absorbed light energy, plants emit chlorophyll (Chl) fluorescence (650-850 nm) from the same location where photosynthesis takes place. Hence, it provides information on the efficiency of primary energy conversion. From this knowledge, many applications on vegetation and crop stress monitoring could be developed, a necessity for our planet under threat of a changing global climate. Even though the Chl fluorescence signal is weak against the intense reflected radiation background, methods for retrieving the solar-induced Chl fluorescence have been refined over the last years, both at leaf and airborne scale. However, a lack of studies on solar-induced Chl fluorescence gives difficulties for the interpretation of the signal. Within this thesis, hyperspectral upward and downward solar-induced Chl fluorescence is measured at leaf level. Fluorescence yield (FY) is calculated as well as different ratios characterizing the emitted Chl fluorescence shape. The research in this PhD dissertation illustrates the influence of several factors on the solar-induced Chl fluorescence signal. For instance, both the intensity of FY and its spectral shape of urban tree leaves are able to change under influence of stress factors such as traffic air pollution. This shows how solar-induced Chl fluorescence could function as an early stress indicator for vegetation. Further, it is shown that the signal contains information on the ultrastructure of the photosynthetic apparatus. Also, it is proven that the leaf anatomical structure and related light scattering properties play a role in the partitioning between upward and downward Chl fluorescence emission. All these findings indicate how the Chl fluorescence spectrum is influenced by factors which also influence

  1. Novel nanocarriers for topical drug delivery: investigating delivery efficiency and distribution in skin using two-photon microscopy

    Science.gov (United States)

    Kirejev, Vladimir; Guldbrand, Stina; Bauer, Brigitte; Smedh, Maria; Ericson, Marica B.

    2011-03-01

    The complex structure of skin represents an effective barrier against external environmental factors, as for example, different chemical and biochemical compounds, yeast, bacterial and viral infections. However, this impermeability prevents efficient transdermal drug delivery which limits the number of drugs that are able to penetrate the skin efficiently. Current trends in drug application through skin focus on the design and use of nanocarriers for transport of active compounds. The transport systems applied so far have several drawbacks, as they often have low payload, high toxicity, a limited variability of inclusion molecules, or long degradation times. The aim of these current studies is to investigate novel topical drug delivery systems, e.g. nanocarriers based on cyclic oligosaccharides - cyclodextrins (CD) or iron (III)-based metal-organic frameworks (MOF). Earlier studies on cell cultures imply that these drug nanocarriers show promising characteristics compared to other drug delivery systems. In our studies, we use two-photon microscopy to investigate the ability of the nanocarriers to deliver compounds through ex-vivo skin samples. Using near infrared light for excitation in the so called optical window of skin allows deep-tissue visualization of drug distribution and localization. In addition, it is possible to employ two-photon based fluorescence correlation spectroscopy for quantitative analysis of drug distribution and concentrations in different cell layers.

  2. Determination of absolute Ba densities during dimming operation of fluorescent lamps by laser-induced fluorescence measurements

    International Nuclear Information System (INIS)

    Hadrath, S; Beck, M; Garner, R C; Lieder, G; Ehlbeck, J

    2007-01-01

    Investigations of fluorescent lamps (FL) are often focused on the electrodes, since the lifetime of the lamps is typically limited by the electrode lifetime and durability. During steady state operation, the work function lowering emitter material, in particular, barium, is lost. Greater barium losses occur under dimming conditions, in which reduced discharge currents lead to increased cathode falls, the result of the otherwise diminished heating of the electrode by the bombarding plasma ions. In this work the barium density near the electrodes of (FL), operating in high frequency dimming mode is investigated using the high-sensitivity method of laser-induced fluorescence. From these measurements we infer barium loss for a range of discharge currents and auxiliary coil heating currents. We show that the Ba loss can very easily be reduced by moderate auxiliary coil heating

  3. Two-photon excited whispering-gallery mode ultraviolet laser from an individual ZnO microneedle

    Science.gov (United States)

    Zhu, G. P.; Xu, C. X.; Zhu, J.; Lv, C. G.; Cui, Y. P.

    2009-02-01

    Wurtzite structural ZnO microneedles with hexagonal cross section were fabricated by vapor-phase transport method and an individual microneedle was employed as a lasing microcavity. Under excitation of a femtosecond pulse laser with 800 nm wavelength, the ultraviolet (UV) laser emission was obtained, which presented narrow linewidth and high Q value. The UV emission, resonant mechanism, and laser mode characteristics were discussed in detail. The results demonstrated that the UV laser originated from the whispering-gallery mode induced by two-photon absorption assisted by Rabi oscillation.

  4. Spatial variability of oceanic phycoerythrin spectral types derived from airborne laser-induced fluorescence emissions

    Science.gov (United States)

    Hoge, Frank E.; Wright, C. Wayne; Kana, Todd M.; Swift, Robert N.; Yungel, James K.

    1998-07-01

    We report spatial variability of oceanic phycoerythrin spectral types detected by means of a blue spectral shift in airborne laser-induced fluorescence emission. The blue shift of the phycoerythrobilin fluorescence is known from laboratory studies to be induced by phycourobilin chromophore substitution at phycoerythrobilin chromophore sites in some strains of phycoerythrin-containing marine cyanobacteria. The airborne 532-nm laser-induced phycoerythrin fluorescence of the upper oceanic volume showed distinct segregation of cyanobacterial chromophore types in a flight transect from coastal water to the Sargasso Sea in the western North Atlantic. High phycourobilin levels were restricted to the oceanic (oligotrophic) end of the flight transect, in agreement with historical ship findings. These remotely observed phycoerythrin spectral fluorescence shifts have the potential to permit rapid, wide-area studies of the spatial variability of spectrally distinct cyanobacteria, especially across interfacial regions of coastal and oceanic water masses. Airborne laser-induced phytoplankton spectral fluorescence observations also further the development of satellite algorithms for passive detection of phytoplankton pigments. Optical modifications to the NASA Airborne Oceanographic Lidar are briefly described that permitted observation of the fluorescence spectral shifts.

  5. Development of laser-induced fluorescence detection to assay DNA damage

    International Nuclear Information System (INIS)

    Sharma, M.; Freund, H.G.

    1991-01-01

    A precolumn derivation method has been developed for high performance liquid chromatographic (HPLC) analysis of DNA damage using fluorescence detection. The modified nucleotide, having excised enzymatically from the exposed DNA, is enriched from the normal nucleotides and labeled with a fluorescent reagent. The labeling procedure involves phosphoramidation of the nucleotide with ethylenediamine (EDA) followed by conjugation of the free amino end of the phosphoramidate with 5-dimethylaminonaphthalene 1-sulfonyl chloride, commonly known as Dansyl chloride. The dansylated nucleotide can be analyzed with a sub-picomole limit of detection (LOD) by conventional HPLC using a conventional fluorescence detector. By combining microbore HPLC with laser-induced fluorescence (LIF) detection, the authors present the development of an analytical system that has sub-femtomole LOD for real-time analysis of the dansylated nucleotide. In this paper the application of the developed system in fluorescence postlabeling assay of a small alkyl-modified nucleotide (5-methyl CMP) in calf-thymus DNA is discussed

  6. Effects of autoionizing states on two-photon double ionization of the H2 molecule

    International Nuclear Information System (INIS)

    Guan, Xiaoxu; Bartschat, Klaus; Koesterke, Lars; Schneider, Barry I

    2014-01-01

    We report angle-resolved and angle-integrated cross sections for two-photon double-ionization of H by a strong laser pulse. The effect of doubly excited states on the predicted cross sections is addressed.

  7. Mitochondria-targeted cationic porphyrin-triphenylamine hybrids for enhanced two-photon photodynamic therapy.

    Science.gov (United States)

    Hammerer, Fabien; Poyer, Florent; Fourmois, Laura; Chen, Su; Garcia, Guillaume; Teulade-Fichou, Marie-Paule; Maillard, Philippe; Mahuteau-Betzer, Florence

    2018-01-01

    The proof of concept for two-photon activated photodynamic therapy has already been achieved for cancer treatment but the efficiency of this approach still heavily relies on the availability of photosensitizers combining high two-photon absorption and biocompatibility. In this line we recently reported on a series of porphyrin-triphenylamine hybrids which exhibit high singlet oxygen production quantum yield as well as high two-photon absorption cross-sections but with a very poor cellular internalization. We present herein new photosensitizers of the same porphyrin-triphenylamine hybrid series but bearing cationic charges which led to strongly enhanced water solubility and thus cellular penetration. In addition the new compounds have been found localized in mitochondria that are preferential target organelles for photodynamic therapy. Altogether the strongly improved properties of the new series combined with their specific mitochondrial localization lead to a significantly enhanced two-photon activated photodynamic therapy efficiency. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Two-photon Microscopy and Polarimetry for Assessment of Myocardial Tissue Organization

    Science.gov (United States)

    Archambault-Wallenburg, Marika

    Optical methods can provide useful tissue characterization tools. For this project, two-photon microscopy and polarized light examinations (polarimetry) were used to assess the organizational state of myocardium in healthy, infarcted, and stem-cell regenerated states. Two-photon microscopy visualizes collagen through second-harmonic generation and myocytes through two-photon excitation autofluorescence, providing information on the composition and structure/organization of the tissue. Polarimetry measurements yield a value of linear retardance that can serve as an indicator of tissue anisotropy, and with a dual-projection method, information about the anisotropy axis orientation can also be extracted. Two-photon microscopy results reveal that stem-cell treated tissue retains more myocytes and structure than infarcted myocardium, while polarimetry findings suggest that the injury caused by temporary ligation of a coronary artery is less severe and more diffuse that than caused by a permanent ligation. Both these methods show potential for tissue characterization.

  9. A compact two photon light sheet microscope for applications in neuroscience

    DEFF Research Database (Denmark)

    Piksarv, Peeter; Marti, Dominik; Le, Tuan

    2016-01-01

    We present a compact setup for two photon light sheet microscopy. By using pulsed Airy beam illumination we demonstrate eight-fold increase of the FOV compared to Gaussian light sheet with the same axial resolution....

  10. Efficient non-linear two-photon effects from the Cesium 6D manifold

    Science.gov (United States)

    Haluska, Nathan D.; Perram, Glen P.; Rice, Christopher A.

    2018-02-01

    We report several non-linear process that occur when two-photon pumping the cesium 6D states. Cesium vapor possess some of the largest two-photon pump cross sections in nature. Pumping these cross sections leads to strong amplified spontaneous emission that we observe on over 17 lasing lines. These new fields are strong enough to couple with the pump to create additional tunable lines. We use a heat pipe with cesium densities of 1014 to 1016 cm-3 and 0 to 5 Torr of helium buffer gas. The cesium 6D States are interrogated by both high energy pulses and low power CW sources. We observe four-wave mixing, six-wave mixing, potential two-photon lasing, other unknown nonlinear processes, and the persistence of some processes at low thresholds. This system is also uniquely qualified to support two-photon lasing under the proper conditions.

  11. Suitable photo-resists for two-photon polymerization using femtosecond fiber lasers

    KAUST Repository

    Rajamanickam, V.P.; Ferrara, L.; Toma, A.; Proietti Zaccaria, R.; Das, G.; Di Fabrizio, Enzo M.; Liberale, Carlo

    2014-01-01

    We present suitable materials with good optical and mechanical properties, simple processing, efficient and optimized for two-photon polymerization (TPP) with femtosecond fiber lasers. We selected readily available acrylic monomer Bisphenol A

  12. Simultaneous control of emission localization and two-photon absorption efficiency in dissymmetrical chromophores

    International Nuclear Information System (INIS)

    Tretiak, Sergei

    2009-01-01

    The aim of the present work is to demonstrate that combined spectral tuning of fluorescence and two-photon absorption (TPA) properties of multipolar chromophores can be achieved by introduction of slight electronic chemical dissymmetry. In that perspective, two novel series of structurally related chromophores have been designed and studied: a first series based on rod-like quadrupolar chromophores bearing different electron-donating (D) end groups and a second series based on three-branched octupolar chromophores built from a trigonal donating moiety and bearing various acceptor (A) peripheral groups. The influence of the electronic dissymmetry is investigated by combined experimental and theoretical studies of the linear and nonlinear optical properties of dissymmetric chromophores compared to their symmetrical counterparts. In both types of systems (i.e. quadrupoles and octupoles) experiments and theory reveal that excitation is essentially delocalized and that excitation involves synchronized charge redistribution between the different D and A moieties within the multipolar structure (i.e. concerted intramolecular charge transfer). In contrast, the emission stems only from a particular dipolar subunit bearing the strongest D or A moieties due to fast excitation localization after excitation prior to emission. Hence control of emission characteristics (polarization and emission spectrum) in addition to localization can be achieved by controlled introduction of electronic dissymmetry (i.e. replacement of one of the D or A end-groups by a slightly stronger D(prime) or A(prime) units). Interestingly dissymmetrical functionalization of both quadrupolar and octupolar compounds does not lead to significant loss in TPA responses and can even be beneficial due to the spectral broadening and peak position tuning that it allows. This study thus reveals an original molecular engineering route strategy allowing major TPA enhancement in multipolar structures due to concerted

  13. Two-photon cooperative emission in the presence of athermal electromagnetic field

    International Nuclear Information System (INIS)

    Enaki, N.A.; Mihalache, D.

    1997-01-01

    The possibility of cooperative spontaneous two-photon emission of an extended radiators system and the influence of the external thermal electromagnetic field on the spontaneous emission rate, in such a system, are investigated. It is concluded that, in an external electromagnetic field, the two-photon cooperative emission rate increases significantly. The importance of this effect on the emission of gamma rays from inverted long-lived isomers triggered by X-ray thermal fields, is emphasized

  14. Saturated two-photon absorption by atoms in a perturber gas

    International Nuclear Information System (INIS)

    Nienhuis, G.

    1980-01-01

    We derive a general expression for the two-photon absorption spectrum of a three-state atom excited by two mono-chromatic radiation fields. Collisional line-broadening effects are incorporated, and the result allows inclusion of profiles with a validity outside the impact limit. Results of previous work are recovered in the appropriate limits. Saturation affects the different lines in the two-photon absorption spectrum in a different fashion. (orig.)

  15. Selection rules for the dematerialization of a particle into two photons

    International Nuclear Information System (INIS)

    Yang, C.N.

    1983-01-01

    Selection rules governing the disintegration of a particle into two photons are derived from the general principle of invariance under rotation and inversion. The polarization state of the photons is completely fixed by the selection rules for initial particles with spin less than 2. These results which are independent of any specific assumption about the interactions may possibly offer a method of deciding the symmetry nature of mesons which decay into two photons. 4 tables

  16. Suppression of two-photon resonantly enhanced nonlinear processes in extended media

    International Nuclear Information System (INIS)

    Garrett, W.R.; Moore, M.A.; Payne, M.G.; Wunderlich, R.K.

    1988-11-01

    On the basis of combined experimental and theoretical studies of nonlinear processes associated with two-photon excitations near 3d and 4d states in Na, we show how resonantly enhanced stimulated hyper-Raman emission, parametric four-wave mixing processes and total resonant two-photon absorption can become severely suppressed through the actions of internally generated fields on the total atomic response in extended media. 7 refs., 3 figs

  17. Influence of Two Photon Absorption on Soliton Self-Frequency Shift

    DEFF Research Database (Denmark)

    Steffensen, Henrik; Rottwitt, Karsten; Jepsen, Peter Uhd

    2011-01-01

    The creation of mid-infrared supercontinua necessitates the use of soft-glass fibers. However, some materials, like chalcogenide, have a substantial two photon absorption. We introduce a model for soliton self-frequency shift that successfully includes this effect.......The creation of mid-infrared supercontinua necessitates the use of soft-glass fibers. However, some materials, like chalcogenide, have a substantial two photon absorption. We introduce a model for soliton self-frequency shift that successfully includes this effect....

  18. Two-Photon Entanglement and EPR Experiments Using Type-2 Spontaneous Parametric Down Conversion

    Science.gov (United States)

    Sergienko, A. V.; Shih, Y. H.; Pittman, T. B.; Rubin, M. H.

    1996-01-01

    Simultaneous entanglement in spin and space-time of a two-photon quantum state generated in type-2 spontaneous parametric down-conversion is demonstrated by the observation of quantum interference with 98% visibility in a simple beam-splitter (Hanburry Brown-Twiss) anticorrelation experiment. The nonlocal cancellation of two-photon probability amplitudes as a result of this double entanglement allows us to demonstrate two different types of Bell's inequality violations in one experimental setup.

  19. Time-synchronized continuous wave laser-induced fluorescence on an oscillatory xenon discharge.

    Science.gov (United States)

    MacDonald, N A; Cappelli, M A; Hargus, W A

    2012-11-01

    A novel approach to time-synchronizing laser-induced fluorescence measurements to an oscillating current in a 60 Hz xenon discharge lamp using a continuous wave laser is presented. A sample-hold circuit is implemented to separate out signals at different phases along a current cycle, and is followed by a lock-in amplifier to pull out the resulting time-synchronized fluorescence trace from the large background signal. The time evolution of lower state population is derived from the changes in intensity of the fluorescence excitation line shape resulting from laser-induced fluorescence measurements of the 6s(')[1/2](1)(0)-6p(')[3/2](2) xenon atomic transition at λ = 834.68 nm. Results show that the lower state population oscillates at twice the frequency of the discharge current, 120 Hz.

  20. Time-synchronized continuous wave laser-induced fluorescence on an oscillatory xenon discharge

    Energy Technology Data Exchange (ETDEWEB)

    MacDonald, N. A.; Cappelli, M. A. [Stanford Plasma Physics Laboratory, Stanford University, Stanford, California 94305 (United States); Hargus, W. A. Jr. [Air Force Research Laboratory, Edwards AFB, California 93524 (United States)

    2012-11-15

    A novel approach to time-synchronizing laser-induced fluorescence measurements to an oscillating current in a 60 Hz xenon discharge lamp using a continuous wave laser is presented. A sample-hold circuit is implemented to separate out signals at different phases along a current cycle, and is followed by a lock-in amplifier to pull out the resulting time-synchronized fluorescence trace from the large background signal. The time evolution of lower state population is derived from the changes in intensity of the fluorescence excitation line shape resulting from laser-induced fluorescence measurements of the 6s{sup Prime }[1/2]{sub 1}{sup 0}-6p{sup Prime }[3/2]{sub 2} xenon atomic transition at {lambda}= 834.68 nm. Results show that the lower state population oscillates at twice the frequency of the discharge current, 120 Hz.

  1. Disruption of the hydrogen bonding network determines the pH-induced non-fluorescent state of the fluorescent protein ZsYellow by protonation of Glu221.

    Science.gov (United States)

    Bae, Ji-Eun; Kim, In Jung; Nam, Ki Hyun

    2017-11-04

    Many fluorescent proteins (FPs) exhibit fluorescence quenching at a low pH. This pH-induced non-fluorescent state of an FP serves as a useful indicator of the cellular pH. ZsYellow is widely used as an optical marker in molecular biology, but its pH-induced non-fluorescent state has not been characterized. Here, we report the pH-dependent spectral properties of ZsYellow, which exhibited the pH-induced non-fluorescence state at a pH below 4.0. We determined the crystal structures of ZsYellow at pH 3.5 (non-fluorescence state) and 8.0 (fluorescence state), which revealed the cis-configuration of the chromophore without pH-induced isomerization. In the non-fluorescence state, Arg95, which is involved in stabilization of the exited state of the chromophore, was found to more loosely interact with the carbonyl oxygen atom of the chromophore when compared to the interaction at pH 8.0. In the fluorescence state, Glu221, which is involved in the hydrogen bonding network around the chromophore, stably interacted with Gln42 and His202. By contrast, in the non-fluorescence state, the protonated conserved Glu221 residue exhibited a large conformational change and was separated from His202 by 5.46 Å, resulting in breakdown of the hydrogen bond network. Our results provide insight into the critical role of the conserved Glu221 residue for generating the pH-induced non-fluorescent state. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Self-assembly of Terbium(III)-based metal-organic complexes with two-photon absorbing active

    Science.gov (United States)

    Li, Dandan; Shao, Nanqi; Sun, Xianshun; Zhang, Guocui; Li, Shengli; Zhou, Hongping; Wu, Jieying; Tian, Yupeng

    2014-12-01

    Hybrid complexes based on D-π-A type dyes p-aminostyryl-pyridinum and Terbium(III) complex anion (1, 2) have been synthesized by ionic exchange reaction. Meanwhile two different alkyl-substituted amino groups were used as electron donors in organic dyes cations. The synthesized complexes were characterized by element analysis. In addition, the structural features of them were systematic studied by single crystal X-ray diffraction analysis. Their linear properties have been systematically investigated by absorption spectra and fluorescence, the results show that the energy transfer takes place from the trans-4-[4‧-(N,N-diethylamino)styryl]-N-methyl pyridinium (2‧) cation to Tb(III). In addition, complex 2 exhibit a large two-photon absorption coefficient β: 0.044 cm/GW at 710 nm.

  3. Two-photon autofluorescence/FLIM/SHG endoscopy to study the oral cavity and wound healing in humans (Conference Presentation)

    Science.gov (United States)

    König, Karsten

    2016-03-01

    Monitoring the oral cavity noninvasively with superior 3D resolution is realized by clinical multiphoton tomography and high NA two-photon endoscopy without the need of additional contrast agents. The technology behind this investigation is based on nonlinear optical contrast of the multiphoton tomograph MPTflex®. Furthermore, the miniaturized GRIN endoscope was used to realize more accessibility for more demanding wound conditions in skin. The MPTflex® distinguishes autofluorescence (AF) signals from second harmonic generation (SHG) signals simultaneously. Fluorescence lifetime imaging (FLIM) based on time correlated single photon counting (TCSPC) technology offers additional information on the functional level of the intratissue fluorophores, their binding status, and the contribution of SHG signals in chronic wounds.

  4. Cascaded two-photon nonlinearity in a one-dimensional waveguide with multiple two-level emitters

    Science.gov (United States)

    Roy, Dibyendu

    2013-01-01

    We propose and theoretically investigate a model to realize cascaded optical nonlinearity with few atoms and photons in one-dimension (1D). The optical nonlinearity in our system is mediated by resonant interactions of photons with two-level emitters, such as atoms or quantum dots in a 1D photonic waveguide. Multi-photon transmission in the waveguide is nonreciprocal when the emitters have different transition energies. Our theory provides a clear physical understanding of the origin of nonreciprocity in the presence of cascaded nonlinearity. We show how various two-photon nonlinear effects including spatial attraction and repulsion between photons, background fluorescence can be tuned by changing the number of emitters and the coupling between emitters (controlled by the separation). PMID:23948782

  5. Quantitative evaluation of redox ratio and collagen characteristics during breast cancer chemotherapy using two-photon intrinsic imaging.

    Science.gov (United States)

    Wu, Shulian; Huang, Yudian; Tang, Qinggong; Li, Zhifang; Horng, Hannah; Li, Jiatian; Wu, Zaihua; Chen, Yu; Li, Hui

    2018-03-01

    Preoperative neoadjuvant treatment in locally advanced breast cancer is recognized as an effective adjuvant therapy, as it improves treatment outcomes. However, the potential complications remain a threat, so there is an urgent clinical need to assess both the tumor response and changes in its microenvironment using non-invasive and precise identification techniques. Here, two-photon microscopy was employed to detect morphological alterations in breast cancer progression and recession throughout chemotherapy. The changes in structure were analyzed based on the autofluorescence and collagen of differing statuses. Parameters, including optical redox ratio, the ratio of second harmonic generation and auto-fluorescence signal, collagen density, and collagen shape orientation, were studied. Results indicate that these parameters are potential indicators for evaluating breast tumors and their microenvironment changes during progression and chemotherapy. Combined analyses of these parameters could provide a quantitative, novel method for monitoring tumor therapy.

  6. A fully-automated multiscale kernel graph cuts based particle localization scheme for temporal focusing two-photon microscopy

    Science.gov (United States)

    Huang, Xia; Li, Chunqiang; Xiao, Chuan; Sun, Wenqing; Qian, Wei

    2017-03-01

    The temporal focusing two-photon microscope (TFM) is developed to perform depth resolved wide field fluorescence imaging by capturing frames sequentially. However, due to strong nonignorable noises and diffraction rings surrounding particles, further researches are extremely formidable without a precise particle localization technique. In this paper, we developed a fully-automated scheme to locate particles positions with high noise tolerance. Our scheme includes the following procedures: noise reduction using a hybrid Kalman filter method, particle segmentation based on a multiscale kernel graph cuts global and local segmentation algorithm, and a kinematic estimation based particle tracking method. Both isolated and partial-overlapped particles can be accurately identified with removal of unrelated pixels. Based on our quantitative analysis, 96.22% isolated particles and 84.19% partial-overlapped particles were successfully detected.

  7. Two-photon decay of K-shell vacancy states in heavy atoms

    International Nuclear Information System (INIS)

    Ilakovac, K.; Uroic, M.; Majer, M.; Pasic, S.; Vukovic, B.

    2006-01-01

    Two-photon decay has been extensively studied in atomic, nuclear and particle physics since the 1930s when the problem of stability of the 2s state of the hydrogen atom emerged. Since then, many theoretical and experimental investigations have been made on hydrogen and one-electron (H-like) ions and on helium and two-electron (He-like) ions. The work on two-photon decay in many-electron systems involving inner shells started about 30 years ago and, in the meantime, two-photon decay of the K-shell vacancy state has been the subject of many theoretical and experimental studies. Experimental results have been obtained for 2s->1s and higher-state electron ->1s two-photon transitions in molybdenum, and for 2s -> 1s, 3s -> 1s, 3d -> 1s and 4sd -> 1s two-photon transitions in silver, xenon, hafnium and mercury. Nonrelativistic and relativistic calculations of the processes have been made. The relativistic calculations for transitions in molybdenum, silver and xenon atoms are in a reasonable agreement with the experimental results, but some problems remain to be solved. A review of investigations of two-photon transitions in atomic systems is presented

  8. Chlorophyll fluorescence lifetime imaging provides new insight into the chlorosis induced by plant virus infection.

    Science.gov (United States)

    Lei, Rong; Jiang, Hongshan; Hu, Fan; Yan, Jin; Zhu, Shuifang

    2017-02-01

    Leaf chlorosis induced by plant virus infection has a short fluorescence lifetime, which reflects damaged photosynthetic complexes and degraded chloroplasts. Plant viruses often induce chlorosis and necrosis, which are intimately related to photosynthetic functions. Chlorophyll fluorescence lifetime measurement is a valuable noninvasive tool for analyzing photosynthetic processes and is a sensitive indicator of the environment surrounding the fluorescent molecules. In this study, our central goal was to explore the effect of viral infection on photosynthesis by employing chlorophyll fluorescence lifetime imaging (FLIM), steady-state fluorescence, non-photochemical quenching (NPQ), transmission electron microscopy (TEM), and pigment analysis. The data indicated that the chlorophyll fluorescence lifetime of chlorotic leaves was significantly shorter than that of healthy control leaves, and the fitted short lifetime component of chlorophyll fluorescence of chlorotic leaves was dominant. This dominant short lifetime component may result from damage to the structure of thylakoid, which was confirmed by TEM. The NPQ value of chlorotic leaves was slightly higher than that of healthy green leaves, which can be explained by increased neoxanthin, lutein and violaxanthin content relative to chlorophyll a. The difference in NPQ is slight, but FLIM can provide simple and direct characterization of PSII structure and photosynthetic function. Therefore, this technique shows great potential as a simple and rapid method for studying mechanisms of plant virus infection.

  9. Visualization of Two-Phase Fluid Distribution Using Laser Induced Exciplex Fluorescence

    Science.gov (United States)

    Kim, J. U.; Darrow, J.; Schock, H.; Golding, B.; Nocera, D.; Keller, P.

    1998-03-01

    Laser-induced exciplex (excited state complex) fluorescence has been used to generate two-dimensional images of dispersed liquid and vapor phases with spectrally resolved two-color emissions. In this method, the vapor phase is tagged by the monomer fluorescence while the liquid phase is tracked by the exciplex fluorescence. A new exciplex visualization system consisting of DMA and 1,4,6-TMN in an isooctane solvent was developed.(J.U. Kim et al., Chem. Phys. Lett. 267, 323-328 (1997)) The direct ca

  10. Fluorescence and Four-Wave Mixing in Electromagnetically Induced Transparency Windows

    International Nuclear Information System (INIS)

    Wang Zhi-Guo; Li Cheng; Zhang Zhao-Yang; Che Jun-Ling; Qin Meng-Zhe; He Jia-Nan; Zhang Yan-Peng

    2013-01-01

    We simultaneously compare the probe transmission, Four-Wave Mixing (FWM) and fluorescence signals with dressing effects in a four-level atomic system. The variation rules of three types of signals are exhibited by changing the frequency detuning and power of incident laser beams. The interplay between two ladder subsystems is investigated in the Y-type atomic system. In particular, the fluorescence signal with ultra-narrow linewidth is obtained due to being sheared twice by the electromagnetically induced transparency window. Such fluorescence with very high coherence and monochromaticity can be used for the quantum correlation and narrow linewidth laser

  11. Porous Porphyrin-Based Organosilica Nanoparticles for NIR Two-Photon Photodynamic Therapy and Gene Delivery in Zebrafish

    KAUST Repository

    Mauriello Jimenez, Chiara; Aggad, Dina; Croissant, Jonas G.; Tresfield, Karen; Laurencin, Danielle; Berthomieu, Dorothé e; Cubedo, Nicolas; Rossel, Mireille; Alsaiari, Shahad K.; Anjum, Dalaver H.; Sougrat, Rachid; Roldan-Gutierrez, Manuel A.; Richeter, Sé bastien; Oliviero, Erwan; Raehm, Laurence; Charnay, Clarence; Cattoë n, Xavier; Clé ment, Sé bastien; Wong Chi Man, Michel; Maynadier, Marie; Chaleix, Vincent; Sol, Vincent; Garcia, Marcel; Gary-Bobo, Magali; Khashab, Niveen M.; Bettache, Nadir; Durand, Jean-Olivier

    2018-01-01

    functionalization of the nanoparticles with aminopropyltriethoxysilane, two-photon-excited photodynamic therapy in zebrafish is successfully achieved. Two-photon photochemical internalization in cancer cells of the nanoparticles loaded with siRNA is also performed

  12. Solvent induced fluorescence enhancement of graphene oxide studied by ultrafast spectroscopy

    Science.gov (United States)

    Zhao, Litao; Chen, Jinquan; He, Xiaoxiao; Yu, Xiantong; Yan, Shujun; Zhang, Sanjun; Pan, Haifeng; Xu, Jianhua

    2018-05-01

    Femtosecond transient absorption (TA) spectroscopy combined with picosecond time resolved fluorescence (TRF) were used to reveal the fluorescence kinetics of graphene oxide (GO) in water, ethanol and water-ethanol mixtures. Size-independent fluorescence of GO were observed in water, and pH-dependent fluorescence spectra could be fitted well by a triple emission relaxation with peaks around 440 nm, 500 nm, and 590 nm respectively. The results indicate that polycyclic aromatic hydrocarbons (PAHs) linked by oxygen-containing functional groups dominate GO's fluorescence emission. GO's fluorescence quantum yield was measured to be 2.8% in ethanol but 1.2% in water. The three decay components fluorescence decay, as well as the transient absorption dynamics with an offset, confirmed this solvent induced fluorescence enhancement. GO's Raman spectral signals showed that GO in ethanol has a smaller average size of PAHs than that of GO in water. Therefore, besides other enhancement effects reported in literatures, we proposed that solvents could also change the size of PAHs, resulting in a photoluminescence enhancement. Our experimental data demonstrates that GO's quantum yield could be up to 2.8% in water and 8.4% in ethanol and this observation may help ones to improve GO's photoluminescence efficiency as well as its applications in solution.

  13. Two-photon spectral amplitude of entangled states resolved in separable Schmidt modes

    International Nuclear Information System (INIS)

    Avella, A; Brida, G; Gramegna, M; Shurupov, A; Genovese, M; Chekhova, M

    2015-01-01

    The ability to access high dimensionality in Hilbert spaces represents a demanding key-stone for state-of-the-art quantum information. The manipulation of entangled states in continuous variables, wavevector as well frequency, represents a powerful resource in this sense. The number of dimensions of the Hilbert space that can be used in practical information protocols can be determined by the number of Schmidt modes that it is possible to address one by one. In the case of wavevector variables, the Schmidt modes can be losslessly selected using single-mode fibre and a spatial light modulator, but no similar procedure exists for the frequency space. The aim of this work is to present a technique to engineer the spectral properties of biphoton light, emitted via ultrafast spontaneous parametric down conversion, in such a way that the two-photon spectral amplitude (TPSA) contains several non-overlapping Schmidt modes, each of which can be filtered losslessly in frequency variables. Such TPSA manipulation is operated by a fine balancing of parameters like the pump frequency, the shaping of pump pulse spectrum, the dispersion dependence of spontaneous parametric down-conversion crystals as well as their length. Measurements have been performed exploiting the group velocity dispersion induced by the passage of optical fields through dispersive media, operating a frequency-to-time two-dimensional Fourier transform of the TPSA. Exploiting this kind of measurement we experimentally demonstrate the ability to control the Schmidt modes structure in TPSA through the pump spectrum manipulation. (paper)

  14. Interim report on modeling studies of two-photon isotope separation

    International Nuclear Information System (INIS)

    Hwang, W.C.; Badcock, C.C.; Kamada, R.F.

    1975-01-01

    The two-photon or two-step dissociation method of laser induced isotope enrichment is being modeled for the HBrNO photochemical system. In the model, H 79 Br is selectively excited by resonance IR laser radiation and then dissociated by uv radiation. Selectively dissociated Br atoms are scavenged to form isotopically enriched BrNO and Br 2 . This model includes all kinetic and absorption processes found to be significant and the time-varying concentrations of any species involved in a significant process. Among these processes are vibrational energy transfer reactions (including isotopic exchange) involving HBr v = 0 - 3, rotational and translational (velocity) relaxation processes, dissociation of HBr in the v = 0 - 3 levels, and secondary chemical reactions of the dissociation products. The absorption and kinetic processes that are most important to 79 Br enrichment have been identified and the study of the effects on enrichment upon variation of external parameters (such as reactant pressure, ir or uv source intensity, and temperature) is in progress. Some preliminary results are: (1) intensity of the ir source is usually more important than the uv intensity; (2) chemical reactions are the dominant kinetic processes at lower pressures while energy transfer reactions dominate at higher pressures; (3) kinetic processes usually have greater effect on the absolute amount of enriched products; (4) isotopic abundance of 79 Br in the products can range from 0.55 to 0.80 for the conditions used in the model

  15. Electron-electron correlation in two-photon double ionization of He-like ions

    Science.gov (United States)

    Hu, S. X.

    2018-01-01

    Electron correlation plays a crucial role in quantum many-body physics ranging from molecular bonding and strong-field-induced multielectron ionization, to superconducting in materials. Understanding the dynamic electron correlation in the photoionization of relatively simple quantum three-body systems, such as He and He-like ions, is an important step toward manipulating complex systems through photoinduced processes. Here we have performed ab initio investigations of two-photon double ionization (TPDI) of He and He-like ions (L i+,B e2 + , and C4 +) exposed to intense attosecond x-ray pulses. Results from such fully correlated quantum calculations show weaker and weaker electron correlation effects in TPDI spectra as the ionic charge increases, which is opposite to the intuition that the absolute increase of correlation in the ground state should lead to more equal energy sharing in photoionization. These findings indicate that the final-state electron-electron correlation ultimately determines the energy sharing of the two ionized electrons in TPDI.

  16. Linking Neurons to Network Function and Behavior by Two-Photon Holographic Optogenetics and Volumetric Imaging.

    Science.gov (United States)

    Dal Maschio, Marco; Donovan, Joseph C; Helmbrecht, Thomas O; Baier, Herwig

    2017-05-17

    We introduce a flexible method for high-resolution interrogation of circuit function, which combines simultaneous 3D two-photon stimulation of multiple targeted neurons, volumetric functional imaging, and quantitative behavioral tracking. This integrated approach was applied to dissect how an ensemble of premotor neurons in the larval zebrafish brain drives a basic motor program, the bending of the tail. We developed an iterative photostimulation strategy to identify minimal subsets of channelrhodopsin (ChR2)-expressing neurons that are sufficient to initiate tail movements. At the same time, the induced network activity was recorded by multiplane GCaMP6 imaging across the brain. From this dataset, we computationally identified activity patterns associated with distinct components of the elicited behavior and characterized the contributions of individual neurons. Using photoactivatable GFP (paGFP), we extended our protocol to visualize single functionally identified neurons and reconstruct their morphologies. Together, this toolkit enables linking behavior to circuit activity with unprecedented resolution. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Ratiometric two-photon excited photoluminescence of quantum dots triggered by near-infrared-light for real-time detection of nitric oxide release in situ

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Hui [Shandong Sino-Japanese Center for Collaborative Research of Carbon Nanomaterials, Collaborative Innovation Center for Marine Biomass Fiber Materials and Textiles, College of Chemistry and Chemical Engineering, Laboratory of Fiber Materials and Modern Textile, The Growing Base for State Key Laboratory, Qingdao University, Shandong 266071 (China); Gui, Rijun, E-mail: guirijun@qdu.edu.cn [Shandong Sino-Japanese Center for Collaborative Research of Carbon Nanomaterials, Collaborative Innovation Center for Marine Biomass Fiber Materials and Textiles, College of Chemistry and Chemical Engineering, Laboratory of Fiber Materials and Modern Textile, The Growing Base for State Key Laboratory, Qingdao University, Shandong 266071 (China); Sun, Jie; Wang, Yanfeng [Institute of Materia Medica, Shandong Academy of Medical Sciences, Jinan 250062 (China)

    2016-05-30

    Probe-donor integrated nanocomposites were developed from conjugating silica-coated Mn{sup 2+}:ZnS quantum dots (QDs) with MoS{sub 2} QDs and photosensitive nitric oxide (NO) donors (Fe{sub 4}S{sub 3}(NO){sub 7}{sup −}, RBS). Under excitation with near-infrared (NIR) light at 808 nm, the Mn{sup 2+}:ZnS@SiO{sub 2}/MoS{sub 2}-RBS nanocomposites showed the dual-emissive two-photon excited photoluminescence (TPEPL) that induced RBS photolysis to release NO in situ. NO caused TPEPL quenching of Mn{sup 2+}:ZnS QDs, but it produced almost no impact on the TPEPL of MoS{sub 2} QDs. Hence, the nanocomposites were developed as a novel QDs-based ratiometric TPEPL probe for real-time detection of NO release in situ. The ratiometric TPEPL intensity is nearly linear (R{sup 2} = 0.9901) with NO concentration in the range of 0.01∼0.8 μM, which corresponds to the range of NO release time (0∼15 min). The detection limit was calculated to be approximately 4 nM of NO. Experimental results confirmed that this novel ratiometric TPEPL probe possessed high selectivity and sensitivity for the detection of NO against potential competitors, and especially showed high detection performance for NIR-light triggered NO release in tumor intracellular microenvironments. These results would promote the development of versatile probe-donor integrated systems, also providing a facile and efficient strategy to real-time detect the highly controllable drug release in situ, especially in physiological microenvironments. - Highlights: • Mn{sup 2+}:ZnS@SiO{sub 2}/MoS{sub 2}-RBS nanocomposites were developed as a novel ratiometric two-photon excited fluorescence probe. • This probe could conduct real-time detection of nitric oxide release in situ. • High feasibility of this probe was confirmed in tumor intracellular microenvironments.

  18. Single-Shot, Volumetrically Illuminated, Three-Dimensional, Tomographic Laser-Induced-Fluorescence Imaging in a Gaseous Free Jet

    Science.gov (United States)

    2016-04-28

    Single-shot, volumetrically illuminated, three- dimensional, tomographic laser-induced- fluorescence imaging in a gaseous free jet Benjamin R. Halls...acquisition; (110.6955) Tomographic imaging ; (110.6960) Tomography; (280.2490) Flow diagnostics; (300.2530) Fluorescence , laser-induced...84 (1983). 2. I. van Cruyningen, A. Lozano, and R. K. Hanson, “Quantitative imaging of concentration by planar laser-induced fluorescence ,” Exp

  19. Two photon absorption energy transfer in the light-harvesting complex of photosystem II (LHC-II) modified with organic boron dye

    Science.gov (United States)

    Chen, Li; Liu, Cheng; Hu, Rui; Feng, Jiao; Wang, Shuangqing; Li, Shayu; Yang, Chunhong; Yang, Guoqiang

    2014-07-01

    The plant light-harvesting complexes of photosystem II (LHC-II) play important roles in collecting solar energy and transferring the energy to the reaction centers of photosystems I and II. A two photon absorption compound, 4-(bromomethyl)-N-(4-(dimesitylboryl)phenyl)-N-phenylaniline (DMDP-CH2Br), was synthesized and covalently linked to the LHC-II in formation of a LHC-II-dye complex, which still maintained the biological activity of LHC-II system. Under irradiation with femtosecond laser pulses at 754 nm, the LHC-II-dye complex can absorb two photons of the laser light effectively compared with the wild type LHC-II. The absorbed excitation energy is then transferred to chlorophyll a with an obvious fluorescence enhancement. The results may be interesting and give potentials for developing hybrid photosystems.

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

    International Nuclear Information System (INIS)

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

    2011-01-01

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

  1. Two-photon excitation laser scanning microscopy of rabbit nasal septal cartilage following Nd:YAG-laser-mediated stress relaxation

    Science.gov (United States)

    Kim, Charlton C.; Wallace, Vincent P.; Coleno, Mariah L.; Dao, Xavier; Tromberg, Bruce J.; Wong, Brian J.

    2000-04-01

    Laser irradiation of hyaline cartilage result in stable shape changes due to temperature dependent stress relaxation. In this study, we determined the structural changes in chondrocytes within rabbit nasal septal cartilage tissue over a 12-day period using a two-photon laser scanning microscope (TPM) following Nd:YAG laser irradiation. During laser irradiation surface temperature, stress relaxation, and diffuse reflectance, were measured dynamically. Each specimen received one or two sequential laser exposures. The cartilage reached a peak surface temperature of about 61 degrees C during irradiation. Cartilage denatured in 50 percent EtOH was used as a positive control. TPM was performed to detect the fluorescence emission from the chondrocytes. Images of chondrocytes were obtained at depths up to 150 microns, immediately following laser exposure, and also following 12 days in culture. Few differences in the pattern or intensity of fluorescence was observed between controls and irradiated specimens imaged immediately following exposure, regardless of the number of laser pulses. However, following twelve days in tissue culture, the irradiated specimens increase, whereas the native tissue diminishes, in intensity and distribution of fluorescence in the cytoplasm. In contrast, the positive control shows only extracellular matrices and empty lacuna, feature consistent with cell membrane lysis.

  2. Development of Laser-Induced Fluorescence Diagnostic for the Paul Trap Simulator Experiment

    CERN Document Server

    Chung, Moses; Efthimion, Philip; Gilson, Erik P; Majeski, Richard; Startsev, Edward

    2005-01-01

    The Paul Trap Simulator Experiment (PTSX) is a cylindrical Paul trap whose purpose is to simulate the nonlinear dynamics of intense charged particle beam propagation in alternating-gradient magnetic transport systems. For the in-situ measurement of the transverse ion density profile in the PTSX device, which is essential for the study of beam mismatch and halo particle production, a laser-induced fluorescence diagnostic system is being developed. Instead of cesium, which has been used in the initial phase of the PTSX experiment, barium has been selected as the preferred ion for the laser-induced fluorescence diagnostic. The installation of the barium ion source and the characterization of the tunable dye laser system are discussed. The design of the collection optics with an intensified CCD camera system is also discussed. Finally, initial test results using the laser-induced fluorescence diagnostic will be presented.

  3. Laser-induced fluorescence line narrowing in atomic vapors

    International Nuclear Information System (INIS)

    Meier, T.; Schuessler, H.A.

    1983-01-01

    The use of highly monochromatic light allows the selective excitation of atoms in vapors if excitation and detection of the fluorescence is carried out collinearly. The atoms capable of absorbing light then form an atomic beam of well defined velocity along the direction of the laser beam, but no velocity selection occurs perpendicular to it. The potential of the technique for Doppler-free atomic spectroscopy and for the study of excited atom collisions is demonstrated using the Na D 1 line as an example

  4. Pathological diagnosis of bladder cancer by image analysis of hypericin induced fluorescence cystoscopic images

    Science.gov (United States)

    Kah, James C. Y.; Olivo, Malini C.; Lau, Weber K. O.; Sheppard, Colin J. R.

    2005-08-01

    Photodynamic diagnosis of bladder carcinoma based on hypericin fluorescence cystoscopy has shown to have a higher degree of sensitivity for the detection of flat bladder carcinoma compared to white light cystoscopy. The potential of the photosensitizer hypericin-induced fluorescence in performing non-invasive optical biopsy to grade bladder cancer in vivo using fluorescence cystoscopic image analysis without surgical resection for tissue biopsy is investigated in this study. The correlation between tissue fluorescence and histopathology of diseased tissue was explored and a diagnostic algorithm based on fluorescence image analysis was developed to classify the bladder cancer without surgical resection for tissue biopsy. Preliminary results suggest a correlation between tissue fluorescence and bladder cancer grade. By combining both the red-to-blue and red-to-green intensity ratios into a 2D scatter plot yields an average sensitivity and specificity of around 70% and 85% respectively for pathological cancer grading of the three different grades of bladder cancer. Therefore, the diagnostic algorithm based on colorimetric intensity ratio analysis of hypericin fluorescence cystoscopic images developed in this preliminary study shows promising potential to optically diagnose and grade bladder cancer in vivo.

  5. Laser-induced fluorescence imaging of acetone inside evaporating and burning fuel droplets

    Science.gov (United States)

    Shringi, D. S.; Shaw, B. D.; Dwyer, H. A.

    2009-01-01

    Laser-induced fluorescence was used to visualize acetone fields inside individual droplets of pure acetone as well as droplets composed of methanol or 1-propanol initially mixed with acetone. Droplets were supported on a horizontal wire and two vaporization conditions were investigated: (1) slow evaporation in room air and (2) droplet combustion, which leads to substantially faster droplet surface regression rates. Acetone was preferentially gasified, causing its concentration in droplets to drop in time with resultant decreases in acetone fluorescence intensities. Slowly vaporizing droplets did not exhibit large spatial variations of fluorescence within droplets, indicating that these droplets were relatively well mixed. Ignition of droplets led to significant variations in fluorescence intensities within droplets, indicating that these droplets were not well mixed. Ignited droplets composed of mixtures of 1-propanol and acetone showed large time-varying changes in shapes for higher acetone concentrations, suggesting that bubble formation was occurring in these droplets.

  6. Controllable ultra-narrow fluorescence and six-wave mixing under double electromagnetically induced transparency

    International Nuclear Information System (INIS)

    Wang, Z G; Zhang, Z Y; Che, J L; Zhang, Y Z; Li, C B; Zheng, H B; Zhang, Y P

    2013-01-01

    We report the first observation of six-wave mixing (SWM) and fluorescence signals in an electromagnetically induced transparency (EIT) window. Several remarkable advantages are described. First, multiple bright and dark states are simultaneously observed due to enhancement or suppression of the SWM signal. Second, ultra-narrow fluorescence, much narrower than the EIT window, is experimentally obtained. Third, the ultra-narrow fluorescence can also generate Autler–Townes splitting on scanning the coupling beam. Fourth, a double-peak EIT window is obtained using the nest-dressing scheme. Such studies concerning SWM and fluorescence have applications in optical switching, multi-channel communication and narrowband and long-range quantum communication. (letter)

  7. Pseudopotential calculations and photothermal lensing measurements of two-photon absorption in solids

    International Nuclear Information System (INIS)

    White, W.T. III.

    1985-01-01

    We have studied two-photon absorption in solids theoretically and experimentally. We have shown that it is possible to use accurate band structure techniques to compute two-photon absorption spectra within 15% of measured values in a wide band-gap material, ZnS. The empirical pseudopotential technique that we used is significantly more accurate than previous models of two-photon absorption in zinc blende materials, including present tunneling theories (which are essentially parabolic-band results in disguise) and the nonparabolic-band formalism of Pidgeon et al. and Weiler. The agreement between our predictions and previous measurements allowed us to use ZnS as a reference material in order to validate a technique for measuring two-photon absorption that was previously untried in solids, pulsed dual-beam thermal lensing. With the validated technique, we examined nonlinear absorption in one other crystal (rutile) and in several glasses, including silicates, borosilicates, and one phosphate glass. Initially, we believed that the absorption edges of all the materials were comparable; however, subsequent evidence suggested that the effective band-gap energies of the glasses were above the energy of two photons in our measurement. Therefore, we attribute the nonlinear absorption that we observed in glasses to impurities or defects. The measured nonlinear absorption coefficients were of the order of a few cm/TW in the glasses and of the order of 10 cm/GW in the crystals, four orders of magnitude higher than in glasses. 292 refs

  8. Pseudopotential calculations and photothermal lensing measurements of two-photon absorption in solids

    Energy Technology Data Exchange (ETDEWEB)

    White, W.T. III

    1985-11-04

    We have studied two-photon absorption in solids theoretically and experimentally. We have shown that it is possible to use accurate band structure techniques to compute two-photon absorption spectra within 15% of measured values in a wide band-gap material, ZnS. The empirical pseudopotential technique that we used is significantly more accurate than previous models of two-photon absorption in zinc blende materials, including present tunneling theories (which are essentially parabolic-band results in disguise) and the nonparabolic-band formalism of Pidgeon et al. and Weiler. The agreement between our predictions and previous measurements allowed us to use ZnS as a reference material in order to validate a technique for measuring two-photon absorption that was previously untried in solids, pulsed dual-beam thermal lensing. With the validated technique, we examined nonlinear absorption in one other crystal (rutile) and in several glasses, including silicates, borosilicates, and one phosphate glass. Initially, we believed that the absorption edges of all the materials were comparable; however, subsequent evidence suggested that the effective band-gap energies of the glasses were above the energy of two photons in our measurement. Therefore, we attribute the nonlinear absorption that we observed in glasses to impurities or defects. The measured nonlinear absorption coefficients were of the order of a few cm/TW in the glasses and of the order of 10 cm/GW in the crystals, four orders of magnitude higher than in glasses. 292 refs.

  9. Enhanced Size Selection in Two-Photon Excitation for CsPbBr3 Perovskite Nanocrystals.

    Science.gov (United States)

    Chen, Junsheng; Chábera, Pavel; Pascher, Torbjörn; Messing, Maria E; Schaller, Richard; Canton, Sophie; Zheng, Kaibo; Pullerits, Tõnu

    2017-10-19

    Cesium lead bromide (CsPbBr 3 ) perovskite nanocrystals (NCs), with large two-photon absorption (TPA) cross-section and bright photoluminescence (PL), have been demonstrated as stable two-photon-pumped lasing medium. With two-photon excitation, red-shifted PL spectrum and increased PL lifetime is observed compared with one-photon excitation. We have investigated the origin of such difference using time-resolved laser spectroscopies. We ascribe the difference to the enhanced size selection of NCs by two-photon excitation. Because of inherent nonlinearity, the size dependence of absorption cross-section under TPA is stronger. Consequently, larger size NCs are preferably excited, leading to longer excited-state lifetime and red-shifted PL emission. In a broad view, the enhanced size selection in two-photon excitation of CsPbBr 3 NCs is likely a general feature of the perovskite NCs and can be tuned via NC size distribution to influence their performance within NC-based nonlinear optical materials and devices.

  10. A mitochondrial targeted two-photon iridium(III) phosphorescent probe for selective detection of hypochlorite in live cells and in vivo.

    Science.gov (United States)

    Li, Guanying; Lin, Qian; Sun, Lingli; Feng, Changsheng; Zhang, Pingyu; Yu, Bole; Chen, Yu; Wen, Ya; Wang, Hui; Ji, Liangnian; Chao, Hui

    2015-01-01

    Endogenous hypochlorite ion (ClO(-)) is a highly reactive oxygen species (ROS) that is produced from hydrogen peroxide and chloride ions catalyzed by myeloperoxidase (MPO). And mitochondrion is one of the major sources of ROS including ClO(-). In the present work, a two-photon phosphorescent probe for ClO(-) in mitochondria was developed. An iridium(III) complex bearing a diaminomaleonitrile group as ClO(-) reactive moiety specifically responded to ClO(-) over other ions and ROSs. When the probe was reacted with ClO(-) to form an oxidized carboxylate product, a significant enhancement in phosphorescence intensity was observed under one-photon (402 nm) and two-photon (750 nm) excitation, with a two-photon absorption cross-section of 78.1 GM at 750 nm. More importantly, ICP-MS results and cellular images co-stained with Mito-tracker Green demonstrated that this probe possessed high specificity for mitochondria. This probe was applied in the one- and two-photon imaging of ClO(-) in vitro and in vivo. The results suggested endotoxin lipopolysaccharide (LPS) induced ClO(-) mostly generated in the liver of zebrafish. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Detection of bacterial infection of agave plants by laser-induced fluorescence

    Science.gov (United States)

    Cervantes-Martinez, Jesus; Flores-Hernandez, Ricardo; Rodriguez-Garay, Benjamin; Santacruz-Ruvalcaba, Fernando

    2002-05-01

    Greenhouse-grown plants of Agave tequilana Weber var. azul were inoculated with Erwinia carotovora, the causal agent of stem soft rot. We investigated the laser-induced fluorescence (LIF) of agave plants to determine whether LIF can be used as a noninvasive sensing tool for pathological studies. The LIF technique was also investigated as a means of detecting the effect of the polyamine biosynthesis inhibitor beta-hydroxyethylhydrazine as a bactericide against the pathogenic bacterium Erwinia carotovora. A He-Ne laser at 632.8 nm was used as the excitation source, and in vivo fluorescence emission spectra were recorded in the 660-790-range. Fluorescence maxima were at 690 and 740 nm. The infected plants that were untreated with the bactericide showed a definite increase in fluorescence intensity at both maxima within the first three days after infection. Beginning on the fifth day, a steady decrease in fluorescence intensity was observed, with a greater effect at 740 than at 690 nm. After 30 days there was no fluorescence. The infected plants that had been treated with the bactericide showed no significant change in fluorescence compared with that of the uninfected plants. The ratio of fluorescence intensities was determined to be F 690 nm/F 740 nm for all treatments. These studies indicate that LIF measurements of agave plants may be used for the early detection of certain types of disease and for determining the effect of a bactericide on bacteria. The results also showed that fluorescence intensity ratios can be used as a reliable indicator of the progress of disease.

  12. Experimental Studies of Laser-Induced Fluorescence Spectra of Plants Immunity to the Kind of Ground

    Directory of Open Access Journals (Sweden)

    Yu. V. Fedotov

    2015-01-01

    Full Text Available Various external factors (pollutants available in the soil, a lack or insufficient amount of water and nutrients, etc. lead to stressful conditions of plants and impossibility of their normal development. At the early stages it is difficult to identify visually the stressful situations of plants. Therefore development of methods and devices to detect stressful states is important.A method of the laser-induced fluorescence is one of perspective methods for detection of stressful conditions of plants.In spite of quite a great number of work presenting results of the pilot studies of fluorescence spectra of vegetation, there are some important issues, which are unclear.The paper gives results of pilot studies of stability of a spectrum form of the laser-induced fluorescence of plants for different types of soil at the wavelength of excitation fluorescence of 532 nm.Results of processing fluorescence spectra of plants show:- fluorescence spectra of plants grown up under similar conditions have good repeatability of a spectra form for different samples of plants and different measurement time for each type of studied soil. The ratio value R of the fluorescence intensity at the wavelength of 685 nm to the fluorescence intensity at the wavelength of 740 nm has high stability. The standard deviation in sampling of the ratio R of different samples of a plant for one type of soil (for width of spectral ranges of recording fluorescent radiation of 10 nm lies in the range ~ 0.055 - ~ 0.12;- a difference in plant fluorescence spectra between themselves for different types of soil has the same order as a difference in fluorescence spectra of different samples of a plant for one type of soil. Difference in average value of the ratio R for different types of soil lies in the range ~ 0.01 - ~ 0.15.Thus, the value of the ratio R is steady against a type of soil and can be used to control a condition of plants.

  13. Crossed-coil detection of two-photon excited nuclear quadrupole resonance

    Science.gov (United States)

    Eles, Philip T.; Michal, Carl A.

    2005-08-01

    Applying a recently developed theoretical framework for determining two-photon excitation Hamiltonians using average Hamiltonian theory, we calculate the excitation produced by half-resonant irradiation of the pure quadrupole resonance of a spin-3/2 system. This formalism provides expressions for the single-quantum and double-quantum nutation frequencies as well as the Bloch-Siegert shift. The dependence of the excitation strength on RF field orientation and the appearance of the free-induction signal along an axis perpendicular to the excitation field provide an unmistakable signature of two-photon excitation. We demonstrate single- and double-quantum excitation in an axially symmetric system using 35Cl in a single crystal of potassium chlorate ( ωQ = 28 MHz) with crossed-coil detection. A rotation plot verifies the orientation dependence of the two-photon excitation, and double-quantum coherences are observed directly with the application of a static external magnetic field.

  14. Highly selective population of two excited states in nonresonant two-photon absorption

    International Nuclear Information System (INIS)

    Zhang Hui; Zhang Shi-An; Sun Zhen-Rong

    2011-01-01

    A nonresonant two-photon absorption process can be manipulated by tailoring the ultra-short laser pulse. In this paper, we theoretically demonstrate a highly selective population of two excited states in the nonresonant two-photon absorption process by rationally designing a spectral phase distribution. Our results show that one excited state is maximally populated while the other state population is widely tunable from zero to the maximum value. We believe that the theoretical results may play an important role in the selective population of a more complex nonlinear process comprising nonresonant two-photon absorption, such as resonance-mediated (2+1)-three-photon absorption and (2+1)-resonant multiphoton ionization. (atomic and molecular physics)

  15. Selective two-photon excitation of a vibronic state by correlated photons.

    Science.gov (United States)

    Oka, Hisaki

    2011-03-28

    We theoretically investigate the two-photon excitation of a molecular vibronic state by correlated photons with energy anticorrelation. A Morse oscillator having three sets of vibronic states is used, as an example, to evaluate the selectivity and efficiency of two-photon excitation. We show that a vibrational mode can be selectively excited with high efficiency by the correlated photons, without phase manipulation or pulse-shaping techniques. This can be achieved by controlling the quantum correlation so that the photon pair concurrently has two pulse widths, namely, a temporally narrow width and a spectrally narrow width. Though this concurrence is seemingly contradictory, we can create such a photon pair by tailoring the quantum correlation between two photons.

  16. Two-photon patterning of a polymer containing Y-shaped azochromophores

    International Nuclear Information System (INIS)

    Ambrosio, A.; Orabona, E.; Maddalena, P.; Camposeo, A.; Polo, M.; Neves, A. A. R.; Pisignano, D.; Carella, A.; Borbone, F.; Roviello, A.

    2009-01-01

    We report on the patterning of the free surface of azo-based polymer films by means of mass migration driven by one- or two-photon absorption. A symmetric donor-acceptor-donor structured Y-shaped azochromophore is specifically synthesized to enhance two-photon absorption in the polymer. The exposure of the polymer film to a focused laser beam results in light-driven mass migration for both one- and two-photon absorptions. Features with subdiffraction resolution (250 nm) are realized and the patterning dynamics is investigated as a function of the light dose. Furthermore, functional photonic structures, such as diffraction gratings with periods ranging between 0.5 and 2.0 μm, have been realized

  17. Absolute atomic hydrogen density distribution in a hollow cathode discharge by two-photon polarization spectroscopy

    International Nuclear Information System (INIS)

    Gonzalo, A B; Rosa, M I de la; Perez, C; Mar, S; Gruetzmacher, K

    2004-01-01

    We report on quantitative measurements of ground-state atomic hydrogen densities in a stationary plasma far off thermodynamic equilibrium, generated in a hollow cathode discharge, by two-photon polarization spectroscopy via the 1S-2S transition. Absolute densities are obtained using a well established calibration method based on the non-resonant two-photon polarization signal of xenon gas at room temperature, which serves as the reference at the wavelength of the hydrogen transition. This study is dedicated to demonstrating the capability of two-photon polarization spectroscopy close to the detection limit. Therefore, it requires single-longitudinal mode UV-laser radiation provided by an advanced UV-laser spectrometer

  18. Exact analysis of the response of quantum systems to two-photons using a QSDE approach

    International Nuclear Information System (INIS)

    Pan, Yu; Dong, Daoyi; Zhang, Guofeng

    2016-01-01

    We introduce the quantum stochastic differential equation (QSDE) approach to exactly analyze the response of quantum systems to a continuous-mode two-photon input. The QSDE description of the two-photon process allows us to integrate the input–output analysis with the quantum network theory, and so the analytical computability of the output state of a general quantum system can be addressed within this framework. We show that the time-domain two-photon output states can be exactly calculated for a large class of quantum systems including passive linear networks, optomechanical oscillators and two-level emitter in waveguide systems. In particular, we propose to utilise the results for the exact simulation of the stimulated emission as well as the study of the scattering of two-mode photon wave packets. (paper)

  19. Scanless functional imaging of hippocampal networks using patterned two-photon illumination through GRIN lenses

    KAUST Repository

    Moretti, Claudio

    2016-09-12

    Patterned illumination through the phase modulation of light is increasingly recognized as a powerful tool to investigate biological tissues in combination with two-photon excitation and light-sensitive molecules. However, to date two-photon patterned illumination has only been coupled to traditional microscope objectives, thus limiting the applicability of these methods to superficial biological structures. Here, we show that phase modulation can be used to efficiently project complex two-photon light patterns, including arrays of points and large shapes, in the focal plane of graded index (GRIN) lenses. Moreover, using this approach in combination with the genetically encoded calcium indicator GCaMP6, we validate our system performing scanless functional imaging in rodent hippocampal networks in vivo ~1.2 mm below the brain surface. Our results open the way to the application of patterned illumination approaches to deep regions of highly scattering biological tissues, such as the mammalian brain.

  20. THz/Infrared Double Resonance Two-Photon Spectroscopy of HD+ for Determination of Fundamental Constants

    Directory of Open Access Journals (Sweden)

    Florin Lucian Constantin

    2017-10-01

    Full Text Available A double resonance two-photon spectroscopy scheme is discussed to probe jointly rotational and rovibrational transitions of ensembles of trapped HD+ ions. The two-photon transition rates and lightshifts are calculated with the two-photon tensor operator formalism. The rotational lines may be observed with sub-Doppler linewidth at the hertz level and good signal-to-noise ratio, improving the resolution in HD+ spectroscopy beyond the 10−12 level. The experimental accuracy, estimated at the 10−12 level, is comparable with the accuracy of theoretical calculations of HD+ energy levels. An adjustment of selected rotational and rovibrational HD+ lines may add clues to the proton radius puzzle, may provide an independent determination of the Rydberg constant, and may improve the values of proton-to-electron and deuteron-to-proton mass ratios beyond the 10−11 level.

  1. Two-Photon Rabi Splitting in a Coupled System of a Nanocavity and Exciton Complexes

    Science.gov (United States)

    Qian, Chenjiang; Wu, Shiyao; Song, Feilong; Peng, Kai; Xie, Xin; Yang, Jingnan; Xiao, Shan; Steer, Matthew J.; Thayne, Iain G.; Tang, Chengchun; Zuo, Zhanchun; Jin, Kuijuan; Gu, Changzhi; Xu, Xiulai

    2018-05-01

    Two-photon Rabi splitting in a cavity-dot system provides a basis for multiqubit coherent control in a quantum photonic network. Here we report on two-photon Rabi splitting in a strongly coupled cavity-dot system. The quantum dot was grown intentionally large in size for a large oscillation strength and small biexciton binding energy. Both exciton and biexciton transitions couple to a high-quality-factor photonic crystal cavity with large coupling strengths over 130 μ eV . Furthermore, the small binding energy enables the cavity to simultaneously couple with two exciton states. Thereby, two-photon Rabi splitting between the biexciton and cavity is achieved, which can be well reproduced by theoretical calculations with quantum master equations.

  2. Quantum-correlated two-photon transitions to excitons in semiconductor quantum wells.

    Science.gov (United States)

    Salazar, L J; Guzmán, D A; Rodríguez, F J; Quiroga, L

    2012-02-13

    The dependence of the excitonic two-photon absorption on the quantum correlations (entanglement) of exciting biphotons by a semiconductor quantum well is studied. We show that entangled photon absorption can display very unusual features depending on space-time-polarization biphoton parameters and absorber density of states for both bound exciton states as well as for unbound electron-hole pairs. We report on the connection between biphoton entanglement, as quantified by the Schmidt number, and absorption by a semiconductor quantum well. Comparison between frequency-anti-correlated, unentangled and frequency-correlated biphoton absorption is addressed. We found that exciton oscillator strengths are highly increased when photons arrive almost simultaneously in an entangled state. Two-photon-absorption becomes a highly sensitive probe of photon quantum correlations when narrow semiconductor quantum wells are used as two-photon absorbers.

  3. One-dimensional modulation instability in biased two-photon photorefractive-photovoltaic crystals

    International Nuclear Information System (INIS)

    Zhan Kaiyun; Hou Chunfeng; Li Xin

    2010-01-01

    The one-dimensional modulation instability of broad optical beams in biased two-photon photorefractive-photovoltaic crystals is investigated under steady-state conditions. Our analysis indicates that the modulation instability growth rate depends on the external bias field, the bulk photovoltaic effect and the ratio of the intensity of the incident beam to that of the dark irradiance. Moreover, our results show that this modulation instability growth rate is the same as that in two-photon photorefractive-photovoltaic crystals under open circuit conditions in the absence of an external bias field, and the modulation instability growth rate in two-photon biased photorefractive-nonphotovoltaic crystals can be predicted when the bulk photovoltaic effect is neglected.

  4. In-vivo optical detection of cancer using chlorin e6 – polyvinylpyrrolidone induced fluorescence imaging and spectroscopy

    International Nuclear Information System (INIS)

    Chin, William WL; Thong, Patricia SP; Bhuvaneswari, Ramaswamy; Soo, Khee Chee; Heng, Paul WS; Olivo, Malini

    2009-01-01

    Photosensitizer based fluorescence imaging and spectroscopy is fast becoming a promising approach for cancer detection. The purpose of this study was to examine the use of the photosensitizer chlorin e6 (Ce6) formulated in polyvinylpyrrolidone (PVP) as a potential exogenous fluorophore for fluorescence imaging and spectroscopic detection of human cancer tissue xenografted in preclinical models as well as in a patient. Fluorescence imaging was performed on MGH human bladder tumor xenografted on both the chick chorioallantoic membrane (CAM) and the murine model using a fluorescence endoscopy imaging system. In addition, fiber optic based fluorescence spectroscopy was performed on tumors and various normal organs in the same mice to validate the macroscopic images. In one patient, fluorescence imaging was performed on angiosarcoma lesions and normal skin in conjunction with fluorescence spectroscopy to validate Ce6-PVP induced fluorescence visual assessment of the lesions. Margins of tumor xenografts in the CAM model were clearly outlined under fluorescence imaging. Ce6-PVP-induced fluorescence imaging yielded a specificity of 83% on the CAM model. In mice, fluorescence intensity of Ce6-PVP was higher in bladder tumor compared to adjacent muscle and normal bladder. Clinical results confirmed that fluorescence imaging clearly captured the fluorescence of Ce6-PVP in angiosarcoma lesions and good correlation was found between fluorescence imaging and spectral measurement in the patient. Combination of Ce6-PVP induced fluorescence imaging and spectroscopy could allow for optical detection and discrimination between cancer and the surrounding normal tissues. Ce6-PVP seems to be a promising fluorophore for fluorescence diagnosis of cancer

  5. Investigation of the two-photon polymerisation of a Zr-based inorganic-organic hybrid material system

    International Nuclear Information System (INIS)

    Bhuian, B.; Winfield, R.J.; O'Brien, S.; Crean, G.M.

    2006-01-01

    Two-photon polymerisation of photo-sensitive materials allows the fabrication of three dimensional micro- and nano-structures for photonic, electronic and micro-system applications. However the usable process window and the applicability of this fabrication technique is significantly determined by the properties of the photo-sensitive material employed. In this study investigation of a custom inorganic-organic hybrid system, cross-linked by a two-photon induced process, is described. The material was produced by sol-gel synthesis using a silicon alkoxide species that also possessed methacrylate functionality. Stabilized zirconium alkoxide precursors were added to the precursor solution in order to reduce drying times and impart enhanced mechanical stability to deposited films. This enabled dry films to be used in the polymerisation process. A structural, optical and mechanical analysis of the optimised sol-gel material is presented. A Ti:sapphire laser with 80 MHz repetition rate, 100 fs pulse duration and 795 nm is used. The influence of both material system and laser processing parameters including: laser power, photo-initiator concentration and zirconium loading, on achievable micro-structure and size is presented

  6. Serum Protein Profile Study of Clinical Samples Using High Performance Liquid Chromatography-Laser Induced Fluorescence

    DEFF Research Database (Denmark)

    Karemore, Gopal Raghunath; Ukendt, Sujatha; Rai, Lavanya

    2009-01-01

    The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested for estab...

  7. Laser-induced fluorescence of metal-atom impurities in a neutral beam

    International Nuclear Information System (INIS)

    Burrell, C.F.; Pyle, R.V.; Sabetimani, Z.; Schlachter, A.S.

    1984-10-01

    The need to limit impurities in fusion devices to low levels is well known. We have investigated, by the technique of laser-induced fluorescence, the concentration of heavy-metal atoms in a neutral beam caused by their evaporation from the hot filaments in a conventional high-current multifilament hydrogen-ion source

  8. Speciation of actinides in aqueous solution by time-resolved laser-induced fluorescence spectroscopy (TRLFS)

    International Nuclear Information System (INIS)

    Kimura, Takaumi; Kato, Yoshiharu; Meinrath, G.; Yoshida, Zenko; Choppin, G.R.

    1995-01-01

    Time-resolved laser-induced fluorescence spectroscopy (TRLFS) as a sensitive and selective method has been applied to the speciation of actinides in aqueous solution. Studies on hydrolysis and carbonate complexation of U(VI) and on determination of hydration number of Cm(III) are reported. (author)

  9. Detection of organic residues on poultry processing equipment surfaces by LED-induced fluorescence imaging

    Science.gov (United States)

    Organic residues on equipment surfaces in poultry processing plants can generate cross- contamination and increase the risk of unsafe food for consumers. This research was aimed to investigate the potential of LED-induced fluorescence imaging technique for rapid inspection of stainless steel proces...

  10. Formation of nitric oxide in an industrial burner measured by 2-D laser induced fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Arnold, A; Bombach, R; Kaeppeli, B [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1997-06-01

    We have performed two-dimensional Laser Induced Fluorescence (2-D LIF) measurements of nitric oxide and hydroxyl radical distributions in an industrial burner at atmospheric pressure. The relative 2-D LIF data of NO were set to an absolute scale by calibration with probe sampling combined with gas analysis. (author) 3 figs., 7 refs.

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

    Czech Academy of Sciences Publication Activity Database

    Rossini, P. M.; Nedbal, L.; Guanter, L.; Ač, Alexander; Alonso, L.; Burkart, A.; Cogliati, S.; Colombo, R.; Damm, A.; Drusch, M.; Hanuš, Jan; Janoutová, Růžena; Julitta, T.; Kokkalis, P.; Moreno, J.; Novotný, Jan; Panigada, C.; Pinto, F.; Schickling, A.; Schuettemeyer, D.; Zemek, František; Rascher, U.

    2015-01-01

    Roč. 42, č. 6 (2015), s. 1632-1639 ISSN 0094-8276 Institutional support: RVO:67179843 Keywords : sun-induced fluorescence * remote sensing * stress detection * airborne images * HyPlant Subject RIV: EH - Ecology, Behaviour Impact factor: 4.212, year: 2015

  12. Laser resonant ionization spectroscopy and laser-induced resonant fluorescence spectra of samarium atom

    International Nuclear Information System (INIS)

    Jin, Changtai

    1995-01-01

    We have measured new high-lying levels of Sm atom by two-colour resonant photoionisation spectroscopy; we have observed the isotope shifts of Sm atom by laser-induced resonant fluorescence spectroscopy; the lifetime of eight low-lying levels of Sm atom were measured by using pulsed laser-Boxcar technique in atomic beam.

  13. Characterization of scintillating CaWO{sub 4} crystals for the CRESST experiment using two-photon excitation

    Energy Technology Data Exchange (ETDEWEB)

    Hampf, Raphael; Dandl, Thomas; Muenster, Andrea; Oberauer, Lothar; Roth, Sabine; Schoenert, Stefan; Ulrich, Andreas [Physik-Department and Excellence Cluster Universe, Technische Universitaet Muenchen, D-85747 Garching (Germany)

    2016-07-01

    In the CRESST experiment for direct dark matter search, phonon and photon signals from cryogenic CaWO{sub 4} crystals are used to search for WIMP-induced nuclear recoil events. We present a novel table-top setup in which the scintillation of CaWO{sub 4} is induced by 0.7 ns laser pulses of 355 nm wavelength. The excitation occurs via two-photon absorption in the bulk material. The scintillation light is observed by time resolved optical spectroscopy. By varying the focusing of the laser-beam the excitation density can be made high enough to study quenching effects due to exciton-exciton annihilation. This allows to perform experiments to test models for the quenching factors of different ionizing projectiles in CaWO{sub 4} which are used to identify these projectiles on an event by event basis.

  14. Search for a Higgs Boson Decaying into Two Photons at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery P.; Anselmo, F.; Arefev, A.; Azemoon, T.; Aziz, T.; Bagnaia, P.; Bajo, A.; Baksay, G.; Baksay, L.; Baldew, S.V.; Banerjee, S.; Banerjee, Sw.; Barczyk, A.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Echenard, B.; Eline, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Ewers, A.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hakobyan, R.S.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Krenz, W.; Kruger, A.; Kunin, A.; Ladron de Guevara, P.; Laktineh, I.; Landi, G.; Lebeau, M.; Lebedev, A.; Lebrun, P.; Lecomte, P.; Lecoq, P.; Le Coultre, P.; Le Goff, J.M.; Leiste, R.; Levtchenko, M.; Levtchenko, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Lubelsmeyer, K.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mangeol, D.; Mans, J.; Martin, J.P.; Marzano, F.; Mazumdar, K.; McNeil, R.R.; Mele, S.; Merola, L.; Meschini, M.; Metzger, W.J.; Mihul, A.; Milcent, H.; Mirabelli, G.; Mnich, J.; Mohanty, G.B.; Muanza, G.S.; Muijs, A.J.M.; Musicar, B.; Musy, M.; Nagy, S.; Natale, S.; Napolitano, M.; Nessi-Tedaldi, F.; Newman, H.; Niessen, T.; Nisati, A.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Palomares, C.; Pandoulas, D.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.O.; Prokofiev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, M.A.; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Roux, B.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Sanders, M.P.; Schafer, C.; Schegelsky, V.; Schmidt-Kaerst, S.; Schmitz, D.; Schopper, H.; Schotanus, D.J.; Schwering, G.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Siedenburg, T.; Son, D.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, I.; Vogel, H.; Vogt, H.; Vorobev, I.; Vorobyov, A.A.; Wadhwa, M.; Wallraff, W.; Wang, X.L.; Wang, Z.M.; Weber, M.; Wienemann, P.; Wilkens, H.; Wynhoff, S.; Xia, L.; Xu, Z.Z.; Yamamoto, J.; Yang, B.Z.; Yang, C.G.; Yang, H.J.; Yang, M.; Yeh, S.C.; Zalite, A.; Zalite, Yu.; Zhang, Z.P.; Zhao, J.; Zhu, G.Y.; Zhu, R.Y.; Zhuang, H.L.; Zichichi, A.; Zilizi, G.; Zimmermann, B.; Zoller, M.

    2002-01-01

    A Higgs particle produced in association with a Z boson and decaying into two photons is searched for in the data collected by the L3 experiment at LEP. All possible decay modes of the Z boson are investigated. No signal is observed in 447.5 pb^-1 of data recorded at centre-of-mass energies up to 209 GeV. Limits on the branching fraction of the Higgs boson decay into two photons as a function of the Higgs mass are derived. A lower limit on the mass of a fermiophobic Higgs boson is set at 105.4 GeV at 95% confidence level.

  15. Closed form for two-photon free-free transition matrix elements

    Energy Technology Data Exchange (ETDEWEB)

    Karule, Erna E-mail: karule@latnet.lv

    2000-08-01

    Two-photon free-free transitions happen in the multiphoton ionization with more than one excess photon and in Bremsstrahlung. Up to now, the configuration space free-free transition amplitudes have not been written in closed form. We propose a modified Coulomb Green's function (CGF) Sturm ian expansion which allows one to obtain expressions for two-photon radial transition matrix elements in the closed form which are easy to continue analytically to calculate free-free transitions in H.

  16. Virtual-pion and two-photon production in pp scattering

    International Nuclear Information System (INIS)

    Scholten, O.; Korchin, A.Yu.

    2002-01-01

    Two-photon production in pp scattering is proposed as a means of studying virtual-pion emission. Such a process is complementary to real-pion emission in pp scattering. The virtual-pion signal is embedded in a background of double-photon bremsstrahlung. We have developed a model to describe this background process and show that in certain parts of phase space the virtual-pion signal gives significant contributions. In addition, through interference with the two-photon bremsstrahlung background, one can determine the relative phase of the virtual-pion process

  17. Comments on the observations of two-photon decay in n-p capture

    International Nuclear Information System (INIS)

    Alburger, D.E.

    1975-01-01

    Two-photon decay in n-p capture has been reported as having a branching ratio of 10 -3 relative to single 2.23-MeV γ-ray emission. It is suggested that this result can be explained by crosstalk effects between detectors due to single γ rays, principally that associated with annihiliation in flight following pair production. The shape of the observed γ-γ coincidence spectrum is consistent with this type of crosstalk and the apparent two-photon branch is about what is expected from a rough calculation of the magnitude of the crosstalk effect

  18. Enhancement of intermediate-field two-photon absorption by rationally shaped femtosecond pulses

    International Nuclear Information System (INIS)

    Chuntonov, Lev; Rybak, Leonid; Gandman, Andrey; Amitay, Zohar

    2008-01-01

    We extend the powerful frequency-domain analysis of femtosecond two-photon absorption to the intermediate-field regime of considerable absorption yields, where additionally to the weak-field nonresonant two-photon transitions also four-photon transitions play a role. Consequently, we rationally find that the absorption is enhanced over the transform-limited pulse by any shaped pulse having a spectral phase that is antisymmetric around one-half of the transition frequency and a spectrum that is asymmetric around it (red or blue detuned according to the system). The enhancement increases as the field strength increases. The theoretical results for Na are verified experimentally

  19. Two-photon transitions in hydrogen atoms embedded in weakly coupled plasmas

    International Nuclear Information System (INIS)

    Paul, S.; Ho, Y. K.

    2008-01-01

    The pseudostate method has been applied to calculate energy eigenvalues and corresponding eigenfunctions of the hydrogen atom in Debye plasma environments. Resonant two-photon transition rates from the ground state of atomic hydrogen to 2s and 3s excited states have been computed as a function of photon frequency in the length and velocity gauges for different Debye lengths. A two-photon transparency is found in correspondence to each resonance for 1s-3s. The transparency frequency and resonance enhancement frequency vary significantly with the Debye length.

  20. Two-photon processes of π0, η, η', ηc and ηb

    International Nuclear Information System (INIS)

    Klabucar, D.

    1997-01-01

    Two-photon processes of π 0 , η, η', η c and η b are studied in the consistently coupled Schwinger-Dyson (SD) and Bethe-Salpeter (BS) approach, where dynamical chiral symmetry breaking (DχSB) is obtained through the SD equation for the quark propagator which is then used in the BS equation. It is shown that the coupled SD-BS approach is similarly successful in the description of two-photon processes of pseudoscalar mesons over a wide range of masses. (K.A.)