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Sample records for infrared multiphoton excitation

  1. Infrared multiphoton dissociation of vinyltrifluorosilane

    NASA Astrophysics Data System (ADS)

    Koshliakov, P. V.; Gorelik, S. R.; Chesnokov, E. N.; Vorobiev, A. V.; Petrov, A. K.

    2006-09-01

    Infrared multiphoton absorption and dissociation of vinyltrifluorosilane molecules under the action of pulsed TEA CO2-laser were experimentally studied. The composition of the end dissociation products has been analyzed. Using quantum chemical calculations, we have identified the dissociation pathways that lead to the observed end products. Transition state geometries, enthalpies and activation energies for the dissociation pathways have been calculated. The dissociation channel ratio was determined under used experimental conditions. The silicon isotope selective infrared multiphoton dissociation has been performed at different wavelengths of the CO2-laser radiation.

  2. Multi-photon excitation microscopy

    PubMed Central

    Diaspro, Alberto; Bianchini, Paolo; Vicidomini, Giuseppe; Faretta, Mario; Ramoino, Paola; Usai, Cesare

    2006-01-01

    Multi-photon excitation (MPE) microscopy plays a growing role among microscopical techniques utilized for studying biological matter. In conjunction with confocal microscopy it can be considered the imaging workhorse of life science laboratories. Its roots can be found in a fundamental work written by Maria Goeppert Mayer more than 70 years ago. Nowadays, 2PE and MPE microscopes are expected to increase their impact in areas such biotechnology, neurobiology, embryology, tissue engineering, materials science where imaging can be coupled to the possibility of using the microscopes in an active way, too. As well, 2PE implementations in noninvasive optical bioscopy or laser-based treatments point out to the relevance in clinical applications. Here we report about some basic aspects related to the phenomenon, implications in three-dimensional imaging microscopy, practical aspects related to design and realization of MPE microscopes, and we only give a list of potential applications and variations on the theme in order to offer a starting point for advancing new applications and developments. PMID:16756664

  3. Controllable infrared continuum source for multiphoton imaging

    NASA Astrophysics Data System (ADS)

    de Mauro, C.; Alfieri, D.; Arrigoni, M.; Armstrong, D.; Pavone, F. S.

    2010-02-01

    We report on multiphoton imaging of biological samples performed with continuum infrared source generated in photonic crystal fibers (PCFs). We studied the spectra generated in PCFs with dispersion profiles designed to maximize the power density in the 700-1000 nm region, where the two-photon absorption cross sections of the most common dyes lie. Pumping in normal dispersion region, with <140 femtosecond pulses delivered by a tunable Ti:Sa laser (Chameleon Ultra II by Coherent Inc.), results in a limitation of nonlinear broadening up to a mean power density above 2 mW/nm. Axial and lateral resolution obtained with a scanning multiphoton system has been measureed to be near the theoretical limit. The possibility of simultaneous two-photon excitation of different dyes in the same sample and high image resolution are demonstrated at tens of microns in depth. Signal-to-noise ratio and general performances are found to be comparable with those of a single wavelength system, used for comparison.

  4. Routes to multiphoton double ionization in combined extreme ultraviolet and infrared laser pulses

    SciTech Connect

    Boettcher, M.; Rottke, H.; Zhavoronkov, N.; Sandner, W.; Agostini, P.; Gisselbrecht, M.; Huetz, A.

    2007-03-15

    Xenon multiphoton double ionization pathways are studied in a reaction microscope using a pump-probe arrangement of extreme ultraviolet high harmonic and infrared laser radiation. The momentum of photoelectrons is recorded in coincidence with singly or doubly charged ions. Among all possible routes to multiphoton double ionization, sequential processes using ionic excited states as intermediate steps are clearly identified.

  5. Multiphoton excitation of fluorescent DNA base analogs.

    PubMed

    Katilius, Evaldas; Woodbury, Neal W

    2006-01-01

    Multiphoton excitation was used to investigate properties of the fluorescent DNA base analogs, 2-aminopurine (2AP) and 6-methylisoxanthopterin (6MI). 2-aminopurine, a fluorescent analog of adenine, was excited by three-photon absorption. Fluorescence correlation measurements were attempted to evaluate the feasibility of using three-photon excitation of 2AP for DNA-protein interaction studies. However, high excitation power and long integration times needed to acquire high signal-to-noise fluorescence correlation curves render three-photon excitation FCS of 2AP not very useful for studying DNA base dynamics. The fluorescence properties of 6-methylisoxanthopterin, a guanine analog, were investigated using two-photon excitation. The two-photon absorption cross-section of 6MI was estimated to be about 2.5 x 10(-50) cm(4)s (2.5 GM units) at 700 nm. The two-photon excitation spectrum was measured in the spectral region from 700 to 780 nm; in this region the shape of the two-photon excitation spectrum is very similar to the shape of single-photon excitation spectrum in the near-UV spectral region. Two-photon excitation of 6MI is suitable for fluorescence correlation measurements. Such measurements can be used to study DNA base dynamics and DNA-protein interactions over a broad range of time scales. PMID:16965161

  6. Multiphoton excitation of fluorescent DNA base analogs

    NASA Astrophysics Data System (ADS)

    Katilius, Evaldas; Woodbury, Neal W.

    2006-07-01

    Multiphoton excitation was used to investigate properties of the fluorescent DNA base analogs, 2-aminopurine (2AP) and 6-methylisoxanthopterin (6MI). 2-aminopurine, a fluorescent analog of adenine, was excited by three-photon absorption. Fluorescence correlation measurements were attempted to evaluate the feasibility of using three-photon excitation of 2AP for DNA-protein interaction studies. However, high excitation power and long integration times needed to acquire high signal-to-noise fluorescence correlation curves render three-photon excitation FCS of 2AP not very useful for studying DNA base dynamics. The fluorescence properties of 6-methylisoxanthopterin, a guanine analog, were investigated using two-photon excitation. The two-photon absorption cross-section of 6MI was estimated to be about 2.510-50 cm4s (2.5 GM units) at 700 nm. The two-photon excitation spectrum was measured in the spectral region from 700 to 780 nm; in this region the shape of the two-photon excitation spectrum is very similar to the shape of single-photon excitation spectrum in the near-UV spectral region. Two-photon excitation of 6MI is suitable for fluorescence correlation measurements. Such measurements can be used to study DNA base dynamics and DNA-protein interactions over a broad range of time scales.

  7. Multiphoton Excitation of Upconverting Nanoparticles in Pulsed Regime.

    PubMed

    Hodak, Jose; Chen, Zhijun; Wu, Si; Etchenique, Roberto

    2016-01-19

    Upconverting nanoparticles (UCNPs) present emission in the visible region upon irradiation with NIR light through a multiphoton mechanism. However, the long characteristic time of their emission has prevented the use of this kind of entities as multiphoton probes. We present a study on the use of erbium-containing UCNPs under pulsed excitation, showing that both the power density and the duration of the excitation pulse are key factors to understand the emission behavior. By adjusting power and excitation rate, we can obtain typical multiphoton z-axis focal exclusive excitation. These findings open the possibility of using UCNPs as probes for controlled localization of uncaging and imaging with multiphoton z-axis sectioning. We show that this can be achieved even at power densities several orders of magnitude lower than traditional multiphoton microscopies. PMID:26703425

  8. Widefield multiphoton excited fluorescence microscopy for animal study in vivo

    NASA Astrophysics Data System (ADS)

    Cheng, L.-C.; Chang, C.-Y.; Lin, C.-H.; Su, Y.-D.; Huang, T.-Y.; Chen, S.-J.

    2010-08-01

    Unlike conventional multiphoton excited microscopy according to pixel-by-pixel point scanning, a widefield multiphoton excited microscopy based on spatiotemporal focusing has been developed to construct three-dimensional (3D) multiphoton fluorescence images only with the need of an axial scanning. By implementing a 4.0 W 10 kHz femtosecond laser amplifier with an instant strong peak power and a fast TE-cooled EMCCD camera with an ultra-sensitive fluorescence detection, the multiphoton excited fluorescence images with the excitation area over 100 ?m x 100 ?m can be achieved at a frame rate up to 80 Hz. A mechanical shutter is utilized to control the exposure time of 1 ms, i.e. average ten laser pulses reach the fluorescent specimen, and hence an uniform enough multiphoton excited fluorescence image can be attained with less photobleaching. The Brownian motion of microbeads and 3D neuron cells of a rat cerebellum have been observed with a lateral spatial resolution of 0.24 ?m and an axial resolution of 2.5 ?m. Therefore, the developed widefield multiphoton microscopy can provide fast and high-resolution multiphoton excited fluorescence images for animal study in vivo.

  9. Relaxation channels of multi-photon excited xenon clusters

    NASA Astrophysics Data System (ADS)

    Serdobintsev, P. Yu.; Rakcheeva, L. P.; Murashov, S. V.; Melnikov, A. S.; Lyubchik, S.; Timofeev, N. A.; Pastor, A. A.; Khodorkovskii, M. A.

    2015-09-01

    The relaxation processes of the xenon clusters subjected to multi-photon excitation by laser radiation with quantum energies significantly lower than the thresholds of excitation of atoms and ionization of clusters were studied. Results obtained by means of the photoelectron spectroscopy method showed that desorption processes of excited atoms play a significant role in the decay of two-photon excited xenon clusters. A number of excited states of xenon atoms formed during this process were discovered and identified.

  10. Relaxation channels of multi-photon excited xenon clusters.

    PubMed

    Serdobintsev, P Yu; Rakcheeva, L P; Murashov, S V; Melnikov, A S; Lyubchik, S; Timofeev, N A; Pastor, A A; Khodorkovskii, M A

    2015-09-21

    The relaxation processes of the xenon clusters subjected to multi-photon excitation by laser radiation with quantum energies significantly lower than the thresholds of excitation of atoms and ionization of clusters were studied. Results obtained by means of the photoelectron spectroscopy method showed that desorption processes of excited atoms play a significant role in the decay of two-photon excited xenon clusters. A number of excited states of xenon atoms formed during this process were discovered and identified. PMID:26395700

  11. Multiphoton interband excitations of quantum gases in driven optical lattices

    NASA Astrophysics Data System (ADS)

    Weinberg, M.; lschlger, C.; Strter, C.; Prelle, S.; Eckardt, A.; Sengstock, K.; Simonet, J.

    2015-10-01

    We report on the observation of multiphoton interband absorption processes for quantum gases in shaken light crystals. Periodic inertial forcing, induced by a spatial motion of the lattice potential, drives multiphoton interband excitations of up to the ninth order. The occurrence of such excitation features is systematically investigated with respect to the potential depth and the driving amplitude. Ab initio calculations of resonance positions as well as numerical evaluation of their strengths exhibit good agreement with experimental data. In addition our findings could make it possible to reach novel phases of quantum matter by tailoring appropriate driving schemes.

  12. The effects of spherical aberration on multiphoton fluorescence excitation microscopy

    PubMed Central

    Young, P.A.; Clendenon, S.G.; Byars, J.M.; Decca, R.S.; Dunn, K.W.

    2015-01-01

    Summary Multiphoton fluorescence excitation microscopy is almost invariably conducted with samples whose refractive index differ from that of the objective immersion medium, conditions that cause spherical aberration. Due to the quadratic nature of multiphoton fluorescence excitation, spherical aberration is expected to profoundly affect the depth dependence of fluorescence excitation. In order to determine the effect of refractive index mismatch in multiphoton fluorescence excitation microscopy, we measured signal attenuation, photobleaching rates and resolution degradation with depth in homogeneous samples with minimal light scattering and absorption over a range of refractive indices. These studies demonstrate that signal levels and resolution both rapidly decline with depth into refractive index mismatched samples. Analyses of photobleaching rates indicate that the preponderance of signal attenuation with depth results from decreased rates of fluorescence excitation, even in a system with a descannd emission collection pathway. Similar results were obtained in analyses of fluorescence microspheres embedded in rat kidney tissue, demonstrating that spherical aberration is an important limiting factor in multiphoton fluorescence excitation microscopy of biological samples. PMID:21118240

  13. Multiphoton electron detachment studies of C 7H 7- using a low-power infrared laser. Competition of radiative and collisional relaxation with vibrational excitation

    NASA Astrophysics Data System (ADS)

    Wight, Charles A.; Beauchamp, J. L.

    1989-07-01

    Infrared laser-induced electron detachment from C 6H 5CH 2-, C 6D 5CH 2- and cyclo-C 7H 7- has been investigated by ion cyclotron resonance spectroscopy. Anions are trapped at low pressure for up to several seconds and are uniformly irradiated with the unfocused output of a cw CO 2 laser. Electron detachment is monitored as a function of irradiation period, laser intensity, wave-length and neutral gas pressure. Infrared photodetachment spectra of C 6H 5CH 2- and C 6D 5CH 2- exhibit bands at 945 and 932 cm -1, respectively, while the spectrum of cyclo-C 7H 7- shows only slight dependence on laser frequency between 920 and 1080 cm -1. Modeling of the C 6H 5CH 2- photodetachment kinetics suggests that one step in the laser excitation process is rate limiting. Radiative relaxation does not appear to be significant at laser intensities greater than 10 W cm -2 and collisions with neutral molecules are only moderately effective in deactivating vibrationally excited anions.

  14. Multiphoton excitation in spin split two-dimensional electron gas

    NASA Astrophysics Data System (ADS)

    Gumber, Sukirti; Kumar, Manoj; Jha, Pradip Kumar; Mohan, Man

    2015-12-01

    The terahertz radiation induced multiphoton excitation processes have been studied in a two dimensional electron gas where the mixing of spin-up and spin-down states of neighbouring Landau levels by Rashba spin orbit interaction results in two new, unequally spaced energy branches. The multiphoton transitions between different quasi-spin branches of electron gas are found to be dominant over transitions involving the states from the same branch. It has been observed that enhancement and power-broadening of absorption spectrum can be effectively tuned by strength and frequency of the laser field as well as the spin-orbit coupling.

  15. Capillary electrophoresis based on continuous-wave multiphoton excitation fluorescence

    NASA Astrophysics Data System (ADS)

    Chen, Sheng; Luo, Qingming; Zeng, Shaoqun; Chen, Tongsheng; Gong, Hui; Fu, Ling

    2003-12-01

    High performance capillary electrophoresis(HPCE) has been widely applied to the field of chemistry and life science for its advantages of high separation efficiency, short analysis times and low sample volume requirements. But how to improve the detection capability of capillary electrophoresis was ever a key problem. Multi-photon excitation (MPE) fluorescence detection could solve this problem, however this method needed expensive femtosecond mod-locked laser, which restricted the improvement of capillary electrophoresis. In this paper, we implemented a set of MPE-CE system based on a continuous wave (CW), successfully exploited the CW multi-photon excitation to achieve detection of capillary electrophoresis. In our current MPE-CE system we used Ti:sapphire laser to prove MPE-CE could work in CW mode. From this point, we would substitute the laser with laser diode (LD) in future. This is really a new but also practical idea in MPE-CE system based on CW.

  16. Infrared multiphoton ignition and enhancement of combustion. Final report

    SciTech Connect

    Lavid, M.; Poulos, A.T.

    1986-08-01

    The research demonstrated the feasibility of obtaining reliable ignition and enhancement of combustion by a novel concept, Infrared Multiphoton Absorption/Dissociation (IRMPA/IRMPD). A methanol-oxygen system was chosen to test this concept because methanol is a potential alternative fuel susceptible to IRMPD. Multiphoton absorption, dissociation, and ignition experiments were conducted in closed pyrex reaction cells with IR-transmitting NaCl windows. Various pressures of pure methanol and methanol-oxygen mixtures were irradiated with single pulses from a TEA CO/sub 2/ laser usually tuned at 10.33 micrometers and focused to fluences in the range 1-80 J/cm2. Temporal and spatial ignition behaviors were investigated and successful ignitions were obtained by multiphoton absorption processes. The research scope was expanded to include a second alcohol, 2-propanol.

  17. Individual bioaerosol particle discrimination by multi-photon excited fluorescence.

    PubMed

    Kiselev, Denis; Bonacina, Luigi; Wolf, Jean-Pierre

    2011-11-21

    Femtosecond laser induced multi-photon excited fluorescence (MPEF) from individual airborne particles is tested for the first time for discriminating bioaerosols. The fluorescence spectra, analysed in 32 channels, exhibit a composite character originating from simultaneous two-photon and three-photon excitation at 790 nm. Simulants of bacteria aggregates (clusters of dyed polystyrene microspheres) and different pollen particles (Ragweed, Pecan, Mulberry) are clearly discriminated by their MPEF spectra. This demonstration experiment opens the way to more sophisticated spectroscopic schemes like pump-probe and coherent control. PMID:22109478

  18. Multi-photon excited luminescence of magnetic FePt core-shell nanoparticles.

    PubMed

    Seemann, K M; Kuhn, B

    2014-07-01

    We present magnetic FePt nanoparticles with a hydrophilic, inert, and biocompatible silico-tungsten oxide shell. The particles can be functionalized, optically detected, and optically manipulated. To show the functionalization the fluorescent dye NOPS was bound to the FePt core-shell nanoparticles with propyl-triethoxy-silane linkers and fluorescence of the labeled particles were observed in ethanol (EtOH). In aqueous dispersion the NOPS fluorescence is quenched making them invisible using 1-photon excitation. However, we observe bright luminescence of labeled and even unlabeled magnetic core-shell nanoparticles with multi-photon excitation. Luminescence can be detected in the near ultraviolet and the full visible spectral range by near infrared multi-photon excitation. For optical manipulation, we were able to drag clusters of particles, and maybe also single particles, by a focused laser beam that acts as optical tweezers by inducing an electric dipole in the insulated metal nanoparticles. In a first application, we show that the luminescence of the core-shell nanoparticles is bright enough for in vivo multi-photon imaging in the mouse neocortex down to cortical layer 5. PMID:25071977

  19. Multi-photon excited luminescence of magnetic FePt core-shell nanoparticles

    PubMed Central

    Seemann, K.M.; Kuhn, B.

    2014-01-01

    We present magnetic FePt nanoparticles with a hydrophilic, inert, and biocompatible silico-tungsten oxide shell. The particles can be functionalized, optically detected, and optically manipulated. To show the functionalization the fluorescent dye NOPS was bound to the FePt core-shell nanoparticles with propyl-triethoxy-silane linkers and fluorescence of the labeled particles were observed in ethanol (EtOH). In aqueous dispersion the NOPS fluorescence is quenched making them invisible using 1-photon excitation. However, we observe bright luminescence of labeled and even unlabeled magnetic core-shell nanoparticles with multi-photon excitation. Luminescence can be detected in the near ultraviolet and the full visible spectral range by near infrared multi-photon excitation. For optical manipulation, we were able to drag clusters of particles, and maybe also single particles, by a focused laser beam that acts as optical tweezers by inducing an electric dipole in the insulated metal nanoparticles. In a first application, we show that the luminescence of the core-shell nanoparticles is bright enough for in vivo multi-photon imaging in the mouse neocortex down to cortical layer 5. PMID:25071977

  20. vuv fluorescence from selective high-order multiphoton excitation of N{sub 2}

    SciTech Connect

    Coffee, Ryan N.; Gibson, George N.

    2004-05-01

    Recent fluorescence studies suggest that ultrashort pulse laser excitation may be highly selective. Selective high-intensity laser excitation holds important consequences for the physics of multiphoton processes. To establish the extent of this selectivity, we performed a detailed comparative study of the vacuum ultraviolet fluorescence resulting from the interaction of N{sub 2} and Ar with high-intensity infrared ultrashort laser pulses. Both N{sub 2} and Ar reveal two classes of transitions, inner-valence ns <- np and Rydberg np <- n{sup '}l{sup '}. From their pressure dependence, we associate each transition with either plasma or direct laser excitation. Furthermore, we qualitatively confirm such associations with the time dependence of the fluorescence signal. Remarkably, only N{sub 2} presents evidence of direct laser excitation. This direct excitation produces ionic nitrogen fragments with inner-valence (2s) holes, two unidentified transitions, and one molecular transition, the N{sub 2}{sup +}:X {sup 2}{sigma}{sub g}{sup +} <- C {sup 2}{sigma}{sub u}{sup +}. We discuss these results in the light of a recently proposed model for multiphoton excitation.

  1. Evidence for excitation of fluorescence in RPE melanin by multiphoton absorption

    NASA Astrophysics Data System (ADS)

    Glickman, Randolph D.; Rockwell, Benjamin A.; Noojin, Gary D.; Stolarski, David J.; Denton, Michael L.

    2002-06-01

    Previously, we reported that ultrashort, near infrared (NIR) laser pulses caused more DNA breakage in cultured retinal pigment epithelial (RPE) cells than did CW, NIR laser radiation delivering a similar radiant exposure. We hypothesized that this difference was due to multiphoton absorption in an intracellular chromophore such as the RPE melanin. We investigated two-photon excitation of fluorescence in a suspension of isolated bovine RPE melanosomes exposed to a 1-KHz train of approximately 50- fsec laser pulses at 810 nm from a Ti:Sapphire laser, and compared this to the fluorescence excited by CW exposures at 406 nm from a Krypton ion laser. Fluorescence was measured with a PC-based spectrometer. The CW sources excited fluorescence with a peak at 525 nm. The fluorescence intensity depended on the irradiance of the sample, as well as the melanosome concentration. Peak fluorescence was obtained with a suspension of ~2 x 107 melanin granules/ml. The 810-nm, ultrashort pulses also excited fluorescence, but with a broader, lower-amplitude peak. The weaker fluorescence signal excited by the 810-nm ultrashort pulse laser for a given melanosome concentration, compared to 406-nm CW excitation, is possibly due to the smaller two- photon absorption cross-section. These results indicate the involvement of multiphoton absorption in DNA damage.

  2. Selective multiphoton excitation by parametrically shaped laser pulses

    NASA Astrophysics Data System (ADS)

    Lindinger, A.

    2015-05-01

    Laser pulse shaping is reported for applications on multiphoton processes in dye molecules. Particularly phase-tailored pulse shapes are employed for two-photon excited fluorescence of dyes in a liquid environment, also at the distal end of an optical fiber, in order to improve the contrast between dye markers having similar excitation spectra. Precompensation of the optical fiber properties is utilized by analytical pulse shaping in order to receive specific parametric pulse forms after the fiber. This will lead to new endoscopic imaging applications with an increased fluorescence contrast. Moreover, selective excitation is also demonstrated for three-photon transitions of the two dyes, p-Terphenyl (PTP) and BM-Terphenyl (BMT), in solution by using shaped pulses without a fiber. A good agreement between experiment and theoretical simulation is obtained. With this approach it is possible to achieve a considerable change of the fluorescence contrast between the two dyes which is relevant for imaging applications of biological molecules.

  3. Infrared multiphoton ignition and combustion enhancement of natural gas. Final report, November 1989-October 1992

    SciTech Connect

    Lavid, M.; Poulos, A.T.; Gulati, S.K.; Stevens, J.G.; Lempert, W.R.

    1993-11-30

    This research program has demonstrated the feasibility of using Infrared Multiphoton (IRMP) processes for reliable ignition and combustion enhancement of natural gas. Hydrocarbon constituents of natural gas are excited with focused, pulsed infrared lasers tuned to discrete resonant frequencies causing their dissociation to very reactive radicals. These radicals participate in chain-branching and chain-propagating reactions significantly improving the combustion of natural gas. Experimental and computational tasks were performed to discern IR laser conditions needed to reliably obtain IRMP ignition and reduction in ignition delay time.

  4. Infrared multiphoton resummation in quantum electrodynamics

    NASA Astrophysics Data System (ADS)

    Mati, P.

    2016-02-01

    Infrared singularities in massless gauge theories are known since the foundation of quantum field theories. The root of this problem can be tracked back to the very definition of these long-range interacting theories such as QED. It can be shown that singularities are caused by the massless degrees of freedom (i.e. the photons in the case of QED). In the Bloch-Nordsieck model the absence of the infrared catastrophe can be shown exactly by the complete summation of the radiative corrections. In this paper we will give the idea of the derivation of the Bloch-Nordsieck propagators, that describes the infrared structure of the electron propagation, at zero and finite temperatures. Some ideas of the possible applications are also mentioned.

  5. Multiphoton dissociation and thermal unimolecular reactions induced by infrared lasers. [REAMPA code

    SciTech Connect

    Dai, H.L.

    1981-04-01

    Multiphoton dissociation (MPD) of ethyl chloride was studied using a tunable 3.3 ..mu..m laser to excite CH stretches. The absorbed energy increases almost linearly with fluence, while for 10 ..mu..m excitation there is substantial saturation. Much higher dissociation yields were observed for 3.3 ..mu..m excitation than for 10 ..mu..m excitation, reflecting bottlenecking in the discrete region of 10 ..mu..m excitation. The resonant nature of the excitation allows the rate equations description for transitions in the quasicontinuum and continuum to be extended to the discrete levels. Absorption cross sections are estimated from ordinary ir spectra. A set of cross sections which is constant or slowly decreasing with increasing vibrational excitation gives good fits to both absorption and dissociation yield data. The rate equations model was also used to quantitatively calculate the pressure dependence of the MPD yield of SF/sub 6/ caused by vibrational self-quenching. Between 1000-3000 cm/sup -1/ of energy is removed from SF/sub 6/ excited to approx. > 60 kcal/mole by collision with a cold SF/sub 6/ molecule at gas kinetic rate. Calculation showed the fluence dependence of dissociation varies strongly with the gas pressure. Infrared multiphoton excitation was applied to study thermal unimolecular reactions. With SiF/sub 4/ as absorbing gas for the CO/sub 2/ laser pulse, transient high temperature pulses were generated in a gas mixture. IR fluorescence from the medium reflected the decay of the temperature. The activation energy and the preexponential factor of the reactant dissociation were obtained from a phenomenological model calculation. Results are presented in detail. (WHK)

  6. Multiphoton excitation fluorescence correlation spectroscopy of fluorescent DNA base analogs

    NASA Astrophysics Data System (ADS)

    Katilius, Evaldas; Woodbury, Neal W.

    2004-06-01

    Two- and three-photon excitation was used to investigate the properties of two fluorescent DNA base analogs: 2-aminopurine and 6-methylisoxanthopterin. 2-aminopurine is a widely used fluorescent analog of the DNA base adenine. Three-photon excitation of 2-aminopurine is achievable by using intense femtosecond laser pulses in 850-950 nm spectral region. Interestingly, the three-photon excitation spectrum is blue-shifted relative to the three-times-wavelength single-photon excitation spectrum. The maximum of the absorbance band in the UV is at 305 nm, while the three-photon excitation spectrum has a maximum at around 880 nm. Fluorescence correlation measurements were attempted to evaluate the feasibility of using three-photon excitation of 2-aminopurine for DNA-protein interaction studies. However, due to relatively small three-photon absorption cross-section, a good signal-to-noise fluorescence correlation curves take very long time to obtain. Fluorescence properties of 6-methylisoxanthopterin, the fluorescent analog of guanine, were investigated using two-photon excitation. This molecule has the lowest energy absorption band centered around 350 nm, thus, two-photon excitation is attainable using 700 to 760 nm output of Ti-sapphire laser. The excitation spectrum of this molecule in the infrared well matches the doubled-wavelength single-photon excitation spectrum in the UV. The high fluorescence quantum yield of 6-methylisoxanthopterin allows efficient fluorescence correlation measurements and makes this molecule a very good candidate for using in in vitro DNA-protein interaction studies.

  7. ELECTRON DETACHMENT DISSOCIATION AND INFRARED MULTIPHOTON DISSOCIATION OF HEPARIN TETRASACCHARIDES

    PubMed Central

    Leach, Franklin E.; Xiao, Zhongping; Laremore, Tatiana N.; Linhardt, Robert J.; Amster, I. Jonathan

    2011-01-01

    Heparin glycosaminoglycans (GAGs) present the most difficult glycoform for analytical characterization due to high levels of sulfation and structural heterogeneity. Recent contamination of the clinical heparin supply and subsequent fatalities has highlighted the need for sensitive methodologies of analysis. In the last decade, tandem mass spectrometry has been increasingly applied for the analysis of GAGs, but developments in the characterization of highly sulfated compounds have been minimal due to the low number of cross-ring cleavages generated by threshold ion activation by collisional induced dissociation (CID). In the current work, electron detachment dissociation (EDD) and infrared multiphoton dissociation (IRMPD) are applied to a series of heparin tetrasaccharides. With both activation methods, abundant glycosidic and cross-ring cleavages are observed. The concept of Ionized Sulfate Criteria (ISC) is presented as a succinct method for describing the charge state, degree of ionization and sodium/proton exchange in the precursor ion. These factors contribute to the propensity for useful fragmentation during MS/MS measurements. Precursors with ISC values of 0 are studied here, and shown to yield adequate structural information from ion activation by EDD or IRMPD. PMID:22247649

  8. The origin of the infrared multiphoton induced luminescence of chromyl chloride

    NASA Astrophysics Data System (ADS)

    Tsao, Jeffrey Y.; Bloembergen, N.; Burak, Itamar

    1981-07-01

    We have studied systematically the luminescence induced when CrO2Cl2 is infrared multiphoton excited by a moderate fluence (0-15 J/cm2) CO2 laser. This includes the dependence of the full luminescence spectrum on CO2 laser fluence, and for various spectral regions of the luminescence their dependence on CO2 laser wavelength and fluence, their intramolecular decay rates, and using a time-of-flight technique the translational temperatures associated with their sources. These measurements show that two species contribute to the luminescence. In addition, a chemical identification of these species is made by preceding the IR laser pulse with a UV laser pulse to photochemically prepare the CrO2Cl fragment, and isolate its contribution to the luminescence. Strong evidence is obtained in support of both CrO2Cl fragment and CrO2Cl2 parent contributions to the luminescence. The relative contributions of the two to the luminescence and an absolute yield for the IR multiphoton induced dissociation of CrO2Cl2 into CrO2Cl as functions of CO2 laser fluence are estimated.

  9. The effects of refractive index heterogeneity within kidney tissue on multiphoton fluorescence excitation microscopy.

    PubMed

    Young, P A; Clendenon, S G; Byars, J M; Dunn, K W

    2011-05-01

    Although multiphoton fluorescence excitation microscopy has improved the depth at which useful fluorescence images can be collected in biological tissues, the reach of multiphoton fluorescence excitation microscopy is nonetheless limited by tissue scattering and spherical aberration. Scattering can be reduced in fixed samples by mounting in a medium whose refractive index closely matches that of the fixed material. Using optical 'clearing', the effects of refractive index heterogeneity on signal attenuation with depth are investigated. Quantitative measurements show that by mounting kidney tissue in a high refractive index medium, less than 50% of signal attenuates in 100 μm of depth. PMID:21118239

  10. Fermi-coupled spherically adapted effective states in the collisionless multiphoton excitation of SF 6

    NASA Astrophysics Data System (ADS)

    Di Lauro, C.; Lattanzi, F.

    1982-10-01

    A calculation method for the collisionless multiphoton excitation of SF 6 by intense CO 2 laser light up to a chain of parallel nv3, ( n - 1) v3 + v2 + v6 … vibrational-rotational ladders linked by Fermi interaction is described. Spherically adapted effective states suitable to the purpose are defined, and matrix elements for multiphoton excitation in the rotatingwave approximation effective hamiltonian formalism are given in this basis. The method is aimed at the investigation of population transfer between the cited parallel vibrational ladders, and is suitable for computer-calculation programmation.

  11. Photodynamic therapy and knocking out of single tumor cells by multiphoton excitation processes

    NASA Astrophysics Data System (ADS)

    Riemann, Iris; Fischer, Peter; Koenig, Karsten

    2004-09-01

    Near infrared (NIR) ultrashort laser pulses of 780 nm have been used to induce intracellular photodynamic reactions by nonlinear excitation of porphyrin photosensitizers. Intracellular accumulation and photobleaching of the fluorescent photosensitizers protoporphyrin IX and Photofrin (PF) have been studied by non-resonant two-photon fluorescence excitation of PF and aminolevulinic acid (ALA)-labeled Chinese hamster ovary (CHO) cells. To testify the efficacy of both substrates to induce irreversible destructive effects, the cloning efficiency (CE) of cells exposed to femtosecond pulses of a multiphoton laser scanning microscope (40x/1.3) was determined. In the case of Photofrin accumulation, CEs of 50% and 0% were obtained after 17 laserscans (2 mW?, 16 s/ frame) and 50 scans, respectively. All cells exposed to 50 scans died within 48h after laser exposure. 100 scans were required to induce lethal effects in ALA labeled cells. Sensitizer-free control cells could be scanned 250 times (1.1 h) and more without impact on the reproduction behavior, morphology, and vitality. In addition to the slow phototoxic effect by photooxidation processes, another destructive but immediate effect based on optical breakdown was induced when employing high intense NIR femtosecond laser beams. This was used to optically knock out single tumor cells in living mice (solid Ehrlich-Carcinoma) in a depth of 10 to 100 ?m.

  12. Investigations of multiphoton excitation and ionization in a short range potential

    SciTech Connect

    Susskind, S.M.; Cowley, S.C.; Valeo, E.J.

    1989-02-01

    We introduce an approach to the study of excitation and ionization for a system with a short range potential. In particular, analytical and numerical results are presented for the multiphoton ionization rate, under strong field conditions, of an electron confined by a delta-function potential. 9 refs., 3 figs.

  13. Arbitrary two-dimensional multiphoton excitation patterns with temporally focused digital holograms

    NASA Astrophysics Data System (ADS)

    Oron, Dan; Papagiakoumou, Eirini; de-Sars, Vincent; Emiliani, Valentina

    2009-02-01

    Multiphoton excitation has recently found application in the fields of bioimaging, uncaging and lithography. In order to fully exploit the advantages of nonlinear excitation, in particular the axial resolution due to nonlinearity, most systems to date operate with point or multipoint excitation, while scanning either the laser beam or the sample to generate the illumination pattern. Here we combine the recently introduced technique of scanningless multiphoton excitation by temporal focusing with recent advances in digital holography to generate arbitrarily shaped, depth resolved, two-dimensional excitation patterns completely without scanning. This is of particular importance in applications requiring uniform excitation of large areas over short time scales, such as neuronal activation by multiphoton uncaging of neurotransmitters. We present an experimental and theoretical analysis of the effect of spatial patterning on the depth resolution achieved in temporal focusing microscopy. It is shown that the depth resolution for holographic excitation is somewhat worse than that achieved for uniform illumination. This is also accompanied by the appearance of a speckle pattern at the temporal focal plane. The origin of the two effects, as well as means to overcome them, are discussed.

  14. Chirped multiphoton adiabatic passage for a four-level ladder-type Rydberg excitation

    NASA Astrophysics Data System (ADS)

    Qian, Jing; Zhai, Jingjing; Zhang, Lu; Zhang, Weiping

    2015-01-01

    We develop a multiphoton adiabatic passage to realize a highly efficient Rydberg excitation in a four-level ladder-type atomic system. The adiabatic passage is based on the existence of a novel quasidark state in the cascade excitation system where the frequencies of the lasers are appropriately chirped with time. We also investigate the influence of the interatomic Rydberg interaction on the passage and extend its application to the preparation of antiblockade Rydberg atom pairs in the Rydberg blockade regime.

  15. Infrared multiphoton dissociation of SF6 at long wavelengths

    NASA Astrophysics Data System (ADS)

    Del Bello, U.; Fuss, W.; Kompa, K. L.; Tablas, F. M. G.

    1989-03-01

    The dissociation probability Pd of 32 SF6 was investigated between 908 and 875 cm-1 at room temperature and at 140 K. In the region of the expected Pd maximum of 36SF6 (?905 cm-1), we found a ten times higher Pd(32) (implying a ten times smaller selectivity for 36S) than expected from extrapolation of shorter wavelength data. The Pd spectrum in this and in other regions point to an important role of direct multiphoton transitions to levels containing quanta of the neighboring combination mode ?5+?6 (870 cm-1) besides ?3 quanta. On the short wavelength side of ?3, the analogous transitions involve the ?2+?6 mode (991 cm-1). The wings, and thus the spectral shape of Pd, in the ?3 region are also governed by them. They depend more strongly on temperature than stepwise transitions, because the intermediate near-resonances favor the high rotational states.

  16. Multi-photon excitation microscopy in intact animals.

    PubMed

    Rothstein, Emily C; Nauman, Michael; Chesnick, Scott; Balaban, Robert S

    2006-04-01

    Two-photon excitation fluorescence microscopy and backscattered-second harmonic generation microscopy permit the investigation of the subcellular events within living animals but numerous aspects of these experiments need to be optimized to overcome the traditional microscope geometry, motion and optical coupling to the subject. This report describes a stable system for supporting a living instrumented mouse or rabbit during endogenous reduced nicotinamide adenine dinucleotide and exogenous dye two-photon excitation fluorescence microscopy measurements, and backscattered-second harmonic generation microscopy measurements. The system was a modified inverted LSM510 microscope (Carl Zeiss, Inc., Thornwood, NY, U.S.A.) with a rotating periscope that converted the inverted scope to an upright format, with the objective located approximately, 15 cm from the centre of the microscope base, allowing easy placement of an instrumented animal. An Olympus 20x water immersion objective was optically coupled to the tissue, without a cover glass, via a saline bath or custom hydrated transparent gel. The instrumented animals were held on a specially designed holder that poised the animal under the objective as well as permitted different ventilation schemes to minimize motion. Using this approach, quality images were routinely collected in living animals from both the peripheral and body cavity organs. The remaining most significant issue for physiological studies using this approach is motion on the micrometre scale. Several strategies for motion compensation are described and discussed. PMID:16734715

  17. Microfluorometric Detection of Catecholamines with Multiphoton-Excited Fluorescence

    NASA Astrophysics Data System (ADS)

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

    2004-04-01

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

  18. Modification of a tandem mass-spectrometer for infrared multi-photon dissociation (IRMPD) of gas-phase ions

    NASA Astrophysics Data System (ADS)

    Gillis, Julie M.; Osburn, Sandra M.; van Stipdonk, Michael J.; Corcovilos, Theodore A.

    2015-05-01

    Infrared multi-photon dissociation (IRMPD) is a method of fragmenting molecular ions for structural analysis of the parent molecule. The target ions absorb many photons, increasing the vibrational state of the excited bonds until the dissociation occurs. We have modified a commercial linear quadrupole trap tandem mass spectrometer (Thermo-Fisher LTQ) by installing a removable high-vacuum window in the rear accessory plate of the mass spectrometer. The window allows us to inject laser light into the ion trap. The shape of the injected laser beam is optimized to match the volume of the ion cloud within the ion trap, improving IRMPD efficiency. We present preliminary data of the IRMPD of weakly bound uranyl-acetone and uranyl-dimethyl sulfoxide clusters using a 20-W pulsed CO2 laser (wavelength 10 . 6 μm), showing previously undetected fragmentation products.

  19. Multiphoton stimulation of neurons.

    PubMed

    Hirase, Hajime; Nikolenko, Volodymyr; Goldberg, Jesse H; Yuste, Rafael

    2002-06-01

    We pulsed the activation of neurons using a femtosecond laser. Pyramidal neurons are depolarized and fire action potentials when high intensity mode-locked infrared light irradiates somatic membranes and axon initial segments. This depolarization is reversible, does not occur with CW laser light, and appears to be due to multiphoton excitation. We describe two regimes of multiphoton optical stimulation. Low intensity, long duration laser irradiation produces a sustained depolarization, insensitive to sodium channel blockers yet sensitive to antioxidants. On the other hand, high intensity, short duration irradiation can induce fast depolarizations, which appear due to different mechanism. The combination of multiphoton stimulation and optical probing could enable systematic analysis of circuits. PMID:11984845

  20. Multiphoton imaging of quantum dot bioconjugates in cultured cells following Nd:YLF laser excitation

    NASA Astrophysics Data System (ADS)

    Serrano, Elba E.; Knight, V. B.

    2005-04-01

    Quantum dot bioconjugates offer unprecedented opportunities for monitoring biological processes and molecular interactions in cells, tissues, and organs. We are interested in developing applications that permit investigation of physiological processes and cytoskeletal organization in live cells, and allow imaging of complex organs, such as the auditory and vestibular sensory structures of the inner ear. Multiphoton microscopy is a powerful technique for acquiring images from deep within a sample while reducing phototoxic effects of laser light exposure on cells. Previous studies have established that a solid-state Nd:YLF laser can be used to acquire two-photon and three-photon images from live cells while minimizing phototoxic side effects (Wokosin et al., 1996, Bioimaging, 4:208-214; Squirrell et al., 1999, Nature Biotechnology, 8:763-767). We present here the results of experiments using an all-solid-state Nd:YLF 1047 nm femtosecond laser (Microlase DPM1000) source to excite quantum dot bioconjugates. Cells were labeled with Qdot (Quantum Dot Corporation) bioconjugates or with Alexa Fluor (Molecular Probes) bioconjugates and then imaged with a BioRad 1024 confocal microscope configured for multiphoton imaging using internal or external (non-descanned) detectors. Results demonstrate that the Nd:YLF laser can be used to stimulate fluorescence emission of quantum dots and Alexa Fluor bioconjugates in cultured amphibian (Xenopus) and mammalian (rat, chinese hamster) cells. We conclude that the Nd:YLF laser is a viable excitation source that extends the applicability of quantum dots for investigation of biological processes using multiphoton microscopy.

  1. Nonlinear structured-illumination enhanced temporal focusing multiphoton excitation microscopy with a digital micromirror device

    PubMed Central

    Cheng, Li-Chung; Lien, Chi-Hsiang; Da Sie, Yong; Hu, Yvonne Yuling; Lin, Chun-Yu; Chien, Fan-Ching; Xu, Chris; Dong, Chen Yuan; Chen, Shean-Jen

    2014-01-01

    In this study, the light diffraction of temporal focusing multiphoton excitation microscopy (TFMPEM) and the excitation patterning of nonlinear structured-illumination microscopy (NSIM) can be simultaneously and accurately implemented via a single high-resolution digital micromirror device. The lateral and axial spatial resolutions of the TFMPEM are remarkably improved through the second-order NSIM and projected structured light, respectively. The experimental results demonstrate that the lateral and axial resolutions are enhanced from 397 nm to 168 nm (2.4-fold) and from 2.33 ?m to 1.22 ?m (1.9-fold), respectively, in full width at the half maximum. Furthermore, a three-dimensionally rendered image of a cytoskeleton cell featuring ~25 nm microtubules is improved, with other microtubules at a distance near the lateral resolution of 168 nm also able to be distinguished. PMID:25136483

  2. Freeform multiphoton excited microfabrication for biological applications using a rapid prototyping CAD-based approach

    NASA Astrophysics Data System (ADS)

    Cunningham, Lawrence P.; Veilleux, Matthew P.; Campagnola, Paul J.

    2006-09-01

    Multiphoton excited polymerization has attracted increasing attention as a powerful 3 dimensional nano/microfabrication tool. The nonlinear excitation confines the fabrication region to the focal volume allowing the potential to achieve freeform fabrication with submicron capabilities. We report the adaptation and use of a computer aided design (CAD) approach, based on rapid prototyping software, which exploits this potential for fabricating with protein and polymers in biologically compatible aqueous environments. 3D structures are drawn in the STL format creating a solid model that can be sliced, where the individual sections are then serially fabricated without overwriting previous layers. The method is shown for potential biological applications including microfluidics, cell entrapment, and tissue engineering.

  3. Freeform multiphoton excited microfabrication for biological applications using a rapid prototyping CAD-based approach.

    PubMed

    Cunningham, Lawrence P; Veilleux, Matthew P; Campagnola, Paul J

    2006-09-18

    Multiphoton excited polymerization has attracted increasing attention as a powerful 3 dimensional nano/microfabrication tool. The nonlinear excitation confines the fabrication region to the focal volume allowing the potential to achieve freeform fabrication with submicron capabilities. We report the adaptation and use of a computer aided design (CAD) approach, based on rapid prototyping software, which exploits this potential for fabricating with protein and polymers in biologically compatible aqueous environments. 3D structures are drawn in the STL format creating a solid model that can be sliced, where the individual sections are then serially fabricated without overwriting previous layers. The method is shown for potential biological applications including microfluidics, cell entrapment, and tissue engineering. PMID:19529241

  4. Analysis of microparticle penetration into human and porcine skin: non-invasive imaging with multiphoton excitation microscopy

    NASA Astrophysics Data System (ADS)

    Mulholland, William J.; Kendall, Mark A.; Bellhouse, Brian J.; White, Nick

    2002-06-01

    At the University of Oxford and PowderJect Pharmaceuticals plc, a unique form of needle-free injection technology has been developed. Powdered vaccines and drugs in micro-particle form are accelerated in a high-speed gas flow to sufficient velocity to enter the skin, subsequently achieving a pharmaceutical effect. To optimize the delivery of vaccines and drugs with this method a detailed understanding of the interactive processes that occur between the microparticles and the skin is necessary. Investigations to date of micro-particle delivery into excised human and animal tissue have involved image analyses of histology sections. In the present study, a series of investigations were conducted on excised human and porcine skin using the technique of Multi-Photon fluorescence excitation Microscopy (MPM) to image particles and skin structures post-penetration. Micro-particles of various size and composition were imaged with infrared laser excitation. Three-dimensional images of stratum corneum and epidermal cell deformation due to micro-particle penetration were obtained. Measurements of micro-particle penetration depth taken from z-scan image stacks were used to successfully quantify micro-particle distribution within the skin, without invasively disrupting the skin target. This study has shown that MPM has great potential for the non-invasive imaging of particle skin interactive processes that occur with the transdermal delivery of powdered micro-particle vaccines and drugs.

  5. Imaging NO elimination in the infrared multiphoton dissociation of nitroalkanes and alkyl nitrites

    NASA Astrophysics Data System (ADS)

    Fernando, Ravin; Ariyasingha, Nuwandi M.; Suits, Arthur G.

    2016-02-01

    We present a DC slice imaging study of the decomposition of C2, C3 and C4 nitroalkanes and alkyl nitrites, focusing on the NO elimination channel, possibly a minor pathway. Infrared multiphoton dissociation (IRMPD) is used to induce dissociation on the ground electronic state under collisionless conditions. The channels that produced NO as a product were studied and compared among the target molecules to gain a better understanding of the isomerization of the nitroalkanes prior to dissociation. Trends in the total translational energy and NO rotational temperatures obtained from the images are discussed.

  6. Infrared multiphoton dissociation of anisole: Production and dissociation of phenoxy radical

    SciTech Connect

    Schmoltner, A.M.; Anex, D.S.; Lee, Y.T.

    1992-02-06

    The infrared multiphoton dissociation of anisole in a molecular beam was studied using pulsed CO{sub 2} laser radiation, with the product recoil energy distributions measured using the time-of-flight technique. The only primary process identified was the dissociation into phenoxy and methyl radicals, with the shape of the translational energy distribution of the products revealing a small exit barrier. Under conditions of higher laser fluence, secondary dissociation of the phenoxy radical into carbon monoxide and C{sub 5}H{sub 5} was observed. 28 refs., 9 figs.

  7. Multiphoton resonant excitation of Fermi-Dirac sea in graphene at the interaction with strong laser fields

    NASA Astrophysics Data System (ADS)

    Avetissian, Hamlet K.; Avetissian, Ara K.; Mkrtchian, Garnik F.; Sedrakian, Khachik V.

    2012-01-01

    In order to develop the microscopic theory of the multiphoton interaction of graphene with strong laser fields of arbitrary polarization, the analytical solution of established evolutionary equation for graphene single-particle density matrix was obtained revealing the generalized Rabi oscillations of Fermi-Dirac sea in the graphene. Creation of coherent superposition states of the particle-hole pairs in the graphene at the multiphoton resonant excitation by laser pulses of arbitrary polarizations and harmonics generation at the particle-hole annihilation from these superposition states are shown.

  8. Influence of multi-photon excitation on the atomic above-threshold ionization

    NASA Astrophysics Data System (ADS)

    Tian, Yuan-Ye; Wang, Chun-Cheng; Li, Su-Yu; Guo, Fu-Ming; Ding, Da-Jun; Wim-G, Roeterdink; Chen, Ji-Gen; Zeng, Si-Liang; Liu, Xue-Shen; Yang, Yu-Jun

    2015-04-01

    Using the time-dependent pseudo-spectral scheme, we solve the time-dependent Schrdinger equation of a hydrogen-like atom in a strong laser field in momentum space. The intensity-resolved photoelectron energy spectrum in above-threshold ionization is obtained and further analyzed. We find that with the increase of the laser intensity, the above-threshold ionization emission spectrum exhibits periodic resonance structure. By analyzing the population of atomic bound states, we find that it is the multi-photon excitation of bound state that leads to the occurrence of this phenomenon, which is in fairly good agreement with the experimental results. Project supported by the National Basic Research Program of China (Grant No. 2013CB922200), the National Natural Science Foundation of China (Grants Nos. 11274141, 11034003, 11304116, 11274001, and 11247024), and the Jilin Provincial Research Foundation for Basic Research, China (Grant No. 20140101168JC).

  9. Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation

    NASA Astrophysics Data System (ADS)

    Zipfel, Warren R.; Williams, Rebecca M.; Christie, Richard; Nikitin, Alexander Yu; Hyman, Bradley T.; Webb, Watt W.

    2003-06-01

    Multicolor nonlinear microscopy of living tissue using two- and three-photon-excited intrinsic fluorescence combined with second harmonic generation by supermolecular structures produces images with the resolution and detail of standard histology without the use of exogenous stains. Imaging of intrinsic indicators within tissue, such as nicotinamide adenine dinucleotide, retinol, indoleamines, and collagen provides crucial information for physiology and pathology. The efficient application of multiphoton microscopy to intrinsic imaging requires knowledge of the nonlinear optical properties of specific cell and tissue components. Here we compile and demonstrate applications involving a range of intrinsic molecules and molecular assemblies that enable direct visualization of tissue morphology, cell metabolism, and disease states such as Alzheimer's disease and cancer.

  10. Feasibility of using multiphoton excited tissue autofluorescence for in vivo human histopathology.

    PubMed Central

    Dela Cruz, Johanna M.; McMullen, Jesse D.; Williams, Rebecca M.; Zipfel, Warren R.

    2010-01-01

    Rapid and direct imaging of microscopic tissue morphology and pathology can be achieved by multiphoton imaging of intrinsic tissue fluorophores and second harmonic signals. Engineering parameters for developing this technology for clinical applications include excitation levels and collection efficiencies required to obtain diagnostic quality images from different tissue types and whether these levels are mutagenic. Here we provide data on typical average powers required for high signal-to-noise in vivo tissue imaging and assess the risk potential of these irradiance levels using a mammalian cell gene mutation assay. Exposure times of ~16 milliseconds per cell to 760 nm, ~200 fs raster-scanned laser irradiation delivered through a 0.75 NA objective produced negligible mutagenicity at powers up to about 50 mW. PMID:21258552

  11. Nonperturbative analysis of the two-level atom: Applications to multiphoton excitation

    SciTech Connect

    Duvall, R.E.; Valeo, E.J.; Oberman, C.R.

    1987-08-01

    Selective excitation in an atomic system subjected to a slowly varying external electromagnetic field is studied using a two-level model. Time evolution of the system is found using an approach which is nonperturbative in the field strength. There is no constraint to small values of the applied field, that is, the field (in appropriate energy units) need not be small compared to the difference in energies of the two levels. Rather, we prey upon the fact that the situation of interest to us is where the frequency of the exciting field is small compared to the frequency associated with the level difference. Transition probabilities and resonance conditions are found which circumscribe both the large and small field limits. In the weak field limit the previous results of high-order perturbation theory are readily recovered. For a monochromatic field the characteristic features of resonance excitation at high harmonic number of the applied field are (a) extremely narrow resonance widths and (b) shifts in resonance positions which are strong functions of field intensity. Because of this sensitivity, we are able to demonstrate that when slow temporal evolution of the field amplitude is taken into account (e.g., due to finite pulse duration) the appropriate mean excitation rate is that due to the uncorrelated contribution of many resonances. The results of this analysis are used to estimate excitation rates in a specific atomic system, Cd/sup 12 +/, which are then compared to multiphoton ionization rates. Our calculations suggest that the ionization rate exceeds the excitation rate by several orders of magnitude. 15 refs., 3 figs.

  12. Infrared multiphoton ignition and combustion enhancement of natural gas. Final report, February 1988-May 1989

    SciTech Connect

    Lavid, M.; Poulos, A.T.

    1990-08-14

    The Phase I feasibility study has demonstrated the potential of using Infrared Multiphoton (IRMP) processes for ignition and combustion enhancement of natural gas. Experiments with IR lasers were conducted to investigate the effects of IRMP techniques on methane, ethane, isotopically substituted surrogates, and sensitized mixtures. Major findings include: (1) Identification of HF/DF laser as an appropriate source for IRMP; (2) Demonstration of IRMP decomposition of deuterated methane with CO{sub 2} laser; (3) Successful IRMP ignition of deuterated methane and ethane mixtures; (4) Successful IRMP ignition of methane mixtures using photo-sensitizers. These experimental findings support the concept that IRMP processes are capable of dissociating methane, other minor constituents of natural gas, or sensitizers to reactive, combustion-enhancing radicals. A comprehensive Phase II contract to bring the concept closer to commercial implementation is underway.

  13. Ultrafast Multiphoton Pump-probe Photoemission Excitation Pathways in Rutile TiO2(110)

    SciTech Connect

    Argondizzo, Adam; Cui, Xuefeng; Wang, Cong; Sun, Huijuan; Shang, Honghui; Zhao, Jin; Petek, Hrvoje

    2015-04-27

    We investigate the spectroscopy and photoinduced electron dynamics within the conduction band of reduced rutile TiO2(110) surface by multiphoton photoemission (mPP) spectroscopy with wavelength tunable ultrafast (!20 fs) laser pulse excitation. Tuning the mPP photon excitation energy between 2.9 and 4.6 eV reveals a nearly degenerate pair of new unoccupied states located at 2.73 ± 0.05 and 2.85 ± 0.05 eV above the Fermi level, which can be analyzed through the polarization and sample azimuthal orientation dependence of the mPP spectra. Based on the calculated electronic structure and optical transition moments, as well as related spectroscopic evidence, we assign these resonances to transitions between Ti 3d bands of nominally t2g and eg symmetry, which are split by crystal field. The initial states for the optical transition are the reduced Ti3+ states of t2g symmetry populated by formation oxygen vacancy defects, which exist within the band gap of TiO2. Furthermore,we studied the electron dynamics within the conduction band of TiO2 by three-dimensional time-resolved pump-probe interferometric mPP measurements. The spectroscopic and time-resolved studies reveal competition between 2PP and 3PP processes where the t2g-eg transitions in the 2PP process saturate, and are overtaken by the 3PP process initiated by the band-gap excitation from the valence band of TiO2.

  14. Cell Adhesion on Micro-Structured Fibronectin Gradients Fabricated by Multiphoton Excited Photochemistry

    PubMed Central

    Chen, Xiyi; Su, Yuan-Deng; Ajeti, Visar; Chen, Shean-Jen; Campagnola, Paul J.

    2013-01-01

    Concentration gradients of ECM proteins play active roles in many areas of cell biology including wound healing and metastasis. They may also form the basis of tissue engineering scaffolds, as these can direct cell adhesion and migration and promote new matrix synthesis. To better understand cellmatrix interactions on attractive gradients, we have used multiphoton excited (MPE) photochemistry to fabricate covalently linked micro-structured gradients from fibronectin (FN). The gradient design is comprised of a parallel series of individual linear gradients with overall dimensions of approximately 800 800 ?m, where a linear dynamic range of nearly 10-fold in concentration was achieved. The adhesion dynamics of 3T3 fibroblasts were investigated, where the cell morphology and actin cytoskeleton became increasingly elongated and aligned with the direction of the gradient at increasing protein concentration. Moreover, the cell morphologies are distinct when adhered to regions of differing FN concentration but with similar topography. These results show that the fabrication approach allows investigating the roles of contact guidance and ECM cues on the cellmatrix interactions. We suggest this design overcomes some of the limitations with other fabrication methods, especially in terms of 3D patterning capabilities, and will serve as a new tool to study cellmatrix interactions. PMID:23710258

  15. Nonperturbative methods in the problem of multiphoton excitation of atom by squeezed light

    NASA Technical Reports Server (NTRS)

    Belousov, A. V.; Kovarsky, V. A.

    1993-01-01

    Multiphoton detectors for the strong squeezed light vacuum are considered. The result is compared with the perturbation theory. It is shown that as the degree of squeezing is increased the statistical factor decreases.

  16. First in vivo animal studies on intraocular nanosurgery and multiphoton tomography with low-energy 80-MHz near-infrared femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Konig, Karsten; Wang, Bagui; Krauss, Oliver; Riemann, Iris; Schubert, Harald; Kirste, Sigrun; Fischer, Peter

    2004-07-01

    We report on a method for refractive laser surgery based on low-energy femtosecond laser pulses provided by ultracompact turn-key non-amplified laser systems. An additional excimer laser is not required for ablation of the stroma. The novel method has the potential to be used for (i) optical flap creation as well as stroma ablation and (ii) for non-invasive flap-free intrastromal ablation. In addition, 3D multiphoton imaging of the cornea can be performed. In particular, we used sub-nanojoule near infrared 80 MHz femtosecond laser pulses for multiphoton imaging of corneal structures with ultrahigh resolution (< 1?m) as well as for highly precise intraocular refractive surgery. Imaging based on two-photon excited cellular autofluorescence and SHG formation in collagen structures was performed at GW/cm2 intensities, whereas destructive optical breakdown for nanoprocessing occurred at TW/cm2 light intensities. These high intensities were realized with sub-nJ pulses within a subfemtoliter intrastromal volume by diffraction-limited focussing with high NA objectives and beam scanning 50 to 140 ?m below the epithelial surface. Multiphoton tomography of the cornea was used to determine the target of interest and to visualize intraocular post-laser effects. Histological examination with light- and electron microscopes of laser-exposed porcine and rabbit eyes reveal a minimum intratissue cut size below 1 ?m without destructive effects to surrounding collagen structures. LASIK flaps and intracorneal cavities could be realized with high precision using 200 fs, 80 MHz, sub-nanojoule pulses at 800 nm. First studies on 80 MHz femtosecond laser surgery on living rabbits have been performed.

  17. Charge-state selective fragmentation analysis for protonated peptides in infrared multiphoton dissociation

    NASA Astrophysics Data System (ADS)

    Fukui, Kazuhiko; Naito, Yasuhide; Akiyama, Yutaka; Takahashi, Katsutoshi

    2004-06-01

    We investigate the charge-state selective cleavage in gas-phase protonated peptides by using electrospray ionization (ESI) Fourier-transform ion cyclotron resonance (FTICR) mass spectrometer (MS). The singly and multiply protonated peptides (Angiotensin II, Angiotensin I, Urotensin II, Bradykinin, Substance P) at the selected charge state were cleaved with the techniques of infrared multiphoton dissociation (IRMPD) in FTICR MS. Systematically changing IR laser power, the fragment ions were assessed to determine the cleaved amino bonds in the peptides at the selected charge state, then to quantitatively obtain the dissociation efficiency for each fragment. The results show that the fragment ions are observed at the selective cleavage of Asp-Xaa in singly charged Angiotensin I, Angiotensin II and Urotensin II ions that contain an acidic residue (Asp) and a basic residue (Arg or Lys), while the fragment ions arising from cleavage at Xaa-Pro dominate in those doubly charged peptides. It is observed that the dissociation channel of Asp-Xaa requires higher energy than that of Xaa-Pro. The charge selective fragmentation is not seen for Bradykinin and Substance P containing strong basic amino residues without an acidic residue.

  18. In vivo manipulation of fluorescently labeled organelles in living cells by multiphoton excitation.

    PubMed

    Watanabe, Wataru; Matsunaga, Sachihiro; Higashi, Tsunehito; Fukui, Kiichi; Itoh, Kazuyoshi

    2008-01-01

    Femtosecond laser pulses in the near-infrared region have potential applications in the imaging and manipulation of intracellular organelles. We report on the manipulation of intracellular organelles by two-photon excitation. The dynamics of green fluorescent protein (GFP)-histone are investigated by two-photon fluorescence recovery after photobleaching (FRAP). Intracellular ablation of fluorescently labeled organelles in living cells is performed by focusing femtosecond laser pulses. We report on the selective marking of individual organelles by using two-photon conversion of a photoconvertible fluorescent protein. PMID:18601537

  19. Isotopically selective multiphoton dissociation of CF/sub 2/HCl in collision conditions of excitation

    SciTech Connect

    Avatkov, O.N.; Vetsko, V.M.; Dzneladze, D.I.; Esadze, G.G.; Kaminskii, A.V.; Kudziev, A.G.; Kudziev, T.A.

    1988-09-01

    The effect of the energy density, laser radiation frequency, and resonance and buffer gas pressure on the process of multiphoton dissociation of CF/sub 2/HCl molecules was experimentally studied. The dependence of the threshold value of the energy density on the resonance gas pressure in long-wave tuning of the radiation frequency on the maxima of the spectra of linear and multiphoton absorption of /sup 13/CF/sub 2/HCl molecules was studied. A model which qualitatively explains the threshold character of dissociation and the increase in selectivity with an increase in the gas pressure and radiation pulse duration was proposed.

  20. Multiphoton spectroscopy of human skin in vivo

    NASA Astrophysics Data System (ADS)

    Breunig, Hans G.; Weinigel, Martin; Knig, Karsten

    2012-03-01

    In vivo multiphoton-intensity images and emission spectra of human skin are reported. Optical sections from different depths of the epidermis and dermis have been measured with near-infrared laser-pulse excitation. While the intensity images reveal information on the morphology, the spectra show emission characteristics of main endogenous skin fluorophores like keratin, NAD(P)H, melanin, elastin and collagen as well as of second harmonic generation induced by the excitation-light interaction with the dermal collagen network.

  1. COMPACT NON-CONTACT TOTAL EMISSION DETECTION FOR IN-VIVO MULTI-PHOTON EXCITATION MICROSCOPY

    PubMed Central

    Glancy, Brian; Karamzadeh, Nader S.; Gandjbakhche, Amir H.; Redford, Glen; Kilborn, Karl; Knutson, Jay R.; Balaban, Robert S.

    2014-01-01

    Summary We describe a compact, non-contact design for a Total Emission Detection (c-TED) system for intra-vital multi-photon imaging. To conform to a standard upright two-photon microscope design, this system uses a parabolic mirror surrounding a standard microscope objective in concert with an optical path that does not interfere with normal microscope operation. The non-contact design of this device allows for maximal light collection without disrupting the physiology of the specimen being examined. Tests were conducted on exposed tissues in live animals to examine the emission collection enhancement of the c-TED device compared to heavily optimized objective-based emission collection. The best light collection enhancement was seen from murine fat (5×-2× gains as a function of depth), while murine skeletal muscle and rat kidney showed gains of over two and just under two-fold near the surface, respectively. Gains decreased with imaging depth (particularly in the kidney). Zebrafish imaging on a reflective substrate showed close to a two-fold gain throughout the entire volume of an intact embryo (approximately 150 μm deep). Direct measurement of bleaching rates confirmed that the lower laser powers (enabled by greater light collection efficiency) yielded reduced photobleaching in vivo. The potential benefits of increased light collection in terms of speed of imaging and reduced photo-damage, as well as the applicability of this device to other multi-photon imaging methods is discussed. PMID:24251437

  2. Carcinogenic damage to deoxyribonucleic acid is induced by near-infrared laser pulses in multiphoton microscopy via combination of two- and three-photon absorption

    NASA Astrophysics Data System (ADS)

    Nadiarnykh, Oleg; Thomas, Giju; Van Voskuilen, Johan; Sterenborg, Henricus J. C. M.; Gerritsen, Hans C.

    2012-11-01

    Nonlinear optical imaging modalities (multiphoton excited fluorescence, second and third harmonic generation) applied in vivo are increasingly promising for clinical diagnostics and the monitoring of cancer and other disorders, as they can probe tissue with high diffraction-limited resolution at near-infrared (IR) wavelengths. However, high peak intensity of femtosecond laser pulses required for two-photon processes causes formation of cyclobutane-pyrimidine-dimers (CPDs) in cellular deoxyribonucleic acid (DNA) similar to damage from exposure to solar ultraviolet (UV) light. Inaccurate repair of subsequent mutations increases the risk of carcinogenesis. In this study, we investigate CPD damage that results in Chinese hamster ovary cells in vitro from imaging them with two-photon excited autofluorescence. The CPD levels are quantified by immunofluorescent staining. We further evaluate the extent of CPD damage with respect to varied wavelength, pulse width at focal plane, and pixel dwell time as compared with more pronounced damage from UV sources. While CPD damage has been expected to result from three-photon absorption, our results reveal that CPDs are induced by competing two- and three-photon absorption processes, where the former accesses UVA absorption band. This finding is independently confirmed by nonlinear dependencies of damage on laser power, wavelength, and pulse width.

  3. Current developments in clinical multiphoton tomography

    NASA Astrophysics Data System (ADS)

    Knig, Karsten; Weinigel, Martin; Breunig, Hans Georg; Gregory, Axel; Fischer, Peter; Kellner-Hfer, Marcel; Bckle, Rainer

    2010-02-01

    Two-photon microscopy has been introduced in 1990 [1]. 13 years later, CE-marked clinical multiphoton systems for 3D imaging of human skin with subcellular resolution have been launched by the JenLab company with the tomograph DermaInspectTM. In 2010, the second generation of clinical multiphoton tomographs was introduced. The novel mobile multiphoton tomograph MPTflexTM, equipped with a flexible articulated optical arm, provides an increased flexibility and accessibility especially for clinical and cosmetical examinations. The multiphoton excitation of fluorescent biomolecules like NAD(P)H, flavins, porphyrins, elastin, and melanin as well as the second harmonic generation of collagen is induced by picojoule femtosecond laser pulses from an tunable turn-key near infrared laser system. The ability for rapid highquality image acquisition, the user-friendly operation of the system, and the compact and flexible design qualifies this system to be used for melanoma detection, diagnostics of dermatological disorders, cosmetic research, and skin aging measurements as well as in situ drug monitoring and animal research. So far, more than 1,000 patients and volunteers have been investigated with the multiphoton tomographs in Europe, Asia, and Australia.

  4. Multi-photon ionization and fragmentation of uracil: Neutral excited-state ring opening and hydration effects

    SciTech Connect

    Barc, B.; Ryszka, M.; Spurrell, J.; Dampc, M.; Limão-Vieira, P.; Parajuli, R.; Mason, N. J.; Eden, S.

    2013-12-28

    Multi-photon ionization (MPI) of the RNA base uracil has been studied in the wavelength range 220–270 nm, coinciding with excitation to the S{sub 2}(ππ*) state. A fragment ion at m/z = 84 was produced by 2-photon absorption at wavelengths ≤232 nm and assigned to C{sub 3}H{sub 4}N{sub 2}O{sup +} following CO abstraction. This ion has not been observed in alternative dissociative ionization processes (notably electron impact) and its threshold is close to recent calculations of the minimum activation energy for a ring opening conical intersection to a σ(n-π)π* closed shell state. Moreover, the predicted ring opening transition leaves a CO group at one end of the isomer, apparently vulnerable to abstraction. An MPI mass spectrum of uracil-water clusters is presented for the first time and compared with an equivalent dry measurement. Hydration enhances certain fragment ion pathways (particularly C{sub 3}H{sub 3}NO{sup +}) but represses C{sub 3}H{sub 4}N{sub 2}O{sup +} production. This indicates that hydrogen bonding to water stabilizes uracil with respect to neutral excited-state ring opening.

  5. Multi-photon ionization and fragmentation of uracil: Neutral excited-state ring opening and hydration effects

    NASA Astrophysics Data System (ADS)

    Barc, B.; Ryszka, M.; Spurrell, J.; Dampc, M.; Limo-Vieira, P.; Parajuli, R.; Mason, N. J.; Eden, S.

    2013-12-01

    Multi-photon ionization (MPI) of the RNA base uracil has been studied in the wavelength range 220-270 nm, coinciding with excitation to the S2(??*) state. A fragment ion at m/z = 84 was produced by 2-photon absorption at wavelengths ?232 nm and assigned to C3H4N2O+ following CO abstraction. This ion has not been observed in alternative dissociative ionization processes (notably electron impact) and its threshold is close to recent calculations of the minimum activation energy for a ring opening conical intersection to a ?(n-?)?* closed shell state. Moreover, the predicted ring opening transition leaves a CO group at one end of the isomer, apparently vulnerable to abstraction. An MPI mass spectrum of uracil-water clusters is presented for the first time and compared with an equivalent dry measurement. Hydration enhances certain fragment ion pathways (particularly C3H3NO+) but represses C3H4N2O+ production. This indicates that hydrogen bonding to water stabilizes uracil with respect to neutral excited-state ring opening.

  6. Multi-photon ionization and fragmentation of uracil: neutral excited-state ring opening and hydration effects.

    PubMed

    Barc, B; Ryszka, M; Spurrell, J; Dampc, M; Limão-Vieira, P; Parajuli, R; Mason, N J; Eden, S

    2013-12-28

    Multi-photon ionization (MPI) of the RNA base uracil has been studied in the wavelength range 220-270 nm, coinciding with excitation to the S2(ππ*) state. A fragment ion at m/z = 84 was produced by 2-photon absorption at wavelengths ≤232 nm and assigned to C3H4N2O(+) following CO abstraction. This ion has not been observed in alternative dissociative ionization processes (notably electron impact) and its threshold is close to recent calculations of the minimum activation energy for a ring opening conical intersection to a σ(n-π)π* closed shell state. Moreover, the predicted ring opening transition leaves a CO group at one end of the isomer, apparently vulnerable to abstraction. An MPI mass spectrum of uracil-water clusters is presented for the first time and compared with an equivalent dry measurement. Hydration enhances certain fragment ion pathways (particularly C3H3NO(+)) but represses C3H4N2O(+) production. This indicates that hydrogen bonding to water stabilizes uracil with respect to neutral excited-state ring opening. PMID:24387374

  7. Laser-driven electron dynamics for circular dichroism in mass spectrometry: from one-photon excitations to multiphoton ionization.

    PubMed

    Krner, Dominik

    2015-07-15

    The distinction of enantiomers is a key aspect of chemical analysis. In mass spectrometry the distinction of enantiomers has been achieved by ionizing the sample with circularly polarized laser pulses and comparing the ion yields for light of opposite handedness. While resonant excitation conditions are expected to be most efficient, they are not required for the detection of a circular dichroism (CD) in the ion yield. However, the prediction of the size and sign of the circular dichroism becomes challenging if non-resonant multiphoton excitations are used to ionize the sample. Employing femtosecond laser pulses to drive electron wavepacket dynamics based on ab initio calculations, we attempt to reveal underlying mechanisms that determine the CD under non-resonant excitation conditions. Simulations were done for (R)-1,2-propylene oxide, using time-dependent configuration interaction singles with perturbative doubles (TD-CIS(D)) and the aug-cc-pVTZ basis set. Interactions between the electric field and the electric dipole and quadrupole as well as between the magnetic field and the magnetic dipole were explicitly accounted for. The ion yield was determined by treating states above the ionization potential as either stationary or non-stationary with energy-dependent lifetimes based on an approved heuristic approach. The observed population dynamics do not allow for a simple interpretation, because of highly non-linear interactions. Still, the various transition pathways are governed by resonant enantiospecific n-photon excitation, with preferably high transition dipole moments, which eventually dominate the CD in the ionized population. PMID:26151731

  8. Multi-photon excited coherent random laser emission in ZnO powders

    NASA Astrophysics Data System (ADS)

    Tolentino Dominguez, Christian; Gomes, Maria De A.; Macedo, Zélia S.; de Araújo, Cid B.; Gomes, Anderson S. L.

    2014-11-01

    We report the observation and analysis of anti-Stokes coherent random laser (RL) emission from zinc oxide (ZnO) powders excited by one-, two- or three-photon femtosecond laser radiation. The ZnO powders were produced via a novel proteic sol-gel, low-cost and environmentally friendly route using coconut water in the polymerization step of the metal precursor. One- and two-photon excitation at 354 nm and 710 nm, respectively, generated single-band emissions centred at about 387 nm. For three-photon excitation, the emission spectra showed a strong ultraviolet (UV) band (380-396 nm) attributed to direct three-photon absorption from the valence band to the conduction band. The presence of an intensity threshold and a bandwidth narrowing of the UV band from about 20 to 4 nm are clear evidence of RL action. The observation of multiple sub-nanometre narrow peaks in the emission spectra for excitation above the RL threshold is consistent with random lasing by coherent feedback.

  9. Infrared multiphoton dissociation in two-channel systems. Pulse-duration effects

    SciTech Connect

    Brenner, D.M.

    1982-01-07

    Both ethyl vinyl ether (EVE) and dihydrofuran (DHF) dissociate by two reaction channels which differ by approx.25 kcal mol/sup -1/ in activation energies. In DHF, the branching ratio between these channels produced by IR-multiphoton-induced dissociation shows no power dependence, by contrast to previous work with EVE. In view of the similarity of the Arrhenius activation parameters in the two systems, this is surprising. Therefore, a comparison of the peak-power dependence of the branching ratio in the two systems has been studied at h..nu..CO/sub 2/ = 1076 cm/sup -1/ where the average energy absorbed is the same in both molecules. Results on the power dependence of branching ratio and yield suggest that differences in absorption coefficients (pumping rates) in the quasicontinuum may account for the differences in power dependence.

  10. Precise fluorophore lifetime mapping in live-cell, multi-photon excitation microscopy

    PubMed Central

    Chang, Ching-Wei; Mycek, Mary-Ann

    2010-01-01

    Fluorophore excited state lifetime is a useful indicator of micro-environment in cellular optical molecular imaging. For quantitative sensing, precise lifetime determination is important, yet is often difficult to accomplish when using the experimental conditions favored by live cells. Here we report the first application of temporal optimization and spatial denoising methods to two-photon time-correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) to improve lifetime precision in live-cell images. The results demonstrated a greater than five-fold improvement in lifetime precision. This approach minimizes the adverse effects of excitation light on live cells and should benefit FLIM applications to high content analysis and bioimage informatics. PMID:20588712

  11. Dynamics of Cell Migration for cells embedded in Collagen using a multimodal platform of Optical Coherence Tomography, Multi-Photon excitation and Second Harmonic Generation

    NASA Astrophysics Data System (ADS)

    Tanner, Kandice; Tang, Shuo; Gratton, Enrico

    2007-03-01

    We developed Raster Image Correlation Spectroscopy (RICS) to analyze the dynamics of cell migration from data obtained on a confocal multi-photon microscope. We assembled a microscope that can simultaneously measure the scattering signal from optical coherence tomography (OCT), multi-photon excited emission (TPEF) and second harmonic signals (SHG) with comparable spatial resolution and the same time resolution. We present data here showing the combined 3-D images of the cells embedded in a collagen matrix. The OCT signal adds fine structural information of the cellular morphology and collagen which is enhanced by the SHG image. The RICS analysis of the TPEF signal gives the dynamics of the GFP --style proteins. We show that the cell morphology and the distribution of cell organelles are different in the collagen matrix than what is observed in cells growing on flat surfaces. Using the three modalities of cell imaging we could reach a more realistic interpretation of cell dynamics in tissue.

  12. Wavelength dependence of nanosecond infrared laser-induced breakdown in water: Evidence for multiphoton initiation via an intermediate state

    NASA Astrophysics Data System (ADS)

    Linz, Norbert; Freidank, Sebastian; Liang, Xiao-Xuan; Vogelmann, Hannes; Trickl, Thomas; Vogel, Alfred

    2015-04-01

    Investigation of the wavelength dependence (725-1025 nm) of the threshold for nanosecond optical breakdown in water revealed steps consistent with breakdown initiation by multiphoton ionization, with an initiation energy of about 6.6 eV. This value is considerably smaller than the autoionization threshold of about 9.5 eV, which can be regarded as band gap relevant for avalanche ionization. Breakdown initiation is likely to occur via excitation of a valence band electron into a solvated state, followed by rapid excitation into the conduction band. Theoretical analysis based on these assumptions suggests that the seed electron density required for initiating avalanche ionization amounts to 2.5 ×1015c m-3 at 725 nm and drops to 1.1 ×1012c m-3 at 1025 nm. These results demand changes of future breakdown modeling for water, including the use of a larger band gap than previously employed, the introduction of an intermediate energy level for initiation, and consideration of the wavelength dependence of seed electron density.

  13. Multiphoton interactions in the strong field regime: Microwave excitation and ionization of Rydberg eigenstates

    NASA Astrophysics Data System (ADS)

    Griffith, Warren Morton

    We present here an experimental study and theoretical exposition of two-level atoms in the strong field regime, i.e. a regime where we must resort to quantum mechanical techniques other than perturbation theory to properly describe the interactions that take place, as well as an experimental investigation of the many-level problem of microwave ionization. All involve the interaction of Rydberg atoms, atoms with one highly excited electron, with short pulses of radio-frequency or microwave radiation. We also show the relationship between the single cycle time propagator (SCTP), which is simply a time integral of the Schrodinger equation over one cycle of a periodic field, and Floquet theory. We then go on to apply the SCTP to calculate Rabi frequencies, show the dramatic effect of phase in a pulse, and describe the effects of frequency modulation of a strong field.

  14. Differentiation and Distributions of DNA/Cisplatin Crosslinks by Liquid Chromatography-Electrospray Ionization-Infrared Multiphoton Dissociation Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Xu, Zhe; Brodbelt, Jennifer S.

    2014-01-01

    Liquid chromatography-electrospray ionization-infrared multiphoton dissociation (IRMPD) mass spectrometry was developed to investigate the distributions of intrastrand crosslinks formed between cisplatin and two oligodeoxynucleotides (ODNs), d(A1T2G3G4G5T6A7C8C9C10A11T12) (G3-D) and its analog d(A1T2G3G4G5T6T7C8C9C10A11T12) (G3-H), which have been reported to adopt different secondary structures in solution. Based on the formation of site-specific fragment ions upon IRMPD, two isobaric crosslink products were differentiated for each ODN. The preferential formation of G3G4 and G4G5 crosslinks was determined as a function of reaction conditions, including incubation temperature and presence of metal ions. G3-D consistently exhibited a greater preference for formation of the G4G5 crosslink compared with the G3-H ODN. The ratio of G3G4:G4G5 crosslinks increased for both G3-D and G3-H at higher incubation temperatures or when metal salts were added. Comparison of the IRMPD fragmentation patterns of the unmodified ODNs and the intramolecular platinated crosslinks indicated that backbone cleavage was significantly suppressed near the crosslink.

  15. VIBRATIONALLY EXCITED HCN IN THE LUMINOUS INFRARED GALAXY NGC 4418

    SciTech Connect

    Sakamoto, Kazushi; Aalto, Susanne; Evans, Aaron S.; Wiedner, Martina C.; Wilner, David J.

    2010-12-20

    Infrared pumping and its effect on the excitation of HCN molecules can be important when using rotational lines of HCN to probe dense molecular gas in galaxy nuclei. We report the first extragalactic detection of (sub)millimeter rotational lines of vibrationally excited HCN, in the dust-enshrouded nucleus of the luminous infrared galaxy NGC 4418. We estimate the excitation temperature of T{sub vib} {approx} 230 K between the vibrational ground and excited (v{sub 2} = 1) states. This excitation is most likely due to infrared radiation. At this high vibrational temperature the path through the v{sub 2} = 1 state must have a strong impact on the rotational excitation in the vibrational ground level, although it may not be dominant for all rotational levels. Our observations also revealed nearly confusion-limited lines of CO, HCN, HCO{sup +}, H{sup 13}CN, HC{sup 15}N, CS, N{sub 2}H{sup +}, and HC{sub 3}N at {lambda} {approx} 1 mm. Their relative intensities may also be affected by the infrared pumping.

  16. Infrared light excites cells by changing their electrical capacitance

    PubMed Central

    Shapiro, Mikhail G.; Homma, Kazuaki; Villarreal, Sebastian; Richter, Claus-Peter; Bezanilla, Francisco

    2012-01-01

    Optical stimulation has enabled important advances in the study of brain function and other biological processes, and holds promise for medical applications ranging from hearing restoration to cardiac pace making. In particular, pulsed laser stimulation using infrared wavelengths >1.5 μm has therapeutic potential based on its ability to directly stimulate nerves and muscles without any genetic or chemical pre-treatment. However, the mechanism of infrared stimulation has been a mystery, hindering its path to the clinic. Here we show that infrared light excites cells through a novel, highly general electrostatic mechanism. Infrared pulses are absorbed by water, producing a rapid local increase in temperature. This heating reversibly alters the electrical capacitance of the plasma membrane, depolarizing the target cell. This mechanism is fully reversible and requires only the most basic properties of cell membranes. Our findings underscore the generality of pulsed infrared stimulation and its medical potential. PMID:22415827

  17. Infrared imaging results of an excited planar jet

    NASA Astrophysics Data System (ADS)

    Farrington, R. B.

    1991-12-01

    Planar jets are used for many applications including heating, cooling, and ventilation. Generally such a jet is designed to provide good mixing within an enclosure. In building applications, the jet provides both thermal comfort and adequate indoor air quality. Increased mixing rates may lead to lower short-circuiting of conditioned air, elimination of dead zones within the occupied zone, reduced energy costs, increased occupant comfort, and higher indoor air quality. This paper discusses using an infrared imaging system to show the effect of excitation of a jet on the spread angle and on the jet mixing efficiency. Infrared imaging captures a large number of data points in real time (over 50,000 data points per image) providing significant advantages over single-point measurements. We used a screen mesh with a time constant of approximately 0.3 seconds as a target for the infrared camera to detect temperature variations in the jet. The infrared images show increased jet spread due to excitation of the jet. Digital data reduction and analysis show change in jet isotherms and quantify the increased mixing caused by excitation.

  18. Infrared imaging results of an excited planar jet

    SciTech Connect

    Farrington, R.B.

    1991-12-01

    Planar jets are used for many applications including heating, cooling, and ventilation. Generally such a jet is designed to provide good mixing within an enclosure. In building applications, the jet provides both thermal comfort and adequate indoor air quality. Increased mixing rates may lead to lower short-circuiting of conditioned air, elimination of dead zones within the occupied zone, reduced energy costs, increased occupant comfort, and higher indoor air quality. This paper discusses using an infrared imaging system to show the effect of excitation of a jet on the spread angle and on the jet mixing efficiency. Infrared imaging captures a large number of data points in real time (over 50,000 data points per image) providing significant advantages over single-point measurements. We used a screen mesh with a time constant of approximately 0.3 seconds as a target for the infrared camera to detect temperature variations in the jet. The infrared images show increased jet spread due to excitation of the jet. Digital data reduction and analysis show change in jet isotherms and quantify the increased mixing caused by excitation. 17 refs., 20 figs.

  19. Near-Infrared Emitting Squaraine Dyes with High 2PA Cross Sections for Multiphoton Fluorescence Imaging

    PubMed Central

    Ahn, Hyo-Yang; Yao, Sheng; Wang, Xuhua; Belfield, Kevin D.

    2012-01-01

    Designed to achieve high two-photon absorptivity, new near infrared (NIR) emitting squaraine dyes, (E)-2-(1-(2-(2-methoxyethoxy)ethyl)-5-(3,4,5-trimethoxystyryl)-1H-pyrrol-2-yl)-4-(1-(2-(2-methoxyethoxy)ethyl)-5-(3,4,5-trimethoxystyryl)-2H-pyrrolium-2-ylidene)-3-oxocyclobut-1-enolate (1) and (Z)-2-(4-(dibutylamino)-2-hydroxyphenyl)-4-(4-(dibutyliminio)-2-hydroxycyclohexa-2,5-dienylidene)-3-oxocyclobut-1-enolate (2) were synthesized and characterized. Their linear photophysical properties were investigated via UV-visible absorption spectroscopy and fluorescence spectroscopy in various solvents, while their nonlinear photophysical properties were investigated using a combination of two-photon induced fluorescence and open aperture z-scan methods. Squaraine 1 exhibited a high two-photon absorption (2PA) cross section (δ2PA), ~ 20,000 GM at 800 nm, and high photostability with the photochemical decomposition quantum yield one order of magnitude lower than Cy 5, a commercially available pentamethine cyanine NIR dye. The cytotoxicity of the squaraine dyes were evaluated in HCT 116 and COS 7 cell lines to assess the potential of these probes for biomedical imaging. The viability of both cell lines was maintained above 80% at dye concentrations up to 30 μM, indicating good biocompatibility of the probes. Finally, one-photon fluorescence microscopy (1PFM) and two-photon fluorescence microscopy (2PFM) imaging was accomplished after incubation of micelle-encapsulated squaraine probes with HCT 116 and COS 7 cells, demonstrating their potential in 2PFM bioimaging. PMID:22591003

  20. Detecting cracks in teeth using ultrasonic excitation and infrared imaging

    NASA Astrophysics Data System (ADS)

    Han, Xiaoyan; Favro, Lawrence D.; Thomas, Robert L.

    2001-06-01

    We describe a new technique, Thermosonics, that can be used to detect cracks in teeth. This technique was initially invented and developed for finding cracks in industrial and aerospace applications. The thermosonics technique employs a single short pulse (typically tens of milliseconds) of ultrasound excitation combined with infrared imaging. Ultrasonic waves vibrate the target material. This vibration causes rubbing and clapping between faying surfaces of any cracks which are present, resulting in a temperature rise around the cracks. An infrared camera is used to image the temperature distribution during and after the ultrasound excitation. Thus, cracks in teeth can be detected. Although this technique is still under development, it shows promise for clinical use by dentists.

  1. TEA CO[sub 2] laser-induced infrared multiphoton dissociation of zirconium tetra-tert-butoxide; Reaction mechanism and dissociation products

    SciTech Connect

    Okada, Y.; Kato, S.; Satooka, S.; Takeuchi, K. )

    1992-01-01

    Experiments of laser irradiation were performed to investigate the reaction mechanism of TEA CO[sub 2] laser-induced dissociation of zirconium tetra-tert-butoxide (Zr(t-OC[sub 4]H[sub 9])[sub 4]). The chemical structure of the dissociation product was analyzed by fast atom bombardment mass spectrometry (FAB-MS), infrared absorption spectroscopy, Raman spectroscopy, and [sup 1]H and [sup 13]C nuclear magnetic resonance (NMR) measurements. In this paper, from the results, the authors surmise a reaction mechanism of the infrared multiphoton dissociation of Zr(t-OC[sub 4]H[sub 9])[sub 4]. The dissociation product is hydrolyzed in moist air to zirconium hydroxide (Zr(OH)[sub 4]).

  2. Multifocal multiphoton microscopy with adaptive optical correction

    NASA Astrophysics Data System (ADS)

    Coelho, Simao; Poland, Simon; Krstajic, Nikola; Li, David; Monypenny, James; Walker, Richard; Tyndall, David; Ng, Tony; Henderson, Robert; Ameer-Beg, Simon

    2013-02-01

    Fluorescence lifetime imaging microscopy (FLIM) is a well established approach for measuring dynamic signalling events inside living cells, including detection of protein-protein interactions. The improvement in optical penetration of infrared light compared with linear excitation due to Rayleigh scattering and low absorption have provided imaging depths of up to 1mm in brain tissue but significant image degradation occurs as samples distort (aberrate) the infrared excitation beam. Multiphoton time-correlated single photon counting (TCSPC) FLIM is a method for obtaining functional, high resolution images of biological structures. In order to achieve good statistical accuracy TCSPC typically requires long acquisition times. We report the development of a multifocal multiphoton microscope (MMM), titled MegaFLI. Beam parallelization performed via a 3D Gerchberg-Saxton (GS) algorithm using a Spatial Light Modulator (SLM), increases TCSPC count rate proportional to the number of beamlets produced. A weighted 3D GS algorithm is employed to improve homogeneity. An added benefit is the implementation of flexible and adaptive optical correction. Adaptive optics performed by means of Zernike polynomials are used to correct for system induced aberrations. Here we present results with significant improvement in throughput obtained using a novel complementary metal-oxide-semiconductor (CMOS) 1024 pixel single-photon avalanche diode (SPAD) array, opening the way to truly high-throughput FLIM.

  3. Optical visualization of Alzheimer’s pathology via multiphoton-excited intrinsic fluorescence and second harmonic generation

    PubMed Central

    Kwan, Alex C.; Duff, Karen; Gouras, Gunnar K.; Webb, Watt W.

    2010-01-01

    Intrinsic optical emissions, such as autofluorescence and second harmonic generation (SHG), are potentially useful for functional fluorescence imaging and biomedical disease diagnosis for neurodegenerative diseases such as Alzheimer’s disease (AD). Here, using multiphoton and SHG microscopy, we identified sources of intrinsic emissions in ex vivo, acute brain slices from AD transgenic mouse models. We observed autofluorescence and SHG at senile plaques as well as characterized their emission spectra. The utility of intrinsic emissions was demonstrated by imaging senile plaque autofluorescence in conjunction with SHG from microtubule arrays to assess the polarity of microtubules near pathological lesions. Our results suggest that tissues from AD transgenic models contain distinct intrinsic emissions, which can provide valuable information about the disease mechanisms. PMID:19259208

  4. Formation of infrared solitons in graphene ensemble under Raman excitation

    NASA Astrophysics Data System (ADS)

    Ding, Chunling; Yu, Rong; Yang, Xiaoxue; Zhang, Duo; Huang, Mingju

    2015-11-01

    The formation of infrared solitons in graphene under Raman excitation is investigated using density-matrix approach. We find that the unique band structure and selection rules for the optical transitions near the Dirac point can result in extremely strong optical nonlinearity. Theoretical investigations with the aid of slowly varying envelope approximation and perturbation theory clearly indicate the existence of bright and dark solitons in Landau-quantized graphene. Actually, the formation of spatial soliton in such a material is the consequence of the balance between nonlinear effects and the dispersion properties. Also, the corresponding carrier frequency is tunable in the infrared range. These results can make us know better the crossover between optical solitons and graphene metamaterials. The predicted nonlinear optical effect in graphene may provide a new possibility for designing high-fidelity graphene-based information processing device.

  5. Studies of protonated and anionic artemisinin in the gas-phase by infrared multi-photon dissociation and by negative ion photoelectron spectroscopies

    NASA Astrophysics Data System (ADS)

    Seydou, M.; Gillet, J. C.; Li, X.; Wang, H.; Posner, G. H.; Grgoire, G.; Schermann, J. P.; Bowen, K. H.; Desfranois, C.

    2007-12-01

    Protonated and anionic artemisinin in the gas phase have respectively been studied by infrared multi-photon dissociation (IRMPD) spectroscopy and by anion photoelectron spectroscopy. Comparison of the measured IRMPD spectrum with calculated spectra of various conformations showed that the two lowest-energy protonated structures, both corresponding to protonation at the C dbnd O 14 carbonyl site, were observed experimentally. The calculations also indicated that the peroxide bridge in artemisinin is only slightly modified by protonation. Additionally, stable, intact (parent) artemisinin radical anions have been obtained for the first time in the gas phase and the photoelectron spectrum supports the computational finding that the excess electron is mainly localized on the ? ? orbital of the peroxide bond. The vertical detachment energy and adiabatic electron affinity, calculated at the MP2/6-31+G ? level, are in good agreement with the experimental data and the O-O distance is calculated to be stretched by more than 50% in the anion.

  6. Infrared multiphoton induced isomerization and dissociation of FCN, ClCN, and BrCN in liquid Ar: A classical simulation study

    SciTech Connect

    Zhang Ming; Gong Jiangbin; Ma Ao; Rice, Stuart A.

    2007-10-14

    We report the results of classical mechanics simulations of infrared multiphoton induced control of isomerization of FCN, ClCN, and BrCN in liquid Ar, using ab initio potential energy and dipole moment surfaces for the XCN molecules. The field induced isomerization and fragmentation dynamics of these molecules are found to be different from that of HCN in liquid Ar. In particular, the scheme that provides complete controlled conversion of HCN to CNH in liquid Ar fails to generate complete conversion of XCN to CNX in liquid Ar for X=F,Cl,Br. It is suggested that the sources of the differences in behavior arise from differences in the spectra of vibrational nonlinear resonances in HCN and XCN and to the occurrence of monodromy in the dynamics of the XCN molecules.

  7. Multiphoton multicolor FISH

    NASA Astrophysics Data System (ADS)

    Koenig, Karsten; Riemann, Iris; Goehlert, Axel; Fischer, Peter; Liehr, Thomas; Loncarevic, Ivan F.; Claussen, Uwe; Halbhuber, Karl-Juergen

    2000-12-01

    We describe a novel method of 3D imaging of specific regions of DNA in interphase nuclei and tissues based on multiphoton microscopy and multicolor fluorescence in situ hybridization (M-FISH). Multiphoton Multicolor FISH (MM-FISH) combines the advantages of (i) using a single NIR excitation wavelength for the simultaneous excitation of multiple FISH fluorophores, (ii) absence of fading in out-of-focus regions, (iii) intrinsic 3D imaging capability and (iv) high light penetration depth. Detection of chromosomal aberrations in amniocytes and tumor cells as well as imaging of FISH fluorophores in biopsies using femtosecond laser pulses at 780 nm and 800 nm are described. First two-photon excited fluorescence decay curves of FISH fluorophores are presented. The fluorophores have been excited via non- resonant two-photon excitation with 150 fs pulses of 0.1 to 8 mW mean laser power of a frequency doubled ultra compact 50 MHz fiber laser and with 80 fs pulses of a compact 80 MHz Ti:sapphire laser. MM-FISH may become an interesting tool in preimplantation diagnosis and molecular pathology.

  8. Clinical multiphoton FLIM tomography

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2012-03-01

    This paper gives an overview on current clinical high resolution multiphoton fluorescence lifetime imaging in volunteers and patients. Fluorescence lifetime imaging (FLIM) in Life Sciences was introduced in Jena/Germany in 1988/89 based on a ZEISS confocal picosecond dye laser scanning microscope equipped with a single photon counting unit. The porphyrin distribution in living cells and living tumor-bearing mice was studied with high spatial, temporal, and spectral resolution. Ten years later, time-gated cameras were employed to detect dental caries in volunteers based on one-photon excitation of autofluorescent bacteria with long fluorescence lifetimes. Nowadays, one-photon FLIM based on picosecond VIS laser diodes are used to study ocular diseases in humans. Already one decade ago, first clinical twophoton FLIM images in humans were taken with the certified clinical multiphoton femtosecond laser tomograph DermaInspectTM. Multiphoton tomographs with FLIM modules are now operating in hospitals at Brisbane, Tokyo, Berlin, Paris, London, Modena and other European cities. Multiple FLIM detectors allow spectral FLIM with a temporal resolution down to 20 ps (MCP) / 250 ps (PMT) and a spectral resolution of 10 nm. Major FLIM applications include the detection of intradermal sunscreen and tattoo nanoparticles, the detection of different melanin types, the early diagnosis of dermatitis and malignant melanoma, as well as the measurement of therapeutic effects in pateints suffering from dermatitis. So far, more than 1,000 patients and volunteers have been investigated with the clinical multiphoton FLIM tomographs DermaInspectTM and MPTflexTM.

  9. Multiphoton microspectroscopy of biological specimens

    NASA Astrophysics Data System (ADS)

    Lin, Bai-Ling; Kao, Fu-Jen; Cheng, Ping C.; Sun, Chi-Kuang; Chen, RangWu; Wang, YiMin; Chen, JianCheng; Wang, Yung-Shun; Liu, Tzu-Ming; Huang, Mao-Kuo

    2000-07-01

    The non-linear nature of multi-photon fluorescence excitation restricts the fluorescing volume to the vicinity of the focal point. As a result, the technology has the capacity for micro- spectroscopy of biological specimen at high spatial resolution. Chloroplasts in mesophyll protoplast of Arabidopsis thaliana and maize stem sections were used to demonstrate the feasibility of multi-photon fluorescence micro-spectroscopy at subcellular compartments. Time-lapse spectral recording provides a means for studying the response of cell organelles to high intensity illumination.

  10. Quantum dot infrared photodetector enhanced by surface plasma wave excitation.

    PubMed

    Lee, S C; Krishna, S; Brueck, S R J

    2009-12-01

    Up to a thirty-fold detectivity enhancement is achieved for an InAs quantum dot infrared photodetector (QDIP) by the excitation of surface plasma waves (SPWs) using a metal photonic crystal (MPC) integrated on top of the detector absorption region. The MPC is a 100 nm-thick gold film perforated with a 3.6 microm period square array of circular holes. A bare QDIP shows a bias-tunable broadband response from approximately 6 to 10 microm associated with the quantum confined Stark (QCS) effect. On the other hand, an MPC-integrated QDIP exhibits a dominant peak at 11.3 microm with a approximately 1 microm full width at half maximum and the highly enhanced detectivity at the bias polarity optimized for long wavelength. This is very different from the photoresponse of the bare QDIP but fully consistent with the direct coupling of the QDs in the detector absorption region to the SPWs excited at the MPC/detector interface by incident photons. The SPW resonance wavelength, lambda, for the smallest coupling wavevector of the array in the MPC is close to 11.3 microm. The response also shows other SPW-coupled peaks: a significant peak at 8.1 microm (approximately lambda/radical2) and noticeable peaks at 5.8 microm (approximately lambda/2) and 5.4 microm (approximately lambda/ radical5) which correspond to higher-order coupling wavevectors. For the opposite bias, the MPC-integrated QDIP shows the highest response at 8.1 microm, providing a dramatic voltage tunability that is associated with QCS effect. SPWs propagate with TM (x, z) polarization along the MPC/detector interface. The enhanced detectivity is explained by these characteristics which increase both the effective absorption cross section with propagation and the interaction strength with TM polarization in the coupling to the QDs. Simulations show good qualitative agreement with the observed spectral behavior. PMID:20052244

  11. Multiphoton fluorescence microspectroscopy

    NASA Astrophysics Data System (ADS)

    Kao, Fu-Jen; Lin, Bai-Ling; Cheng, Ping C.

    2000-05-01

    The intrinsic confined photo-interacting volume in multi- photon fluorescence microscopy provides the possibility of obtaining fluorescence spectrum from specific cellular structure in a tissue. In this article, we demonstrated that it is feasible to obtain useful two-photon pumped fluorescence spectrum from cell wall and single chloroplast. The difference in fluorescence spectra obtained with single- and two-photon excitation indicates that a significant shift in fluorescence maximum may occur due to the non-linear nature of excitation. Therefore, in order to properly interpret two-photon fluorescence micrographs, it is important to characterize the fluorescence spectrum of the specimen and the commonly used fluorescence probes. The fluorescence spectra will in turn be useful in the selection of filter sets in multi-photon fluorescence microscopy.

  12. Control of multiphoton and avalanche ionization using an ultraviolet-infrared pulse train in femtosecond laser micro/nano-machining of fused silica

    NASA Astrophysics Data System (ADS)

    Yu, Xiaoming; Bian, Qiumei; Chang, Zenghu; Corkum, P. B.; Lei, Shuting

    2014-03-01

    We report on the experimental results of micro- and nanostructures fabricated on the surface of fused silica by a train of two femtosecond laser pulses, a tightly focused 266 nm (ultraviolet, UV) pulse followed by a loosely focused 800 nm (infrared, IR) pulse. By controlling the fluence of each pulse below the damage threshold, micro- and nanostructures are fabricated using the combined beams. The resulting damage size is defined by the UV pulse, and a reduction of UV damage threshold is observed when the two pulses are within ~ 1 ps delay. The effects of IR pulse duration on the UV damage threshold and shapes are investigated. These results suggest that the UV pulse generates seed electrons through multiphoton absorption and the IR pulse utilizes these electrons to cause damage by avalanche process. A single rate equation model based on electron density can be used to explain these results. It is further demonstrated that structures with dimensions of 124 nm can be fabricated on the surface of fused silica using 0.5 NA objective. This provides a possible route to XUV (or even shorter wavelength) laser nano-machining with reduced damage threshold.

  13. Three-dimensional imaging and uptake of the anticancer drug combretastatin in cell spheroids and photoisomerization in gels with multiphoton excitation

    NASA Astrophysics Data System (ADS)

    Scherer, Kathrin M.; Bisby, Roger H.; Botchway, Stanley W.; Hadfield, John A.; Haycock, John W.; Parker, Anthony W.

    2015-07-01

    The uptake of E-combretastatins, potential prodrugs of the anticancer Z-isomers, into multicellular spheroids has been imaged by intrinsic fluorescence in three dimensions using two-photon excited fluorescence lifetime imaging with 625-nm ultrafast femtosecond laser pulses. Uptake is initially observed at the spheroid periphery but extends to the spheroid core within 30 min. Using agarose gels as a three-dimensional model, the conversion of Z(trans)→E(cis) via two-photon photoisomerization is demonstrated and the location of this photochemical process may be precisely selected within the micron scale in all three dimensions at depths up to almost 2 mm. We discuss these results for enhanced tissue penetration at longer near-infrared wavelengths for cancer therapy and up to three-photon excitation and imaging using 930-nm laser pulses with suitable combretastatin analogs.

  14. Three-dimensional imaging and uptake of the anticancer drug combretastatin in cell spheroids and photoisomerization in gels with multiphoton excitation.

    PubMed

    Scherer, Kathrin M; Bisby, Roger H; Botchway, Stanley W; Hadfield, John A; Haycock, John W; Parker, Anthony W

    2015-07-01

    The uptake of E -combretastatins, potential prodrugs of the anticancer Z -isomers, into multicellular spheroids has been imaged by intrinsic fluorescence in three dimensions using two-photon excited fluorescence lifetime imaging with 625-nm ultrafast femtosecond laser pulses. Uptake is initially observed at the spheroid periphery but extends to the spheroid core within 30 min. Using agarose gels as a three-dimensional model, the conversion of Z(trans)?E(cis) via two-photon photoisomerization is demonstrated and the location of this photochemical process may be precisely selected within the micron scale in all three dimensions at depths up to almost 2 mm. We discuss these results for enhanced tissue penetration at longer near-infrared wavelengths for cancer therapy and up to three-photon excitation and imaging using 930-nm laser pulses with suitable combretastatin analogs. PMID:26146878

  15. Optical biopsy of liver fibrosis by use of multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Lee, Hsuan-Shu; Liu, Yuan; Chen, Hsiao-Ching; Chiou, Ling-Ling; Huang, Guan-Tarn; Lo, Wen; Dong, Chen-Yuan

    2004-11-01

    We demonstrate the application of multiphoton microscopy in diagnosing toxin- CCl4 - induced liver fibrosis in mice. Although hepatocyte autofluorescence does not vary significantly, different degrees of necrosis and stellate cell proliferation at necrotic sites in livers with fibrosis (ex vivo) can be detected easily from multiphoton-induced autofluorescence images by use of 780-nm excitation. Our result suggests that multiphoton microscopy can be developed into an effective technique for the detection and diagnosis of liver fibrosis in vivo.

  16. Time-Resolved Resonance Raman Spectroscopy of Vibrational Populations Monitored after Electronic and Infrared Excitation

    SciTech Connect

    Werncke, W.; Kozich, V.; Dreyer, J.

    2008-11-14

    Pathways of vibrational energy flow in molecules with an intramolecular hydrogen bond are studied after intramolecular proton transfer reactions as well as after infrared excitation of the O-H stretching vibration which is coupled to this hydrogen bond.

  17. Infrared emission from photo-excited gaseous benzene: detection with a new home-made spectrometer

    NASA Astrophysics Data System (ADS)

    Fraud, G.; Carpentier, Y.; Pino, T.; Longval, Y.; Dartois, E.; Chamaill, T.; Vasquez, R.; Vincent, J.; Parneix, P.; Falvo, C.; Brchignac, Ph.

    2012-02-01

    The infrared fluorescence decay and the dispersed emission spectrum are presented for gaseous benzene following 193 nm laser excitation. They were measured with FIREFLY (Fluorescence in the InfraRed from Excited FLYing molecules), a new home-made spectrometer. Redshift and redtail in the CH stretch emission spectra (3.3 ?m region) demonstrate that anharmonicity plays a key role when dealing with high internal energies, as it is the case in the interstellar medium.

  18. Combined infrared multiphoton dissociation and electron capture dissociation with a hollow electron beam in Fourier transform ion cyclotron resonance mass spectrometry.

    PubMed

    Tsybin, Youri O; Witt, Matthias; Baykut, Gkhan; Kjeldsen, Frank; Hkansson, Per

    2003-01-01

    An electron injection system based on an indirectly heated ring-shaped dispenser cathode has been developed and installed in a 7 Tesla Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. This new hardware design allows high-rate electron capture dissociation (ECD) to be carried out by a hollow electron beam coaxial with the ion cyclotron resonance (ICR) trap. Infrared multiphoton dissociation (IRMPD) can also be performed with an on-axis IR-laser beam passing through a hole at the centre of the dispenser cathode. Electron and photon irradiation times of the order of 100 ms are required for efficient ECD and IRMPD, respectively. As ECD and IRMPD generate fragments of different types (mostly c, z and b, y, respectively), complementary structural information that improves the characterization of peptides and proteins by FTICR mass spectrometry can be obtained. The developed technique enables the consecutive or simultaneous use of the ECD and IRMPD methods within a single FTICR experimental sequence and on the same ensemble of trapped ions in multistage tandem (MS/MS/MS or MS(n)) mass spectrometry. Flexible changing between ECD and IRMPD should present advantages for the analysis of protein digests separated by liquid chromatography prior to FTICRMS. Furthermore, ion activation by either electron or laser irradiation prior to, as well as after, dissociation by IRMPD or ECD increases the efficiency of ion fragmentation, including the w-type fragment ion formation, and improves sequencing of peptides with multiple disulfide bridges. The developed instrumental configuration is essential for combined ECD and IRMPD on FTICR mass spectrometers with limited access into the ICR trap. PMID:12872281

  19. Combined electron capture and infrared multiphoton dissociation for multistage MS/MS in a Fourier transform ion cyclotron resonance mass spectrometer.

    PubMed

    Hkansson, Kristina; Chalmers, Michael J; Quinn, John P; McFarland, Melinda A; Hendrickson, Christopher L; Marshall, Alan G

    2003-07-01

    We have mounted a permanent on-axis dispenser cathode electron source inside the magnet bore of a 9.4-T Fourier transform ion cyclotron resonance mass spectrometer. This configuration allows electron capture dissociation (ECD) to be performed reliably on a millisecond time scale. We have also implemented an off-axis laser geometry that enables simultaneous access to ECD and infrared multiphoton dissociation (IRMPD). Optimum performance of both fragmentation techniques is maintained. The analytical utility of performing either ECD or IRMPD on a given precursor ion population is demonstrated by structural characterization of several posttranslationally modified peptides: IRMPD of phosphorylated peptides results in few backbone (b- and y-type) cleavages, and product ion spectra are dominated by neutral loss of H3PO4. In contrast, ECD provides significantly more backbone (c- and z*-type) cleavages without loss of H3PO4. For N-glycosylated tryptic peptides, IRMPD causes extensive cleavage of the glycosidic bonds, providing structural information about the glycans. ECD cleaves all backbone bonds (except the N-terminal side of proline) in a 3-kDa glycopeptide with no saccharide loss. However, only a charge-reduced radical species and some side chain losses are observed following ECD of a 5-kDa glycopeptide from the same protein. An MS3 experiment involving IR laser irradiation of the charge-reduced species formed by electron capture results in extensive dissociation into c- and z-type fragment ions. Mass-selective external ion accumulation is essential for the structural characterization of these low-abundance (modified) peptides. PMID:12964777

  20. Microbeam-integrated multiphoton imaging system

    NASA Astrophysics Data System (ADS)

    Bigelow, Alan W.; Geard, Charles R.; Randers-Pehrson, Gerhard; Brenner, David J.

    2008-12-01

    Multiphoton microscopy has been added to the array of imaging techniques at the endstation for the Microbeam II cell irradiator at Columbia University's Radiological Research Accelerator Facility (RARAF). This three-dimensional (3D), laser-scanning microscope functions through multiphoton excitation, providing an enhanced imaging routine during radiation experiments with tissuelike samples, such as small living animals and organisms. Studies at RARAF focus on radiation effects; hence, this multiphoton microscope was designed to observe postirradiation cellular dynamics. This multiphoton microscope was custom designed into an existing Nikon Eclipse E600-FN research fluorescence microscope on the irradiation platform. Design details and biology applications using this enhanced 3D-imaging technique at RARAF are reviewed.

  1. Calculating singlet excited states: Comparison with fast time-resolved infrared spectroscopy of coumarins

    NASA Astrophysics Data System (ADS)

    Hanson-Heine, Magnus W. D.; Wriglesworth, Alisdair; Uroos, Maliha; Calladine, James A.; Murphy, Thomas S.; Hamilton, Michelle; Clark, Ian P.; Towrie, Michael; Dowden, James; Besley, Nicholas A.; George, Michael W.

    2015-04-01

    In contrast to the ground state, the calculation of the infrared (IR) spectroscopy of molecular singlet excited states represents a substantial challenge. Here, we use the structural IR fingerprint of the singlet excited states of a range of coumarin dyes to assess the accuracy of density functional theory based methods for the calculation of excited state IR spectroscopy. It is shown that excited state Kohn-Sham density functional theory provides a high level of accuracy and represents an alternative approach to time-dependent density functional theory for simulating the IR spectroscopy of singlet excited states.

  2. Multiphoton spectroscopy of Rydberg states of small molecules

    SciTech Connect

    Pratt, Stephen T.; McCormack, E. F.; Dehmer, Joseph L.; Dehmer, Patricia M.

    1990-09-01

    Multiphoton ionization techniques provide a versatile means for studying highly excited states of atoms and molecules and provide a valuable complement to traditional techniques based on single-photon absorption and ionization studies. In this paper we present the results of new multiphoton ionization studies of molecular nitrogen and molecular oxygen that serve to illustrate the power of these techniques. 30 refs., 3 figs.

  3. High multi-photon visible upconversion emissions of Er{sup 3+} singly doped BiOCl microcrystals: A photon avalanche of Er{sup 3+} induced by 980?nm excitation

    SciTech Connect

    Li, Yongjin; Song, Zhiguo Li, Chen; Wan, Ronghua; Qiu, Jianbei; Yang, Zhengwen; Yin, Zhaoyi; Yang, Yong; Zhou, Dacheng; Wang, Qi

    2013-12-02

    Under 980?nm excitation, high multi-photon upconversion (UC) emission from the {sup 2}H{sub 11/2}/{sup 4}S{sub 3/2} (green) and {sup 4}F{sub 9/2} (red) levels of Er{sup 3+} ions were observed from Er{sup 3+} singly doped BiOCl microcrystals. These high-energy excited states were populated by a three to ten photon UC process conditionally, which depended on the pump power density and the Er{sup 3+} ion doping concentration, characterizing as a hetero-looping enhanced energy transfer avalanche UC process. UC emission lifetime and Raman analysis suggest that the unusual UC phenomena are initiated by the new and intense phonon vibration modes of BiOCl lattices due to Er{sup 3+} ions doping.

  4. Quasi white light multiphoton imaging

    NASA Astrophysics Data System (ADS)

    de Mauro, C.; Alfieri, D.; Arrigoni, M.; Armstrong, D.; Pavone, F. S.

    2009-07-01

    We describe the realization and characterization of a broadband, high power density and fully spectrally controllable source, suitable for multiphoton imaging of biological samples. We used a photonic crystal fiber (PCF) with selected dispersive and non-linear properties, in order to generate, when pumped with <140 femtosecond pulses delivered by a tunable Ti:Sa laser (Chameleon Ultra II by Coherent Inc.), a smooth continuum in the 700nm-950nm region, with average power density grater than 2mW/nm. Time distribution of the generated spectrum has been measured with autocorrelation technique. Axial and lateral resolution obtained with a scanning multiphoton system has been determined to be near the theoretical limit. The possibility of two-photon excitation of different dyes in the same sample and high image resolution are demonstrated at tens of microns in depth. Future developments and different applications are also discussed.

  5. Probing the Infrared Quark Mass from Highly Excited Baryons

    SciTech Connect

    Bicudo, P.; Cardoso, M.; Van Cauteren, T.; Llanes-Estrada, Felipe J.

    2009-08-28

    We argue that three-quark excited states naturally group into quartets, split into two parity doublets, and that the mass splittings between these parity partners decrease higher up in the baryon spectrum. This decreasing mass difference can be used to probe the running quark mass in the midinfrared power-law regime. A measurement of masses of high-partial-wave DELTA* resonances should be sufficient to unambiguously establish the approximate degeneracy. We test this concept with the first computation of excited high-j baryon masses in a chirally invariant quark model.

  6. Probing the infrared quark mass from highly excited baryons.

    PubMed

    Bicudo, P; Cardoso, M; Van Cauteren, T; Llanes-Estrada, Felipe J

    2009-08-28

    We argue that three-quark excited states naturally group into quartets, split into two parity doublets, and that the mass splittings between these parity partners decrease higher up in the baryon spectrum. This decreasing mass difference can be used to probe the running quark mass in the midinfrared power-law regime. A measurement of masses of high-partial-wave Delta* resonances should be sufficient to unambiguously establish the approximate degeneracy. We test this concept with the first computation of excited high-j baryon masses in a chirally invariant quark model. PMID:19792789

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

    NASA Technical Reports Server (NTRS)

    Williams, Richard M.; Leone, Stephen R.

    1994-01-01

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

  8. Three-dimensional multiphoton autofluorescence spectral imaging of live tissues

    NASA Astrophysics Data System (ADS)

    Palero, Jonathan A.; de Bruijn, Henritte S.; van der Ploeg van den Heuvel, Anglique; Sterenborg, Henricus J. C. M.; Gerritsen, Hans C.

    2006-04-01

    We combined a homebuilt multiphoton microscope and a prism-CCD based spectrograph to develop a spectral imaging system capable of imaging deep into live tissues. The spectral images originate from the two-photon autofluorescence of the tissue and second harmonic signal from the collagen fibers. A highly penetrating near-infrared light is used to excite the endogenous fluorophores via multiphoton excitation enabling us to produce high quality images deep into the tissue. We were able to produce 100-channel (330 nm to 600 nm) autofluorescence spectral images of live skin tissues in less than 2 minutes for each xy-section. The spectral images rendered in RGB (real) colors showed green hair shafts, blue cells, and purple collagen. Analysis on the optical signal degradation with increasing depth of the collagen second-harmonic signal showed 1) exponential decay behavior of the intensity and 2) linear broadening of the spectrum. This spectral imaging system is a promising tool for both in biological applications and biomedical applications such as optical biopsy.

  9. Multiphoton processes: conference proceedings

    SciTech Connect

    Lambropoulos, P.; Smith, S.J.

    1984-01-01

    The chapters of this volume represent the invited papers delivered at the conference. They are arranged according to thermatic proximity beginning with atoms and continuing with molecules and surfaces. Section headings include multiphoton processes in atoms, field fluctuations and collisions in multiphoton process, and multiphoton processes in molecules and surfaces. Abstracts of individual items from the conference were prepared separately for the data base. (GHT)

  10. Excitation of far-infrared lines of OH and maser pumping efficiency in circumstellar envelopes

    NASA Astrophysics Data System (ADS)

    Thai-Q-Tung; Dinh-v-Trung; Nguyen-Q-Rieu; Bujarrabal, V.; Le Bertre, T.; Gerard, E.

    1998-03-01

    We present the results obtained with a radiative transfer model to investigate the excitation of OH molecules. Fluxes of far-infrared rotational lines of OH are calculated in terms of a radiatively excited envelope and of a radiative transfer model including collisional and radiative excitation. The agreement with the OH far-infrared lines observed with ISO is satisfactory. The 34.6 mu m line is found to be in absorption as observed. We also predict that the 53.3 mu m line should be seen in absorption and also contributes to the maser pumping process. Maser pumping efficiencies are estimated from the model and compared to the values determined from recent OH far-infrared observations with ISO and OH maser data.

  11. Stepwise multi-photon activation fluorescence reveals a new method of melanoma imaging for dermatologists

    NASA Astrophysics Data System (ADS)

    Lai, Zhenhua; Lian, Christine; Ma, Jie; Yu, Jingyi; Gu, Zetong; Rajadhyaksha, Milind; DiMarzio, Charles A.

    2014-02-01

    Previous research has shown that the stepwise multi-photon activated fluorescence (SMPAF) of melanin, activated by a continuous-wave (CW) mode near infrared (NIR) laser, is a low cost and reliable method of detecting melanin. SMPAF images of melanin in a mouse hair and a formalin fixed mouse melanoma were compared with conventional multiphoton fluorescence microscopy (MPFM) images and confocal reflectance microscopy (CRM) images, all of which were acquired at an excitation wavelength of 920 nm, to further prove the effectiveness of SMPAF in detecting melanin. SMPAF images add specificity for melanin detection to MPFM images and CRM images. Melanin SMPAF can be a promising technology to enable melanoma imaging for dermatologists.

  12. Multiphoton microscopy of atheroslcerotic plaques

    NASA Astrophysics Data System (ADS)

    Lilledahl, Magnus B.; de Lange Davies, Catharina; Haugen, Olav A.; Svaasand, Lars O.

    2007-02-01

    Multiphoton microscopy is a techniques that fascilitates three dimensional imaging of intact, unstained tissue. Especially connective tissue has a relatively strong nonlinear optical response and can easily be imaged. Atherosclerosis is a disease where lipids accumulate in the vessel wall and there is a thickening of the intima by growth of a cap of connective tissue. The mechanical strength of this fibrous cap is of clinically importance. If the cap ruptures a thrombosis forms which can block a coronary vessel and therby causing myocardial infarction. Multiphoton microscopy can be used to image the fibrous cap and thereby determine the thickness of the cap and the structure of the connective fibres. This could possibly be developed into a diagnostic and clincal tool to monitor the vulnerability of a plaque and also to better understand the development of a plaque and effects of treatment. We have collected multiphoton microscopy images from atherosclerotic plaque in human aorta, both two photon excited fluorescens and second harmonic generated signal. The feasability of using this technique to determine the state of the plaque is explored.

  13. MULTIPHOTON IMAGING CAN BE USED FOR MICROSCOPIC EXAMINATION OF INTACT HUMAN GASTROINTESTINAL MUCOSA EX VIVO

    PubMed Central

    Rogart, Jason N.; Nagata, Jun; Loeser, Caroline S.; Roorda, Robert D.; Aslanian, Harry; Robert, Marie E.; Zipfel, Warren R.; Nathanson, Michael H.

    2008-01-01

    Background & Aims The ability to observe cellular and subcellular detail during routine endoscopy is a major goal in the development of new endoscopic imaging techniques. Multiphoton microscopy, which relies on nonlinear infared optical processes, has the potential to identify cellular details by excitation of endogenous fluorescent molecules. We examined the feasibility of using multiphoton microscopy to characterize mucosal histology in the human gastrointestinal tract. Methods A multiphoton microscope was used to determine the optimal excitation wavelength for examination of gastrointestinal mucosa. Fresh, unfixed, and unstained biopsy specimens obtained during routine endoscopy in human subjects were then examined by confocal microscopy and multiphoton microscopy. Multiphoton images also were compared to standard H&E images obtained from paired biopsy specimens. A prototype miniaturized multiphoton probe was used to examine intact rat colon. Results Peak multiphoton autofluorescence intensity was detected in mucosa excited at 735 nm. Multiphoton microscopic examination of unstained biopsy specimens revealed improved cellular detail relative to either unstained or stained specimens examined by confocal imaging. Resolution of structures such as epithelial nuclei, goblet cells, and interstitial fibers and cells was comparable to what was obtained using standard H&E histology. Similar findings were observed when using a prototype miniaturized multiphoton probe. Conclusions Multiphoton microscopy can be used to examine gastrointestinal mucosa at the cellular level, without the need for fluorescent dyes. The construction of a multiphoton endomicroscope could therefore provide a practical means of performing virtual biopsies during the course of routine endoscopy, with advantages over currently available endomicroscopy technologies. PMID:18065276

  14. Non-destructive Testing by Infrared Thermography Under Random Excitation and ARMA Analysis

    NASA Astrophysics Data System (ADS)

    Bodnar, J. L.; Nicolas, J. L.; Candor, J. C.; Detalle, V.

    2012-11-01

    Photothermal thermography is a non-destructive testing (NDT) method, which has many applications in the field of control and characterization of thin materials. This technique is usually implemented under CW or flash excitation. Such excitations are not adapted for control of fragile materials or for multi-frequency analysis. To allow these analyses, in this article, the use of a new control mode is proposed: infrared thermography under random excitation and auto regressive moving average analysis. First, the principle of this NDT method is presented. Then, the method is shown to permit detection, with low energy constraints, of detachments situated in mural paintings.

  15. Excitation of Far-Infrared Lines of OH and Maser Pumping Efficiency in Circumstellar Envelopes

    NASA Astrophysics Data System (ADS)

    Thai-Q-Tung; Dinh-v-Trung; Nguyen-Q-Rieu; Bujarrabal, V.; Le Bertre, T.; Gerard, E.

    We present the results obtained with a radiative transfer model to investigate the excitation of OH molecules. Maser pumping efficiencies are estimated from the model and compared to the value calculated using recent OH far-infrared (FIR) observations with ISO and OH maser data.

  16. Pulse shaping multiphoton FRET microscopy

    NASA Astrophysics Data System (ADS)

    Brenner, Meredith H.; Cai, Dawen; Nichols, Sarah R.; Straight, Samuel W.; Hoppe, Adam D.; Swanson, Joel A.; Ogilvie, Jennifer P.

    2012-03-01

    Fluorescence Resonance Energy Transfer (FRET) microscopy is a commonly-used technique to study problems in biophysics that range from uncovering cellular signaling pathways to detecting conformational changes in single biomolecules. Unfortunately, excitation and emission spectral overlap between the fluorophores create challenges in quantitative FRET studies. It has been shown previously that quantitative FRET stoichiometry can be performed by selective excitation of donor and acceptor fluorophores. Extending this approach to two-photon FRET applications is difficult when conventional femtosecond laser sources are used due to their limited bandwidth and slow tuning response time. Extremely broadband titanium:sapphire lasers enable the simultaneous excitation of both donor and acceptor for two-photon FRET, but do so without selectivity. Here we present a novel two-photon FRET microscopy technique that employs pulse-shaping to perform selective excitation of fluorophores in live cells and detect FRET between them. Pulse-shaping via multiphoton intrapulse interference can tailor the excitation pulses to achieve selective excitation. This technique overcomes the limitation of conventional femtosecond lasers to allow rapid switching between selective excitation of the donor and acceptor fluorophores. We apply the method to live cells expressing the fluorescent proteins mCerulean and mCherry, demonstrating selective excitation of fluorophores via pulse-shaping and the detection of twophoton FRET. This work paves the way for two-photon FRET stoichiometry.

  17. In vivo stepwise multi-photon activation fluorescence imaging of melanin in human skin

    NASA Astrophysics Data System (ADS)

    Lai, Zhenhua; Gu, Zetong; Abbas, Saleh; Lowe, Jared; Sierra, Heidy; Rajadhyaksha, Milind; DiMarzio, Charles

    2014-03-01

    The stepwise multi-photon activated fluorescence (SMPAF) of melanin is a low cost and reliable method of detecting melanin because the activation and excitation can be a continuous-wave (CW) mode near infrared (NIR) laser. Our previous work has demonstrated the melanin SMPAF images in sepia melanin, mouse hair, and mouse skin. In this study, we show the feasibility of using SMPAF to detect melanin in vivo. in vivo melanin SMPAF images of normal skin and benign nevus are demonstrated. SMPAF images add specificity for melanin detection than MPFM images and CRM images. Melanin SMPAF is a promising technology to enable early detection of melanoma for dermatologists.

  18. Multiphoton absorption in amyloid protein fibres

    NASA Astrophysics Data System (ADS)

    Hanczyc, Piotr; Samoc, Marek; Norden, Bengt

    2013-12-01

    Fibrillization of peptides leads to the formation of amyloid fibres, which, when in large aggregates, are responsible for diseases such as Alzheimer's and Parkinson's. Here, we show that amyloids have strong nonlinear optical absorption, which is not present in native non-fibrillized protein. Z-scan and pump-probe experiments indicate that insulin and lysozyme ?-amyloids, as well as ?-synuclein fibres, exhibit either two-photon, three-photon or higher multiphoton absorption processes, depending on the wavelength of light. We propose that the enhanced multiphoton absorption is due to a cooperative mechanism involving through-space dipolar coupling between excited states of aromatic amino acids densely packed in the fibrous structures. This finding will provide the opportunity to develop nonlinear optical techniques to detect and study amyloid structures and also suggests that new protein-based materials with sizable multiphoton absorption could be designed for specific applications in nanotechnology, photonics and optoelectronics.

  19. FDTD/TDSE study of surface-enhanced infrared absorption by metal nanoparticles.

    SciTech Connect

    Chang, S.-H.; Schatz, G. C.; Gray, S. K.; Chemistry; Northwestern Univ.; National Cheng-Kung Univ.

    2006-01-01

    We study surface-enhanced infrared absorption, including multiphoton processes, due to the excitation of surface plasmons on metal nanoparticles. The time-dependent Schroedinger equation and finite-difference time-domain method are self-consistently coupled to treat the problem.

  20. Thermal imaging method to visualize a hidden painting thermally excited by far infrared radiations

    NASA Astrophysics Data System (ADS)

    Davin, T.; Wang, X.; Chabane, A.; Pawelko, R.; Guida, G.; Serio, B.; Hervé, P.

    2015-06-01

    The diagnosis of hidden painting is a major issue for cultural heritage. In this paper, a non-destructive active infrared thermographic technique was considered to reveal paintings covered by a lime layer. An extended infrared spectral range radiation was used as the excitation source. The external long wave infrared energy source delivered to the surface is then propagated through the material until it encounters a painting zone. Due to several thermal effects, the sample surface then presents non-uniformity patterns. Using a high sensitive infrared camera, the presence of covered pigments can thus be highlighted by the analysis of the non-stationary phenomena. Reconstituted thermal contrast images of mural samples covered by a lime layer are shown.

  1. Infrared multiple photon dissociation of chloromethyltrifluorosilane.

    PubMed

    Koshlyakov, Pavel V; Gorelik, Sergey R; Chesnokov, Evgeniy N; Aseev, Oleg S; Rakhymzhan, Asylkhan A; Petrov, Alexander K

    2009-01-01

    Infrared multiphoton absorption and dissociation of chloromethyltrifluorosilane molecules under the action of pulsed transversely excited atmospheric pressure CO2 laser were experimentally studied. Dissociation products were analyzed. The dissociation proceeds via chlorine atom transfer from carbon to silicone. High degrees of silicon isotope separation were achieved. The presence of alpha-chlorine atom in a silicon organic compound brings about a significant improvement in multiple photon dissociation characteristics and an essential increase in isotopic selectivity. PMID:19320849

  2. Controlling electron transfer in condensed phase with bond-specific infrared excitation

    NASA Astrophysics Data System (ADS)

    Delor, Milan; Scattergood, Paul A.; Sazanovich, Igor V.; Keane, Theo; Greetham, Gregory M.; Meijer, Anthony J. H. M.; Towrie, Michael; Parker, Anthony W.; Weinstein, Julia A.

    2015-08-01

    Using an ultrafast electronic-vibrational pulse-sequence, we show that the outcome of light-induced ET can be radically altered by mode-specific infrared (IR) excitation of vibrations which are coupled to the ET-pathway. IR-control is particularly challenging in condensed phase systems due to the ultrafast timescales involved, in particular rapid intramolecular vibrational redistribution. We demonstrate how an IR-pulse following UV-excitation perturbs nuclear-electronic (vibronic) interactions within a donor-bridge-acceptor system similar in design to those utilized in (bio)chemical light-harvesting, and alters charge-transport pathways and product state yields.

  3. Clinical multiphoton and CARS microscopy

    NASA Astrophysics Data System (ADS)

    Breunig, H. G.; Weinigel, M.; Darvin, M. E.; Lademann, J.; König, K.

    2012-03-01

    We report on clinical CARS imaging of human skin in vivo with the certified hybrid multiphoton tomograph CARSDermaInspect. The CARS-DermaInspect provides simultaneous imaging of non-fluorescent intradermal lipid and water as well as imaging of two-photon excited fluorescence from intrinsic molecules. Two different excitation schemes for CARS imaging have been realized: In the first setup, a combination of fs oscillator and optical parametric oscillator provided fs-CARS pump and Stokes pulses, respectively. In the second setup a fs oscillator was combined with a photonic crystal fiber which provided a broadband spectrum. A spectral range out of the broadband-spectrum was selected and used for CARS excitation in combination with the residual fs-oscillator output. In both setups, in addition to CARS, single-beam excitation was used for imaging of two-photon excited fluorescence and second harmonic generation signals. Both CARS-excitation systems were successfully used for imaging of lipids inside the skin in vivo.

  4. Mid-infrared ultrafast laser pulses induced third harmonic generation in nitrogen molecules on an excited state.

    PubMed

    Xie, Hongqiang; Li, Guihua; Yao, Jinping; Chu, Wei; Li, Ziting; Zeng, Bin; Wang, Zhanshan; Cheng, Ya

    2015-01-01

    We report on generation of third harmonic from nitrogen molecules on the excited state with a weak driver laser pulse at a mid-infrared wavelength. The excited nitrogen molecules are generated using a circularly polarized intense femtosecond pulse which produces energetic electrons by photoionization to realize collisional excitation of nitrogen molecules. Furthermore, since the third harmonic is generated using a pump-probe scheme, it enables investigation of the excited-state dynamics of nitrogen molecules produced under different conditions. We also perform a comparative investigation in excited argon atoms, revealing different decay dynamics of the molecules and atoms from the excited states in femtosecond laser induced filaments. PMID:26522886

  5. Mid-infrared ultrafast laser pulses induced third harmonic generation in nitrogen molecules on an excited state

    PubMed Central

    Xie, Hongqiang; Li, Guihua; Yao, Jinping; Chu, Wei; Li, Ziting; Zeng, Bin; Wang, Zhanshan; Cheng, Ya

    2015-01-01

    We report on generation of third harmonic from nitrogen molecules on the excited state with a weak driver laser pulse at a mid-infrared wavelength. The excited nitrogen molecules are generated using a circularly polarized intense femtosecond pulse which produces energetic electrons by photoionization to realize collisional excitation of nitrogen molecules. Furthermore, since the third harmonic is generated using a pump-probe scheme, it enables investigation of the excited-state dynamics of nitrogen molecules produced under different conditions. We also perform a comparative investigation in excited argon atoms, revealing different decay dynamics of the molecules and atoms from the excited states in femtosecond laser induced filaments. PMID:26522886

  6. Multiphoton excitation and ionization by elliptically polarized, intense short laser pulses: Recognizing multielectron dynamics and doorway states in C{sub 60} vs Xe

    SciTech Connect

    Shchatsinin, I.; Ritze, H.-H.; Schulz, C. P.; Hertel, I. V.

    2009-05-15

    Ionization and fragmentation of C{sub 60} fullerenes are studied by time-of-flight mass spectrometry, in elliptically polarized femtosecond laser fields at 797 nm of intensities I{sub 0}=(0.5-4.3)x10{sup 14} W cm{sup -2}. Xe atoms serve as a test case. We derive a qualitative theory describing such polarization studies. It turns out that polarization dependence can very sensitively distinguish single active electron (SAE) and multiple active electrons dynamics. In the case of Xe a clear signature of SAE dynamics is observed, with very pronounced changes in the ion yield as a function of ellipticity, indicative of N=5-8 and 18-22 photon processes for Xe{sup +} and Xe{sup 2+}, respectively. In contrast, only a moderate polarization dependence is observed in the C{sub 60} case, although at least 5 h{nu} photons at 797 nm are needed to generate C{sub 60}{sup +} and additional 11 for C{sub 60}{sup 2+}. At lower intensities, a moderate reduction in the ion yield for circular polarization establishes a two-photon SAE absorption process, connected with the key role of the lowest unoccupied molecular orbital (LUMO)+1(t{sub 1g}) as 'doorway state'. The absence of any polarization effect at 399 nm corroborates this finding. At high intensities enhanced fragmentation is observed, which is tentatively attributed to returning loops of electron trajectories by the combined action of the C{sub 60}{sup +} field and the circularly polarized laser field - in contrast to conventional wisdom that linear polarization should lead to an enhanced recolliding electron yield. No sign of a pronounced multiphoton polarization signature with five and more photons is seen for C{sub 60} which would be predicted by the SAE picture - although the slopes of the ion yield as a function of intensity are given by the corresponding power laws {proportional_to}I{sub 0}{sup N}. This is taken as clear evidence of multielectron dynamics after reaching the doorway state.

  7. Multiphoton harvesting metal-organic frameworks

    NASA Astrophysics Data System (ADS)

    Quah, Hong Sheng; Chen, Weiqiang; Schreyer, Martin K.; Yang, Hui; Wong, Ming Wah; Ji, Wei; Vittal, Jagadese J.

    2015-08-01

    Multiphoton upconversion is a process where two or more photons are absorbed simultaneously to excite an electron to an excited state and, subsequently, the relaxation of electron gives rise to the emission of a photon with frequency greater than those of the absorbed photons. Materials possessing such property attracted attention due to applications in biological imaging, photodynamic therapy, three-dimensional optical data storage, frequency-upconverted lasing and optical power limiting. Here we report four-photon upconversion in metal-organic frameworks containing the ligand, trans, trans-9,10-bis(4-pyridylethenyl)anthracene. The ligand has a symmetrical acceptor-π-donor-π-acceptor structure and a singlet biradical electronic ground state, which boosted its multiphoton absorption cross-sections. We demonstrate that the upconversion efficiency can be enhanced by Förster resonance energy transfer within host-guest metal-organic frameworks consisting of encapsulated high quantum yielding guest molecules. Using these strategies, metal-organic framework materials, which can exhibit frequency-upconverted photoluminescence excited by simultaneous multiphoton absorption, can be rationally designed and synthesized.

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

    NASA Astrophysics Data System (ADS)

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

    2008-02-01

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

  9. Multi-photon processes in alkali metal vapors

    NASA Astrophysics Data System (ADS)

    Gai, Baodong; Hu, Shu; Li, Hui; Shi, Zhe; Cai, Xianglong; Guo, Jingwei; Tan, Yannan; Liu, Wanfa; Jin, Yuqi; Sang, Fengting

    2015-02-01

    Achieving population inversion through multi-photon cascade pumping is almost always difficult, and most laser medium work under 1-photon excitation mechanism. But for alkali atoms such as cesium, relatively large absorption cross sections of several low, cascading energy levels enable them properties such as up conversion. Here we carried out research on two-photon excitation alkali fluorescence. Two photons of near infrared region are used to excite alkali atoms to n 2 D5/2, n 2 D3/2 or higher energy levels, then the blue fluorescence of (n+1) 2 P3/2,(n+1) 2 P1/2-->n 2 S1/2 are observed. Different pumping paths are tried and by the recorded spectra, transition routes of cesium are deducted and concluded. Finally the possibility of two-photon style DPALs (diode pumped alkali laser) are discussed, such alkali lasers can give output wavelengths in the shorter end of visual spectroscopy (400-460 nm) and are expected to get application in underwater communication and material laser processing.

  10. Determining two-photon absorption cross sections via nonresonant multiphoton photoacoustic spectroscopy

    NASA Astrophysics Data System (ADS)

    Kiser, John B.; Chandrasekharan, Nirmala; Cullum, Brian M.

    2007-09-01

    Multiphoton excitation of exogenous dyes and endogenous biochemical species has been used extensively for tissue diagnosis by fluorescence spectroscopy. Unfortunately, the majority of endogenous biochemical chromophores have low quantum yields, less than 0.2, therefore determining two-photon cross sections of weakly luminescencing molecules is difficult using two-photon fluorescence spectroscopy. Accurate determination of two-photon cross sections of these biochemicals could provide insight into fluorescence signal reduction caused by the absorption of excitation energy by non-target intracellular species. Non-resonant multiphoton photoacoustic spectroscopy (NMPPAS) is a novel technique we have developed for condensed matter measurements that has the potential for accurately determining two-photon absorption cross-sections of chemicals with small or non-existant fluorescence quantum yields. In this technique, near infrared light is used to generate an ultrasonic signal following a non-resonant two-photon excitation process. This ultrasonic wave is directly related to the non-radative relaxation of the chromophore of interest and is measured using a contact piezoelectric ultrasonic transducer. The signal from the ultrasonic transducer can then be used to calculate two-photon absorption cross sections. This paper will describe the validation of this technique by measuring the two-photon absorption cross- sections of well characterized chromophores such as rhodamine B and coumarin 1 in solution as well as riboflavin in a gelatin tissue phantom.

  11. Surface plasmon excitation at near-infrared wavelengths in polymer optical fibers.

    PubMed

    Hu, Xuehao; Mgret, Patrice; Caucheteur, Christophe

    2015-09-01

    We report the first excitation of surface plasmon waves at near-infrared telecommunication wavelengths using polymer optical fibers (POFs) made of poly(methyl methacrylate) (PMMA). For this, weakly tilted fiber-Bragg gratings (TFBGs) have been photo-inscribed in the core of step-index POFs and the fiber coated with a thin gold layer. Surface plasmon resonance is excited with radially polarized modes and is spectrally observed as a singular extinction of some cladding-mode resonances in the transmitted amplitude spectrum of gold-coated TFBGs. The refractometric sensitivity can reach ?550??nm/RIU (refractive index unit) with a figure of merit of more than 2000 and intrinsic temperature self-compensation. This kind of sensor is particularly relevant to in situ operation. PMID:26368696

  12. Multiphoton ionization and multiphoton resonances in the tunneling regime

    SciTech Connect

    Potvliege, R. M.; Mese, E.; Vucic, Svetlana

    2010-05-15

    The rate of ionization of an atom of helium, argon, or hydrogen exposed to an intense monochromatic laser field and the quasienergy spectrum of their dressed states are studied for values of the Keldysh parameter between 1 and 0.6 and wavelengths between 390 and 1300 nm. The calculations are carried out within the non-Hermitian Floquet theory. Resonances with intermediate excited states significantly affect ionization from the dressed ground state at all the intensities and all the wavelengths considered. The dressed excited states responsible for these structures are large-{alpha}{sub 0} states akin to the Kramers-Henneberger states of the high-frequency Floquet theory. Within the single-active-electron approximation, these large-{alpha}{sub 0} states become species independent at sufficiently high intensity or sufficiently long wavelength. Apart for the resonance structures arising from multiphoton coupling with excited states, the ab initio Floquet ionization rate is in excellent agreement with the predictions of two different calculations in the strong field approximation, one based on a length-gauge formulation of this approximation and one based on a velocity-gauge formulation. The calculations also confirm the validity of the {omega}{sup 2} expansion as an alternative to the strong field approximation for taking into account the nonadiabaticity of the ionization process in intense low-frequency laser fields.

  13. Multiphoton ionization and multiphoton resonances in the tunneling regime

    NASA Astrophysics Data System (ADS)

    Potvliege, R. M.; Me?e, E.; Vu?i?, Svetlana

    2010-05-01

    The rate of ionization of an atom of helium, argon, or hydrogen exposed to an intense monochromatic laser field and the quasienergy spectrum of their dressed states are studied for values of the Keldysh parameter between 1 and 0.6 and wavelengths between 390 and 1300 nm. The calculations are carried out within the non-Hermitian Floquet theory. Resonances with intermediate excited states significantly affect ionization from the dressed ground state at all the intensities and all the wavelengths considered. The dressed excited states responsible for these structures are large-?0 states akin to the Kramers-Henneberger states of the high-frequency Floquet theory. Within the single-active-electron approximation, these large-?0 states become species independent at sufficiently high intensity or sufficiently long wavelength. Apart for the resonance structures arising from multiphoton coupling with excited states, the ab initio Floquet ionization rate is in excellent agreement with the predictions of two different calculations in the strong field approximation, one based on a length-gauge formulation of this approximation and one based on a velocity-gauge formulation. The calculations also confirm the validity of the ?2 expansion as an alternative to the strong field approximation for taking into account the nonadiabaticity of the ionization process in intense low-frequency laser fields.

  14. In vivo multiphoton tomography of skin cancer

    NASA Astrophysics Data System (ADS)

    Knig, Karsten; Riemann, Iris; Ehlers, Alexander; Buckle, Rainer; Dimitrow, Enrico; Kaatz, Martin; Fluhr, Joachim; Elsner, Peter

    2006-02-01

    The multiphoton tomograph DermaInspect was used to perform first clinical studies on the early non-invasive detection of skin cancer based on non-invasive optical sectioning of skin by two-photon autofluorescence and second harmonic generation. In particular, deep-tissue pigmented lesions -nevi- have been imaged with intracellular resolution using near infrared (NIR) femtosecond laser radiation. So far, more than 250 patients have been investigated. Cancerous tissues showed significant morphological differences compared to normal skin layers. In the case of malignant melanoma, the occurrence of luminescent melanocytes has been detected. Multiphoton tomography will become a novel non-invasive method to obtain high-resolution 3D optical biopsies for early cancer detection, treatment control, and in situ drug screening.

  15. Multiphoton tomography to detect chemo- and biohazards

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2015-03-01

    In vivo high-resolution multiphoton/CARS tomography provides optical biopsies with 300 nm lateral resolution with chemical fingerprints. Thousands of volunteers and patients have been investigated for early cancer diagnosis, evaluation of anti-ageing cosmetic products, and changes of cellular metabolism by UV exposure and decreased oxygen supply. The skin as the outermost and largest organ is also the major target of CB agents. Current UV-based sensors are useful for bio-aerosol sensing but not for evaluating exposed in vivo skin. Here we evaluate the use of 4D multiphoton/CARS tomographs based on near infrared femtosecond laser radiation, time-correlated single photon counting (FLIM) and white light generation by photonic crystal fibers to detect bio- and chemohazards in human in vivo skin using twophoton fluorescence, SHG, and Raman signals.

  16. Multiphoton tomography of intratissue tattoo nanoparticles

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2012-02-01

    Most of today's intratissue tattoo pigments are unknown nanoparticles. So far, there was no real control of their use due to the absence of regulations. Some of the tattoo pigments contain carcinogenic amines e.g. azo pigment Red 22. Nowadays, the European Union starts to control the administration of tattoo pigments. There is an interest to obtain information on the intratissue distribution, their interaction with living cells and the extracellular matrix, and the mechanisms behind laser tattoo removal. Multiphoton tomographs are novel biosafety and imaging tools that can provide such information non-invasively and without further labeling. When using the spectral FLIM module, spatially-resolved emission spectra, excitation spectra, and fluorescence lifetimes can pr provided. Multiphoton tomographs are used by all major cosmetic comapanies to test the biosafety of sunscreen nanoparticles.

  17. Identification of glucosinolates in capers by LC-ESI-hybrid linear ion trap with Fourier transform ion cyclotron resonance mass spectrometry (LC-ESI-LTQ-FTICR MS) and infrared multiphoton dissociation.

    PubMed

    Bianco, Giuliana; Lelario, Filomena; Battista, Fabio Giuseppe; Bufo, Sabino A; Cataldi, Tommaso R I

    2012-09-01

    An liquid chromatography-mass spectrometry method using electrospray ionization in negative ion mode coupled with a hybrid quadrupole linear ion trap and Fourier transform ion cyclotron resonance (FTICR) mass spectrometer was applied to characterize of intact glucosinolates (GLSs) in crude sample extracts of wild bud flowers of Capparis spinosa (Capparis species, family Capparaceae). Structural information of GLSs was obtained upon precursor ions' isolation within the FTICR trapping cell and subsequent fragmentation induced by infrared multiphoton dissociation (IRMPD). Such a fragmentation was found very useful in terms of chemical identification of all precursor ions [M-H](-) including sulfur-rich GLSs reported here for the first time. Along with most common GLSs already found in capers such as glucocapparin, isopropyl/n-propyl-GLS, mercapto-glucocapparin, and two indolic GLS, i.e., 4-hydroxyglucobrassicin and glucobrassicin, the occurrence of the uncommon glycinyl-glucocapparin as well as two sulfur-rich GLSs is reported. IRMPD showed an increased selectivity towards disulfide bond cleavages with thiol migration, suggesting the side chain structure of non-targeted compounds, i.e., disulfanyl-glucocapparin and trisulfanyl-glucocapparin. Glucocapparin [2.05??0.25?mg/g, dry weight (dw)] was the most abundant GLS, followed by glucobrassicin (232??18?g/g, dw) and 4-hydroxyglucobrassicin (89??12?g/g, dw). All other compounds were present at very low content ranging from 0.5 to 1.5?g/g dw. PMID:22972784

  18. Studies of atmospheric molecules by multiphoton spectroscopy

    SciTech Connect

    Johnson, P.M.

    1991-10-01

    Carbon dioxide presents a great challenge to spectroscopy because of its propensity toward dissociation in all of its excited states. Multiphoton ionization spectroscopy is usually not applicable to the study of dissociating molecules because the dissociation competes effectively with ionization, resulting in no signal. We reasoned, however, that with high enough laser fluence, ionization could compete with dissociation in the longer lived states, exposing them for study from the continuous spectral background resulting from rapidly dissociating states. We describe the various spectroscopic and photophysical effects found through the multiphoton ionization and multiphoton photoelectron spectra. A recently developed variant of threshold ionization spectroscopy, usually called ZEKE, has shown a great deal of usefulness in providing the same information as traditional photoelectron spectroscopy but with higher resolution and much better signal-to-noise when using standard laboratory lasers. Threshold ionization techniques locate the states of an ion by scanning a light source across the ionization continuum of a neutral and somehow detecting when electrons are produced with no kinetic energy. We chose to develop our capabilities in threshold ionization spectroscopy using aromatic molecules because of their importance and because their electronic structure allows a pump-probe type of excitation scheme which avoids the use of vacuum ultraviolet laser beams. Among aromatics, the azines are noted for their small S{sub 1}-T{sub 1} energy gap which give them unique and interesting photophysical properties. We have continued our work on the multiphoton spectrum of metastable nitrogen produced by an electric discharge in supersonic beam. We have been able to assign more of the lines and simulated their rotational structure but many peaks remain unassigned.

  19. Excited-state dynamics of protochlorophyllide revealed by subpicosecond infrared spectroscopy.

    PubMed

    Colindres-Rojas, Miriam; Wolf, Matthias M N; Gross, Ruth; Seidel, Sonja; Dietzek, Benjamin; Schmitt, Michael; Popp, Jrgen; Hermann, Gudrun; Diller, Rolf

    2011-01-01

    To gain a better understanding of the light-induced reduction of protochlorophyllide (PChlide) to chlorophyllide as a key regulatory step in chlorophyll synthesis, we performed transient infrared absorption measurements on PChlide in d4-methanol. Excitation in the Q-band at 630 nm initiates dynamics characterized by three time constants: ?? = 3.6 0.2, ?? = 38 2, and ?? = 215 8 ps. As indicated by the C13'=O carbonyl stretching mode in the electronic ground state at 1686 cm?, showing partial ground-state recovery, and in the excited electronic state at 1625 cm?, showing excited-state decay, ?? describes the formation of a state with a strong change in electronic structure, and ?? represents the partial recovery of the PChlide electronic ground state. Furthermore, ?? corresponds with vibrational energy relaxation. The observed kinetics strongly suggest a branched reaction scheme with a branching ratio of 0.5 for the path leading to the PChlide ground state on the 200 ps timescale and the path leading to a long-lived state (>700 ps). The results clearly support a branched reaction scheme, as proposed previously, featuring the formation of an intramolecular charge transfer state with ?25 ps, its decay into the PChlide ground state with 200 ps, and a parallel reaction path to the long-lived PChlide triplet state. PMID:21190679

  20. Excited-State Dynamics of Protochlorophyllide Revealed by Subpicosecond Infrared Spectroscopy

    PubMed Central

    Colindres-Rojas, Miriam; Wolf, Matthias M.N.; Gro, Ruth; Seidel, Sonja; Dietzek, Benjamin; Schmitt, Michael; Popp, Jrgen; Hermann, Gudrun; Diller, Rolf

    2011-01-01

    To gain a better understanding of the light-induced reduction of protochlorophyllide (PChlide) to chlorophyllide as a key regulatory step in chlorophyll synthesis, we performed transient infrared absorption measurements on PChlide in d4-methanol. Excitation in the Q-band at 630nm initiates dynamics characterized by three time constants: ?1= 3.6 0.2, ?2= 38 2, and ?3= 215 8 ps. As indicated by the C13?=O carbonyl stretching mode in the electronic ground state at 1686cm?1, showing partial ground-state recovery, and in the excited electronic state at 1625cm?1, showing excited-state decay, ?2 describes the formation of a state with a strong change in electronic structure, and ?3 represents the partial recovery of the PChlide electronic ground state. Furthermore, ?1 corresponds with vibrational energy relaxation. The observed kinetics strongly suggest a branched reaction scheme with a branching ratio of 0.5 for the path leading to the PChlide ground state on the 200 ps timescale and the path leading to a long-lived state (>>700 ps). The results clearly support a branched reaction scheme, as proposed previously, featuring the formation of an intramolecular charge transfer state with ?25 ps, its decay into the PChlide ground state with 200 ps, and a parallel reaction path to the long-lived PChlide triplet state. PMID:21190679

  1. Compact fixed wavelength femtosecond oscillators for multi-photon imaging

    NASA Astrophysics Data System (ADS)

    Hakulinen, T.; Klein, J.; Zadoyan, R.; Baldacchini, T.; Franke, T.

    2015-03-01

    In recent years two-photon microscopy with fixed-wavelength has raised increasing interest in life-sciences: Two-photon (2P) absorption spectra of common dyes are broader than single-photon ones. Therefore, excitation of several dyes simultaneously with a single IR laser wavelength is feasible and could be seen as an advantage in 2P microscopy. We used pulsed fixed-wavelength infrared lasers with center wavelength at 1040 nm, for two-photon microscopy in a variety of biologically relevant samples, among these a mouse brain sample, a mouse artery (within the animal, acute preparation), and a preparation of mouse bladder. The 1040 nm laser proved to be efficient not only in exciting fluorescence from yellow fluorescent protein (YFP) and red fluorescent dyes, but also for second harmonic generation (SHG) signals from muscle tissue and collagen. With this work we demonstrate that economical, small-footprint fixedwavelength lasers can present an interesting alternative to tunable lasers that are commonly used in multiphoton microscopy.

  2. Characterizing lamina propria of human gastric mucosa by multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Liu, Y. C.; Yang, H. Q.; Chen, G.; Zhuo, S. M.; Chen, J. X.; Yan, J.

    2011-01-01

    Lamina propria (LP) of gastric mucosa plays an important role in progression of gastric cancer because of the site at where inflammatory reactions occur. Multiphoton imaging has been recently employed for microscopic examination of intact tissue. In this paper, using multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG), high resolution multiphoton microscopic images of lamina propria (LP) are obtained in normal human gastric mucosa at excitation wavelength λex = 800 nm. The main source of tissue TPEF originated from the cells of gastric glands, and loose connective tissue, collagen, produced SHG signals. Our results demonstrated that MPM can be effective for characterizing the microstructure of LP in human gastric mucosa. The findings will be helpful for diagnosing and staging early gastric cancer in the clinics.

  3. Enhanced eumelanin emission by stepwise three-photon excitation

    NASA Astrophysics Data System (ADS)

    Kerimo, Josef; Rajadhyaksha, Milind; DiMarzio, Charles A.

    2011-03-01

    Eumelanin fluorescence from Sepia officinalis and black human hair was activated with near-infrared radiation and multiphoton excitation. A third order multiphoton absorption by a step-wise process appears to be the underlying mechanism. The activation was caused by a photochemical process since it could not be reproduced by simple heating. Both fluorescence and brightfield imaging indicate the near-infrared irradiation caused photodamage to the eumelanin and the activated emission originated from the photodamaged region. At least two different components with about thousand-fold enhanced fluorescence were activated and could be distinguished by their excitation properties. One component was excited with wavelengths in the visible region and exhibited linear absorption dependence. The second component could be excited with near-infrared wavelengths and had a third order dependence on the laser power. The third order dependence is explained by a step-wise excited state absorption (ESA) process since it could be observed equally with the CW and femtosecond lasers. The new method for photoactivating the eumelanin fluorescence was used to map the melanin content in human hair.

  4. Free electron laser-Fourier transform ion cyclotron resonance mass spectrometry facility for obtaining infrared multiphoton dissociation spectra of gaseous ions

    SciTech Connect

    Valle, Jose J.; Eyler, John R.; Oomens, Jos; Moore, David T.; Meer, A.F.G. van der; Helden, Gert von; Meijer, Gerard; Hendrickson, Christopher L.; Marshall, Alan G.; Blakney, Gregory T.

    2005-02-01

    A Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer has been installed at a free electron laser (FEL) facility to obtain infrared absorption spectra of gas phase ions by infrared multiple photon dissociation (IRMPD). The FEL provides continuously tunable infrared radiation over a broad range of the infrared spectrum, and the FT-ICR mass spectrometer, utilizing a 4.7 Tesla superconducting magnet, permits facile formation, isolation, trapping, and high-mass resolution detection of a wide range of ion classes. A description of the instrumentation and experimental parameters for these experiments is presented along with preliminary IRMPD spectra of the singly-charged chromium-bound dimer of diethyl ether (Cr(C{sub 4}H{sub 10}O){sub 2}{sup +}) and the fluorene molecular ion (C{sub 13}H{sub 10}{sup +}). Also presented is a brief comparison of the fluorene cation spectrum obtained by the FT-ICR-FEL with an earlier spectrum recorded using a quadrupole ion trap (QIT)

  5. Multispot multiphoton Ca2+ imaging in acute myocardial slices

    NASA Astrophysics Data System (ADS)

    Borile, Giulia; de Mauro, Claudio; Urbani, Andrea; Alfieri, Domenico; Pavone, Francesco S.; Mongillo, Marco

    2015-05-01

    Multiphoton microscopy has become essential for dynamic imaging in thick living tissues. High-rate, full-field image acquisition in multiphoton microscopy is achievable by parallelization of the excitation and detection pathways. We developed our approach via a diffractive optical element which splits a pulsed laser into 16 beamlets and exploits a descanned detection system consisting of an array of beamlet-associated photomultiplier tubes. The optical performance of the multiphoton multispot system (MCube) has been characterized in cardiac tissue sections and subsequently used for the first time for fluorescence imaging of cardiomyocyte Ca2+ dynamics in viable acute cardiac slices. Multispot multiphoton microscopy (MMM) has never been used before to monitor Ca2+ dynamics in thick, viable tissue samples. Acute heart slices are a powerful close-to-in vivo model of Ca2+ imaging allowing the simultaneous observation of several cells in their own tissue environment, exploiting the multiphoton excitation ability to penetrate scattering tissues. Moreover, we show that the concurrent high spatial and temporal resolutions afforded by the parallel scanning in MMM can be exploited to simultaneously assess subcellular Ca2+ dynamics in different cells in the tissue. We recorded local Ca2+ release events including macrosparks, travelling waves, and rotors.

  6. Interaction of an infrared surface plasmon with an excited molecular vibration.

    PubMed

    Rodriguez, Kenneth R; Tian, Hong; Heer, Joseph M; Teeters-Kennedy, Shannon; Coe, James V

    2007-04-21

    The interaction of an infrared surface plasmon and an excited molecular vibration was investigated by using a square array of subwavelength holes in a Ni film which supports propagating, surface-plasmon-mediated, transmission resonances. The largest transmission resonance [the (1,0)(-)] was tuned through the rocking vibration of the hexadecane molecule (at 721 cm(-1)) in a hexadecane film on the mesh by varying the thickness of the film. The interaction of the rocking vibration and surface plasmon is characterized spectroscopically by an increase in the intensity of the vibrational band by more than a factor of 2, variation of the vibrational line shape relative to the spectrum on a nonmetallic surface, and shifts in vibrational peak position by as much as 3.0 cm(-1). Relationships are developed between the transmission resonance position and the thickness and dielectric properties of the coating. PMID:17461607

  7. Visible and near-infrared excitation spectra from the neptunyl ion doped into a uranyl tetrachloride lattice

    NASA Astrophysics Data System (ADS)

    Barker, Beau J.; Berg, John M.; Kozimor, Stosh A.; Wozniak, Nicholas R.; Wilkerson, Marianne P.

    2016-03-01

    Visible and near-infrared illumination induces 5f-5f and ligand-to-metal charge-transfer (LMCT) transitions of the neptunyl tetrachloride anion in polycrystalline Cs2U(Np)O2Cl4, and results in near-infrared luminescence from the second electronically excited state to the ground state. This photoluminescence is used as a detection method to collect excitation spectra throughout the near-infrared and visible regions. The excitation spectra of LMCT transitions in excitation spectra were identified in previous work. Here the measurement and analysis is extended to include both LMCT and intra-5f transitions. The results manifest variation in structural properties of the neptunium-oxo bond among the low-lying electronic states. Vibronic intensity patterns and energy spacings are used to compare bond lengths and vibrational frequencies in the excited states, confirming significant characteristic differences between those excited by 5f-5f transitions from those due to LMCT transitions. Results are compared with recently published RASPT2/SO calculations of [NpO2Cl4]2-.

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

    PubMed

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

    2013-09-01

    We report a new type of UV-excitable red/NIR-emissive fluorescent dyads (PKF series). Conjugation of a pyrene and a novel bright red/near-infrared (NIR) fluorophore resulted in large quasi-Stokes shift while retaining intense fluorescence emission and sharp spectral bands. Labeling of PKF dyads to biomolecules was performed by means of introduction of a succinimidyl ester. Simultaneous Ca(2+)/albumin dual-color intracellular imaging by PKF in combination with fura-2 (UV-excitable/VIS-emissive Ca(2+) indicator) reveals its usefulness as a new bioimaging tool. PMID:23700213

  9. Ultrafast excited-state dynamics of nanoscale near-infrared emissive polymersomes.

    PubMed

    Duncan, Timothy V; Ghoroghchian, P Peter; Rubtsov, Igor V; Hammer, Daniel A; Therien, Michael J

    2008-07-30

    Formed through cooperative self-assembly of amphiphilic diblock copolymers and electronically conjugated porphyrinic near-infrared (NIR) fluorophores (NIRFs), NIR-emissive polymersomes (50 nm to 50 microm diameter polymer vesicles) define a family of organic-based, soft-matter structures that are ideally suited for deep-tissue optical imaging and sensitive diagnostic applications. Here, we describe magic angle and polarized pump-probe spectroscopic experiments that: (i) probe polymersome structure and NIRF organization and (ii) connect emitter structural properties and NIRF loading with vesicle emissive output at the nanoscale. Within polymersome membrane environments, long polymer chains constrain ethyne-bridged oligo(porphinato)zinc(II) based supermolecular fluorophore (PZn n ) conformeric populations and disperse these PZn n species within the hydrophobic bilayer. Ultrafast excited-state transient absorption and anisotropy dynamical studies of NIR-emissive polymersomes, in which the PZn n fluorophore loading per nanoscale vesicle is varied between 0.1-10 mol %, enable the exploration of concentration-dependent mechanisms for nonradiative excited-state decay. These experiments correlate fluorophore structure with its gross spatial arrangement within specific nanodomains of these nanoparticles and reveal how compartmentalization of fluorophores within reduced effective dispersion volumes impacts bulk photophysical properties. As these factors play key roles in determining the energy transfer dynamics between dispersed fluorophores, this work underscores that strategies that modulate fluorophore and polymer structure to optimize dispersion volume in bilayered nanoscale vesicular environments will further enhance the emissive properties of these sensitive nanoscale probes. PMID:18611010

  10. Multi-Photon Nanosurgery in Live Brain

    PubMed Central

    Mascaro, Anna Letizia Allegra; Sacconi, Leonardo; Pavone, Francesco S.

    2010-01-01

    In the last few years two-photon microscopy has been used to perform in vivo high spatial resolution imaging of neurons, glial cells and vascular structures in the intact neocortex. Recently, in parallel to its applications in imaging, multi-photon absorption has been used as a tool for the selective disruption of neural processes and blood vessels in living animals. In this review we present some basic features of multi-photon nanosurgery and we illustrate the advantages offered by this novel methodology in neuroscience research. We show how the spatial localization of multi-photon excitation can be exploited to perform selective lesions on cortical neurons in living mice expressing fluorescent proteins. This methodology is applied to disrupt a single neuron without causing any visible collateral damage to the surrounding structures. The spatial precision of this method allows to dissect single processes as well as individual dendritic spines, preserving the structural integrity of the main neuronal arbor. The same approach can be used to breach the blood-brain barrier through a targeted photo-disruption of blood vessels walls. We show how the vascular system can be perturbed through laser ablation leading toward two different models of stroke: intravascular clot and extravasation. Following the temporal evolution of the injured system (either a neuron or a blood vessel) through time lapse in vivo imaging, the physiological response of the target structure and the rearrangement of the surrounding area can be characterized. Multi-photon nanosurgery in live brain represents a useful tool to produce different models of neurodegenerative disease. PMID:20725602

  11. Highly Emitting Near-Infrared Lanthanide Encapsulated Sandwich Metallacrown Complexes with Excitation Shifted Toward Lower Energy

    PubMed Central

    2015-01-01

    Near-infrared (NIR) luminescent lanthanide complexes hold great promise for practical applications, as their optical properties have several complementary advantages over organic fluorophores and semiconductor nanoparticles. The fundamental challenge for lanthanide luminescence is their sensitization through suitable chromophores. The use of the metallacrown (MC) motif is an innovative strategy to arrange several organic sensitizers at a well-controlled distance from a lanthanide cation. Herein we report a series of lanthanide encapsulated sandwich MC complexes of the form Ln3+[12-MCZn(II),quinHA-4]2[24-MCZn(II),quinHA-8] (Ln3+[Zn(II)MCquinHA]) in which the MC framework is formed by the self-assembly of Zn2+ ions and tetradentate chromophoric ligands based on quinaldichydroxamic acid (quinHA). A first-generation of luminescent MCs was presented previously but was limited due to excitation wavelengths in the UV. We report here that through the design of the chromophore of the MC assembly, we have significantly shifted the absorption wavelength toward lower energy (450 nm). In addition to this near-visible inter- and/or intraligand charge transfer absorption, Ln3+[Zn(II)MCquinHA] exhibits remarkably high quantum yields, long luminescence lifetimes (CD3OD; Yb3+, QLnL = 2.88(2)%, ?obs = 150.7(2) ?s; Nd3+, QLnL = 1.35(1)%, ?obs = 4.11(3) ?s; Er3+, QLnL = 3.60(6)102%, ?obs = 11.40(3) ?s), and excellent photostability. Quantum yields of Nd3+ and Er3+ MCs in the solid state and in deuterated solvents, upon excitation at low energy, are the highest values among NIR-emitting lanthanide complexes containing CH bonds. The versatility of the MC strategy allows modifications in the excitation wavelength and absorptivity through the appropriate design of the ligand sensitizer, providing a highly efficient platform with tunable properties. PMID:24432702

  12. Highly emitting near-infrared lanthanide "encapsulated sandwich" metallacrown complexes with excitation shifted toward lower energy.

    PubMed

    Trivedi, Evan R; Eliseeva, Svetlana V; Jankolovits, Joseph; Olmstead, Marilyn M; Petoud, Stphane; Pecoraro, Vincent L

    2014-01-29

    Near-infrared (NIR) luminescent lanthanide complexes hold great promise for practical applications, as their optical properties have several complementary advantages over organic fluorophores and semiconductor nanoparticles. The fundamental challenge for lanthanide luminescence is their sensitization through suitable chromophores. The use of the metallacrown (MC) motif is an innovative strategy to arrange several organic sensitizers at a well-controlled distance from a lanthanide cation. Herein we report a series of lanthanide encapsulated sandwich MC complexes of the form Ln3+ [12-MC(Zn(II),quinHA)-4]2[24-MC(Zn(II),quinHA)-8] (Ln3+ [Zn(II)MC(quinHA)]) in which the MC framework is formed by the self-assembly of Zn2+ ions and tetradentate chromophoric ligands based on quinaldichydroxamic acid (quinHA). A first-generation of luminescent MCs was presented previously but was limited due to excitation wavelengths in the UV. We report here that through the design of the chromophore of the MC assembly, we have significantly shifted the absorption wavelength toward lower energy (450 nm). In addition to this near-visible inter- and/or intraligand charge transfer absorption, Ln3+ [Zn(II)MC(quinHA)] exhibits remarkably high quantum yields, long luminescence lifetimes (CD3OD; Yb3+, QLn(L) = 2.88(2)%, ?obs = 150.7(2) ?s; Nd3+, QLn(L) = 1.35(1)%, ?obs = 4.11(3) ?s; Er3+, QLn(L) = 3.60(6)102%, ?obs = 11.40(3) ?s), and excellent photostability. Quantum yields of Nd3+ and Er3+ MCs in the solid state and in deuterated solvents, upon excitation at low energy, are the highest values among NIR-emitting lanthanide complexes containing CH bonds. The versatility of the MC strategy allows modifications in the excitation wavelength and absorptivity through the appropriate design of the ligand sensitizer, providing a highly efficient platform with tunable properties. PMID:24432702

  13. Infrared Multiphoton Dissociation Spectroscopy of a Gas-Phase Complex of Uranyl and 3-Oxa-Glutaramide: An Extreme Red-Shift of the [O=U=O]²⁺ Asymmetric Stretch

    SciTech Connect

    Gibson, John K.; Hu, Hanshi; Van Stipdonk, Michael J.; Berden, Giel; Oomens, Jos; Li, Jun

    2015-04-09

    The gas-phase complex UO₂(TMOGA)₂²⁺ (TMOGA = tetramethyl-3-oxa-glutaramide) prepared by electrospray ionization was characterized by infrared multiphoton dissociation (IRMPD) spectroscopy. The IRMPD spectrum from 700–1800 cm⁻¹ was interpreted using a computational study based on density functional theory. The predicted vibrational frequencies are in good agreement with the measured values, with an average deviation of only 8 cm⁻¹ (<1%) and a maximum deviation of 21 cm⁻¹ (<2%). The only IR peak assigned to the linear uranyl moiety was the asymmetric ν₃ mode, which appeared at 965 cm⁻¹ and was predicted by DFT as 953 cm⁻¹. This ν₃ frequency is red-shifted relative to bare uranyl, UO₂²⁺, by ca. 150 cm⁻¹ due to electron donation from the TMOGA ligands. Based on the degree of red-shifting, it is inferred that two TMOGA oxygen-donor ligands have a greater effective gas basicity than the four monodentate acetone ligands in UO₂(acetone)₄²⁺. The uranyl ν₃ frequency was also computed for uranyl coordinated by two TMGA ligands, in which the central Oether of TMOGA has been replaced by CH₂. The computed ν₃ for UO₂(TMGA)₂²⁺, 950 cm⁻¹, is essentially the same as that for UO₂(TMOGA)₂²⁺, suggesting that electron donation to uranyl from the Oether of TMOGA is minor. The computed ν₃ asymmetric stretching frequencies for the three actinyl complexes, UO₂(TMOGA)₂²⁺, NpO₂(TMOGA)₂²⁺ and PuO₂(TMOGA)₂²⁺, are comparable. This similarity is discussed in the context of the relationship between ν₃ and intrinsic actinide-oxygen bond energies in actinyl complexes.

  14. Femtosecond transient infrared and stimulated Raman spectroscopy shed light on the relaxation mechanisms of photo-excited peridinin

    NASA Astrophysics Data System (ADS)

    Di Donato, Mariangela; Ragnoni, Elena; Lapini, Andrea; Foggi, Paolo; Hiller, Roger G.; Righini, Roberto

    2015-06-01

    By means of one- and two-dimensional transient infrared spectroscopy and femtosecond stimulated Raman spectroscopy, we investigated the excited state dynamics of peridinin, a carbonyl carotenoid occurring in natural light harvesting complexes. The presence of singly and doubly excited states, as well as of an intramolecular charge transfer (ICT) state, makes the behavior of carbonyl carotenoids in the excited state very complex. In this work, we investigated by time resolved spectroscopy the relaxation of photo-excited peridinin in solvents of different polarities and as a function of the excitation wavelength. Our experimental results show that a characteristic pattern of one- and two-dimensional infrared bands in the C=C stretching region allows monitoring the relaxation pathway. In polar solvents, moderate distortions of the molecular geometry cause a variation of the single/double carbon bond character, so that the partially ionic ICT state is largely stabilized by the solvent reorganization. After vertical photoexcitation at 400 nm of the S2 state, the off-equilibrium population moves to the S1 state with ca. 175 fs time constant; from there, in less than 5 ps, the non-Franck Condon ICT state is reached, and finally, the ground state is recovered in 70 ps. That the relevant excited state dynamics takes place far from the Franck Condon region is demonstrated by its noticeable dependence on the excitation wavelength.

  15. Non Destructive Testing by active infrared thermography coupled with shearography under same optical heat excitation

    NASA Astrophysics Data System (ADS)

    Theroux, Louis-Daniel; Dumoulin, Jean; Maldague, Xavier

    2014-05-01

    As infrastructures are aging, the evaluation of their health is becoming crucial. To do so, numerous Non Destructive Testing (NDT) methods are available. Among them, thermal shearography and active infrared thermography represent two full field and contactless methods for surface inspection. The synchronized use of both methods presents multiples advantages. Most importantly, both NDT are based on different material properties. Thermography depend on the thermal properties and shearography on the mechanical properties. The cross-correlation of both methods result in a more accurate and exact detection of the defects. For real site application, the simultaneous use of both methods is simplified due to the fact that the excitation method (thermal) is the same. Active infrared thermography is the measure of the temperature by an infrared camera of a surface subjected to heat flux. Observation of the variation of temperature in function of time reveal the presence of defects. On the other hand, shearography is a measure of out-of-plane surface displacement. This displacement is caused by the application of a strain on the surface which (in our case) take the form of a temperature gradient inducing a thermal stress To measure the resulting out-of-plane displacement, shearography exploit the relation between the phase difference and the optical path length. The phase difference is measured by the observation of the interference between two coherent light beam projected on the surface. This interference is due to change in optical path length as the surface is deformed [1]. A series of experimentation have been conducted in laboratory with various sample of concrete reinforced with CFRP materials. Results obtained reveal that with both methods it was possible to detect defects in the gluing. An infrared lamp radiating was used as the active heat source. This is necessary if measurements with shearography are to be made during the heating process. A heating lamp in the visible spectrum would hinder the projected light beam since a laser with wavelength of 532 nm was used as the coherent light source. Experimentations were successful, but only with mitigated efficiency for shearography [2]. The thermal response was the fastest and it was possible to fully locate all defects. For shearography, the available equipment forced us to restrict the area of observation to only one defect at a time (roughly 100 cm). Numerical models were designed based on the multiple sample tested in the experimental step of the study. Using the COMSOL finite elements modeling software, numerous simulations yielded results in accordance with experimental data. Different types of defect could be modeled and showed that both shearography and thermography have different sensibility in function of the nature of the defect. Furthermore, analysis of the simulated results demonstrated a relation between the contrast evolution of the temperature and displacement field. In the near future, we expect to make several improvement to our experimental setup. As for the numerical model, some small disparities between the theoretical and experimental results still remain to be addressed. The numerical model could be improved but to do so it requires to raise the shearographic measurements sampling rate close to the one used for infrared thermography. Once this issue will be resolved, it will be possible to use experimental data to refine the numerical model. So, accurate models will be helpful to optimize the overall efficiency of the coupling of thermal shearography and active infrared thermography for in situ NDT application. References [1] Y.Y. Hung, C.Y. Liand, Image-shearing camera for direct measurement of surface strains, Applied Optics, Vol. 18, n7, pages 1046-1051, 1979 [2] L-D. Throux, J. Dumoulin, X. Maldague, Square heating applied to shearography and active infrared thermography measurements coupling: form feasibility test in laboratory to numerical study of pultruded CFRP plates glued on concrete specimen, STRAIN journal, in press

  16. Fibre-coupled multiphoton microscope with adaptive motion compensation

    PubMed Central

    Sherlock, Ben; Warren, Sean; Stone, James; Neil, Mark; Paterson, Carl; Knight, Jonathan; French, Paul; Dunsby, Chris

    2015-01-01

    To address the challenge of sample motion during in vivo imaging, we present a fibre-coupled multiphoton microscope with active axial motion compensation. The position of the sample surface is measured using optical coherence tomography and fed back to a piezo actuator that adjusts the axial location of the objective to compensate for sample motion. We characterise the system’s performance and demonstrate that it can compensate for axial sample velocities up to 700 µm/s. Finally we illustrate the impact of motion compensation when imaging multiphoton excited autofluorescence in ex vivo mouse skin. PMID:26137387

  17. Fibre-coupled multiphoton microscope with adaptive motion compensation.

    PubMed

    Sherlock, Ben; Warren, Sean; Stone, James; Neil, Mark; Paterson, Carl; Knight, Jonathan; French, Paul; Dunsby, Chris

    2015-05-01

    To address the challenge of sample motion during in vivo imaging, we present a fibre-coupled multiphoton microscope with active axial motion compensation. The position of the sample surface is measured using optical coherence tomography and fed back to a piezo actuator that adjusts the axial location of the objective to compensate for sample motion. We characterise the system's performance and demonstrate that it can compensate for axial sample velocities up to 700 m/s. Finally we illustrate the impact of motion compensation when imaging multiphoton excited autofluorescence in ex vivo mouse skin. PMID:26137387

  18. Experimental Resonance Enhanced Multiphoton Ionization (REMPI) studies of small molecules

    NASA Technical Reports Server (NTRS)

    Dehmer, J. L.; Dehmer, P. M.; Pratt, S. T.; Ohalloran, M. A.; Tomkins, F. S.

    1987-01-01

    Resonance enhanced multiphoton ionization (REMPI) utilizes tunable dye lasers to ionize an atom or molecule by first preparing an excited state by multiphoton absorption and then ionizing that state before it can decay. This process is highly selective with respect to both the initial and resonant intermediate states of the target, and it can be extremely sensitive. In addition, the products of the REMPI process can be detected as needed by analyzing the resulting electrons, ions, fluorescence, or by additional REMPI. This points to a number of exciting opportunities for both basic and applied science. On the applied side, REMPI has great potential as an ultrasensitive, highly selective detector for trace, reactive, or transient species. On the basic side, REMPI affords an unprecedented means of exploring excited state physics and chemistry at the quantum-state-specific level. An overview of current studies of excited molecular states is given to illustrate the principles and prospects of REMPI.

  19. Performances of high numerical aperture water and oil immersion objective in deep-tissue, multi-photon microscopic imaging of excised human skin.

    PubMed

    Dong, Chen-Yuan; Yu, Betty; Kaplan, Peter D; So, Peter T C

    2004-01-01

    Multi-photon fluorescence microscopy (MPFM) is a powerful technique for imaging scattering, biological specimens in depth. In addition to the sectioning effect generated by the point-like excitation volume, the near-infrared wavelengths used for multi-photon excitation allow deeper penetration into optically turbid specimens. In physiological specimens, the optical properties such as the scattering coefficients and refractive indices are often heterogeneous. In these specimens, it is not clear which type of immersion objective can provide optimized images in-depth. In particular, in-depth dermatological imaging applications using MPFM requires such optimization to obtain qualitative and quantitative information from the skin specimens. In this work, we address this issue by comparing the performances of two common types of high numerical aperture (NA) objectives: water-immersion and oil-immersion. A high-quality water-immersion objective (Zeiss, 40 x C-Apochromat, NA 1.2) and a comparable oil-immersion objective (Zeiss, 40 x Fluar, NA 1.25) were used for in-depth imaging of autofuorescent excised human skin and sulforhodamine B treated human skin specimens. Our results show that in the epidermal layers, the two types of immersion objectives perform comparably. However, in the dermis, multi-photon imaging using the oil immersion objective results in stronger fluorescence detection. These observations are most likely due to the degraded point-spread-function (PSF) caused by refractive index mismatch between the epidermis and the dermis. PMID:14677137

  20. Multiphoton FLIM: a reliable FRET detection tool in cell biological applications

    NASA Astrophysics Data System (ADS)

    Krishnan, Ramanujan V.; Biener, Eva; Centonze, Victoria E.; Gertler, Arieh; Herman, Brian A.

    2004-06-01

    Fluorescence lifetime imaging microscopy (FLIM) using multiphoton excitation is emerging as a reliable quantitative tool for measuring fluorescence resonance energy transfer (FRET) in living cells. By virtue of being free from spectroscopic artifacts encountered in conventional FRET detection methods, multiphoton FLIM methods offer the advantages of high spatial and temporal resolution, faster data acquisition and data analysis. We compare the FRET results obtained by two different methods namely (i) multiphoton excitation lifetime-based FRET and (ii) single photon excitation intensity-based acceptor photobleaching FRET. Using the same biological samples, we apply these two different methods in understanding the growth hormone receptor dimerization kinetics at the cell surface of human embryonic kidney cells. We conclude that the multiphoton FLIM using the streak-camera approach provides the best ability to monitor FRET in dynamic situations where high temporal and spatial resolution are required with minimal photodamage/phototoxicity.

  1. Generalized Multiphoton Quantum Interference

    NASA Astrophysics Data System (ADS)

    Tillmann, Max; Tan, Si-Hui; Stoeckl, Sarah E.; Sanders, Barry C.; de Guise, Hubert; Heilmann, René; Nolte, Stefan; Szameit, Alexander; Walther, Philip

    2015-10-01

    Nonclassical interference of photons lies at the heart of optical quantum information processing. Here, we exploit tunable distinguishability to reveal the full spectrum of multiphoton nonclassical interference. We investigate this in theory and experiment by controlling the delay times of three photons injected into an integrated interferometric network. We derive the entire coincidence landscape and identify transition matrix immanants as ideally suited functions to describe the generalized case of input photons with arbitrary distinguishability. We introduce a compact description by utilizing a natural basis that decouples the input state from the interferometric network, thereby providing a useful tool for even larger photon numbers.

  2. Record Multiphoton Absorption Cross-Sections by Dendrimer Organometalation.

    PubMed

    Simpson, Peter V; Watson, Laurance A; Barlow, Adam; Wang, Genmiao; Cifuentes, Marie P; Humphrey, Mark G

    2016-02-01

    Large increases in molecular two-photon absorption, the onset of measurable molecular three-photon absorption, and record molecular four-photon absorption in organic ?-delocalizable frameworks are achieved by incorporation of bis(diphosphine)ruthenium units with alkynyl linkages. The resultant ruthenium alkynyl-containing dendrimers exhibit strong multiphoton absorption activity through the biological and telecommunications windows in the near-infrared region. The ligated ruthenium units significantly enhance solubility and introduce fully reversible redox switchability to the optical properties. Increasing the ruthenium content leads to substantial increases in multiphoton absorption properties without any loss of optical transparency. This significant improvement in multiphoton absorption performance by incorporation of the organometallic units into the organic ?-framework is maintained when the relevant parameters are scaled by molecular weights or number of delocalizable ?-electrons. The four-photon absorption cross-section of the most metal-rich dendrimer is an order of magnitude greater than the previous record value. PMID:26797727

  3. A novel flexible clinical multiphoton tomograph for early melanoma detection, skin analysis, testing of anti-age products, and in situ nanoparticle tracking

    NASA Astrophysics Data System (ADS)

    Weinigel, Martin; Breunig, Hans Georg; Gregory, Axel; Fischer, Peter; Kellner-Hfer, Marcel; Bckle, Rainer; Knig, Karsten

    2010-02-01

    High-resolution 3D microscopy based on multiphoton induced autofluorescence and second harmonic generation have been introduced in 1990. 13 years later, CE-marked clinical multiphoton systems for 3D imaging of human skin with subcellular resolution have first been launched by JenLab company with the tomography DermaInspect. This year, the second generation of clinical multiphoton tomographs was introduced. The novel multiphoton tomograph MPTflex, equipped with a flexible articulated optical arm, provides an increased flexibility and accessibility especially for clinical and cosmetical examinations. Improved image quality and signal to noise ratio (SNR) are achieved by a very short source-drain spacing, by larger active areas of the detectors and by single photon counting (SPC) technology. Shorter image acquisition time due to improved image quality reduces artifacts and simplifies the operation of the system. The compact folded optical design and the light-weight structure of the optical head eases the handling. Dual channel detectors enable to distinguish between intratissue elastic fibers and collagenous structures simultaneously. Through the use of piezo-driven optics a stack of optical cross-sections (optical sectioning) can be acquired and 3D imaging can be performed. The multiphoton excitation of biomolecules like NAD(P)H, flavins, porphyrins, elastin, and melanin is done by picojoule femtosecond laser pulses from an tunable turn-key femtosescond near infrared laser system. The ability for rapid high-quality image acquisition, the user-friendly operation of the system and the compact and flexible design qualifies this system to be used for melanoma detection, diagnostics of dermatological disorders, cosmetic research and skin aging measurements as well as in situ drug monitoring and animal research.

  4. High-resolution multimodal clinical multiphoton tomography of skin

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2011-03-01

    This review focuses on multimodal multiphoton tomography based on near infrared femtosecond lasers. Clinical multiphoton tomographs for 3D high-resolution in vivo imaging have been placed into the market several years ago. The second generation of this Prism-Award winning High-Tech skin imaging tool (MPTflex) was introduced in 2010. The same year, the world's first clinical CARS studies have been performed with a hybrid multimodal multiphoton tomograph. In particular, non-fluorescent lipids and water as well as mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen has been imaged with submicron resolution in patients suffering from psoriasis. Further multimodal approaches include the combination of multiphoton tomographs with low-resolution wide-field systems such as ultrasound, optoacoustical, OCT, and dermoscopy systems. Multiphoton tomographs are currently employed in Australia, Japan, the US, and in several European countries for early diagnosis of skin cancer, optimization of treatment strategies, and cosmetic research including long-term testing of sunscreen nanoparticles as well as anti-aging products.

  5. A series of flexible design adaptations to the Nikon E-C1 and E-C2 confocal microscope systems for UV, multiphoton and FLIM imaging.

    PubMed

    Botchway, Stanley W; Scherer, Kathrin M; Hook, Steve; Stubbs, Christopher D; Weston, Eleanor; Bisby, Roger H; Parker, Anthony W

    2015-04-01

    Multiphoton microscopy is widely employed in the life sciences using extrinsic fluorescence of low- and high-molecular weight labels with excitation and emission spectra in the visible and near infrared regions. For imaging of intrinsic and extrinsic fluorophores with excitation spectra in the ultraviolet region, multiphoton excitation with one- or two-colour lasers avoids the need for ultraviolet-transmitting excitation optics and has advantages in terms of optical penetration in the sample and reduced phototoxicity. Excitation and detection of ultraviolet emission around 300 nm and below in a typical inverted confocal microscope is more difficult and requires the use of expensive quartz optics including the objective. In this technical note we describe the adaptation of a commercial confocal microscope (Nikon, Japan E-C1 or E-C2) for versatile use with Ti-sapphire and OPO laser sources and the addition of a second detection channel that enables detection of ultraviolet fluorescence and increases detection sensitivity in a typical fluorescence lifetime imaging microscopy experiment. Results from some experiments with this setup illustrate the resulting capabilities. PMID:25664385

  6. The stepwise multi-photon activation fluorescence guided ablation of melanin

    NASA Astrophysics Data System (ADS)

    Lai, Zhenhua; Gu, Zetong; DiMarzio, Charles

    2015-02-01

    Previous research has shown that the stepwise multi-photon activation fluorescence (SMPAF) of melanin, activated and excited by a continuous-wave (CW) mode near infrared (NIR) laser, is a low-cost and reliable method for detecting melanin. We have developed a device utilizing the melanin SMPAF to guide the ablation of melanin with a 975 nm CW laser. This method provides the ability of targeting individual melanin particles with micrometer resolution, and enables localized melanin ablation to be performed without collateral damage. Compared to the traditional selective photothermolysis, which uses pulsed lasers for melanin ablation, this method demonstrates higher precision and lower cost. Therefore, the SMPAF guided selective ablation of melanin is a promising tool of melanin ablation for both medical and cosmetic purposes.

  7. A radiative transfer model to treat infrared molecular excitation in cometary atmospheres

    NASA Astrophysics Data System (ADS)

    Debout, V.; Bockelée-Morvan, D.; Zakharov, V.

    2016-02-01

    The exospheres of small Solar System bodies are now observed with high spatial resolution from space missions. Interpreting infrared spectra of cometary gases obtained with the VIRTIS experiment onboard the Rosetta cometary mission requires detailed modeling of infrared fluorescence emission in optically thick conditions. Efficient computing methods are required since numerous ro-vibrational lines excited by the Sun need to be considered. We propose a new model working in a 3-D environment to compute numerically the local incoming radiation. It uses a new algorithm using pre-defined directions of ray propagation and ray grids to reduce the CPU cost in time with respect to Monte Carlo methods and to treat correctly the sunlight direction. The model is applied to the ν3 bands of CO2 and H2O at 4.3 μ m and 2.7 μ m respectively, and to the CO ∨ (1 → 0) band at 4.7 μ m. The results are compared to the ones obtained by a 1-D algorithm which uses the Escape Probability (EP) method, and by a 3-D "Coupled Escape Probability" (CEP) model, for different levels of optical thickness. Our results suggest that the total band flux may vary strongly with azimuth for optically thick cases whereas the azimuth average total band flux computed is close to the one obtained with EP. Our model globally predicts less intensity reduction from opacity than the CEP model of Gersch and A'Hearn (Gersch, A.M., A'Hearn, M.F. [2014]. Astrophys. J. 787, 36-56). An application of the model to the observation of CO2, CO and H2O bands in 67/P atmosphere with VIRTIS is presented to predict the evolution of band optical thickness along the mission.

  8. Red-emitting upconverting nanoparticles for photodynamic therapy in cancer cells under near-infrared excitation.

    PubMed

    Tian, Gan; Ren, Wenlu; Yan, Liang; Jian, Shan; Gu, Zhanjun; Zhou, Liangjun; Jin, Shan; Yin, Wenyan; Li, Shoujian; Zhao, Yuliang

    2013-06-10

    Upconverting nanoparticles (UCNPs) have attracted considerable attention as potential photosensitizer carriers for photodynamic therapy (PDT) in deep tissues. In this work, a new and efficient NIR photosensitizing nanoplatform for PDT based on red-emitting UCNPs is designed. The red emission band matches well with the efficient absorption bands of the widely used commercially available photosensitizers (Ps), benefiting the fluorescence resonance energy transfer (FRET) from UCNPs to the attached photosensitizers and thus efficiently activating them to generate cytotoxic singlet oxygen. Three commonly used photosensitizers, including chlorine e6 (Ce6), zinc phthalocyanine (ZnPc) and methylene blue (MB), are loaded onto the alpha-cyclodextrin-modified UCNPs to form Ps@UCNPs complexes that efficiently produce singlet oxygen to kill cancer cells under 980 nm near-infrared excitation. Moreover, two different kinds of drugs are co-loaded onto these nanoparticles: chemotherapy drug doxorubicin and PDT agent Ce6. The combinational therapy based on doxorubicin (DOX)-induced chemotherapy and Ce6-triggered PDT exhibits higher therapeutic efficacy relative to the individual means for cancer therapy in vitro. PMID:23239556

  9. Visible-near-infrared luminescent lanthanide ternary complexes based on beta-diketonate using visible-light excitation.

    PubMed

    Sun, Lining; Qiu, Yannan; Liu, Tao; Feng, Jing; Deng, Wei; Shi, Liyi

    2015-11-01

    We used the synthesized dinaphthylmethane (Hdnm) ligand whose absorption extends to the visible-light wavelength, to prepare a family of ternary lanthanide complexes, named as [Ln(dnm)3 phen] (Ln = Sm, Nd, Yb, Er, Tm, Pr). The properties of these complexes were investigated by Fourier transform infrared (FT-IR) spectroscopy, diffuse reflectance (DR) spectroscopy, thermogravimetric analyses, and excitation and emission spectroscopy. Generally, excitation with visible light is much more advantageous than UV excitation. Importantly, upon excitation with visible light (401-460 nm), the complexes show characteristic visible (Sm(3+)) as well as near-infrared (Sm(3+), Nd(3+), Yb(3+), Er(3+), Tm(3+), Pr(3+)) luminescence of the corresponding lanthanide ions, attributed to the energy transfer from the ligands to the lanthanide ions, an antenna effect. Now, using these near-infrared luminescent lanthanide complexes, the luminescent spectral region from 800 to 1650 nm, can be covered completely, which is of particular interest for biomedical imaging applications, laser systems, and optical amplification applications. PMID:25691149

  10. Transverse correlations in multiphoton entanglement

    SciTech Connect

    Wen Jianming; Rubin, Morton H.; Shih Yanhua

    2007-10-15

    We have analyzed the transverse correlation in multiphoton entanglement. The generalization of quantum ghost imaging is extended to the N-photon state. The Klyshko's two-photon advanced-wave picture is generalized to the N-photon case.

  11. Biomedical applications involving multiphoton probes

    NASA Astrophysics Data System (ADS)

    Potasek, M.; Beeson, K.; Parilov, E.

    2015-03-01

    Many techniques in biological and clinical science use multiphoton absorbers for fluorescence. The applications include medical imaging for living cells, diagnostic techniques for disease and spectroscopy. The intrinsic value of the multiphoton absorber coefficients is therefore of the utmost importance. Additionally, the laser intensity at which the absorber saturates can determine which absorber, dye or protein is useful for a particular application. Yet, experimental methods for determining the optical coefficients often yield different results. We describe several common methods of 2PA measurements and describe their features. As an example of the importance of applying the correct analysis to measurements, we fit experimental data and obtain values for multiphoton absorbers and accurately obtain their intrinsic values. Finally, we present the optical properties of several multiphoton materials used in biology.

  12. Maintaining polarization in polarimetric multiphoton microscopy.

    PubMed

    Bélanger, Erik; Turcotte, Raphaël; Daradich, Amy; Sadetsky, Grégory; Gravel, Pierre; Bachand, Karine; De Koninck, Yves; Côté, Daniel C

    2015-11-01

    Polarimetric measurements in multiphoton microscopy can reveal information about the local molecular order of a sample. However, the presence of a dichroic through which the excitation beam propagates will generally scramble its polarization. We propose a simple scheme whereby a second properly-oriented compensation dichroic is used to negate any alteration regardless of the wavelength and the initial polarization. We demonstrate how this robust and rapid approach simplifies polarimetric measurements in second-harmonic generation, two-photon excited fluorescence and coherent anti-Stokes Raman scattering. Illustration of the polarization maintaining strategy with the compensating dichroic oriented such that its s- and p-axes are interchanged with these of the primary dichroic. PMID:25691172

  13. Multiphoton FLIM and spectral imaging of cells and tissues

    NASA Astrophysics Data System (ADS)

    Konig, Karsten; Riemann, Iris; Ehrlich, Gunter; Ulrich, Volker; Fischer, Peter

    2004-06-01

    Five-dimensional (5D) multiphoton measurements with submicron spatial resolution, 270 ps temporal resolution and 5 nm spectral resolution have been performed on living cells and tissues at 750 nm - 850 nm laser excitation. A compact (65x62x48 cm3) multiport laser scanning microscope TauMap (JenLab GmbH) equipped with fast PMT and CCD camera, SPC 830 time-correlated single photon counting board and Sagnac interferometer was used. Laser excitation radiation was provided by a tuneable MaiTai Ti:sapphire femtosecond laser as well as by a 405 nm 50 MHz picosecond laser diode. The spectral and temporal fluorescence behaviour of intratissue chloroplasts of water plant leafs, of a variety of exogenous fluorophores as well as of fluorescent proteins in transfected brain cells have been studied. When calculating fluorescence lifetime images (FLIM) we found differences in intracellular twophoton fluorescence lifetimes vs. one-photon fluorescence lifetimes. Multiphoton FLIM-FRET and multiphoton spectral FRET studies have been performed in living HBMEC brain cells using CFP and YFP fusion proteins. It was shown that FLIM-FRET data depend on laser power due to photodestructive multiphoton effects. This has to be considered in long-term fluorescence resonance energy transfer studies of dynamic protein-protein interactions.

  14. Acousto-optic multiphoton laser scanning microscopy and multiphoton photon counting spectroscopy: Applications and implications for optical neurobiology

    NASA Astrophysics Data System (ADS)

    Iyer, Vijay

    Multiphoton excitation of molecular probes has become an important tool in experimental neurobiology owing to the intrinsic optical sectioning and low light scattering it affords. Using molecular functional indicators, multiphoton excitation allows physiological signals within single neurons to be observed from within living brain tissue. Ideally, it would be possible to record from multiple sites located throughout the elaborately branching dendritic arbors, in order to study the correlations of structure and function both within and across experiments. However, existing multiphoton microscope systems based on scanning mirrors do not allow optical recordings to be obtained from more than a handful of sites simultaneously at the high rates required to capture the fast physiological signals of interest (>100Hz for Ca2+ signals, >1kHz for membrane potential transients). In order to overcome this limitation, two-dimensional acousto-optic deflection was employed, to allow an ultrafast laser beam suited for multiphoton excitation to be rapidly repositioned with low latency (15mus). This supports a random-access scanning mode in which the beam can repeatedly visit a succession of user-selected sites of interest within the microscope's field-of-view at high rates, with minimal sacrifice of pixel dwell time. This technique of acousto-optic multiphoton laser scanning microscope (AO-MPLSM) was demonstrated to allow the spatial profile of signals arising in response to physiological stimulation to be rapidly mapped. Means to compensate or avoid problems of dispersion which have hampered AO-MPLSM in the past are presented, with the latter being implemented. Separately, the combination of photon counting detection with multiphoton excitation, termed generally multiphoton photon counting spectroscopy (MP-PCS), was also considered, with particular emphasis on the technique of fluorescence correlation spectroscopy (FCS). MP-PCS was shown to allow information about molecular numbers and mobility, as well as the focal volume itself, to be obtained. This capability may in the future be employed to study the number and transport of native neuronal signaling molecules. MP-PCS was also found to be a promising off-line tool which can allow the performance of AO-MPLSM to be optimized, with respect to both the instrument and the indicators employed.

  15. Enhanced thermal radiation in terahertz and far-infrared regime by hot phonon excitation in a field effect transistor

    SciTech Connect

    Chung, Pei-Kang; Yen, Shun-Tung

    2014-11-14

    We demonstrate the hot phonon effect on thermal radiation in the terahertz and far-infrared regime. A pseudomorphic high electron mobility transistor is used for efficiently exciting hot phonons. Boosting the hot phonon population can enhance the efficiency of thermal radiation. The transistor can yield at least a radiation power of 13 μW and a power conversion efficiency higher than a resistor by more than 20%.

  16. Pulsed infrared radiation excites cultured neonatal spiral and vestibular ganglion neurons by modulating mitochondrial calcium cycling

    PubMed Central

    Lumbreras, Vicente; Bas, Esperanza; Gupta, Chhavi

    2014-01-01

    Cochlear implants are currently the most effective solution for profound sensorineural hearing loss, and vestibular prostheses are under development to treat bilateral vestibulopathies. Electrical current spread in these neuroprostheses limits channel independence and, in some cases, may impair their performance. In comparison, optical stimuli that are spatially confined may result in a significant functional improvement. Pulsed infrared radiation (IR) has previously been shown to elicit responses in neurons. This study analyzes the response of neonatal rat spiral and vestibular ganglion neurons in vitro to IR (wavelength = 1,863 nm) using Ca2+ imaging. Both types of neurons responded consistently with robust intracellular Ca2+ ([Ca2+]i) transients that matched the low-frequency IR pulses applied (4 ms, 0.251 pps). Radiant exposures of ?637 mJ/cm2 resulted in continual neuronal activation. Temperature or [Ca2+] variations in the media did not alter the IR-evoked transients, ruling out extracellular Ca2+ involvement or primary mediation by thermal effects on the plasma membrane. While blockage of Na+, K+, and Ca2+ plasma membrane channels did not alter the IR-evoked response, blocking of mitochondrial Ca2+ cycling with CGP-37157 or ruthenium red reversibly inhibited the IR-evoked [Ca2+]i transients. Additionally, the magnitude of the IR-evoked transients was dependent on ryanodine and cyclopiazonic acid-dependent Ca2+ release. These results suggest that IR modulation of intracellular calcium cycling contributes to stimulation of spiral and vestibular ganglion neurons. As a whole, the results suggest selective excitation of neurons in the IR beam path and the potential of IR stimulation in future auditory and vestibular prostheses. PMID:24920028

  17. Pulsed infrared radiation excites cultured neonatal spiral and vestibular ganglion neurons by modulating mitochondrial calcium cycling.

    PubMed

    Lumbreras, Vicente; Bas, Esperanza; Gupta, Chhavi; Rajguru, Suhrud M

    2014-09-15

    Cochlear implants are currently the most effective solution for profound sensorineural hearing loss, and vestibular prostheses are under development to treat bilateral vestibulopathies. Electrical current spread in these neuroprostheses limits channel independence and, in some cases, may impair their performance. In comparison, optical stimuli that are spatially confined may result in a significant functional improvement. Pulsed infrared radiation (IR) has previously been shown to elicit responses in neurons. This study analyzes the response of neonatal rat spiral and vestibular ganglion neurons in vitro to IR (wavelength = 1,863 nm) using Ca(2+) imaging. Both types of neurons responded consistently with robust intracellular Ca(2+) ([Ca(2+)]i) transients that matched the low-frequency IR pulses applied (4 ms, 0.25-1 pps). Radiant exposures of ∼637 mJ/cm(2) resulted in continual neuronal activation. Temperature or [Ca(2+)] variations in the media did not alter the IR-evoked transients, ruling out extracellular Ca(2+) involvement or primary mediation by thermal effects on the plasma membrane. While blockage of Na(+), K(+), and Ca(2+) plasma membrane channels did not alter the IR-evoked response, blocking of mitochondrial Ca(2+) cycling with CGP-37157 or ruthenium red reversibly inhibited the IR-evoked [Ca(2+)]i transients. Additionally, the magnitude of the IR-evoked transients was dependent on ryanodine and cyclopiazonic acid-dependent Ca(2+) release. These results suggest that IR modulation of intracellular calcium cycling contributes to stimulation of spiral and vestibular ganglion neurons. As a whole, the results suggest selective excitation of neurons in the IR beam path and the potential of IR stimulation in future auditory and vestibular prostheses. PMID:24920028

  18. Photochemistry of UV-excited trifluoroacetylacetone and hexafluoroacetylacetone I: infrared spectra of fluorinated methylfuranones formed by HF photoelimination.

    PubMed

    Muyskens, Karen J; Alsum, Joel R; Thielke, Timothy A; Boer, Jodi L; Heetderks, Tina R; Muyskens, Mark A

    2012-12-20

    The photochemistry of gas-phase 1,1,1-trifluoroacetylacetone (TFAA) excited with ultraviolet (UV) light involves a significant photoelimination channel that produces hydrogen fluoride and a fluorinated methylfuranone, 2,2-difluoro-5-methyl-3(2H)-furanone (2FMF). This pathway is remarkable because it is a gas-phase unimolecular reaction that forms a five-membered ring product. This report is the first of such a TFAA photoelimination channel, which is similar to one observed with 1,1,1,5,5,5-hexafluoroacetylacetone (HFAA), resulting in 2,2-difluoro-5-trifluoromethyl-3(2H)-furanone. We present infrared spectral observations of 2FMF produced by pulsed, UV-laser excitation of TFAA, along with analogous results from HFAA, supported by density functional theory (DFT) computational studies. DFT results for the infrared spectrum of 5-methyl-3(2H)-furanone, the expected comparable acetylacetone photoelimination product, help suggest that UV excitation of acetylacetone fails to follow a similar type of photoelimination. We use a weighted RMS approach as a figure of merit for comparing calculated infrared frequencies with experimental data. Results from the three acetylacetones reveal how the presence of fluorine atoms in acetylacetone influences the gas-phase molecular photochemistry. PMID:23176295

  19. Development and characterization of non-resonant multiphoton photoacoustic spectroscopy (NMPPAS) for brain tumor margining

    NASA Astrophysics Data System (ADS)

    Dahal, Sudhir

    During tumor removal surgery, due to the problems associated with obtaining high-resolution, real-time chemical images of where exactly the tumor ends and healthy tissue begins (tumor margining), it is often necessary to remove a much larger volume of tissue than the tumor itself. In the case of brain tumor surgery, however, it is extremely unsafe to remove excess tissue. Therefore, without an accurate image of the tumor margins, some of the tumor's finger-like projections are inevitably left behind in the surrounding parenchyma to grow again. For this reason, the development of techniques capable of providing high-resolution real-time images of tumor margins up to centimeters below the surface of a tissue is ideal for the diagnosis and treatment of tumors, as well as surgical guidance during brain tumor excision. A novel spectroscopic technique, non-resonant multiphoton photoacoustic spectroscopy (NMPPAS), is being developed with the capabilities of obtaining high-resolution subsurface chemical-based images of underlying tumors. This novel technique combines the strengths of multiphoton tissue spectroscopy and photoacoustic spectroscopy into a diagnostic methodology that will, ultimately, provide unparalleled chemical information and images to provide the state of sub-surface tissues. The NMPPAS technique employs near-infrared light (in the diagnostic window) to excite ultraviolet and/or visible light absorbing species deep below the tissue's surface. Once a multiphoton absorption event occurs, non-radiative relaxation processes generates a localized thermal expansion and subsequent acoustic wave that can be detected using a piezoelectric transducer. Since NMPPAS employs an acoustic detection modality, much deeper diagnoses can be performed than that is possible using current state of the art high-resolution chemical imaging techniques such as multiphoton fluorescence spectroscopy. NMPPAS was employed to differentiate between excised brain tumors (astrocytoma III) and healthy tissue with over 99% accuracy. NMPPAS spectral features showed evident differences between tumor and healthy tissues, and ratiometric analysis ensured that only a few wavelengths could be used for excitation instead of using numerous wavelength excitations to create spectra. This process would significantly reduce the analysis time while maintaining the same degree of accuracy. Tissue phantoms were fabricated in order to characterize the properties of NMPPAS. Scattering particles were doped into the phantoms to simulate their light scattering properties to real tissues. This allowed for better control over shape, size, reproducibility and doping in the sample while maintaining the light-tissue interaction properties of real tissue. To make NMPPAS viable for clinical applications, the technique was characterized to determine the spatial (lateral and longitudinal) resolution, depth of penetration and its ability to image in three-dimension through layers of tissue. Both resolutions were determined to be near-cellular level resolution (50-70 microm), obtained initially with the aid of the technique of multiphoton fluorescence, and later verified using NMPPAS imaging. Additionally, the maximum depth of penetration and detection was determined to be about 1.4cm, making the technique extremely suitable to margin tumors from underlying tissues in the brain. The capability of NMPPAS to detect and image layers that lie beneath other structures and blood vessels was also investigated. Three-dimensional images were obtained for the first time using NMPPAS. The images were obtained from different depths and structures were imaged through other layers of existing structures in the sample. This verified that NMPPAS was capable of detecting and imaging structures that lie embedded within the tissues. NMPPAS images of embedded structures were also obtained with the presence of hemoglobin, which is potentially the largest source of background in blood-perfused tissues, thus showing that the technique is capable of detecting and differentiating in blood-perfused samples.

  20. Quantitative multiphoton imaging

    NASA Astrophysics Data System (ADS)

    Knig, Karsten; Weinigel, Martin; Breunig, Hans Georg; Uchugonova, Aisada

    2014-02-01

    Certified clinical multiphoton tomographs for label-free multidimensional high-resolution in vivo imaging have been introduced to the market several years ago. Novel tomographs include a flexible 360 scan head attached to a mechanooptical arm for autofluorescence and SHG imaging as well as a CARS module. Non-fluorescent lipids and water, mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen can be imaged in vivo with submicron resolution in human skin. Sensitive and rapid detectors allow single photon counting and the construction of 3D maps where the number of detected photons per voxel is depicted. Intratissue concentration profiles from endogenous as well exogenous substances can be generated when the number of detected photons can be correlated with the number of molecules with respect to binding and scattering behavior. Furthermore, the skin ageing index SAAID based on the ratio elastin/collagen as well as the epidermis depth based on the onset of SHG generation can be determined.

  1. Multiphoton tomography of astronauts

    NASA Astrophysics Data System (ADS)

    König, Karsten; Weinigel, Martin; Pietruszka, Anna; Bückle, Rainer; Gerlach, Nicole; Heinrich, Ulrike

    2015-03-01

    Weightlessness may impair the astronaut's health conditions. Skin impairments belong to the most frequent health problems during space missions. Within the Skin B project, skin physiological changes during long duration space flights are currently investigated on three European astronauts that work for nearly half a year at the ISS. Measurements on the hydration, the transepidermal water loss, the surface structure, elasticity and the tissue density by ultrasound are conducted. Furthermore, high-resolution in vivo histology is performed by multiphoton tomography with 300 nm spatial and 200 ps temporal resolution. The mobile certified medical tomograph with a flexible 360° scan head attached to a mechano-optical arm is employed to measure two-photon autofluorescence and SHG in the volar forearm of the astronauts. Modification of the tissue architecture and of the fluorescent biomolecules NAD(P)H, keratin, melanin and elastin are detected as well as of SHG-active collagen. Thinning of the vital epidermis, a decrease of the autofluoresence intensity, an increase in the long fluorescence lifetime, and a reduced skin ageing index SAAID based on an increased collagen level in the upper dermis have been found. Current studies focus on recovery effects.

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

    PubMed Central

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

    2015-01-01

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

  3. Enhancement of visible He2 emission by infrared laser excitation of He metastable atoms in atmospheric-pressure plasma jet

    NASA Astrophysics Data System (ADS)

    Urabe, Keiichiro; Motomura, Hideki; Sakai, Osamu; Tachibana, Kunihide

    2011-02-01

    We observed bright red emission at an intersection of an infrared (IR) laser beam and an atmospheric-pressure plasma jet (APPJ). The APPJ had a coaxial dielectric barrier discharge configuration and generated a plasma plume in a He gas flow ejected into air from a glass capillary. Although the laser-beam wavelength was 1083 nm corresponding to He I (23P-23S) atomic transition, the enhanced red emission peaked in the visible region at 640 nm and had broad bandwidth corresponding to He2 (d\\,^{3}\\!\\Sigma_u^{+}{{--}}b\\,^{3}\\Pi _g) molecular transition. This result indicates that the IR laser excitation of He metastable (23S) atoms leads to selective formation of excited He2 (d\\,^{3}\\!\\Sigma_u^{+}) molecules from excited He (23P) atoms. The 640 nm emission intensity decreased as the laser beam moved downstream in the plasma plume, because of the quenching effects on the excited He atoms and the excited He2 molecules by air entrainment. We believe that the He2 (d\\,^{3}\\!\\Sigma_u^{+}) molecular formation follows two potential curves of He2 (f 3Σu or f 3Πu) and (d\\,^{3}\\!\\Sigma_u^{+}) states, overcoming a potential hump by three-body collision processes.

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

    NASA Astrophysics Data System (ADS)

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

    2015-08-01

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

  5. Comparison of objective lenses for multiphoton microscopy in turbid samples.

    PubMed

    Singh, Avtar; McMullen, Jesse D; Doris, Eli A; Zipfel, Warren R

    2015-08-01

    Optimization of illumination and detection optics is pivotal for multiphoton imaging in highly scattering tissue and the objective lens is the central component in both of these pathways. To better understand how basic lens parameters (NA, magnification, field number) affect fluorescence collection and image quality, a two-detector setup was used with a specialized sample cell to separate measurement of total excitation from epifluorescence collection. Our data corroborate earlier findings that low-mag lenses can be superior at collecting scattered photons, and we compare a set of commonly used multiphoton objective lenses in terms of their ability to collect scattered fluorescence, providing guidance for the design of multiphoton imaging systems. For example, our measurements of epi-fluorescence beam divergence in the presence of scattering reveal minimal beam broadening, indicating that often-advocated over-sized collection optics are not as advantageous as previously thought. These experiments also provide a framework for choosing objective lenses for multiphoton imaging by relating the results of our measurements to various design parameters of the objectives lenses used. PMID:26309771

  6. Comparison of objective lenses for multiphoton microscopy in turbid samples

    PubMed Central

    Singh, Avtar; McMullen, Jesse D.; Doris, Eli A.; Zipfel, Warren R.

    2015-01-01

    Optimization of illumination and detection optics is pivotal for multiphoton imaging in highly scattering tissue and the objective lens is the central component in both of these pathways. To better understand how basic lens parameters (NA, magnification, field number) affect fluorescence collection and image quality, a two-detector setup was used with a specialized sample cell to separate measurement of total excitation from epifluorescence collection. Our data corroborate earlier findings that low-mag lenses can be superior at collecting scattered photons, and we compare a set of commonly used multiphoton objective lenses in terms of their ability to collect scattered fluorescence, providing guidance for the design of multiphoton imaging systems. For example, our measurements of epi-fluorescence beam divergence in the presence of scattering reveal minimal beam broadening, indicating that often-advocated over-sized collection optics are not as advantageous as previously thought. These experiments also provide a framework for choosing objective lenses for multiphoton imaging by relating the results of our measurements to various design parameters of the objectives lenses used. PMID:26309771

  7. Optimization of silver nanoparticles for surface enhanced Raman spectroscopy of structurally diverse analytes using visible and near-infrared excitation.

    PubMed

    Meyer, Matthew W; Smith, Emily A

    2011-09-01

    Several experimental parameters affecting surface enhanced Raman (SER) signals using 488, 785 and 1064 nm excitation for eight diverse analytes are reported. Citrate reduced silver colloids having average diameters ranging from 40 ± 10 to 100 ± 20 nm were synthesized. The nanoparticles were characterized by transmission electron microscopy, dynamic light scattering and absorbance spectrophotometry before and after inducing nanoparticle aggregation with 0.99% v/v 0.5 M magnesium chloride. The nanoparticle aggregates and SERS signal were stable between 30 and 90 minutes after inducing aggregation. For the analytes 4-mercaptopyridine, 4-methylthiobenzoic acid and the dipeptide phenylalanine-cysteine using all three excitation wavelengths, the highest surface area adjusted SER signal was obtained using 70 ± 20 nm nanoparticles, which generated 290 ± 40 nm aggregates with the addition of magnesium chloride. The decrease in the SER signal using non-optimum colloids was 12 to 42% using 488 nm excitation and larger decreases in signal, up to 92%, were observed using near infrared excitation wavelengths. In contrast, pyridine, benzoic acid, and phenylalanine required 220 ± 30 nm aggregates for the highest SER signal with 785 or 1064 nm excitation, but larger aggregates (290 ± 40 nm) were required with 488 nm excitation. The optimum experimental conditions measured with the small molecule analytes held for a 10 amino acid peptide and hemoglobin. Reproducible SERS measurements with 2 to 9% RSD have been obtained by considering nanoparticle size, aggregation conditions, excitation wavelength and the nature of the analyte-silver interaction. PMID:21301711

  8. Evaluation of Barrett Esophagus by Multiphoton Microscopy

    PubMed Central

    Chen, Jianxin; Wong, Serena; Nathanson, Michael H.; Jain, Dhanpat

    2014-01-01

    Context Multiphoton microscopy (MPM) based on 2-photon excitation fluorescence and second-harmonic generation allows simultaneous visualization of cellular details and extracellular matrix components of fresh, unfixed, and unstained tissue. Portable multiphoton microscopes, which could be placed in endoscopy suites, and multiphoton endomicroscopes are in development, but their clinical utility is unknown. Objectives To examine fresh, unfixed endoscopic biopsies obtained from the distal esophagus and gastroesophageal junction to (1) define the MPM characteristics of normal esophageal squamous mucosa and gastric columnar mucosa, and (2) evaluate whether diagnosis of intestinal metaplasia/Barrett esophagus (BE) could be made reliably with MPM. Design The study examined 35 untreated, fresh biopsy specimens from 25 patients who underwent routine upper endoscopy. A Zeiss LSM 710 Duo microscope (Carl Zeiss, Thornwood, New York) coupled to a Spectra-Physics (Mountain View, California) Tsunami Ti:sapphire laser was used to obtain a MPM image within 4 hours of fresh specimen collection. After obtaining MPM images, the biopsy specimens were placed in 10% buffered formalin and submitted for routine histopathologic examination. Then, the MPM images were compared with the findings in the hematoxylin-eosin–stained, formalin-fixed, paraffin-embedded sections. The MPM characteristics of the squamous, gastric-type columnar and intestinal-type columnar epithelium were analyzed. In biopsies with discrepancy between MPM imaging and hematoxylin-eosin–stained sections, the entire tissue block was serially sectioned and reevaluated. A diagnosis of BE was made when endoscopic and histologic criteria were satisfied. Results Based on effective 2-photon excitation fluorescence of cellular reduced pyridine nucleotides and flavin adenine dinucleotide and lack of 2-photon excitation fluorescence of mucin and cellular nuclei, MPM could readily identify and distinguish among squamous epithelial cells, goblet cells, gastric foveolar-type mucous cells, and parietal cells in the area of gastroesophageal junction. Based on the cell types identified, the mucosa was defined as squamous, columnar gastric type (cardia/fundic-type), and metaplastic columnar intestinal-type/BE. Various types of mucosa seen in the study of 35 biopsies included normal squamous mucosa only (n = 14; 40%), gastric cardia-type mucosa only (n = 2; 6%), gastric fundic mucosa (n = 6; 17%), and both squamous and gastric mucosa (n = 13; 37%). Intestinal metaplasia was identified by the presence of goblet cells in 10 of 25 cases (40%) leading to a diagnosis of BE on MPM imaging and only in 7 cases (28%) by histopathology. In 3 of 35 biopsies (9%), clear-cut goblet cells were seen by MPM imaging but not by histopathology, even after the entire tissue block was sectioned. Based on effective 2-photon excitation fluorescence of elastin and second-harmonic generation of collagen, connective tissue in the lamina propria and the basement membrane was also visualized with MPM. Conclusions Multiphoton microscopy has the ability to accurately distinguish squamous epithelium and different cellular elements of the columnar mucosa obtained from biopsies around the gastroesophageal junction, including goblet cells that are important for the diagnosis of BE. Thus, use of MPM in the endoscopy suite might provide immediate microscopic images during endoscopy, improving screening and surveillance of patients with BE. PMID:24476518

  9. Multiphoton microscopic imaging of rabbit dorsal skin.

    PubMed

    Zhu, Xiaoqin; Xu, Yahao; Hong, Zhipeng; Chen, Jing; Zhuo, Shuangmu; Chen, Jianxin

    2015-01-01

    Rabbits are often preferred to be experimental animals during the skin research. The visualizing and understanding the full-thickness structure of rabbit skin has significance in biology, medicine, and animal husbandry. In this study, multiphoton microscopy (MPM) was employed to examine the rabbit skin on the back, which was based on second harmonic generation and two-photon excited fluorescence. High-resolution images were achieved from the fresh, unfixed, and unstained tissues, showing detailed microstructure of the skin without the administration of exogenous contrast agents. The morphology and distribution of the main components of epidermis and dermis, such as keratin, collagen fibers, elastic fibers, and hair follicles, can be distinctly identified in MPM images. Since the changes in these components are tightly related to skin diseases and wound healing, the noninvasive nature of MPM enables it become a valuable tool in skin research for detecting and monitoring. PMID:25521496

  10. Advances in multiphoton microscopy technology

    PubMed Central

    Hoover, Erich E.; Squier, Jeff A.

    2013-01-01

    Multiphoton microscopy has enabled unprecedented dynamic exploration in living organisms. A significant challenge in biological research is the dynamic imaging of features deep within living organisms, which permits the real-time analysis of cellular structure and function. To make progress in our understanding of biological machinery, optical microscopes must be capable of rapid, targeted access deep within samples at high resolution. In this Review, we discuss the basic architecture of a multiphoton microscope capable of such analysis and summarize the state-of-the-art technologies for the quantitative imaging of biological phenomena. PMID:24307915

  11. Infrared

    NASA Astrophysics Data System (ADS)

    Vollmer, M.

    2013-11-01

    'Infrared' is a very wide field in physics and the natural sciences which has evolved enormously in recent decades. It all started in 1800 with Friedrich Wilhelm Herschel's discovery of infrared (IR) radiation within the spectrum of the Sun. Thereafter a few important milestones towards widespread use of IR were the quantitative description of the laws of blackbody radiation by Max Planck in 1900; the application of quantum mechanics to understand the rotational-vibrational spectra of molecules starting in the first half of the 20th century; and the revolution in source and detector technologies due to micro-technological breakthroughs towards the end of the 20th century. This has led to much high-quality and sophisticated equipment in terms of detectors, sources and instruments in the IR spectral range, with a multitude of different applications in science and technology. This special issue tries to focus on a few aspects of the astonishing variety of different disciplines, techniques and applications concerning the general topic of infrared radiation. Part of the content is based upon an interdisciplinary international conference on the topic held in 2012 in Bad Honnef, Germany. It is hoped that the information provided here may be useful for teaching the general topic of electromagnetic radiation in the IR spectral range in advanced university courses for postgraduate students. In the most general terms, the infrared spectral range is defined to extend from wavelengths of 780 nm (upper range of the VIS spectral range) up to wavelengths of 1 mm (lower end of the microwave range). Various definitions of near, middle and far infrared or thermal infrared, and lately terahertz frequencies, are used, which all fall in this range. These special definitions often depend on the scientific field of research. Unfortunately, many of these fields seem to have developed independently from neighbouring disciplines, although they deal with very similar topics in respect of the underlying physics. There are now at least six different disciplines that deal with infrared radiation in one form or another, and in one or several different spectral portions of the whole IR range. These are spectroscopy, astronomy, thermal imaging, detector and source development and metrology, as well the field of optical data transmission. Scientists working in these fields range from chemists and astronomers through to physicists and even photographers. This issue presents examples from some of these fields. All the papers—though some of them deal with fundamental or applied research—include interesting elements that make them directly applicable to university-level teaching at the graduate or postgraduate level. Source (e.g. quantum cascade lasers) and detector development (e.g. multispectral sensors), as well as metrology issues and optical data transmission, are omitted since they belong to fundamental research journals. Using a more-or-less arbitrary order according to wavelength range, the issue starts with a paper on the physics of near-infrared photography using consumer product cameras in the spectral range from 800 nm to 1.1 µm [1]. It is followed by a series of three papers dealing with IR imaging in spectral ranges from 3 to 14 µm [2-4]. One of them deals with laboratory courses that may help to characterize the IR camera response [2], the second discusses potential applications for nondestructive testing techniques [3] and the third gives an example of how IR thermal imaging may be used to understand cloud cover of the Earth [4], which is the prerequisite for successful climate modelling. The next two papers cover the vast field of IR spectroscopy [5, 6]. The first of these deals with Fourier transform infrared spectroscopy in the spectral range from 2.5 to 25 µm, studying e.g. ro-vibrational excitations in gases or optical phonon interactions within solids [5]. The second deals mostly with the spectroscopy of liquids such as biofuels and special techniques such as attenuated total reflectance [6]. The two final papers deal with what seem to be wholly different scientific fields [7, 8]. One paper describes SOFIA, an aeroplane-based astronomical observatory covering the whole IR range [7], while the other represents a small review of the quite new topic of terahertz physics at the upper end of the IR spectral range, from around 30 µm to 3 mm wavelength, and its many applications in science and industry [8]. Although artificially separated, all these fields use similar kinds of detectors, similar kinds of IR sources and similar technologies, while the instruments use the same physical principles. We are convinced that the field of infrared physics will develop over the next decade in the same dynamic way as during the last, and this special issue may serve as starting point for regular submissions on the topic. At any rate, it shines a light on this fascinating and many-faceted subject, which started more than 200 years ago. References [1] Mangold K, Shaw J A and Vollmer M 2013 The physics of near-infrared photography Eur. J. Phys. 34 S51-71 [2] Vollmer M and Möllmann K-P 2013 Characterization of IR cameras in student labs Eur. J. Phys. 34 S73-90 [3] Ibarra-Castanedo C, Tarpani J R and Maldague X P V 2013 Nondestructive testing with thermography Eur. J. Phys. 34 S91-109 [4] Shaw J A and Nugent P W 2013 Physics principles in radiometric infrared imaging of clouds in the atmosphere Eur. J. Phys. 34 S111-21 [5] Möllmann K-P and Vollmer M 2013 Fourier transform infrared spectroscopy in physics laboratory courses Eur. J. Phys. 34 S123-37 [6] Heise H M, Fritzsche J, Tkatsch H, Waag F, Karch K, Henze K, Delbeck S and Budde J 2013 Recent advances in mid- and near-infrared spectroscopy with applications for research and teaching, focusing on petrochemistry and biotechnology relevant products Eur. J. Phys. 34 S139-59 [7] Krabbe A, Mehlert D, Röser H-P and Scorza C 2013 SOFIA, an airborne observatory for infrared astronomy Eur. J. Phys. 34 S161-77 [8] Zouaghi W, Thomson M D, Rabia K, Hahn R, Blank V and Roskos H G 2013 Broadband terahertz spectroscopy: principles, fundamental research and potential for industrial applications Eur. J. Phys. 34 S179-99

  12. High-throughput multiphoton-induced three-dimensional ablation and imaging for biotissues.

    PubMed

    Lin, Chun-Yu; Li, Pei-Kao; Cheng, Li-Chung; Li, Yi-Cheng; Chang, Chia-Yuan; Chiang, Ann-Shyn; Dong, Chen Yuan; Chen, Shean-Jen

    2015-02-01

    In this study, a temporal focusing-based high-throughput multiphoton-induced ablation system with axially-resolved widefield multiphoton excitation has been successfully applied to rapidly disrupt biotissues. Experimental results demonstrate that this technique features high efficiency for achieving large-area laser ablation without causing serious photothermal damage in non-ablated regions. Furthermore, the rate of tissue processing can reach around 1.6 10(6) ?m(3)/s in chicken tendon. Moreover, the temporal focusing-based multiphoton system can be efficiently utilized in optical imaging through iterating high-throughput multiphoton-induced ablation machining followed by widefield optical sectioning; hence, it has the potential to obtain molecular images for a whole bio-specimen. PMID:25780739

  13. High-throughput multiphoton-induced three-dimensional ablation and imaging for biotissues

    PubMed Central

    Lin, Chun-Yu; Li, Pei-Kao; Cheng, Li-Chung; Li, Yi-Cheng; Chang, Chia-Yuan; Chiang, Ann-Shyn; Dong, Chen Yuan; Chen, Shean-Jen

    2015-01-01

    In this study, a temporal focusing-based high-throughput multiphoton-induced ablation system with axially-resolved widefield multiphoton excitation has been successfully applied to rapidly disrupt biotissues. Experimental results demonstrate that this technique features high efficiency for achieving large-area laser ablation without causing serious photothermal damage in non-ablated regions. Furthermore, the rate of tissue processing can reach around 1.6 106 ?m3/s in chicken tendon. Moreover, the temporal focusing-based multiphoton system can be efficiently utilized in optical imaging through iterating high-throughput multiphoton-induced ablation machining followed by widefield optical sectioning; hence, it has the potential to obtain molecular images for a whole bio-specimen. PMID:25780739

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

    PubMed Central

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

    2009-01-01

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

  15. The excitation of near-infrared H2 emission in NGC 253

    NASA Astrophysics Data System (ADS)

    Rosenberg, M. J. F.; van der Werf, P. P.; Israel, F. P.

    2013-02-01

    Context. Because of its large angular size and proximity to the Milky Way, NGC 253, an archetypal starburst galaxy, provides an excellent laboratory to study the intricacies of this intense episode of star formation. Aims: We aim to characterize the excitation mechanisms driving the emission in NGC 253. Specifically we aim to distinguish between shock excitation and ultraviolet (UV) excitation as the dominant driving mechanism, using Br?, H2 and [FeII] as diagnostic emission line tracers. Methods: Using SINFONI observations, we create linemaps of Br?, [FeII]1.64, and all detected H2 transitions. By using symmetry arguments of the gas and stellar gas velocity field, we find a kinematic center in agreement with previous determinations. The ratio of the 2-1 S(1) to 1-0 S(1) H2 transitions can be used as a diagnostic to discriminate between shock and fluorescent excitation. Results: Using the 1-0 S(1)/2-1 S(1) line ratio as well as several other H2 line ratios and the morphological comparison between H2 and Br? and [FeII], we find that excitation from UV photons is the dominant excitation mechanisms throughout NGC 253. We employ a diagnostic energy level diagram to quantitatively differentiate between mechanisms. We compare the observed energy level diagrams to photon-dominated region (PDR) and shock models and find that in most regions and over the galaxy as a whole, fluorescent excitation is the dominant mechanism exciting the H2 gas. We also place an upper limit of the percentage of shock excited H2 at 29%. Conclusions: We find that UV radiation is the dominant excitation mechanism for the H2 emission. The H2 emission does not correlate well with Br? but closely traces the polycyclic aromatic hydrocarbon emission, showing that not only is H2 fluorescently excited, but it is predominately excited by slightly lower mass stars than O stars which excite Br?, such as B stars. Reduced datacubes and extracted images for each line are only available at the CDS via anonymous ftp to cdsarc.u-strasbg.fr (130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/550/A12

  16. Optical and Near Infrared Study of the Cepheus E Outflow, a Very Low Excitation Object

    NASA Technical Reports Server (NTRS)

    Noreiga-Crespo, A.; Ayala, S.; Garnavich, P.; Curiel, S.; Raga, A.; Bohm, K.; Raymond, J.

    2000-01-01

    In this study, we explore the link between the physical properties of the outflow as determined from optical imaging and spectroscopy, and compare these results with those obtained from observations in the near infrared.

  17. Terahertz-to-infrared emission through laser excitation of surface plasmons in metal films with porous nanostructures.

    PubMed

    Zhang, Liangliang; Zhao, Ji; Wu, Tong; Zhang, Cunlin; Zhang, X-C

    2015-06-29

    We report on the investigation of terahertz-to-infrared (THz-to-IR) thermal emission that relies on the excitation of surface plasmons in metal films deposited on a substrate with randomly ordered nanoscale pore arrays. The THz-to-IR radiation was observed both in the direction of laser beam propagation and the reverse direction. The intensity ratio between backward and forward radiation is exponentially dependent on the nominal thickness of the porous metal films. The findings are discussed in view of the proposed generation mechanism based on propagating surface plasmon polaritons on both air/metal and metal/substrate interfaces. PMID:26191727

  18. Broadband infrared and Raman probes of excited-state vibrational molecular dynamics; Simulation protocols based on loop diagrams

    PubMed Central

    Dorfman, Konstantin E.; Fingerhut, Benjamin P.; Mukamel, Shaul

    2013-01-01

    Vibrational motions in electronically excited states can be observed by either time and frequency resolved infrared absorption or by off resonant stimulated Raman techniques. Multipoint correlation function expressions are derived for both signals. Three representations for the signal which suggest different simulation protocols are developed. These are based on the forward and the backward propagation of the wavefunction, sum over state expansion using an effective vibration Hamiltonian and a semiclassical treatment of a bath. We show that the effective temporal (?t) and spectral (??) resolution of the techniques is not controlled solely by experimental knobs but also depends on the system dynamics being probed. The Fourier uncertainty ???t > 1 is never violated. PMID:23783120

  19. Ultrafast, large-field multiphoton microscopy based on an acousto-optic deflector and a spatial light modulator

    PubMed Central

    Shao, Yonghong; Qin, Wan; Liu, Honghai; Qu, Junle; Peng, Xiang; Niu, Hanben; Gao, Bruce Z.

    2013-01-01

    We present an ultrafast, large-field multiphoton excitation fluorescence microscope with high lateral and axial resolutions based on a two-dimensional (2-D) acousto-optical deflector (AOD) scanner and spatial light modulator (SLM). When a phase-only SLM is used to shape the near-infrared light from a mode-locked titanium:sapphire laser into a multifocus array including the 0-order beam, a 136 ?m 136 ?m field of view is achieved with a 60 objective using a 2-D AOD scanner without any mechanical scan element. The two-photon fluorescence image of a neuronal network that was obtained using this system demonstrates that our microscopy permits observation of dynamic biological events in a large field with high-temporal and -spatial resolution. PMID:22743445

  20. Laser action in chromium-activated forsterite for near infrared excitation

    NASA Technical Reports Server (NTRS)

    Petricevic, V.; Gayen, S. K.; Alfano, R. R.

    1988-01-01

    This paper reports on laser action in chromium-doped forsterite (Cr:Mg2SiO4) for 1064-nm excitation of the crystal's double-hump absorption band spanning the 850-1200-nm wavelength range. The cavity arrangement used for obtaining laser action in Cr:Mg2SiO2 was similar to that described by Petricevic et al. (1988). The fundamental and second harmonic emissions from a Q-switched Nd:YAG laser operating at a 10-Hz repetition rate were used for excitation of the NIR and visible bands, respectively. Pulsed laser action was readily observed for both the 1064-nm and 532-nm pumping at or above the respective thresholds. The laser parameters of the 532-nm and 1064-nm excitations were similar, indicating that the IR band is responsible for laser action for both excitations.

  1. Near-infrared spectroscopy of (proto)-planetary nebulae: molecular hydrogen excitation as an evolutionary tracer

    NASA Astrophysics Data System (ADS)

    Davis, Christopher J.; Smith, Michael D.; Stern, Luke; Kerr, Thomas H.; Chiar, Jean E.

    2003-09-01

    We present an in-depth analysis of molecular excitation in 11 H2-bright planetary and protoplanetary nebulae (PN and PPN). From newly acquired K-band observations, we extract a number of spectra at positions across each source. H2 line intensities are plotted on `column density ratio' diagrams so that we may examine the excitation in and across each region. To achieve this, we combine the shock models of Smith, Khanzadyan & Davis with the photodissociation region (PDR) models of Black & van Dishoeck to yield a shock-plus-fluorescence fit to each data set. Although the combined shock + fluorescence model is needed to explain the low- and high-energy H2 lines in most of the sources observed (fluorescence accounts for much of the emission from the higher-energy H2 lines), the relative importance of shocks over fluorescence does seem to change with evolutionary status. We find that shock excitation may well be the dominant excitation mechanism in the least evolved PPN (CRL 2688 - in both the bipolar lobes and in the equatorial plane) and in the most evolved PN considered (NGC 7048). Fluorescence, on the other hand, becomes more important at intermediate evolutionary stages (i.e. in `young' PN), particularly in the inner core regions and along the inner edges of the expanding post-asymptotic giant branch (AGB) envelope. Since H2 line emission seems to be produced in almost all stages of post-AGB evolution, H2 excitation may prove to be a useful probe of the evolutionary status of PPN and PN alike. Moreover, shocks may play an important role in the molecular gas excitation in (P)PN, in addition to the low- and/or high-density fluorescence usually attributed to the excitation in these sources.

  2. Attosecond x-ray sources generation from pre-excited He+ ions using mid-infrared homogeneous and inhomogeneous fields

    NASA Astrophysics Data System (ADS)

    Feng, Liqiang; Liu, Hang

    2015-10-01

    A promising method is proposed to generate the intense ultrabroadband supercontinuum spectra and the ultrashort attosecond x-ray pulses from pre-excited He+ ion with the homogeneous and inhomogeneous two-color mid-infrared laser field combined with a unipolar pulse. For the homogeneous case, by optimizing the two mid-infrared pulses and the unipolar pulse, not only the harmonic cutoff has been extended, but also the harmonic modulation has been remarkably reduced. Moreover, we found the above harmonic extension scheme can be achieved in the multi-cycle pulse region and by preparing the initial state of He+ ion as the coherent superposition of the ground and the first excited states, the harmonic yield can be enhanced by 4-6 orders of magnitude compared with the case of the single ground initial state. Furthermore, with the introduction of the inhomogeneous of the combined field, the harmonic cutoff can be further enhanced, resulting in a supercontinuum with the bandwidth of 1648?eV. As a result, a series of intense 32as x-ray pulses can be obtained, which are 4-6 orders of magnitude improvement in comparison with the single initial state case.

  3. Single photon infrared emission spectroscopy: a study of IR emission from UV laser excited PAHs between 3 and 15 micrometers

    NASA Technical Reports Server (NTRS)

    Cook, D. J.; Schlemmer, S.; Balucani, N.; Wagner, D. R.; Harrison, J. A.; Steiner, B.; Saykally, R. J.

    1998-01-01

    Single-photon infrared emission spectroscopy (SPIRES) has been used to measure emission spectra from polycyclic aromatic hydrocarbons (PAHs). A supersonic free-jet expansion has been used to provide emission spectra of rotationally cold and vibrationally excited naphthalene and benzene. Under these conditions, the observed width of the 3.3-micrometers (C-H stretch) band resembles the bandwidths observed in experiments in which emission is observed from naphthalene with higher rotational energy. To obtain complete coverage of IR wavelengths relevant to the unidentified infrared bands (UIRs), UV laser-induced desorption was used to generate gas-phase highly excited PAHs. Lorentzian band shapes were convoluted with the monochromator-slit function in order to determine the widths of PAH emission bands under astrophysically relevant conditions. Bandwidths were also extracted from bands consisting of multiple normal modes blended together. These parameters are grouped according to the functional groups mostly involved in the vibration, and mean bandwidths are obtained. These bandwidths are larger than the widths of the corresponding UIR bands. However, when the comparison is limited to the largest PAHs studied, the bandwidths are slightly smaller than the corresponding UIR bands. These parameters can be used to model emission spectra from PAH cations and cations of larger PAHs, which are better candidate carriers of the UIRs.

  4. Spin state transitions upon visible and infrared excitation of ferric MbN3

    NASA Astrophysics Data System (ADS)

    Helbing, Jan

    2012-03-01

    When azide binds to ferric Myoglobin it forms either a low-spin or a high-spin complex, which give rise to two well-separated asymmetric stretch bands of the ligand. Both electronic excitation of the Q-band and vibrational excitation of N3- in the mid-IR lead to a similar ultrafast population redistribution in favor of the high spin configuration, which is characterized by a 8 reorientation of the ligand transition dipole moment. The more stable low spin complex subsequently re-emerges with a 18 ps time-constant. It is argued that the observed spin state changes are caused by the participation of low-lying electronic excitations in the cooling process of heme.

  5. Differentiation of normal and cancerous lung tissues by multiphoton imaging

    NASA Astrophysics Data System (ADS)

    Wang, Chun-Chin; Li, Feng-Chieh; Wu, Ruei-Jhih; Hovhannisyan, Vladimir A.; Lin, Wei-Chou; Lin, Sung-Jan; So, Peter T. C.; Dong, Chen-Yuan

    2009-07-01

    We utilize multiphoton microscopy for the label-free diagnosis of noncancerous, lung adenocarcinoma (LAC), and lung squamous cell carcinoma (SCC) tissues from humans. Our results show that the combination of second-harmonic generation (SHG) and multiphoton excited autofluorescence (MAF) signals may be used to acquire morphological and quantitative information in discriminating cancerous from noncancerous lung tissues. Specifically, noncancerous lung tissues are largely fibrotic in structure, while cancerous specimens are composed primarily of tumor masses. Quantitative ratiometric analysis using MAF to SHG index (MAFSI) shows that the average MAFSI for noncancerous and LAC lung tissue pairs are 0.55+/-0.23 and 0.87+/-0.15, respectively. In comparison, the MAFSIs for the noncancerous and SCC tissue pairs are 0.50+/-0.12 and 0.72+/-0.13, respectively. Our study shows that nonlinear optical microscopy can assist in differentiating and diagnosing pulmonary cancer from noncancerous tissues.

  6. Infrared/ultraviolet quadruple resonance spectroscopy to investigate structures of electronically excited states

    SciTech Connect

    Weiler, M.; Bartl, K.; Gerhards, M.

    2012-03-21

    Molecular beam investigations in combination with IR/UV spectroscopy offer the possibility to obtain structural information on isolated molecules and clusters. One of the demanding tasks is the discrimination of different isomers, e.g., by the use of isomer specific UV excitations. If this discrimination fails due to overlaying UV spectra of different isomers, IR/IR methods offer another possibility. Here, we present a new IR/UV/IR/UV quadruple resonance technique to distinguish between different isomers especially in the electronically excited state. Due to the IR spectra, structural changes and photochemical pathways in excited states can be assigned and identified. The method is applied to the dihydrated cluster of 3-hydroxyflavone which has been investigated as photochemically relevant system and proton wire model in the S{sub 1} state. By applying the new IR/UV/IR/UV technique, we are able to show experimentally that both in the electronic ground (S{sub 0}) and the electronically excited state (S{sub 1}) two isomers have to be assigned.

  7. Infrared/ultraviolet quadruple resonance spectroscopy to investigate structures of electronically excited states

    NASA Astrophysics Data System (ADS)

    Weiler, M.; Bartl, K.; Gerhards, M.

    2012-03-01

    Molecular beam investigations in combination with IR/UV spectroscopy offer the possibility to obtain structural information on isolated molecules and clusters. One of the demanding tasks is the discrimination of different isomers, e.g., by the use of isomer specific UV excitations. If this discrimination fails due to overlaying UV spectra of different isomers, IR/IR methods offer another possibility. Here, we present a new IR/UV/IR/UV quadruple resonance technique to distinguish between different isomers especially in the electronically excited state. Due to the IR spectra, structural changes and photochemical pathways in excited states can be assigned and identified. The method is applied to the dihydrated cluster of 3-hydroxyflavone which has been investigated as photochemically relevant system and proton wire model in the S1 state. By applying the new IR/UV/IR/UV technique, we are able to show experimentally that both in the electronic ground (S0) and the electronically excited state (S1) two isomers have to be assigned.

  8. Controlled Au-Polymer Nanostructures for Multiphoton Imaging, Prodrug Delivery, and Chemo-Photothermal Therapy Platforms.

    PubMed

    Huang, Chih-Chia; Liu, Tzu-Ming

    2015-11-18

    We have successfully introduced a proton-induced controlled reaction of HAuCl4 and poly(styrene-alt-maleic acid) (PSMA) sodium salt to prepare triangular and multicore Au@polymer nanoparticles (NPs). The interparticle interactions in the core gave rise to an absorption band at the near-infrared wavelength. The near-infrared optical properties of the resulting Au-polymer nanostructures are highly stable in a physiological environment, which offered strong photo-to-thermal conversion by a moderate continuous-wave 808 nm laser and exhibited multiphoton fluorescence for imaging using a 1230 nm light excitation (femtosecond laser). Exposure of the carboxylate groups at the polymer shell made the surface structure of the Au multicore @polymer NPs directly conjugate Pt(II)-/Pt(IV)-based drugs, which possessed the elimination of the immediate toxicity over the short time and resulted in an anticancer effect after 3 days. A synergistic effect of the chemo-photothermal therapy showed a moderate hyperthermia assistance (<1 W/cm(2)) and better anticancer performance over time compared with the individual treatments. We demonstrated that such PSMA-based methodology not only enables a broad range of chemical material synthesis in the kinetic control to form Au nano-octahedrons and nanotriangles using Br(-)/I(-) ions additives but also could be extended to form Au/Fe3O4@polymer nanocomposites via proton-assisted PSMA self-assembly. PMID:26501876

  9. Epifluorescence light collection for multiphoton microscopic endoscopy

    NASA Astrophysics Data System (ADS)

    Brown, Christopher M.; Rivera, David R.; Xu, Chris; Webb, Watt W.

    2011-03-01

    Multiphoton microscopic endoscopy (MPM-E) is a promising medical in vivo diagnostic imaging technique because it captures intrinsic fluorescence and second harmonic generation signals to reveal anatomical and histological information about disease states in tissue. However, maximizing light collection from multiphoton endoscopes remains a challenge: weak nonlinear emissions from endogenous structures, miniature optics, large imaging depths, and light scattering in tissue all hamper light collection. The quantity of light that may be collected using a dual-clad fiber system from scattering phantoms that mimic the properties of the in vivo environment is measured. In this experiment, 800nm excitation light from a Ti:Sapphire laser is dispersion compensated and focused through a SM800 optical fiber and lens system into the tissue phantom. Emission light from the phantom passes through the lens system, reflects off the dichroic and is then collected by a second optical fiber actuated by a micromanipulator. The lateral position of the collection fiber varies, measuring the distribution of emitted light 2000?m on either side of the focal point reimaged to the object plane. This spatial collection measurement is performed at depths up to 200?m from the phantom surface. The tissue phantoms are composed of a 15.8 ?M fluorescein solution mixed with microspheres, approximating the scattering properties of human bladder and dermis tissue. Results show that commercially available dual-clad optical fibers collect more than 47% of the total emission returning to the object plane from both phantoms. Based on these results, initial MPM-E devices will image the surface of epithelial tissues.

  10. Multiphoton imaging of excised normal skin and keloid scar: preliminary investigations

    NASA Astrophysics Data System (ADS)

    Brewer, Michael B.; Yeh, Alvin T.; Torkian, Behrooz; Sun, Chung-Ho; Tromberg, Bruce J.; Wong, Brian J.

    2004-07-01

    Wound healing is a physiologic process that acts to repair disruptions in the continuity of tissue caused by injury or surgical incision. Keloids and hypertrophic scars are forms of aberrant wound healing, which are characterized by the overproduction of collagen, resulting in an excessive amount of scar tissue. Keloid tumors, by definition, grow outside the boundary of the original tissue damage. Multiphoton microscopy (MPM) is an imaging technique which allows imaging of living specimens, without the use of fixation or stains. Images of collagen fibers are produced by the second harmonic signal intensity generated by endogenous fluorescence through excitation by infrared laser light. A postauricular keloid tumor was excised from a patient. The tissue was dissected, and a portion was imaged using MPM. Normal skin tissue was isolated from a patient undergoing a facelift. A portion of this tissue was also dissected and imaged using MPM. MPM images were taken using a 63X water immersion objective lens on a two-photon microscope and a titanium-sapphire laser. Images were taken beginning at the surface of the tissue and moving in at intervals of 200 nm to a final depth of 30 ?m. The two-photon images were used to reconstruct three-dimensional representations of the collagen matrix within the tissues, which are readily contrasted. Density of the collagen within each tissue was also ascertained using depth dependant decay of the image intensity. Multiphoton imaging was successfully used to image the collagen matrix of normal skin and a keloid scar, demonstrating differences in their microstructures.

  11. Multiphoton microscopy based cryo-imaging of inflated frozen human lung sections at -60C in healthy and COPD lungs

    NASA Astrophysics Data System (ADS)

    Abraham, Thomas; Kayra, Damian; Zhang, Angela; Suzuki, Masaru; McDonough, John; Elliott, W. M.; Cooper, Joel D.; Hogg, James C.

    2013-02-01

    Lung is a complex gas exchanger with interfacial area (where the gas exchange takes place) is about the size of a tennis court. Respiratory function is linked to the biomechanical stability of the gas exchange or alveolar regions which directly depends on the spatial distributions of the extracellular matrix fibers such fibrillar collagens and elastin fibers. It is very important to visualize and quantify these fibers at their native and inflated conditions to have correct morphometric information on differences between control and diseased states. This can be only achieved in the ex vivo states by imaging directly frozen lung specimens inflated to total lung capacity. Multiphoton microscopy, which uses ultra-short infrared laser pulses as the excitation source, produces multiphoton excitation fluorescence (MPEF) signals from endogenously fluorescent proteins (e.g. elastin) and induces specific second harmonic generation (SHG) signals from non-centrosymmetric proteins such as fibrillar collagens in fresh human lung tissues [J. Struct. Biol. (2010)171,189-196]. Here we report for the first time 3D image data obtained directly from thick frozen inflated lung specimens (~0.7- 1.0 millimeter thick) visualized at -60C without prior fixation or staining in healthy and diseased states. Lung specimens donated for transplantation and released for research when no appropriate recipient was identified served as controls, and diseased lung specimens donated for research by patients receiving lung transplantation for very severe COPD (n=4) were prepared as previously described [N. Engl. J. Med. (2011) 201, 1567]. Lung slices evenly spaced between apex and base were examined using multiphoton microscopy while maintained at -60C using a temperature controlled cold stage with a temperature resolution of 0.1C. Infrared femto-second laser pulses tuned to 880nm, dry microscopic objectives, and non-de-scanned detectors/spectrophotometer located in the reflection geometry were used for generating the 3D images/spectral information. We found that this novel imaging approach can provide spatially resolved 3D images with spectral specificities from frozen inflated lungs that are sensitive enough to identity the micro-structural details of fibrillar collagens and elastin fibers in alveolar walls in both healthy and diseased tissues.

  12. Surface deactivation of vibrationally excited N2 studied using infrared titration combined with quantum cascade laser absorption spectroscopy

    NASA Astrophysics Data System (ADS)

    Marinov, D.; Lopatik, D.; Guaitella, O.; Ionikh, Y.; Röpcke, J.; Rousseau, A.

    2014-01-01

    The wall de-excitation probability \\gamma_{N_{2}} of vibrationally excited nitrogen molecules was determined using infrared (IR) titration with CO, CO2 and N2O. Gas mixtures of N2 with 0.05-0.5% of CO (CO2 or N2O) were excited by a pulsed dc discharge at p = 133 Pa in a cylindrical discharge tube. During the afterglow, the vibrational relaxation of titrating molecules was monitored in situ with quantum cascade laser absorption spectroscopy. The value of \\gamma _{N_{2}} was deduced from measured vibrational relaxation times using a model of vibrational kinetics in N2. It was found that adsorption of IR tracers on the surface may increase the value of \\gamma _{N_{2}} by a factor up to two, depending on the molecule and the surface material. It was demonstrated that N2O is the most inert and reliable tracer and it was used for the determination of \\gamma_{N_{2}} on silica, Pyrex, TiO2, Al2O3 and anodized aluminum. Pretreatment of the silica surface by low-pressure plasma was found to have a strong effect on the vibrational de-excitation. Values of \\gamma_{N_{2}} measured after O2, Ar and N2 plasma pretreatment of the same silica discharge tube were 5.7 × 10-4, 8.2 × 10-4 and 11 × 10-4, respectively. This study clearly demonstrates that the presence of adsorbed atoms and molecules on the surface may significantly alter the value of \\gamma_{N_{2}} .

  13. Population transfer by multiphoton adiabatic rapid passage

    SciTech Connect

    Maeda, H.; Gurian, J. H.; Gallagher, T. F.

    2011-03-15

    The population of atoms in Rydberg states is efficiently transferred with a change in principal quantum number n of up to ten via multiphoton adiabatic rapid passage through a single multiphoton resonance using a frequency-chirped microwave pulse. A quantum-mechanical picture of multiphoton adiabatic rapid passage in a one-dimensional atom using a Floquet approach provides a good description of most, but not all, of the observed phenomena.

  14. Simultaneous time and wavelength resolved spectroscopy under two-colour near infrared and terahertz excitation

    NASA Astrophysics Data System (ADS)

    Bhattacharyya, J.; Wagner, M.; Zybell, S.; Winnerl, S.; Stehr, D.; Helm, M.; Schneider, H.

    2011-10-01

    Time and wavelength resolved spectroscopy requires optical sources emitting very short pulses and a fast detection mechanism capable of measuring the evolution of the output spectrum as a function of time. We use table-top Ti:sapphire lasers and a free-electron laser (FEL) emitting ps pulses as excitation sources and a streak camera coupled to a spectrometer for detection. One of the major aspects of this setup is the synchronization of pulses from the two lasers which we describe in detail. Optical properties of the FEL pulses are studied by autocorrelation and electro-optic sampling measurements. We discuss the advantages of using this setup to perform photoluminescence quenching in semiconductor quantum wells and quantum dots. Carrier redistribution due to pulsed excitation in these heterostructures can be investigated directly. Sideband generation in quantum wells is also studied where the intense FEL pulses facilitate the detection of the otherwise weak nonlinear effect.

  15. Simultaneous time and wavelength resolved spectroscopy under two-colour near infrared and terahertz excitation.

    PubMed

    Bhattacharyya, J; Wagner, M; Zybell, S; Winnerl, S; Stehr, D; Helm, M; Schneider, H

    2011-10-01

    Time and wavelength resolved spectroscopy requires optical sources emitting very short pulses and a fast detection mechanism capable of measuring the evolution of the output spectrum as a function of time. We use table-top Ti:sapphire lasers and a free-electron laser (FEL) emitting ps pulses as excitation sources and a streak camera coupled to a spectrometer for detection. One of the major aspects of this setup is the synchronization of pulses from the two lasers which we describe in detail. Optical properties of the FEL pulses are studied by autocorrelation and electro-optic sampling measurements. We discuss the advantages of using this setup to perform photoluminescence quenching in semiconductor quantum wells and quantum dots. Carrier redistribution due to pulsed excitation in these heterostructures can be investigated directly. Sideband generation in quantum wells is also studied where the intense FEL pulses facilitate the detection of the otherwise weak nonlinear effect. PMID:22047280

  16. Stochastic scanning multiphoton multifocal microscopy.

    PubMed

    Jureller, Justin E; Kim, Hee Y; Scherer, Norbert F

    2006-04-17

    Multiparticle tracking with scanning confocal and multiphoton fluorescence imaging is increasingly important for elucidating biological function, as in the transport of intracellular cargo-carrying vesicles. We demonstrate a simple rapid-sampling stochastic scanning multifocal multiphoton microscopy (SS-MMM) fluorescence imaging technique that enables multiparticle tracking without specialized hardware at rates 1,000 times greater than conventional single point raster scanning. Stochastic scanning of a diffractive optic generated 10x10 hexagonal array of foci with a white noise driven galvanometer yields a scan pattern that is random yet space-filling. SS-MMM creates a more uniformly sampled image with fewer spatio-temporal artifacts than obtained by conventional or multibeam raster scanning. SS-MMM is verified by simulation and experimentally demonstrated by tracking microsphere diffusion in solution. PMID:19516485

  17. Near-infrared long-slit spectra of Seyfert galaxies: gas excitation across the central kiloparsec

    NASA Astrophysics Data System (ADS)

    van der Laan, T. P. R.; Schinnerer, E.; Bker, T.; Armus, L.

    2013-12-01

    Context. The excitation of the gas phase of the interstellar medium can be driven by various mechanisms. In galaxies with an active nucleus, such as Seyfert galaxies, both radiative and mechanical energy from the central black hole, or the stars in the disk surrounding it may play a role. Aims: We investigate the relative importance and range of influence of the active galactic nucleus for the excitation of ionized and molecular gas in the central kiloparsec of its host galaxy. Methods: We present H- and K-band long-slit spectra for a sample of 21 nearby (D < 70 Mpc) Seyfert galaxies obtained with the NIRSPEC instrument on the Keck telescope. For each galaxy, we fit the nebular line emission, stellar continua, and warm molecular gas as a function of distance from the nucleus. Results: Our analysis does not reveal a clear difference between the nucleus proper and off-nuclear environment in terms of excitation mechanisms, suggesting that the influence of an AGN reaches far into the disk of the host galaxy. The radial variations in emission line ratios indicate that, while local mechanisms do affect the gas excitation, they are often averaged out when measuring over extended regions. The data presented herein were obtained at the W. M. Keck Observatory, which is operated as a scientific partnership among the California Institute of Technology, the University of California and the National Aeronautics and Space Administration. The Observatory was made possible by the generous financial support of the W. M. Keck Foundation.Table 4 and Appendix A are available in electronic form at http://www.aanda.orgThe fully calibrated long-slit spectra and fitting are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/560/A99

  18. Resonant Multiphoton Ionization by Short Laser Pulses

    NASA Astrophysics Data System (ADS)

    Adler, Andre Charles

    Motivated by recent experiments a theory of multiphoton ionization of atoms by laser pulses is developed in which dynamical effects due to the finite duration of the laser field are studied in detail. The time-dependent Schrodinger equation is solved analytically in the form of a series for a laser pulse modeled as a hyperbolic secant function of time. The method applies to any number of intermediate resonant bound states and for any number of photons needed for ionization. Recent experiments on the seven-photon ionization of xenon using very short laser pulses have produced results in the photoelectron energy spectrum that have yet to be predicted by any theory. It is hypothesized that the structure is due to different bound excited states being shifted into resonance at different peak intensities. Our theory is able to reproduce the structure seen in the experiments. In addition, by applying the theory to the two-photon ionization of cesium, we have predicted a new structure not seen experimentally. Taking into account a one-photon resonance with two excited states, we have found that in the weak field limit, the single peak in the photoelectron energy spectrum is split into two peaks. We show that one gets a similar result when the pulse is modeled as a square pulse. In the strong field case, we have found new structure due to a resonance with continuum states. We show that this structure is absent when the laser field is modeled as a square pulse.

  19. Multispot multiphoton Ca²⁺ imaging in acute myocardial slices.

    PubMed

    Borile, Giulia; de Mauro, Claudio; Urbani, Andrea; Alfieri, Domenico; Pavone, Francesco S; Mongillo, Marco

    2015-05-01

    Multiphoton microscopy has become essential for dynamic imaging in thick living tissues. High-rate, full-field image acquisition in multiphoton microscopy is achievable by parallelization of the excitation and detection pathways. We developed our approach via a diffractive optical element which splits a pulsed laser into 16 beamlets and exploits a descanned detection system consisting of an array of beamlet-associated photomultiplier tubes. The optical performance of the multiphoton multispot system (MCube) has been characterized in cardiac tissue sections and subsequently used for the first time for fluorescence imaging of cardiomyocyte Ca²⁺ dynamics in viable acute cardiac slices. Multispot multiphoton microscopy (MMM) has never been used before to monitor Ca²⁺ dynamics in thick, viable tissue samples. Acute heart slices are a powerful close-to-in vivo model of Ca²⁺ imaging allowing the simultaneous observation of several cells in their own tissue environment, exploiting the multiphoton excitation ability to penetrate scattering tissues. Moreover, we show that the concurrent high spatial and temporal resolutions afforded by the parallel scanning in MMM can be exploited to simultaneously assess subcellular Ca²⁺ dynamics in different cells in the tissue. We recorded local Ca²⁺ release events including macrosparks, travelling waves, and rotors. PMID:25517401

  20. Infrared imaging of buried objects by thermal step-function excitations.

    PubMed

    Li, P; Maad, A; Moshary, F; Arend, M F; Ahmed, S

    1995-09-01

    Sudden heating or cooling of a surface, by turning on or off radiation flux on the surface, is used as a means of enhancing the detection capability of buried objects by thermal infrared imaging. An experimental analysis of a sand-surface temperature is presented as a function of a buried object's composition and depth, and it is compared with theoretical simulations. Object identification by means of the geometry of isothermal contours and the rate of change of the surface temperature with radiant flux is discussed. PMID:21060414

  1. Vibrational Relaxation Dynamics of AN Infrared Laser-Excited Molecular Impurity Mode in Alkali Halide Lattices.

    NASA Astrophysics Data System (ADS)

    Moerner, William Esco

    1982-03-01

    The vibrational relaxation dynamics of spherical top ReO(,4)('-) molecules have been investigated under conditions of nonequilibrium laser excitation. Previous studies of relaxation mechanisms for molecular impurity modes in crystalline solids have chiefly utilized the techniques of low-power, linear spectroscopy. In this work, the nonequilibrium techniques of incoherent laser saturation and high resolution hole-burning spectroscopy have been used to measure the relaxation times T(,1) and T(,2) for the inhomogeneously broadened (nu)(,3) internal mode of ReO(,4)('-) as functions of temperature and host lattice. In addition, long-lived non-photochemical holes (with lifetimes greater than 10 minutes at 1.4 K) have been observed, indicating for the first time the presence of ground state optical pumping processes for a high symmetry impurity in a cubic crystal. The CO(,2) laser saturation measurements yielded values for the saturation intensity, I(,s), and hence the T(,1) T(,2) product in various hosts. The hole-burning measurements of T(,2) utilized a CO(,2) laser as a saturating pump and either a Pb salt diode laser or another CO(,2) laser as the tunable probe. Holes as narrow as 10 MHz (FWHM) were observed in inhomogeneously broadened lines extending over a frequency interval of several cm('-1). Above 10K, the dominant dephasing (T(,2)) mechanism is acoustic phonon scattering, while below 10K, T(,2) achieves the fundamental upper limit of 2T(,1) signifying that dephasing is lifetime-limited. Study of the alkali halide dependence of the (nu)(,3) excited state decay rate suggests that a combination of molecular internal modes, local modes of the lattice-impurity complex, and band phonons represents the dominant decay channel, rather than the previously expected multiphonon decay. The discovery of long-lived holes for a spherical top molecule at a cubic substitutional site suggests that the molecular ground state consists of at least two quasi-equivalent orientations, which are coupled only by excitation to the (nu)(,3) mode excited state. These studies of the vibrational relaxation dynamics of the ReO(,4)('-) molecule in alkali halides demonstrate the power of nonequilibrium laser techniques and provide a basis for future investigations of molecular impurity modes in solids.

  2. Differentiation of normal and cancerous lung tissues by multiphoton imaging

    NASA Astrophysics Data System (ADS)

    Wang, Chun-Chin; Li, Feng-Chieh; Wu, Ruei-Jr; Hovhannisyan, Vladimir A.; Lin, Wei-Chou; Lin, Sung-Jan; So, Peter T. C.; Dong, Chen-Yuan

    2010-02-01

    In this work, we utilized multiphoton microscopy for the label-free diagnosis of non-cancerous, lung adenocarcinoma (LAC), and lung squamous cell carcinoma (SCC) tissues from human. Our results show that the combination of second harmonic generation (SHG) and multiphoton excited autofluorescence (MAF) signals may be used to acquire morphological and quantitative information in discriminating cancerous from non-cancerous lung tissues. Specifically, non-cancerous lung tissues are largely fibrotic in structure while cancerous specimens are composed primarily of tumor masses. Quantitative ratiometric analysis using MAF to SHG index (MAFSI or SAAID) shows that the average MAFSI for noncancerous and LAC lung tissue pairs are 0.55 +/-0.23 and 0.87+/-0.15 respectively. In comparison, the MAFSIs for the noncancerous and SCC tissue pairs are 0.50+/-0.12 and 0.72+/-0.13 respectively. Intrinsic fluorescence ratio (FAD/NADH) of SCC and non-cancerous tissues are 0.40+/-0.05 and 0.53+/-0.05 respectively, the redox ratio of SCC diminishes significantly, indicating that increased cellular metabolic activity. Our study shows that nonlinear optical microscopy can assist in differentiating and diagnosing pulmonary cancer from non-cancerous tissues. With additional development, multiphoton microscopy may be used for the clinical diagnosis of lung cancers.

  3. Measurement of molecular diffusion in solution by multiphoton fluorescence photobleaching recovery

    PubMed Central

    Brown, EB; Wu, ES; Zipfel, W; Webb, WW

    1999-01-01

    Multiphoton fluorescence photobleaching recovery (MP-FPR) is a technique for measuring the three-dimensional (3D) mobility of fluorescent molecules with 3D spatial resolution of a few microns. A brief, intense flash of mode-locked laser light pulses excites fluorescent molecules via multiphoton excitation in an ellipsoidal focal volume and photobleaches a fraction. Because multiphoton excitation of fluorophores is intrinsically confined to the high-intensity focal volume of the illuminating beam, the bleached region is restricted to a known, three-dimensionally defined volume. Fluorescence in this focal volume is measured with multiphoton excitation, using the attenuated laser beam to measure fluorescence recovery as fresh unbleached dye diffuses in. The time course of the fluorescence recovery signal after photobleaching can be analyzed to determine the diffusion coefficient of the fluorescent species. The mathematical formulas used to fit MP-FPR recovery curves and the techniques needed to properly utilize them to acquire the diffusion coefficients of fluorescently labeled molecules within cells are presented here. MP-FPR is demonstrated on calcein in RBL-2H3 cells, using an anomalous subdiffusion model, as well as in aqueous solutions of wild-type green fluorescent protein, yielding a diffusion coefficient of 8.7 x 10(-7) cm(2)s(-1) in excellent agreement with the results of other techniques. PMID:10545381

  4. Selective excitation of photosystems in chloroplasts inside plant leaves observed by near-infrared laser-based fluorescence spectral microscopy.

    PubMed

    Hasegawa, Makoto; Shiina, Takashi; Terazima, Masahide; Kumazaki, Shigeichi

    2010-02-01

    In this study, we produced selective images of photosystems in plant chloroplasts in situ. We used a spectroimaging microscope, equipped with a near-infrared (NIR) laser that provided light at wavelengths mainly between 800 and 830 nm, to analyze chlorophyll autofluorescence spectra and images from chloroplasts in leaves of Zea mays at room temperature. Femtosecond laser excitation of chloroplasts in mesophyll cells revealed a spectral shape that was attributable to PSII and its antenna in the centers of grana spots. We found that a continuous wave emitted by the NIR laser at a wavelength as long as 820 nm induced chlorophyll autofluorescence with a high contribution from PSI through a one-photon absorption mechanism. A spectral shape attributable to PSI and its antenna was thus obtained using continuous wave laser excitation of chloroplasts in bundle sheath cells. These highly pure spectra of photosystems were utilized for spectral decomposition at every intrachloroplast space to show distributions of PSI and PSII and their associated antenna. A new methodology using an NIR laser to detect the PSI/PSII ratio in single chloroplasts in leaves at room temperature is described. PMID:20022977

  5. Critical evaluation of a handheld Raman spectrometer with near infrared (785nm) excitation for field identification of minerals.

    PubMed

    Jehli?ka, Jan; Culka, Adam; Vandenabeele, Peter; Edwards, Howell G M

    2011-10-01

    Handheld Raman spectrometers (Ahura First Defender XL, Inspector Raman DeltaNu) permit the recording of acceptable and good quality spectra of a large majority of minerals outdoors and on outcrops. Raman spectra of minerals in the current study were obtained using instruments equipped with 785 nm diode lasers. Repetitive measurements carried out under an identical instrumental setup confirmed the reliability of the tested Raman spectrometers. Raman bands are found at correct wavenumber positions within 3 cm(-1) compared to reference values in the literature. Taking into account several limitations such as the spatial resolution and problems with metallic and black and green minerals handheld Raman spectrometers equipped with 785 nm diode lasers can be applied successfully for the detection of minerals from the majority of classes of the mineralogical system. For the detection of biomarkers and biomolecules using Raman spectroscopy, e.g. for exobiological applications, the near infrared excitation can be considered as a preferred excitation. Areas of potential applications of the actual instruments include all kind of common geoscience work outdoors. Modified Raman systems can be proposed for studies of superficial or subsurface targets for Mars or Lunar investigations. PMID:21292545

  6. Femtosecond laser pulse optimization for multiphoton cytometry and control of fluorescence

    NASA Astrophysics Data System (ADS)

    Tkaczyk, Eric Robert

    This body of work encompasses optimization of near infrared femtosecond laser pulses both for enhancement of flow cytometry as well as adaptive pulse shaping to control fluorescence. A two-photon system for in vivo flow cytometry is demonstrated, which allows noninvasive quantification of circulating cell populations in a single live mouse. We monitor fluorescently-labeled red blood cells for more than two weeks, and are also able to noninvasively measure circulation times of two distinct populations of breast cancer cells simultaneously in a single mouse. We build a custom laser excitation source in the form of an extended cavity mode-locked oscillator, which enables superior detection in whole blood or saline of cell lines expressing fluorescent proteins including the green fluorescent protein (GFP), tdTomato and mPlum. A mathematical model explains unique features of the signals. The ability to distinguish different fluorescent species is central to simultaneous measurement of multiple molecular targets in high throughput applications including the multiphoton flow cytometer. We demonstrate that two dyes which are not distinguishable to one-photon measurements can be differentiated and in fact quantified in mixture via phase-shaped two-photon excitation pulses found by a genetic algorithm. We also selectively enhance or suppress two-photon fluorescence of numerous common dyes with tailored pulse shapes. Using a multiplicative (rather than ratiometric) fitness parameter, we are able to control the fluorescence while maintaining a strong signal. With this method, we control the two-photon fluorescence of the blue fluorescent protein (BFP), which is of particular interest in investigations of protein-protein interactions, and has frustrated previous attempts of control. Implementing an acousto-optic interferometer, we use the same experimental setup to measure two-photon excitation cross-sections of dyes and prove that photon-photon interferences are the predominant mechanism of control. This research establishes the basis for molecularly tailored pulse shaping in multiphoton flow cytometry, which will advance our ability to probe the biology of circulating cells during disease progression and response to therapy.

  7. Nitric oxide releasing materials triggered by near-infrared excitation through tissue filters.

    PubMed

    Burks, Peter T; Garcia, John V; GonzalezIrias, Ricardo; Tillman, Jason T; Niu, Mutong; Mikhailovsky, Alexander A; Zhang, Jinping; Zhang, Fan; Ford, Peter C

    2013-12-01

    Novel materials for the phototherapeutic release of the bioregulator nitric oxide (nitrogen monoxide) are described. Also reported is a method for scanning these materials with a focused NIR beam to induce photouncaging while minimizing damage from local heating. The new materials consist of poly(dimethylsiloxane) composites with near-infrared-to-visible upconverting nanoparticles (UCNPs) that are cast into a biocompatible polymer disk (PD). These PDs are then impregnated with the photochemical nitric oxide precursor Roussin's black salt (RBS) to give UCNP_RBS_PD devices that generate NO when irradiated with 980 nm light. When the UCNP_RBS_PD composites were irradiated with NIR light through filters composed of porcine tissue, physiologically relevant NO concentrations were released, thus demonstrating the potential of such devices for minimally invasive phototherapeutic applications. PMID:24245494

  8. Ground and excited state infrared spectroscopy of jet-cooled radicals: Exploring the photophysics of trihydronaphthyl and inden-2-ylmethyl

    NASA Astrophysics Data System (ADS)

    Kidwell, Nathanael M.; Mehta-Hurt, Deepali N.; Korn, Joseph A.; Sibert, Edwin L.; Zwier, Timothy S.

    2014-06-01

    The alkyl and aromatic CH stretch infrared spectra of inden-2-ylmethyl (I2M, C10H9) and trihydronaphthyl (THN, C10H11) radicals have been recorded under jet-cooled conditions in the ground (D0) and first electronically excited (D1) states using resonant ion-dip infrared (RIDIR) spectroscopy. Previously, the vibronic spectroscopy of a series of C10H9 and C10H11 hydronaphthyl radicals were investigated and their thermochemical properties were evaluated with isomer specificity [J. A. Sebree et al., J. Phys. Chem. A 11, 6255-6262 (2010)]. We show here that one of the m/z 129 spectral carriers characterized in that work was misidentified as 2-hydronaphthyl (2-HN) radical, appearing in a discharge of 1,2-dihydronaphthalene in close proximity to 1-hydronaphthyl radical. The D0-RIDIR spectrum in the alkyl CH stretch region positively identifies the m/z 129 isomer as I2M, whose two-color resonant two-photon ionization (2C-R2PI) spectrum was recently reported by Schmidt and co-workers [T. P. Troy et al., Chem. Sci. 2, 1755-1765 (2011)]. Here, we further characterize the I2M and THN radicals by recording their gas phase IR spectra in the alkyl and aromatic CH stretch regions, and explore the spectroscopic consequences of electronic excitation on the CH stretch absorptions. A local-mode CH stretch Hamiltonian incorporating cubic stretch-bend coupling between anharmonic CH stretches and CH2 scissor modes is utilized to describe their Fermi resonance interactions. Excellent agreement between the experimental and theoretical results facilitates the interpretation of the D0- and D1-state RIDIR spectra of I2M, revealing that upon excitation the alkyl CH stretches decrease in frequency by 70 cm-1, while the allyl-like CH stretches experience a modest blueshift. In comparison, the photophysics of THN are strikingly different in that the IR transitions that possess vibrational motion along the CβH and CδH bonds are absent in the D1-RIDIR spectrum yet are predicted to be present from the theoretical model. Several hypotheses are considered to account for the perturbations to these vibrations.

  9. Interpretation of rotationally excited far-infrared OH emission in Orion-KL

    NASA Technical Reports Server (NTRS)

    Melnick, G. J.; Genzel, R.; Lugten, J. B.

    1987-01-01

    The 2Pi(1/2) OH 163-micron J = 3/2-1/2 rotational transitions in Orion-KL were observed and an upper limit was set to the line strength of the 2II(1/2) OH 56-micron J = 9/2-7/2 doublet in this source. The 163-micron line intensities were modeled, along with the previously measured 2II(3/2) 119 and 84-micron rotational line emission and it is found that the gas in the Orion-KL postshocked region can produce OH 119-micron line emission of the same strength as measured; however, the resultant 84 and 163-micron line intensities would be weaker than observed. Shocked gas plus a second component which experiences strong radiative excitation can reproduce the observations.

  10. Mid-infrared Otto excitation of transverse electric modes in doped graphene

    NASA Astrophysics Data System (ADS)

    Ramos-Mendieta, F.

    2015-04-01

    We have studied numerically the excitation of surface modes of transverse electric polarization in doped graphene. Using the prism-based Otto configuration, the electromagnetic resonances were found within ultra-tiny angular windows of width of order of ? ? i = 10-3 degrees or lower, beyond the critical angle where evanescent fields are already available. We obtained absorption peaks of angular position strongly dependent on the prism-graphene separation d, which is larger than dc, the cutoff prism-graphene separation. We prove numerically that dc depends on the graphene parameters and can be written as d c = ? / 2 , where ? ( ? ) is the decay length of the electromagnetic field of the modes. With doping levels of Fermi energy 0.2 eV ? ? ? 0.3 eV, we found resonances within the range of 80-145 THz. The numerical calculations show the mayor role played by the graphene losses that, indeed, can obliterate the mode resonance.

  11. Interstellar polycyclic aromatic hydrocarbons: the infrared emission bands, the excitation/emission mechanism, and the astrophysical implications.

    PubMed

    Allamandola, L J; Tielens, A G; Barker, J R

    1989-12-01

    This article presents a comprehensive treatment of the polycyclic aromatic hydrocarbon (PAH) hypothesis. The interstellar, infrared spectral features which have been attributed to emission from highly vibrationally excited PAHs are discussed in detail. These include major (most intense) bands at 3040, 1615, "1310," 1150, and 885 cm-1 (3.29, 6.2 "7.7," 8.7, and 11.3 micrometers), minor bands and broad features in the 3200-2700 cm-1 [correction of 3200-2700-1] (3.1-3.7 micrometers), 1600-1100 cm-1 (6.0-9 micrometers) and 910-770 cm-1 (11-13 micrometers) regions, as well as the vibrational quasi-continuum spanning the entire mid-IR and the electronic transitions which contribute to the high-frequency IR continuum. All the major and minor bands, as well as the quasi-continuum, can be attributed to vibrational transitions in molecular-sized PAHs. The latter two broad features probably arise from very large PAHs, PAH clusters, and amorphous carbon particles. A precise match of the interstellar spectra with laboratory spectra is not yet possible because laboratory spectra are not available of PAHs in the forms probably present in the interstellar medium (completely isolated, ionized, some completely dehydrogenated, and containing between about 20 and 40 carbon atoms). The method with which one can calculate the IR fluorescence spectrum from a vibrationally excited molecule is also described in detail. Fluorescence band intensities, relaxation rates, and dependence on molecule size and energy content are treated explicitly. Analysis of the interstellar spectra indicates that the PAHs which dominate the infrared spectra contain between about 20 and 40 carbon atoms. The results obtained with this method are compared with the results obtained using a thermal approximation. It is shown that for high levels of vibrational excitation and emission from low-frequency modes, the two methods give similar results. However, at low levels of vibrational excitation and for the high-frequency modes (for example, the 3040 cm-1, 3.3 micrometers band), the thermal approach overestimates the emission intensities. For calculations of molecular reactions (such as H-loss, deuterium enrichment, and carbon skeleton rearrangement) a thermal approximation is invalid. The relationship between PAH molecules and amorphous carbon particles is presented and their production in circumstellar shells is described. The most likely interstellar PAH molecular structures are discussed and the possibility of destructive reactions with interstellar oxygen and hydrogen atoms is considered in detailed and found to be unimportant. Interstellar PAH size and abundance estimates are made. On the order of a few percent of the available interstellar carbon is tied up in the small (20-40 carbon atom) PAHs which are responsible for the sharp features, and a similar amount is tied up in the larger (200-500 carbon atom) PAHs or PAH clusters and amorphous carbon particles which are responsible for the broad components underlying the 1600-1100 and 900-770 cm-1 (6-9 and 11-13 micrometers) regions. It is shown that the spectroscopic structure these PAHs and PAH-related materials produce in the UV portion of the interstellar extinction curve lie just below current detection limits but fall in the range detectable by the Hubble Space Telescope. Finally, the influence of PAH charge on the ultraviolet, visible, and infrared regions is described. PMID:11542189

  12. Line broadening in multiphoton processes with a resonant intermediate transition

    NASA Technical Reports Server (NTRS)

    Wang, C. C.; James, J. V.; Xia, J.-F.

    1983-01-01

    It is found that the line width of the excitation spectrum for multiphoton ionization is broadened much more severely than the cascade fluorescence originating from the resonant intermediate level. These results derive from the mutual effects of the ionizing and resonating transitions, which are not properly accounted for in perturbative treatments. In general, the ionization line shape can be expressed as the sum of two Lorentzian components, each with different degrees of broadening. This line shape reduces to that of the resonating transition in the limit of vanishing intensities. Moreover, the fluorescence line shape may also exhibit a second component whose amplitude may become important under certain conditions.

  13. Spectral-resolved multifocal multiphoton microscopy with multianode photomultiplier tubes

    PubMed Central

    Cha, Jae Won; Tzeranis, Dimitrios; Subramanian, Jaichandar; Yannas, Ioannis V.; Nedivi, Elly; So, Peter T. C.

    2014-01-01

    Multiphoton excitation fluorescence microscopy is the preferred method for in vivo deep tissue imaging. Many biological applications demand both high imaging speed and the ability to resolve multiple fluorophores. One of the successful methods to improve imaging speed in a highly turbid specimen is multifocal multiphoton microscopy (MMM) based on use of multi-anode photomultiplier tubes (MAPMT). This approach improves imaging speed by using multiple foci for parallelized excitation without sacrificing signal to noise ratio (SNR) due to the scattering of emission photons. In this work, we demonstrate that the MAPMT based MMM can be extended with spectral resolved imaging capability. Instead of generating multiple excitation foci in a 2D grid pattern, a linear array of foci is generated. This leaves one axis of the 2D MAPMT available for spectral dispersion and detection. The spectral-resolved MMM can detect several emission signals simultaneously with high imaging speed optimized for high-throughput, high-contents applications. The new procedure is illustrated using imaging data from the kidney, peripheral nerve regeneration and dendritic morphological data from the brain. PMID:25321515

  14. Zero kinetic energy photoelectron spectroscopy of jet cooled benzo[a]pyrene from resonantly enhanced multiphoton ionization

    NASA Astrophysics Data System (ADS)

    Zhang, Jie; Harthcock, Colin; Han, Fangyuan; Kong, Wei

    2011-12-01

    We report zero kinetic energy (ZEKE) photoelectron spectroscopy of benzo[a]pyrene (BaP) via resonantly enhanced multiphoton ionization (REMPI). Our analysis concentrates on the vibrational modes of the first excited state (S1) and those of the ground cationic state (D0). Similar to pyrene, another peri-condensed polycyclic aromatic hydrocarbon we have investigated, the first two electronically excited states of BaP exhibit extensive configuration interactions. However, the two electronic states are of the same symmetry, hence vibronic coupling does not introduce any out-of-plane modes in the REMPI spectrum, and Franck-Condon analysis is qualitatively satisfactory. The ZEKE spectra from the in-plane modes observed in the REMPI spectrum demonstrate strong propensity in preserving the vibrational excitation of the intermediate state. Although several additional bands in combination with the vibrational mode of the intermediate state are identifiable, they are much lower in intensity. This observation implies that the molecular structure of BaP has a tremendous capability to accommodate changes in charge density. All observed bands of the cation are IR active, establishing the role of ZEKE spectroscopy in mapping out far infrared bands for astrophysical applications.

  15. Spread of cochlear excitation during stimulation with pulsed infrared radiation: inferior colliculus measurements

    NASA Astrophysics Data System (ADS)

    Richter, C.-P.; Rajguru, S. M.; Matic, A. I.; Moreno, E. L.; Fishman, A. J.; Robinson, A. M.; Suh, E.; Walsh, J. T., Jr.

    2011-10-01

    Infrared neural stimulation (INS) has received considerable attention over the last few years. It provides an alternative method to artificially stimulate neurons without electrical current or the introduction of exogenous chromophores. One of the primary benefits of INS could be the improved spatial selectivity when compared with electrical stimulation. In the present study, we have evaluated the spatial selectivity of INS in the acutely damaged cochlea of guinea pigs and compared it to stimulation with acoustic tone pips in normal-hearing animals. The radiation was delivered via a 200 m diameter optical fiber, which was inserted through a cochleostomy into the scala tympani of the basal cochlear turn. The stimulated section along the cochlear spiral ganglion was estimated from the neural responses recorded from the central nucleus of the inferior colliculus (ICC). ICC responses were recorded in response to cochlear INS using a multichannel penetrating electrode array. Spatial tuning curves (STCs) were constructed from the responses. For INS, approximately 55% of the activation profiles showed a single maximum, ~22% had two maxima and ~13% had multiple maxima. The remaining 10% of the profiles occurred at the limits of the electrode array and could not be classified. The majority of ICC STCs indicated that the spread of activation evoked by optical stimuli is comparable to that produced by acoustic tone pips.

  16. Plasma membrane nanoporation as a possible mechanism behind infrared excitation of cells

    NASA Astrophysics Data System (ADS)

    Beier, Hope T.; Tolstykh, Gleb P.; Musick, Joshua D.; Thomas, Robert J.; Ibey, Bennett L.

    2014-12-01

    Objective. Short infrared (IR) laser pulses have been used to stimulate action potentials in neurons both in vivo and in vitro. However, the mechanism(s) underlying this phenomenon has remained elusive. In vitro studies have found that pulsed IR exposure generates a nearly instant change in capacitance in the plasma membrane, characterized by inward rectification, a common feature in pore-forming exposures, such as electrical pulses and acoustic shock waves. Based on this similarity, we hypothesize that the mechanism of IR stimulation is the formation of short-lived nanopores in the plasma membrane. These transient, small-diameter pores allow the influx of extracellular ions that lead to action potential generation, possibly through activation of secondary messenger pathways or depolarization of the cell membrane resulting in activation of voltage-gated ion channels. Approach. A variety of fluorescent markers are used to observe the cell response to IR stimulation to monitor for effects indicative of nanoporation in other modalities. Main results. We observe rapid, transient rises in intracellular Ca2+, influx of YO-PRO-1 and propidium iodide into the cell signifying membrane permeabilization, cellular blebbing and swelling, and activation of the intracellular phosphoinositides lipid signaling pathway. Significance. This conclusion better explains the experimental observations and limitations of IR-induced neurological stimulation and represents a distinct theoretical shift in the understanding of the mechanism of IR-induced stimulation.

  17. Multiphoton quantum optics and quantum state engineering

    NASA Astrophysics Data System (ADS)

    Dell'Anno, Fabio; De Siena, Silvio; Illuminati, Fabrizio

    2006-05-01

    We present a review of theoretical and experimental aspects of multiphoton quantum optics. Multiphoton processes occur and are important for many aspects of matter-radiation interactions that include the efficient ionization of atoms and molecules, and, more generally, atomic transition mechanisms; system-environment couplings and dissipative quantum dynamics; laser physics, optical parametric processes, and interferometry. A single review cannot account for all aspects of such an enormously vast subject. Here we choose to concentrate our attention on parametric processes in nonlinear media, with special emphasis on the engineering of nonclassical states of photons and atoms that are relevant for the conceptual investigations as well as for the practical applications of forefront aspects of modern quantum mechanics. We present a detailed analysis of the methods and techniques for the production of genuinely quantum multiphoton processes in nonlinear media, and the corresponding models of multiphoton effective interactions. We review existing proposals for the classification, engineering, and manipulation of nonclassical states, including Fock states, macroscopic superposition states, and multiphoton generalized coherent states. We introduce and discuss the structure of canonical multiphoton quantum optics and the associated one- and two-mode canonical multiphoton squeezed states. This framework provides a consistent multiphoton generalization of two-photon quantum optics and a consistent Hamiltonian description of multiphoton processes associated to higher-order nonlinearities. Finally, we discuss very recent advances that by combining linear and nonlinear optical devices allow to realize multiphoton entangled states of the electromagnetic field, either in discrete or in continuous variables, that are relevant for applications to efficient quantum computation, quantum teleportation, and related problems in quantum communication and information.

  18. Wavefront sensorless adaptive optics temporal focusing-based multiphoton microscopy

    PubMed Central

    Chang, Chia-Yuan; Cheng, Li-Chung; Su, Hung-Wei; Hu, Yvonne Yuling; Cho, Keng-Chi; Yen, Wei-Chung; Xu, Chris; Dong, Chen Yuan; Chen, Shean-Jen

    2014-01-01

    Temporal profile distortions reduce excitation efficiency and image quality in temporal focusing-based multiphoton microscopy. In order to compensate the distortions, a wavefront sensorless adaptive optics system (AOS) was integrated into the microscope. The feedback control signal of the AOS was acquired from local image intensity maximization via a hill-climbing algorithm. The control signal was then utilized to drive a deformable mirror in such a way as to eliminate the distortions. With the AOS correction, not only is the axial excitation symmetrically refocused, but the axial resolution with full two-photon excited fluorescence (TPEF) intensity is also maintained. Hence, the contrast of the TPEF image of a R6G-doped PMMA thin film is enhanced along with a 3.7-fold increase in intensity. Furthermore, the TPEF image quality of 1μm fluorescent beads sealed in agarose gel at different depths is improved. PMID:24940539

  19. Development of infrared sensors using energy transfer/energy upconversion processes: Study of laser excited fluorescence in rare Earth ion doped crystals

    NASA Technical Reports Server (NTRS)

    Nash-Stevenson, S. K.; Reddy, B. R.; Venkateswarlu, P.

    1994-01-01

    A summary is presented of the spectroscopic study of three systems: LaF3:Ho(3+), LaF3:Er(3+) and CaF2:Nd(3+). When the D levels of Ho(3+) in LaF3 were resonantly excited with a laser beam of 640 nm, upconverted emissions were detected from J (416 nm), F (485 nm), and E (546 nm) levels. Energy upconverted emissions were also observed from F and E levels of Ho(3+) when the material was excited with an 800 nm near infrared laser. When the D levels of Er(3+) in LaF3 were resonantly excited with a laser beam of 637 nm, upconverted emissions were detected from the E (540 nm) and P (320, 400, and 468 nm) levels. Energy upconverted emissions were also observed from F, E, and D levels of Er(3+) when the material was resonantly excited with an 804 nm near infrared laser. When the D levels of Nd(3+) in CaF2 were resonantly excited with a laser beam of 577 nm, upconverted emissions were detected from the L (360 and 382 nm), K (418 nm), and I (432 nm) levels. Very weak upconverted emissions were detected when this system was irradiated with a near infrared laser. The numbers in parentheses are the wavelengths of the emissions.

  20. Near-infrared-excited confocal Raman spectroscopy advances in vivo diagnosis of cervical precancer.

    PubMed

    Duraipandian, Shiyamala; Zheng, Wei; Ng, Joseph; Low, Jeffrey J H; Ilancheran, Arunachalam; Huang, Zhiwei

    2013-06-01

    Raman spectroscopy is a unique optical technique that can probe the changes of vibrational modes of biomolecules associated with tissue premalignant transformation. This study evaluates the clinical utility of confocal Raman spectroscopy over near-infrared (NIR) autofluorescence (AF) spectroscopy and composite NIR AF/Raman spectroscopy for improving early diagnosis of cervical precancer in vivo at colposcopy. A rapid NIR Raman system coupled with a ball-lens fiber-optic confocal Raman probe was utilized for in vivo NIR AF/Raman spectral measurements of the cervix. A total of 1240 in vivo Raman spectra [normal (n=993), dysplasia (n=247)] were acquired from 84 cervical patients. Principal components analysis (PCA) and linear discriminant analysis (LDA) together with a leave-one-patient-out, cross-validation method were used to extract the diagnostic information associated with distinctive spectroscopic modalities. The diagnostic ability of confocal Raman spectroscopy was evaluated using the PCA-LDA model developed from the significant principal components (PCs) [i.e., PC4, 0.0023%; PC5, 0.00095%; PC8, 0.00022%, (p<0.05)], representing the primary tissue Raman features (e.g., 854, 937, 1095, 1253, 1311, 1445, and 1654 cm(-1)). Confocal Raman spectroscopy coupled with PCA-LDA modeling yielded the diagnostic accuracy of 84.1% (a sensitivity of 81.0% and a specificity of 87.1%) for in vivo discrimination of dysplastic cervix. The receiver operating characteristic curves further confirmed that the best classification was achieved using confocal Raman spectroscopy compared to the composite NIR AF/Raman spectroscopy or NIR AF spectroscopy alone. This study illustrates that confocal Raman spectroscopy has great potential to improve early diagnosis of cervical precancer in vivo during clinical colposcopy. PMID:23797897

  1. Near-infrared-excited confocal Raman spectroscopy advances in vivo diagnosis of cervical precancer

    NASA Astrophysics Data System (ADS)

    Duraipandian, Shiyamala; Zheng, Wei; Ng, Joseph; Low, Jeffrey J. H.; Ilancheran, Arunachalam; Huang, Zhiwei

    2013-06-01

    Raman spectroscopy is a unique optical technique that can probe the changes of vibrational modes of biomolecules associated with tissue premalignant transformation. This study evaluates the clinical utility of confocal Raman spectroscopy over near-infrared (NIR) autofluorescence (AF) spectroscopy and composite NIR AF/Raman spectroscopy for improving early diagnosis of cervical precancer in vivo at colposcopy. A rapid NIR Raman system coupled with a ball-lens fiber-optic confocal Raman probe was utilized for in vivo NIR AF/Raman spectral measurements of the cervix. A total of 1240 in vivo Raman spectra [normal (n=993), dysplasia (n=247)] were acquired from 84 cervical patients. Principal components analysis (PCA) and linear discriminant analysis (LDA) together with a leave-one-patient-out, cross-validation method were used to extract the diagnostic information associated with distinctive spectroscopic modalities. The diagnostic ability of confocal Raman spectroscopy was evaluated using the PCA-LDA model developed from the significant principal components (PCs) [i.e., PC4, 0.0023% PC5, 0.00095% PC8, 0.00022%, (p<0.05)], representing the primary tissue Raman features (e.g., 854, 937, 1095, 1253, 1311, 1445, and 1654 cm-1). Confocal Raman spectroscopy coupled with PCA-LDA modeling yielded the diagnostic accuracy of 84.1% (a sensitivity of 81.0% and a specificity of 87.1%) for in vivo discrimination of dysplastic cervix. The receiver operating characteristic curves further confirmed that the best classification was achieved using confocal Raman spectroscopy compared to the composite NIR AF/Raman spectroscopy or NIR AF spectroscopy alone. This study illustrates that confocal Raman spectroscopy has great potential to improve early diagnosis of cervical precancer in vivo during clinical colposcopy.

  2. Application of time-resolved, step-scan fourier transform infrared spectroscopy to excited-state electronic structure in polypyridyl complexes of rhenium(I)

    SciTech Connect

    Schoonover, J.R.; Strouse, G.F.; Dyer, R.B.

    1996-01-17

    Recent advances in transient infrared spectroscopy have provided the means for infrared measurements to be made following laser excitation on short time scales. The infrared technique is particularly valuable in the study of metal complexes containing CO or CN{sup -} because v(CO) and v(CN) stretching vibrations have high oscillator strengths and the sensitivities of their energies and bandwidths to electronic and molecular structure are well established. Electronic excitation generally produces significant transient infrared absorption changes which are intense and characteristic of the changes in electron structure between states. For example, in [(phen)-(CO){sub 3}Re(NC)Ru(bpy){sub 2}(CN)]{sup +} (phen is 1,10-phenanthroline; py is 2,2{prime}-bipyridine) Re{sup I}(d{pi}) {yields} phen({pi}*) excitation produces initially a Re{sup II}(phen{sup *-}) metal-to-ligand charge transfer (MLCT) excited state and large shifts in v(CO) (40-80 cm{sup -1}) compared to those in the ground state. Subsequently, rapid cross-bridge energy transfer occurs (k{sub q} {approximately} 2 x 10{sup 11} s{sup -1}) to give [(phen)-(CO){sub 3}Re{sup I}(NC)Ru{sup III}(bpy{sup *-})(CN)]{sup +*} which is accompanied by a shift of +57 cm{sup -1} in the terminal v(CN) band and only a slight shift in v(CO) compared to the ground state. Here, the authors apply step-scan transient infrared to the elucidation of the excited-state electronic structure in complexes.

  3. The multiphoton ionization of uranium hexafluoride

    SciTech Connect

    Armstrong, D.P. . UEO Enrichment Technical Operations Div.)

    1992-05-01

    Multiphoton ionization (MPI) time-of-flight mass spectroscopy and photoelectron spectroscopy studies of UF{sub 6} have been conducted using focused light from the Nd:YAG laser fundamental ({lambda}=1064 nm) and its harmonics ({lambda}=532, 355, or 266 nm), as well as other wavelengths provided by a tunable dye laser. The MPI mass spectra are dominated by the singly and multiply charged uranium ions rather than by the UF{sub x}{sup +} fragment ions even at the lowest laser power densities at which signal could be detected. The laser power dependence of U{sup n+} ions signals indicates that saturation can occur for many of the steps required for their ionization. In general, the doubly-charged uranium ion (U{sup 2+}) intensity is much greater than that of the singly-charged uranium ion (U{sup +}). For the case of the tunable dye laser experiments, the U{sup n+} (n = 1- 4) wavelength dependence is relatively unstructured and does not show observable resonance enhancement at known atomic uranium excitation wavelengths. The dominance of the U{sup 2+} ion and the absence or very small intensities of UF{sub x}{sup +} fragments, along with the unsaturated wavelength dependence, indicate that mechanisms may exist other than ionization of bare U atoms after the stepwise photodissociation of F atoms from the parent molecule.

  4. In Vivo Microscopy of the Mouse Brain Using Multiphoton Laser Scanning Techniques

    PubMed Central

    Yoder, Elizabeth J.

    2008-01-01

    The use of multiphoton microscopy for imaging mouse brain in vivo offers several advantages and poses several challenges. This tutorial begins by briefly comparing multiphoton microscopy with other imaging modalities used to visualize the brain and its activity. Next, an overview of the techniques for introducing fluorescence into whole animals to generate contrast for in vivo microscopy using two-photon excitation is presented. Two different schemes of surgically preparing mice for brain imaging with multiphoton microscopy are reviewed. Then, several issues and problems with in vivo microscopy - including motion artifact, respiratory and cardiac rhythms, maintenance of animal health, anesthesia, and the use of fiducial markers – are discussed. Finally, examples of how these techniques have been applied to visualize the cerebral vasculature and its response to hypercapnic stimulation are provided. PMID:20975841

  5. Application of Multiphoton Microscopy in Dermatological Studies: a Mini-Review

    PubMed Central

    Yew, Elijah; Rowlands, Christopher

    2014-01-01

    This review summarizes the historical and more recent developments of multiphoton microscopy, as applied to dermatology. Multiphoton microscopy offers several advantages over competing microscopy techniques: there is an inherent axial sectioning, penetration depths that compete well with confocal microscopy on account of the use of near-infrared light, and many two-photon contrast mechanisms, such as second-harmonic generation, have no analogue in one-photon microscopy. While the penetration depths of photons into tissue are typically limited on the order of hundreds of microns, this is of less concern in dermatology, as the skin is thin and readily accessible. As a result, multiphoton microscopy in dermatology has generated a great deal of interest, much of which is summarized here. The review covers the interaction of light and tissue, as well as the various considerations that must be made when designing an instrument. The state of multiphoton microscopy in imaging skin cancer and various other diseases is also discussed, along with the investigation of aging and regeneration phenomena, and finally, the use of multiphoton microscopy to analyze the transdermal transport of drugs, cosmetics and other agents is summarized. The review concludes with a look at potential future research directions, especially those that are necessary to push these techniques into widespread clinical acceptance. PMID:25075226

  6. Autofluorescence Imaging With Near-Infrared Excitation:Normalization by Reflectance to Reduce Signal From Choroidal Fluorophores

    PubMed Central

    Cideciyan, Artur V.; Swider, Malgorzata; Jacobson, Samuel G.

    2015-01-01

    Purpose. We previously developed reduced-illuminance autofluorescence imaging (RAFI) methods involving near-infrared (NIR) excitation to image melanin-based fluorophores and short-wavelength (SW) excitation to image lipofuscin-based flurophores. Here, we propose to normalize NIR-RAFI in order to increase the relative contribution of retinal pigment epithelium (RPE) fluorophores. Methods. Retinal imaging was performed with a standard protocol holding system parameters invariant in healthy subjects and in patients. Normalized NIR-RAFI was derived by dividing NIR-RAFI signal by NIR reflectance point-by-point after image registration. Results. Regions of RPE atrophy in Stargardt disease, AMD, retinitis pigmentosa, choroideremia, and Leber congenital amaurosis as defined by low signal on SW-RAFI could correspond to a wide range of signal on NIR-RAFI depending on the contribution from the choroidal component. Retinal pigment epithelium atrophy tended to always correspond to high signal on NIR reflectance. Normalizing NIR-RAFI reduced the choroidal component of the signal in regions of atrophy. Quantitative evaluation of RPE atrophy area showed no significant differences between SW-RAFI and normalized NIR-RAFI. Conclusions. Imaging of RPE atrophy using lipofuscin-based AF imaging has become the gold standard. However, this technique involves bright SW lights that are uncomfortable and may accelerate the rate of disease progression in vulnerable retinas. The NIR-RAFI method developed here is a melanin-based alternative that is not absorbed by opsins and bisretinoid moieties, and is comfortable to view. Further development of this method may result in a nonmydriatic and comfortable imaging method to quantify RPE atrophy extent and its expansion rate. PMID:26024124

  7. Singlet and triplet excitation management in a bichromophoric near-infrared-phosphorescent BODIPY-benzoporphyrin platinum complex

    SciTech Connect

    Whited, M. T.; Djurovich, P. I.; Roberts, Sean T.; Durrell, A. C.; Schlenker, C. W.; Bradforth, Stephen E.; Thompson, Mark E.

    2011-01-12

    Multichromophoric arrays provide one strategy for assembling molecules with intense absorptions across the visible spectrum but are generally focused on systems that efficiently produce and manipulate singlet excitations and therefore are burdened by the restrictions of (a) unidirectional energy transfer and (b) limited tunability of the lowest molecular excited state. In contrast, we present here a multichromophoric array based on four boron dipyrrins (BODIPY) bound to a platinum benzoporphyrin scaffold that exhibits intense panchromatic absorption and efficiently generates triplets. The spectral complementarity of the BODIPY and porphryin units allows the direct observation of fast bidirectional singlet and triplet energy transfer processes (kST(1BDP→1Por) = 7.8 × 1011 s-1, kTT(3Por→3BDP) = 1.0 × 1010 s-1, kTT(3BDP→3Por) = 1.6 × 1010 s-1), leading to a long-lived equilibrated [3BDP][Por]⇌[BDP][3Por] state. This equilibrated state contains approximately isoenergetic porphyrin and BODIPY triplets and exhibits efficient near-infrared phosphorescence (λem = 772 nm, Φ = 0.26). Taken together, these studies show that appropriately designed triplet-utilizing arrays may overcome fundamental limitations typically associated with core-shell chromophores by tunable redistribution of energy from the core back onto the antennae.

  8. Mitigating Phototoxicity during Multiphoton Microscopy of Live Drosophila Embryos in the 1.0–1.2 µm Wavelength Range

    PubMed Central

    Débarre, Delphine; Olivier, Nicolas; Supatto, Willy; Beaurepaire, Emmanuel

    2014-01-01

    Light-induced toxicity is a fundamental bottleneck in microscopic imaging of live embryos. In this article, after a review of photodamage mechanisms in cells and tissues, we assess photo-perturbation under illumination conditions relevant for point-scanning multiphoton imaging of live Drosophila embryos. We use third-harmonic generation (THG) imaging of developmental processes in embryos excited by pulsed near-infrared light in the 1.0–1.2 µm range. We study the influence of imaging rate, wavelength, and pulse duration on the short-term and long-term perturbation of development and define criteria for safe imaging. We show that under illumination conditions typical for multiphoton imaging, photodamage in this system arises through 2- and/or 3-photon absorption processes and in a cumulative manner. Based on this analysis, we derive general guidelines for improving the signal-to-damage ratio in two-photon (2PEF/SHG) or THG imaging by adjusting the pulse duration and/or the imaging rate. Finally, we report label-free time-lapse 3D THG imaging of gastrulating Drosophila embryos with sampling appropriate for the visualisation of morphogenetic movements in wild-type and mutant embryos, and long-term multiharmonic (THG-SHG) imaging of development until hatching. PMID:25111506

  9. Multiphoton microscopy, fluorescence lifetime imaging and optical spectroscopy for the diagnosis of neoplasia

    NASA Astrophysics Data System (ADS)

    Skala, Melissa Caroline

    2007-12-01

    Cancer morbidity and mortality is greatly reduced when the disease is diagnosed and treated early in its development. Tissue biopsies are the gold standard for cancer diagnosis, and an accurate diagnosis requires a biopsy from the malignant portion of an organ. Light, guided through a fiber optic probe, could be used to inspect regions of interest and provide real-time feedback to determine the optimal tissue site for biopsy. This approach could increase the diagnostic accuracy of current biopsy procedures. The studies in this thesis have characterized changes in tissue optical signals with carcinogenesis, increasing our understanding of the sensitivity of optical techniques for cancer detection. All in vivo studies were conducted on the dimethylbenz[alpha]anthracene treated hamster cheek pouch model of epithelial carcinogenesis. Multiphoton microscopy studies in the near infrared wavelength region quantified changes in tissue morphology and fluorescence with carcinogenesis in vivo. Statistically significant morphological changes with precancer included increased epithelial thickness, loss of stratification in the epithelium, and increased nuclear diameter. Fluorescence changes included a statistically significant decrease in the epithelial fluorescence intensity per voxel at 780 nm excitation, a decrease in the fluorescence lifetime of protein-bound nicotinamide adenine dinucleotide (NADH, an electron donor in oxidative phosphorylation), and an increase in the fluorescence lifetime of protein-bound flavin adenine dinucleotide (FAD, an electron acceptor in oxidative phosphorylation) with precancer. The redox ratio (fluorescence intensity of FAD/NADH, a measure of the cellular oxidation-reduction state) did not significantly change with precancer. Cell culture experiments (MCF10A cells) indicated that the decrease in protein-bound NADH with precancer could be due to increased levels of glycolysis. Point measurements of diffuse reflectance and fluorescence spectra in the ultraviolet to visible wavelength range indicated that the most diagnostic optical signals originate from sub-surface tissue layers. Optical properties extracted from these spectroscopy measurements showed a significant decrease in the hemoglobin saturation, absorption coefficient, reduced scattering coefficient and fluorescence intensity (at 400 nm excitation) in neoplastic compared to normal tissues. The results from these studies indicate that multiphoton microscopy and optical spectroscopy can non-invasively provide information on tissue structure and function in vivo that is related to tissue pathology.

  10. Multiphoton cryo microscope with sample temperature control

    NASA Astrophysics Data System (ADS)

    Breunig, H. G.; Uchugonova, A.; Knig, K.

    2013-02-01

    We present a multiphoton microscope system which combines the advantages of multiphoton imaging with precise control of the sample temperature. The microscope provides online insight in temperature-induced changes and effects in plant tissue and animal cells with subcellular resolution during cooling and thawing processes. Image contrast is based on multiphoton fluorescence intensity or fluorescence lifetime in the range from liquid nitrogen temperature up to +600C. In addition, micro spectra from the imaged regions can be recorded. We present measurement results from plant leaf samples as well as Chinese hamster ovary cells.

  11. Excited State Molecular Dynamics in Intense Laser Fields

    NASA Astrophysics Data System (ADS)

    Zipp, Lucas; Natan, Adi; Bucksbaum, Philip

    2015-05-01

    We investigate the dynamics of excited electronic states in molecules created through strong field, multiphoton excitation. Several excited states can come into multiphoton resonance with the ground state during an intense laser pulse due to large AC stark shifts, and while most of the excited state population is subsequently ionized, a fraction of the population remains in these excited states. We probe this excited state population in N2 with a time delayed weak field and collect the angle-resolved photoelectron spectrum. By varying the pump intensity and the probe delay, we gain insight into bound state electron dynamics in molecules in intense laser fields, and the ensuing field free dynamics.

  12. Resonant enhanced multiphoton ionization studies of atomic oxygen

    NASA Technical Reports Server (NTRS)

    Dixit, S. N.; Levin, D.; Mckoy, V.

    1987-01-01

    In resonant enhanced multiphoton ionization (REMPI), an atom absorbs several photons making a transition to a resonant intermediate state and subsequently ionizing out of it. With currently available tunable narrow-band lasers, the extreme sensitivity of REMPI to the specific arrangement of levels can be used to selectively probe minute amounts of a single species (atom) in a host of background material. Determination of the number density of atoms from the observed REMPI signal requires a knowledge of the multiphoton ionization cross sections. The REMPI of atomic oxygen was investigated through various excitation schemes that are feasible with available light sources. Using quantum defect theory (QDT) to estimate the various atomic parameters, the REMPI dynamics in atomic oxygen were studied incorporating the effects of saturation and a.c. Stark shifts. Results are presented for REMPI probabilities for excitation through various 2p(3) (4S sup o) np(3)P and 2p(3) (4S sup o) nf(3)F levels.

  13. Multispectral multiphoton lifetime analysis of human bladder tissue

    NASA Astrophysics Data System (ADS)

    Cicchi, Riccardo; Crisci, Alfonso; Nesi, Gabriella; Cosci, Alessandro; Giancane, Saverio; Carini, Marco; Pavone, Francesco S.

    2009-02-01

    Human tissues intrinsically contain many fluorophores, as such NADH, elastin, collagen, and flavins, that can be excited and imaged using multiphoton microscopy, up to 150 ?m depth. In this work we have used combined two-photon excited fluorescence (TPE), fluorescence lifetime imaging microscopy (FLIM), and multispectral two photon emission detection (MTPE) to investigate different kinds of human ex-vivo fresh biopsies of bladder. Morphological and spectroscopic analyses have allowed to characterize both healthy and pathological tissue samples. In particular, we have examined tissue samples from healthy bladder mucosa, and bladder carcinoma in-situ (CIS), finding both morphological and spectroscopic differences. From the morphological point of view, cancer cells appeared more elongated with respect to corresponding normal cells; they also exhibited a different nucleus to cytoplasm ratio. From the spectroscopic point of view, we have found differences between the two tissue types in both spectral emission and fluorescence lifetime distribution. Even if further analysis, as well as a more significant statistics on a large number of samples would be helpful to discriminate between low and high grade cancer, our method is a promising tool to be used as diagnostic confirmation of histological results, as well as a diagnostic tool in a multiphoton endoscope or cystoscope to be used in in-vivo imaging applications.

  14. Cell-based and in vivo spectral analysis of fluorescent proteins for multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Salomonnson, Emma; Mihalko, Laura Anne; Verkhusha, Vladislav V.; Luker, Kathryn E.; Luker, Gary D.

    2012-09-01

    Multiphoton microscopy of cells and subcellular structures labeled with fluorescent proteins is the state-of-the-art technology for longitudinal imaging studies in tissues and living animals. Successful analysis of separate cell populations or signaling events by intravital microscopy requires optimal pairing of multiphoton excitation wavelengths with spectrally distinct fluorescent proteins. While prior studies have analyzed two photon absorption properties of isolated fluorescent proteins, there is limited information about two photon excitation and fluorescence emission profiles of fluorescent proteins expressed in living cells and intact tissues. Multiphoton microscopy was used to analyze fluorescence outputs of multiple blue, green, and red fluorescent proteins in cultured cells and orthotopic tumor xenografts of human breast cancer cells. It is shown that commonly used orange and red fluorescent proteins are excited efficiently by 750 to 760 nm laser light in living cells, enabling dual color imaging studies with blue or cyan proteins without changing excitation wavelength. It is also shown that small incremental changes in excitation wavelength significantly affect emission intensities from fluorescent proteins, which can be used to optimize multi-color imaging using a single laser wavelength. These data will direct optimal selection of fluorescent proteins for multispectral two photon microscopy.

  15. Cell-based and in vivo spectral analysis of fluorescent proteins for multiphoton microscopy.

    PubMed

    Salomonnson, Emma; Mihalko, Laura Anne; Verkhusha, Vladislav V; Luker, Kathryn E; Luker, Gary D

    2012-09-01

    Multiphoton microscopy of cells and subcellular structures labeled with fluorescent proteins is the state-of-the-art technology for longitudinal imaging studies in tissues and living animals. Successful analysis of separate cell populations or signaling events by intravital microscopy requires optimal pairing of multiphoton excitation wavelengths with spectrally distinct fluorescent proteins. While prior studies have analyzed two photon absorption properties of isolated fluorescent proteins, there is limited information about two photon excitation and fluorescence emission profiles of fluorescent proteins expressed in living cells and intact tissues. Multiphoton microscopy was used to analyze fluorescence outputs of multiple blue, green, and red fluorescent proteins in cultured cells and orthotopic tumor xenografts of human breast cancer cells. It is shown that commonly used orange and red fluorescent proteins are excited efficiently by 750 to 760 nm laser light in living cells, enabling dual color imaging studies with blue or cyan proteins without changing excitation wavelength. It is also shown that small incremental changes in excitation wavelength significantly affect emission intensities from fluorescent proteins, which can be used to optimize multi-color imaging using a single laser wavelength. These data will direct optimal selection of fluorescent proteins for multispectral two photon microscopy. PMID:22975677

  16. Experimental demonstration of mode-selective phonon excitation of 6H-SiC by a mid-infrared laser with anti-Stokes Raman scattering spectroscopy

    SciTech Connect

    Yoshida, Kyohei; Hachiya, Kan; Okumura, Kensuke; Mishima, Kenta; Inukai, Motoharu; Torgasin, Konstantin; Omer, Mohamed; Sonobe, Taro; Zen, Heishun; Negm, Hani; Kii, Toshiteru; Masuda, Kai; Ohgaki, Hideaki

    2013-10-28

    Mode-selective phonon excitation by a mid-infrared laser (MIR-FEL) is demonstrated via anti-Stokes Raman scattering measurements of 6H-silicon carbide (SiC). Irradiation of SiC with MIR-FEL and a Nd-YAG laser at 14 K produced a peak where the Raman shift corresponds to a photon energy of 119 meV (10.4 μm). This phenomenon is induced by mode-selective phonon excitation through the irradiation of MIR-FEL, whose photon energy corresponds to the photon-absorption of a particular phonon mode.

  17. Multiphoton microscopy in brain imaging

    NASA Astrophysics Data System (ADS)

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

    2015-03-01

    Brain imaging is becoming an important field in the frame of the neurophotonics in correlations with other medical ones in neuroscience studying functional and morphological aspects. In this presentation an overview on multi photon imaging of the brain will be presented, together with innovative aspects related to big area imaging and correlative microscopy approaches. Multiphoton imaging applications will be described together with methods to improve the penetration depth and obtain large area detection, or correlating functional aspects in vivo on single neuron with large area, even on whole brain, morphological aspects. Connecting super resolution features at the nanometer level with micro, meso and macroscopic architectures is in fact one of the challenging aspects to understand brain functioning.

  18. Multiphoton dissociative ionization of CS+

    NASA Astrophysics Data System (ADS)

    Rajput, Jyoti; Jochim, Bethany; Zohrabi, M.; Betsch, K. J.; Ablikim, U.; Berry, Ben; Severt, T.; Summers, A. M.; Armstrong, G. S. J.; Esry, B. D.; Carnes, K. D.; Ben-Itzhak, I.

    2015-05-01

    We have studied the dissociative photoionization of a CS+ molecular ion beam in the strong-field regime using <50 fs IR laser pulses (? ~ 790 nm) from a 10 kHz, ~2 mJ (per pulse) Ti:Sapphire laser system. A coincidence three-dimensional momentum imaging method was used to measure all ions and neutrals formed during this multiphoton process. Two prominent channels were observed: charge-symmetric dissociation, yielding C+ + S+, and charge-asymmetric dissociation, yielding C + S2+. The differences between these two channels with reference to their relative production probability, energetics, and angular distributions is the focus of this work. This work was supported by the Chemical Sciences, Geosciences, and Biosciences Division, Office of Basic Energy Sciences, Office of Science, U.S. Department of Energy. BJ is also supported by DOE-SCGF (DE-AC05-06OR23100).

  19. MULTI-PHOTON PHOSPHOR FEASIBILITY RESEARCH

    SciTech Connect

    R. Graham; W. Chow

    2003-05-01

    Development of multi-photon phosphor materials for discharge lamps represents a goal that would achieve up to a doubling of discharge (fluorescent) lamp efficacy. This report reviews the existing literature on multi-photon phosphors, identifies obstacles in developing such phosphors, and recommends directions for future research to address these obstacles. To critically examine issues involved in developing a multi-photon phosphor, the project brought together a team of experts from universities, national laboratories, and an industrial lamp manufacturer. Results and findings are organized into three categories: (1) Multi-Photon Systems and Processes, (2) Chemistry and Materials Issues, and (3) Concepts and Models. Multi-Photon Systems and Processes: This category focuses on how to use our current understanding of multi-photon phosphor systems to design new phosphor systems for application in fluorescent lamps. The quickest way to develop multi-photon lamp phosphors lies in finding sensitizer ions for Gd{sup 3+} and identifying activator ions to red shift the blue emission from Pr{sup 3+} due to the {sup 1}S{sub 0} {yields} {sup 1}I{sub 6} transition associated with the first cascading step. Success in either of these developments would lead to more efficient fluorescent lamps. Chemistry and Materials Issues: The most promising multi-photon phosphors are found in fluoride hosts. However, stability of fluorides in environments typically found in fluorescent lamps needs to be greatly improved. Experimental investigation of fluorides in actual lamp environments needs to be undertaken while working on oxide and oxyfluoride alternative systems for backup. Concepts and Models: Successful design of a multi-photon phosphor system based on cascading transitions of Gd{sup 3+} and Pr{sup 3+} depends critically on how the former can be sensitized and the latter can sensitize an activator ion. Methods to predict energy level diagrams and Judd-Ofelt parameters of multi-photon transitions are needed to help guide the experimental material selection. This report provides the theoretical basis for leading knowledgeable researchers along the path to develop multi-photon phosphor systems. The ultimate goal, a doubling of fluorescent lamp efficacy, is not going to be easily obtained. This report begins the process and should be followed with detailed experimental and theoretical research to continue the development process.

  20. Imaging carious dental tissues with multiphoton fluorescence lifetime imaging microscopy

    PubMed Central

    Lin, Po-Yen; Lyu, Hong-Chou; Hsu, Chin-Ying Stephen; Chang, Chia-Seng; Kao, Fu-Jen

    2011-01-01

    In this study, multiphoton excitation was utilized to image normal and carious dental tissues noninvasively. Unique structures in dental tissues were identified using the available multimodality (second harmonic, autofluorescence, and fluorescence lifetime analysis) without labeling. The collagen in dentin exhibits a strong second harmonic response. Both dentin and enamel emit strong autofluorescence that reveals in detail morphological features (such as dentinal tubules and enamel rods) and, despite their very similar spectral profiles, can be differentiated by lifetime analysis. Specifically, the carious dental tissue exhibits a greatly reduced autofluorescence lifetime, which result is consistent with the degree of demineralization, determined by micro-computed tomography. Our findings suggest that two-photon excited fluorescence lifetime imaging may be a promising tool for diagnosing and monitoring dental caries. PMID:21326645

  1. Detection and Inspection of Steel Bars in Reinforced Concrete Structures Using Active Infrared Thermography with Microwave Excitation and Eddy Current Sensors

    PubMed Central

    Szymanik, Barbara; Frankowski, Paweł Karol; Chady, Tomasz; John Chelliah, Cyril Robinson Azariah

    2016-01-01

    The purpose of this paper is to present a multi-sensor approach to the detection and inspection of steel bars in reinforced concrete structures. In connection with our past experience related to non-destructive testing of different materials, we propose using two potentially effective methods: active infrared thermography with microwave excitation and the eddy current technique. In this article active infrared thermography with microwave excitation is analyzed both by numerical modeling and experiments. This method, based on thermal imaging, due to its characteriatics should be considered as a preliminary method for the assessment of relatively shallowly located steel bar reinforcements. The eddy current technique, on the other hand, allows for more detailed evaluation and detection of deeply located rebars. In this paper a series of measurement results, together with the initial identification of certain features of steel reinforcement bars will be presented. PMID:26891305

  2. Detection and Inspection of Steel Bars in Reinforced Concrete Structures Using Active Infrared Thermography with Microwave Excitation and Eddy Current Sensors.

    PubMed

    Szymanik, Barbara; Frankowski, Paweł Karol; Chady, Tomasz; John Chelliah, Cyril Robinson Azariah

    2016-01-01

    The purpose of this paper is to present a multi-sensor approach to the detection and inspection of steel bars in reinforced concrete structures. In connection with our past experience related to non-destructive testing of different materials, we propose using two potentially effective methods: active infrared thermography with microwave excitation and the eddy current technique. In this article active infrared thermography with microwave excitation is analyzed both by numerical modeling and experiments. This method, based on thermal imaging, due to its characteriatics should be considered as a preliminary method for the assessment of relatively shallowly located steel bar reinforcements. The eddy current technique, on the other hand, allows for more detailed evaluation and detection of deeply located rebars. In this paper a series of measurement results, together with the initial identification of certain features of steel reinforcement bars will be presented. PMID:26891305

  3. Highly-reproducible Raman scattering of NaYF4:Yb,Er@SiO2@Ag for methylamphetamine detection under near-infrared laser excitation.

    PubMed

    Ma, Yongmei; Liu, Honglin; Han, Zhenzhen; Yang, Liangbao; Liu, Jinhuai

    2015-08-01

    This study reported the significantly improved Raman enhancement ability of silver nanoparticles (Ag NPs) by decorating them on single NaYF4:Yb,Er@SiO2 core-shell particles (UC@SiO2@Ag) under a 785 nm excitation. The optimal thickness of the silica shell can be easily obtained by adjusting the amounts of TEOS, which is the crucial element to balance the upconversion and the formation of a hot spot by Ag NP aggregation. This substrate revealed highly reproducible properties, which is crucial to the practical application of SERS technology. This substrate exhibited an excellent sensitivity for methylamphetamine detection under near-infrared excitation. The advantages of NIR excitation in our SERS sensing open up a new application field of UC-noble metal composites, and also promise a new research direction for the synthesis and applications of SERS-active nanostructures. PMID:26090604

  4. Promising new wavelengths for multi-photon microscopy: thinking outside the Ti:Sapphire box

    NASA Astrophysics Data System (ADS)

    Norris, Greg; Amor, Rumelo; Dempster, John; Amos, William B.; McConnell, Gail

    2013-02-01

    Multi-photon excitation (MPE) imaging is dominated by the Ti:Sapphire laser as the source for excitation. However, it is limited when considering 3PE of common fluorophores and efficient 2PE of UV dyes which require wavelengths beyond the range of the Ti:Sapphire. Two ultra-short pulsed sources are presented as alternatives: a novel optical parametric oscillator (OPO) geometry (1400-1600nm) and the sum-frequency mixing of an OPO and Yb-doped fibre laser, providing a tunable output (626-635nm). For long wavelengths, we report three-photon laser scanning microscopy (3PLSM) using a bi-directional pumped optical parametric oscillator (OPO) with signal wavelength output at 1500 nm. This novel laser was used to overcome the high optical loss in the infrared spectral region observed in laser scanning microscopes and objective lenses that renders them otherwise difficult to use for imaging. To test our system, we performed 3PLSM auto-fluorescence imaging of live plant cells at 1500 nm, specifically Spirogyra, and compared performance with two-photon excitation (2PLSM) imaging using a femtosecond pulsed Ti:Sapphire laser at 780 nm. Analysis of cell viability based on cytoplasmic organelle streaming and structural changes of cells revealed that at similar peak powers, 2PLSM caused gross cell damage after 5 minutes but 3PLSM showed little or no interference with cell function after 15 minutes. The 1500 nm OPO was thus shown to be a practical laser source for live cell imaging. For short wavelengths, we report the use of an all-solid-state ultra-short pulsed source specifically for two-photon microscopy at wavelengths shorter than those of the conventional Ti:Sapphire laser. Our approach involved sumfrequency mixing of the output from the long-wavelength OPO described above with residual pump radiation to generate fs-pulsed output in the red spectral region. We demonstrated the performance of our ultra-short pulsed system using fluorescently labelled and autofluorescent tissue, and compared with conventional Ti:Sapphire excitation. We observed a more than 3-fold increase in fluorescence signal intensity using our visible laser source in comparison with the Ti:Sapphire laser for two-photon excitation at equal illumination powers of 22 mW or less.

  5. Ionization delays in few-cycle-pulse multiphoton quantum-beat spectroscopy in helium

    NASA Astrophysics Data System (ADS)

    Pazourek, Renate; Reduzzi, Maurizio; Carpeggiani, Paolo A.; Sansone, Giuseppe; Gaarde, Mette; Schafer, Kenneth

    2016-02-01

    We explore quantum beats in the photoelectron signal produced when a bound electron wave packet created by an isolated attosecond pulse is ionized by a delayed, few-cycle infrared pulse. Our calculations for helium atoms show that the broad bandwidth of the few-cycle pulse creates spectrally overlapping photoelectron peaks that result from one-, two-, or three-photon ionization processes. The beat signals can, in principle, be interferometrically resolved with high resolution, giving access to the relative phase between different multiphoton ionization pathways. For few-cycle near-infrared fields the relative spectral phases can be extracted over a large energy region, and dynamical information becomes available. We find that multiphoton ionization is temporally shifted with respect to one-photon ionization by several hundred attoseconds. Our results also reveal the impact of depletion and resonant pathways on the phase of the quantum beats.

  6. Fiber-based combined optical coherence and multiphoton endomicroscopy.

    PubMed

    Liu, Gangjun; Chen, Zhongping

    2011-03-01

    A fiber-based multimodal imaging system that combines multiphoton microscopy (MPM) with Fourier domain optical coherence microscopy (OCM) is reported. The system uses a fiber-based femtosecond laser, a fiber coupler, and a double-clad fiber (DCF) device. The fiber laser has a central wavelength of 1.04 ?m and bandwidth of 29 nm. Longer excitation wavelength is used to increase penetration depth and increase the excitation efficiency for dyes, such as red fluorescent dyes. A single mode fiber coupler is used to replace the free-space beam splitter and one arm of the coupler is fused with a double-clad fiber device. The MPM and OCM share the same excitation light path in the core of a double-clad fiber, while the OCM and MPM signals were collected by the core and clad of the double-clad fiber, respectively. The performance of the introduced double-clad device is analyzed. The device can confine all the excitation light in the core and has a collection efficiency of 20% for the MPM signal. The efficiency can be further increased by fusing more multimode fibers with the DCF. Simultaneous optical coherence microscopic imaging, second harmonic generation imaging, and two-photon excitation fluorescence imaging are demonstrated in biological samples. PMID:21456873

  7. Multimodal microscopy and the stepwise multi-photon activation fluorescence of melanin

    NASA Astrophysics Data System (ADS)

    Lai, Zhenhua

    The author's work is divided into three aspects: multimodal microscopy, stepwise multi-photon activation fluorescence (SMPAF) of melanin, and customized-profile lenses (CPL) for on-axis laser scanners, which will be introduced respectively. A multimodal microscope provides the ability to image samples with multiple modalities on the same stage, which incorporates the benefits of all modalities. The multimodal microscopes developed in this dissertation are the Keck 3D fusion multimodal microscope 2.0 (3DFM 2.0), upgraded from the old 3DFM with improved performance and flexibility, and the multimodal microscope for targeting small particles (the "Target" system). The control systems developed for both microscopes are low-cost and easy-to-build, with all components off-the-shelf. The control system have not only significantly decreased the complexity and size of the microscope, but also increased the pixel resolution and flexibility. The SMPAF of melanin, activated by a continuous-wave (CW) mode near-infrared (NIR) laser, has potential applications for a low-cost and reliable method of detecting melanin. The photophysics of melanin SMPAF has been studied by theoretical analysis of the excitation process and investigation of the spectra, activation threshold, and photon number absorption of melanin SMPAF. SMPAF images of melanin in mouse hair and skin, mouse melanoma, and human black and white hairs are compared with images taken by conventional multi-photon fluorescence microscopy (MPFM) and confocal reflectance microscopy (CRM). SMPAF images significantly increase specificity and demonstrate the potential to increase sensitivity for melanin detection compared to MPFM images and CRM images. Employing melanin SMPAF imaging to detect melanin inside human skin in vivo has been demonstrated, which proves the effectiveness of melanin detection using SMPAF for medical purposes. Selective melanin ablation with micrometer resolution has been presented using the Target system. Compared to the traditional selective photothermolysis, this method demonstrates higher precision, higher specificity and deeper penetration. Therefore, the SMPAF guided selective ablation of melanin is a promising tool of removing melanin for both medical and cosmetic purposes. Three CPLs have been designed for low-cost linear-motion scanners, low-cost fast spinning scanners and high-precision fast spinning scanners. Each design has been tailored to the industrial manufacturing ability and market demands.

  8. Multiphoton Rabi oscillations of a ringlike three-level system

    NASA Astrophysics Data System (ADS)

    He, Yong Lin; Han, Jiu Ning

    2012-04-01

    We propose a ringlike three-level model and show that it can be realized using Stark states of highly excited potassium interacting with two-mode microwave fields. We demonstrate this by using both analytical model and numerical simulations to study the problems of multiphoton Rabi oscillations. The results show that the Rabi oscillation patterns of the ringlike three-level system exhibit additional envelope and nodes compared to the oscillatory behavior of a three-level cascade system; in other words, there exist collapse and revival phenomena for Rabi oscillation patterns. The origin of the collapse and revival of the population oscillations is the multiphoton two-color resonances at frequency ?=n?1+m?2. Our analytic results are in good agreement with numerical simulations. In addition, all the analytical solutions for the three basic configurations of the three-level system classified as the ?, vee, and cascade systems can be deduced from our analytical solution for the ringlike three-level configuration by setting one of the three coupling strengths equal to 0.

  9. Multimodal optical imaging with multiphoton microscopy and optical coherence tomography.

    PubMed

    Tang, Shuo; Zhou, Yifeng; Ju, Myeong Jin

    2012-05-01

    Two types of combined multiphoton microscopy and optical coherence tomography (MPM/OCT) are compared for multimodal optical imaging. Single-scale multiphoton microscopy and optical coherence microscopy (MPM/OCM) is shown to acquire multiple contrasts from MPM and OCT simultaneously. Multi-scale MPM/OCT is shown to provide multiple field-of-views (FOVs), where OCT provides tissue level imaging and MPM provides cellular level imaging. In both types, the MPM includes two channels which are two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). Representative images using each system are demonstrated on biological specimens. A detailed comparison of the two types of MPM/OCT shows that each system has its own pros and cons. MPM/OCM is high-resolution but with limited FOV, and OCM may or may not provide additional information than MPM depending on the samples. Multi-scale MPM/OCT can change FOV but need both low and high NA objectives. For future development, the two types of MPM/OCT can be further integrated to achieve both functions on a single system. PMID:22461146

  10. Multibeam multifocal multiphoton photon counting imaging in scattering media

    NASA Astrophysics Data System (ADS)

    Hoover, Erich E.

    Multiphoton microscopy is an invaluable technique for the neurological community, allowing for deep explorations within highly scattering tissues such as the brain. However, prior to this research multiphoton microscopy was limited in its ability to rapidly construct volumetric images deep within scattering specimens. This work establishes a technique that permits such exploration through the application of multiple beams separated in both space and time, where signal photons corresponding to those beams are demultiplexed through the use of a field programmable gate array. With this system a number of improvements are provided to research in scattering media, including the coveted ability to perform photon-counting imaging with multiple beams. The ability to perform these measurements with multiple beams permits unique quantitative measurements of fluorophores within living specimens, allowing new research into dynamic three-dimensional behavior occurring within the brain. Additionally, the ability to perform multimodal measurements without filtering allows for unique avenues of research where the harmonic generation is indistinguishable from the two-photon excited fluorescence. These improvements provide neuroscience researchers with a large assortment of technological tools that will permit them to perform numerous novel experiments within the brain and other highly-scattering specimens, which should one day lead to significant advances in our understanding of complex neuronal activity.

  11. Multiphoton polarization imaging of the stratum corneum and the dermis in ex-vivo human skin

    NASA Astrophysics Data System (ADS)

    Sun, Yen; Su, Jiunn-Wen; Lo, Wen; Lin, Sun-Jan; Jee, Shiou-Hwa; Dong, Chen-Yuan

    2003-12-01

    In this work, we demonstrate the application of multiphoton polarization imaging in resolving the structures in surface stratum corneum and dermal layers of ex-vivo human skin. By varying the excitation and emission polarizations, we characterized the structural features in both Laurdan labeled stratum corneum and dermal fibers. The results presented here have important consequences in bioimaging applications of the skin. Both the mechanics of transdermal drug delivery across the skin and physiological significance of the structural changes of the dermis can be monitored. Our results show that the transition dipoles of Laurdan molecules are preferentially oriented normal to the membrane surface. Furthermore, polarization imaging shows that fibrous structures in the dermis generate emission aligned strongly along the excitation polarization. This work shows that multiphoton polarization imaging can be a powerful method in identifying structural orientations in the skin and other biological structures.

  12. DNA Multiphoton Absorption Generates Localized Damage for Studying Repair Dynamics in Live Cells

    PubMed Central

    Daddysman, MatthewK.; Fecko, ChristopherJ.

    2011-01-01

    Investigations into the spatiotemporal dynamics of DNA repair using live-cell imaging are aided by the ability to generate well defined regions of ultravioletlike photolesions in an optical microscope. We demonstrate that multiphoton excitation of DNA in live cells with visible femtosecond pulses produces thymine cyclopyrimidine dimers (CPDs), the primary ultraviolet DNA photoproduct. The CPDs are produced with a cubic to supercubic power dependence using pulses in the wavelength range from at least 400 to 525nm. We show that the CPDs are confined in all three spatial dimensions, making multiphoton excitation of DNA with visible light an ideal technique for generating localized DNA photolesions in a wide variety of samples, from cultured cells to thicker tissues. We demonstrate the utility of this method by applying it to investigate the spatiotemporal recruitment of GFP-tagged topoisomerase I (TopI) to sites of localized DNA damage in polytene chromosomes within live cells of optically thick Drosophila salivary glands. PMID:22067170

  13. Resonance Enhanced Multi-photon Spectroscopy of DNA

    NASA Astrophysics Data System (ADS)

    Ligare, Marshall Robert

    For over 50 years DNA has been studied to better understand its connection to life and evolution. These past experiments have led to our understanding of its structure and function in the biological environment but the interaction of DNA with UV radiation at the molecular level is still not very well understood. Unique mechanisms in nucleobase chromaphores protect us from adverse chemical reactions after UV absorption. Studying these processes can help develop theories for prebiotic chemistry and the possibility of alternative forms of DNA. Using resonance enhanced multi-photon spectroscopic techniques in the gas phase allow for the structure and dynamics of individual nucleobases to be studied in detail. Experiments studying different levels of structure/complexity with relation to their biological function are presented. Resonant IR multiphoton dissociation spectroscopy in conjunction with molecular mechanics and DFT calculations are used to determine gas phase structures of anionic nucleotide clusters. A comparison of the identified structures with known biological function shows how the hydrogen bonding of the nucleotides and their clusters free of solvent create favorable structures for quick incorporation into enzymes such as DNA polymerase. Resonance enhanced multi-photon ionization (REMPI) spectroscopy techniques such as resonant two photon ionization (R2PI) and IR-UV double resonance are used to further elucidate the structure and excited state dynamics of the bare nucleobases thymine and uracil. Both exhibit long lived excited electronic states that have been implicated in DNA photolesions which can ultimately lead to melanoma and carcinoma. Our experimental data in comparison with many quantum chemical calculations suggest a new picture for the dynamics of thymine and uracil in the gas phase. A high probability of UV absorption from a vibrationally hot ground state to the excited electronic state shows that the stability of thymine and uracil comes from its intrinsic molecular properties and possibly a hydrogen bonding solvent capable of dissipating excess vibrational energy. Due to the high specificity and sensitivity of resonant two photon ionization coupled with molecular beam mass spectrometry a new analytical technique for identifying molecular markers in archaeological vessels is presented. The xanthine alkaloids theobromine, theophylline and caffeine are identified in Central American and North American pottery sherds by direct desorption/resonant laser ionization mass spectrometry.

  14. Multi-focal multiphoton lithography.

    PubMed

    Ritschdorff, Eric T; Nielson, Rex; Shear, Jason B

    2012-03-01

    Multiphoton lithography (MPL) provides unparalleled capabilities for creating high-resolution, three-dimensional (3D) materials from a broad spectrum of building blocks and with few limitations on geometry, qualities that have been key to the design of chemically, mechanically, and biologically functional microforms. Unfortunately, the reliance of MPL on laser scanning limits the speed at which fabrication can be performed, making it impractical in many instances to produce large-scale, high-resolution objects such as complex micromachines, 3D microfluidics, etc. Previously, others have demonstrated the possibility of using multiple laser foci to simultaneously perform MPL at numerous sites in parallel, but use of a stage-scanning system to specify fabrication coordinates resulted in the production of identical features at each focal position. As a more general solution to the bottleneck problem, we demonstrate here the feasibility for performing multi-focal MPL using a dynamic mask to differentially modulate foci, an approach that enables each fabrication site to create independent (uncorrelated) features within a larger, integrated microform. In this proof-of-concept study, two simultaneously scanned foci produced the expected two-fold decrease in fabrication time, and this approach could be readily extended to many scanning foci by using a more powerful laser. Finally, we show that use of multiple foci in MPL can be exploited to assign heterogeneous properties (such as differential swelling) to micromaterials at distinct positions within a fabrication zone. PMID:22282105

  15. Measurements of multiphoton action cross sections for multiphoton microscopy

    PubMed Central

    Cheng, Li-Chung; Horton, Nicholas G.; Wang, Ke; Chen, Shean-Jen; Xu, Chris

    2014-01-01

    We report quantitative measurements of two-, three-, and four-photon excitation action cross sections of several commonly used fluorophores and fluorescent proteins at three different excitation wavelengths of 800 nm, 1300 nm, and 1680 nm. The measured cross section values are consistent with simple quantum mechanic estimations. These values indicate that the optimum repetition rate for deep tissue 3-photon microscopy is approximately 1 to 2 MHz. We further demonstrate that it is feasible to perform 4-photon fluorescence microscopy of GFP labeled microglia in mouse brain in vivo at 1700 nm. 4-photon excitation increases the accessibility of fluorophores at the long wavelength spectral window of 1700 nm. PMID:25360361

  16. Fluorescence lifetime imaging with near-infrared dyes

    NASA Astrophysics Data System (ADS)

    Becker, Wolfgang; Shcheslavskiy, Vladislav

    2013-02-01

    Near-infrared (NIR) dyes are used as fluorescence markers in small-animal imaging and in diffuse optical tomography of the human brain. In these applications it is important to know whether the dyes bind to proteins or other tissue constituents, and whether their fluorescence lifetimes depend on the targets they are bound to. Unfortunately, neither the lasers nor the detectors of commonly used confocal and multiphoton laser scanning microscopes allow for excitation and detection of NIR fluorescence. We therefore upgraded existing confocal TCSPC FLIM systems with NIR lasers and NIR sensitive detectors. In multiphoton systems we used the Ti:Sa laser as a one-photon excitation source in combination with an NIR-sensitive detector in the confocal beam path. We tested a number of NIR dyes in biological tissue. Some of them showed clear lifetime changes depending on the tissue structures they are bound to. We therefore believe that NIR FLIM can deliver supplementary information on the tissue constitution and on local biochemical parameters.

  17. Nanoparticle-assisted-multiphoton microscopy for in vivo brain imaging of mice

    NASA Astrophysics Data System (ADS)

    Qian, Jun

    2015-03-01

    Neuro/brain study has attracted much attention during past few years, and many optical methods have been utilized in order to obtain accurate and complete neural information inside the brain. Relying on simultaneous absorption of two or more near-infrared photons by a fluorophore, multiphoton microscopy can achieve deep tissue penetration and efficient light detection noninvasively, which makes it very suitable for thick-tissue and in vivo bioimaging. Nanoparticles possess many unique optical and chemical properties, such as anti-photobleaching, large multiphoton absorption cross-section, and high stability in biological environment, which facilitates their applications in long-term multiphoton microscopy as contrast agents. In this paper, we will introduce several typical nanoparticles (e.g. organic dye doped polymer nanoparticles and gold nanorods) with high multiphoton fluorescence efficiency. We further applied them in two- and three-photon in vivo functional brain imaging of mice, such as brain-microglia imaging, 3D architecture reconstruction of brain blood vessel, and blood velocity measurement.

  18. Infrared, Raman, and Ultraviolet Absorption Spectra and Theoretical Calculations and Structure of 2,6-Difluoropyridine in its Ground and Excited Electronic States

    PubMed Central

    Sheu, Hong-Li; Kim, Sunghwan; Laane, Jaan

    2014-01-01

    The infrared and Raman spectra of 2,6-difluoropyridine (26DFPy) along with ab initio and DFT computations have been used to assign the vibrations of the molecule in its S0 electronic ground state and to calculate its structure. The ultraviolet absorption spectrum showed the electronic transition to the S1(?,?*) state to be at 37,820.2 cm?1. With the aid of ab initio computations the vibrational frequencies for this excited state were also determined. TD-B3LYP and CASSCF computations for the excited states were carried out to calculate the structures for the S1(?,?*) and S2(n,?*) excited states. The CASSCF results predict that the S1(?,?*) state is planar and the S2(n,?*) state has a barrier to planarity of 256 cm?1. The TD-B3LYP computations predict a barrier of 124 cm?1 for the S1(?,?*) states, but the experimental results support the planar structure. Hypothetical models for the ring-puckering potential energy function were calculated for both electronic excited states to show the predicted quantum states. The changes in the vibrational frequencies in the two excited states reflect the weaker ? bonding within the pyridine ring. PMID:24070189

  19. Excitation of the E2 and W1 "Arched" Filaments Near the Galactic Center as Deduced from Far-Infrared Spectroscopy

    NASA Technical Reports Server (NTRS)

    Colgan, Sean W.J.; Erickson, Edwin F.; Simpson, Janet P.; Haas, Michael R.; Morris, Mark

    1994-01-01

    We present measurements of the far-infrared (FIR) fine structure lines [S III] (33 microns), [Si II] (35 microns), [O III] (51, 88 microns), [O I] (63 microns) and [C II] (158 microns) and the adjacent continua along a scan crossing the E2 and W1 thermal radio filaments in the Galactic center 'Arc'. The deduced electron density and excitation vary along the scan by less than factors of two and three, respectively. The properties of the two filaments are similar: the line and continuum fluxes peak at the radio ridge, and the ridge/off-ridge contrast is greatest for the FIR continuum and ionized lines, lower for the single dish radio measurements (Sofue et al. 1986), and smallest for the low excitation lines. The spatial coincidence of the FIR and radio peaks demonstrates that any excitation mechanism for the radio continuum filaments must also account for the FIR line and continuum emission. The FIR luminosity of approx. 3 x 10(exp 5) Solar Luminosity per beam, and the association of [O III] emission with the filaments poses difficulties for shock and MHD models. Photoionization of molecular cloud edges by a random distribution of stars is the most likely excitation mechanism among those proposed. The continuum and the low excitation line fluxes are consistent with an origin in photodissociated molecular material adjacent to the photoionized gas.

  20. Femtosecond multiphoton ionization of free and aggregated NaI via Rydberg states

    NASA Astrophysics Data System (ADS)

    Stibbe, Darian T.; Charron, Eric; Suzor-Weiner, Annick

    2001-12-01

    Using a wave packet simulation on simplified potential curves, we investigate the vibrational dynamics of NaI in its first excited electronic state, both isolated and aggregated with a molecule of acetonitrile. The probe signal is obtained by resonant multiphoton ionization (REMPI) of the diatomic chromophore NaI via Rydberg states. The addition of acetonitrile is found to change greatly the ionization signal, suggesting strong modification of the reaction dynamics.

  1. Fourier transform ion cyclotron resonance mass spectrometry for the characterisation of kavalactones in the kava plant: elemental formulae confirmation by dual spray accurate mass measurement and structural confirmation by infrared multiphoton dissociation and sustained off-resonance irradiation collision induced dissociation.

    PubMed

    Warburton, Emma; Bristow, Tony

    2006-01-01

    Roots and extracts of the kava plant have been used in herbal medicine to treat sleep disturbances, stress and anxiety, although reported cases of liver toxicity led to many countries restricting its sale. The detection of the presence of kava in many medicinal products requires the use of methods capable of identifying the kavalactones with high certainty. Here, we describe the use of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) for the characterisation of six kavalactones (kavain, dihydrokavain, methysticin, dihydromethysticin, yangonin and desmethoxyyangonin) utilising accurate mass measurement for the determination of their elemental formulae and product ion MS (both sustained off-resonance irradiation collision-induced dissociation and infrared multiphoton dissociation (SORI-CID and IRMPD) for structural confirmation. High performance liquid chromatography/FT-ICR-MS with a dual spray system for internal calibration of mass spectra was employed for accurate mass measurement and the determination of elemental formulae of the kavalactones in both standards and a root extract to confirm the presence of the kavalactones in the root powder. Mass accuracy of < 1 ppm was achieved. For structural confirmation, the IRMPD and SORI-CID spectra of the kavalactones in standards and a kava root powder extract were compared. Accurate mass measurement of the product ions was also conducted by external calibration and the elemental formula determined to aid with structural confirmation. The presence of the same fragment ions detected in the standards as in the extract further confirmed the presence of the kavalactones in the kava root powder with high certainty. PMID:17057279

  2. Mitochondria-targeted Triphenylamine Derivatives Activatable by Two-Photon Excitation for Triggering and Imaging Cell Apoptosis.

    PubMed

    Chennoufi, Rahima; Bougherara, Houcine; Gagey-Eilstein, Nathalie; Dumat, Blaise; Henry, Etienne; Subra, Frédéric; Bury-Moné, Stéphanie; Mahuteau-Betzer, Florence; Tauc, Patrick; Teulade-Fichou, Marie-Paule; Deprez, Eric

    2016-01-01

    Photodynamic therapy (PDT) leads to cell death by using a combination of a photosensitizer and an external light source for the production of lethal doses of reactive oxygen species (ROS). Since a major limitation of PDT is the poor penetration of UV-visible light in tissues, there is a strong need for organic compounds whose activation is compatible with near-infrared excitation. Triphenylamines (TPAs) are fluorescent compounds, recently shown to efficiently trigger cell death upon visible light irradiation (458 nm), however outside the so-called optical/therapeutic window. Here, we report that TPAs target cytosolic organelles of living cells, mainly mitochondria, triggering a fast apoptosis upon two-photon excitation, thanks to their large two-photon absorption cross-sections in the 760-860 nm range. Direct ROS imaging in the cell context upon multiphoton excitation of TPA and three-color flow cytometric analysis showing phosphatidylserine externalization indicate that TPA photoactivation is primarily related to the mitochondrial apoptotic pathway via ROS production, although significant differences in the time courses of cell death-related events were observed, depending on the compound. TPAs represent a new class of water-soluble organic photosensitizers compatible with direct two-photon excitation, enabling simultaneous multiphoton fluorescence imaging of cell death since a concomitant subcellular TPA re-distribution occurs in apoptotic cells. PMID:26947258

  3. Mitochondria-targeted Triphenylamine Derivatives Activatable by Two-Photon Excitation for Triggering and Imaging Cell Apoptosis

    PubMed Central

    Chennoufi, Rahima; Bougherara, Houcine; Gagey-Eilstein, Nathalie; Dumat, Blaise; Henry, Etienne; Subra, Frédéric; Bury-Moné, Stéphanie; Mahuteau-Betzer, Florence; Tauc, Patrick; Teulade-Fichou, Marie-Paule; Deprez, Eric

    2016-01-01

    Photodynamic therapy (PDT) leads to cell death by using a combination of a photosensitizer and an external light source for the production of lethal doses of reactive oxygen species (ROS). Since a major limitation of PDT is the poor penetration of UV-visible light in tissues, there is a strong need for organic compounds whose activation is compatible with near-infrared excitation. Triphenylamines (TPAs) are fluorescent compounds, recently shown to efficiently trigger cell death upon visible light irradiation (458 nm), however outside the so-called optical/therapeutic window. Here, we report that TPAs target cytosolic organelles of living cells, mainly mitochondria, triggering a fast apoptosis upon two-photon excitation, thanks to their large two-photon absorption cross-sections in the 760–860 nm range. Direct ROS imaging in the cell context upon multiphoton excitation of TPA and three-color flow cytometric analysis showing phosphatidylserine externalization indicate that TPA photoactivation is primarily related to the mitochondrial apoptotic pathway via ROS production, although significant differences in the time courses of cell death-related events were observed, depending on the compound. TPAs represent a new class of water-soluble organic photosensitizers compatible with direct two-photon excitation, enabling simultaneous multiphoton fluorescence imaging of cell death since a concomitant subcellular TPA re-distribution occurs in apoptotic cells. PMID:26947258

  4. Characterization and application of chirped photonic crystal fiber in multiphoton imaging.

    PubMed

    Yu, Jiali; Zeng, Haishan; Lui, Harvey; Skibina, Julia S; Steinmeyer, Günter; Tang, Shuo

    2014-05-01

    Fiber delivery of ultrashort pulses is important for multiphoton endoscopy. A chirped photonic crystal fiber (CPCF) is first characterized for its transmission bandwidth, propagation loss, and dispersion properties. Its extremely low dispersion (~150 fs(2)/m) enables the delivery of sub-30 fs pulses through a ~1 m-long CPCF. The CPCF is then incorporated into a multiphoton imaging system and its performance is demonstrated by imaging various biological samples including yew leaf, mouse tendon, and human skin. The imaging quality is further compared with images acquired by a multiphoton imaging system with free-space or hollow-core photonic band-gap fiber (PBF) delivery of pulses. Compared with free-space system, the CPCF delivered system maintains the same ultrashort pulsewidth and the image qualities are comparable. Compared with the PBF delivery, CPCF provides a 35 times shorter pulsewidth at the sample location, which results in a ~12 and 50 times improvement in two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG) signals respectively. Our results show that CPCF has great potential for fiber delivery of ultrashort pulses for multiphoton endoscopy. PMID:24921739

  5. Multi-photon laser scanning microscopy using an acoustic optical deflector.

    PubMed Central

    Lechleiter, James D; Lin, Da-Ting; Sieneart, Ilse

    2002-01-01

    Multi-photon laser scanning microscopes have many advantages over single-photon systems. However, the speed and flexibility of currently available multi-photon microscopes are limited by the use of mechanical mirrors to steer pulsed radiation for fluorophore excitation. Here, we describe the multi-photon adaptation of a confocal microscope that uses an acoustic optical deflector (AOD) for beam steering. AODs are capable of very rapid scanning and, in addition, offer the flexibility of zooming, panning, and being adjustable for slow image acquisition. Because of the highly dispersive nature of AODs, pulsed radiation must be temporally compressed by introducing negative dispersion into the beam path. More critically, pulsed radiation must also be spatially compressed by introducing lateral dispersion into the beam path. This was accomplished by using two prisms in the external beam path and by introducing a third prism element subsequent to the AOD. The end result is an AOD-based multi-photon microscope that is capable of rapid imaging of physiological events as well as slow detection of weakly fluorescent biological samples. PMID:12324446

  6. Live-cell multiphoton fluorescence correlation spectroscopy with an improved large Stokes shift fluorescent protein

    PubMed Central

    Guan, Yinghua; Meurer, Matthias; Raghavan, Sarada; Rebane, Aleksander; Lindquist, Jake R.; Santos, Sofia; Kats, Ilia; Davidson, Michael W.; Mazitschek, Ralph; Hughes, Thomas E.; Drobizhev, Mikhail; Knop, Michael; Shah, Jagesh V.

    2015-01-01

    We report an improved variant of mKeima, a monomeric long Stokes shift red fluorescent protein, hmKeima8.5. The increased intracellular brightness and large Stokes shift (?180 nm) make it an excellent partner with teal fluorescent protein (mTFP1) for multiphoton, multicolor applications. Excitation of this pair by a single multiphoton excitation wavelength (MPE, 850 nm) yields well-separable emission peaks (?120-nm separation). Using this pair, we measure homo- and hetero-oligomerization interactions in living cells via multiphoton excitation fluorescence correlation spectroscopy (MPE-FCS). Using tandem dimer proteins and small-molecule inducible dimerization domains, we demonstrate robust and quantitative detection of intracellular proteinprotein interactions. We also use MPE-FCCS to detect drugprotein interactions in the intracellular environment using a Coumarin 343 (C343)-conjugated drug and hmKeima8.5 as a fluorescence pair. The mTFP1/hmKeima8.5 and C343/hmKeima8.5 combinations, together with our calibration constructs, provide a practical and broadly applicable toolbox for the investigation of molecular interactions in the cytoplasm of living cells. PMID:25877871

  7. Polarization phenomena in multiphoton ionization of atoms

    NASA Technical Reports Server (NTRS)

    Jacobs, V. L.

    1973-01-01

    The theory of multiphoton ionization for an atomic system of arbitrary complexity is developed using a density matrix formalism. An expression is obtained which determines the differential N-photon ionization cross section as a function of the polarization states of the target atom and the incident radiation. The parameters which characterize the photoelectron angular distribution are related to the general reduced matrix elements for the N-photon transition. Two-photon ionization of unpolarized atoms is treated as an illustration of the use of the theory. The dependence of the multiphoton ionization cross section on the polarization state of the incident radiation, which has been observed in two- and three-photon ionization of Cs, is accounted for by the theory. Finally, the photoelectron spin polarization produced by the multiphoton ionization of unpolarized atoms, like the analogous polarization resulting from single-photon ionization, is found to depend on the circular polarization of the incident radiation.

  8. Multiphoton microscopy in defining liver function

    NASA Astrophysics Data System (ADS)

    Thorling, Camilla A.; Crawford, Darrell; Burczynski, Frank J.; Liu, Xin; Liau, Ian; Roberts, Michael S.

    2014-09-01

    Multiphoton microscopy is the preferred method when in vivo deep-tissue imaging is required. This review presents the application of multiphoton microscopy in defining liver function. In particular, multiphoton microscopy is useful in imaging intracellular events, such as mitochondrial depolarization and cellular metabolism in terms of NAD(P)H changes with fluorescence lifetime imaging microscopy. The morphology of hepatocytes can be visualized without exogenously administered fluorescent dyes by utilizing their autofluorescence and second harmonic generation signal of collagen, which is useful in diagnosing liver disease. More specific imaging, such as studying drug transport in normal and diseased livers are achievable, but require exogenously administered fluorescent dyes. If these techniques can be translated into clinical use to assess liver function, it would greatly improve early diagnosis of organ viability, fibrosis, and cancer.

  9. Suitable microscope objectives for multiphoton digital imaging

    NASA Astrophysics Data System (ADS)

    Benham, Gerald S.; Schwartz, Stanley

    2002-06-01

    The proper choice of an objective lens can have a profound effect on multiphoton microscopy. Multiphoton requires that the objective have adequate transmission in the IR range. This technology has a requisite to image the specimen near the slide surface next to the cover glass and to of 200 microns or more. Fluor and Plan Apochromat objectives. All lenses suffer from aberrations, which can prevent imaging an exact replica of the object. Most objectives could be used for multiphoton imaging;; however lenses should be selected from Plan Fluors, Plan Apochromats, and Super (S) Fluors. Highly corrected water immersion objectives (WI) permit deep penetration within aqueous environments and living tissues, beyond that of oil immersion objectives. Spectral transmission data is presented for Nikon CFI60 various objective lenses with high Nas, long working distances, and maximum aberration correction.

  10. Kinetic studies following state-selective laser excitation

    SciTech Connect

    Keto, J.W.

    1992-01-01

    We have made measurements of state-to-state deactivation cross sections and radiative lifetimes for Xe*(6p,6p{prime},7p) and Kr*(5p) states in xenon and krypton buffer gases. These results are relevant to kinetic models and both excimer lasers and the infrared xenon laser; and they are a significant improvement in the precision of the known radiative lifetimes. This type of experiment can now be compared with recent calculations of state-to-state collisional relaxation in rare-gases by Hickman, Huestis, and Saxon. We have also made significant progress in the study of the electronic spectra of small molecules of the rare gases. Spectra have been obtained for Xe{sub 2}, Xe{sub 3}, Xe{sub 4}, and larger clusters. As guidance for the larger clusters of the rare gases we have obtained the first multiphoton spectra for excitons in condensed xenon. In collaboration with research on the multiphoton spectra of the rare gases, we have continued experiments using synchrotron radiation in collaboration with the University of Hamburg. In experiments there we have observed excitation and fluorescence spectra for single xenon atoms at the surface, within the second layer, and within the bulk of large argon clusters.

  11. Human bladder cancer diagnosis using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Mukherjee, Sushmita; Wysock, James S.; Ng, Casey K.; Akhtar, Mohammed; Perner, Sven; Lee, Ming-Ming; Rubin, Mark A.; Maxfield, Frederick R.; Webb, Watt W.; Scherr, Douglas S.

    2009-02-01

    At the time of diagnosis, approximately 75% of bladder cancers are non-muscle invasive. Appropriate diagnosis and surgical resection at this stage improves prognosis dramatically. However, these lesions, being small and/or flat, are often missed by conventional white-light cystoscopes. Furthermore, it is difficult to assess the surgical margin for negativity using conventional cystoscopes. Resultantly, the recurrence rates in patients with early bladder cancer are very high. This is currently addressed by repeat cystoscopies and biopsies, which can last throughout the life of a patient, increasing cost and patient morbidity. Multiphoton endoscopes offer a potential solution, allowing real time, noninvasive biopsies of the human bladder, as well as an up-close assessment of the resection margin. While miniaturization of the Multiphoton microscope into an endoscopic format is currently in progress, we present results here indicating that Multiphoton imaging (using a bench-top Multiphoton microscope) can indeed identify cancers in fresh, unfixed human bladder biopsies. Multiphoton images are acquired in two channels: (1) broadband autofluorescence from cells, and (2) second harmonic generation (SHG), mostly by tissue collagen. These images are then compared with gold standard hematoxylin/eosin (H&E) stained histopathology slides from the same specimen. Based on a "training set" and a very small "blinded set" of samples, we have found excellent correlation between the Multiphoton and histopathological diagnoses. A larger blinded analysis by two independent uropathologists is currently in progress. We expect that the conclusion of this phase will provide us with diagnostic accuracy estimates, as well as the degree of inter-observer heterogeneity.

  12. New developments in multimodal clinical multiphoton tomography

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2011-03-01

    80 years ago, the PhD student Maria Goeppert predicted in her thesis in Goettingen, Germany, two-photon effects. It took 30 years to prove her theory, and another three decades to realize the first two-photon microscope. With the beginning of this millennium, first clinical multiphoton tomographs started operation in research institutions, hospitals, and in the cosmetic industry. The multiphoton tomograph MPTflexTM with its miniaturized flexible scan head became the Prism-Award 2010 winner in the category Life Sciences. Multiphoton tomographs with its superior submicron spatial resolution can be upgraded to 5D imaging tools by adding spectral time-correlated single photon counting units. Furthermore, multimodal hybrid tomographs provide chemical fingerprinting and fast wide-field imaging. The world's first clinical CARS studies have been performed with a hybrid multimodal multiphoton tomograph in spring 2010. In particular, nonfluorescent lipids and water as well as mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen have been imaged in patients with dermatological disorders. Further multimodal approaches include the combination of multiphoton tomographs with low-resolution imaging tools such as ultrasound, optoacoustic, OCT, and dermoscopy systems. Multiphoton tomographs are currently employed in Australia, Japan, the US, and in several European countries for early diagnosis of skin cancer (malignant melanoma), optimization of treatment strategies (wound healing, dermatitis), and cosmetic research including long-term biosafety tests of ZnO sunscreen nanoparticles and the measurement of the stimulated biosynthesis of collagen by anti-ageing products.

  13. Suppression of multiphoton intrashell resonances in Li Rydberg atoms

    SciTech Connect

    Waheed, A.; Fregenal, D.; Frette, O.; Foerre, M.; Hjertaker, B. T.; Preclikova, J.; Horsdal, E.; Pilskog, I.

    2011-06-15

    Multiphoton intrashell transitions in strongly driven Li (n=25) Rydberg atoms are studied experimentally. Orthogonal dc electric and magnetic fields lift the degeneracy of the n shell and define the eccentricity e of the initial coherent elliptic states, which are formed by laser excitation and subsequent adiabatic transformation. The intrashell transitions are driven by a time-harmonic electric field linearly polarized parallel to the major axis of the ellipse. N-photon resonances with N=1-9 are studied as a function of e. All resonances with N{>=}3 are suppressed at certain e values in between 0 and 1. A similar system was analyzed by Yabuzaki et al. [Phys. Rev. A 10, 1955 (1974)] who found a simple pattern of suppressions that applies also for the present experiments. The results of these experimentally confirm that each time N is increased by two, an additional suppression is observed.

  14. Label-free multi-photon imaging using a compact femtosecond fiber laser mode-locked by carbon nanotube saturable absorber

    PubMed Central

    Kieu, K.; Mehravar, S.; Gowda, R.; Norwood, R. A.; Peyghambarian, N.

    2013-01-01

    We demonstrate label-free multi-photon imaging of biological samples using a compact Er3+-doped femtosecond fiber laser mode-locked by a single-walled carbon nanotube (CNT). These compact and low cost lasers have been developed by various groups but they have not been exploited for multiphoton microscopy. Here, it is shown that various multiphoton imaging modalities (e.g. second harmonic generation (SHG), third harmonic generation (THG), two-photon excitation fluorescence (TPEF), and three-photon excitation fluorescence (3PEF)) can be effectively performed on various biological samples using a compact handheld CNT mode-locked femtosecond fiber laser operating in the telecommunication window near 1560nm. We also show for the first time that chlorophyll fluorescence in plant leaves and diatoms can be observed using 1560nm laser excitation via three-photon absorption. PMID:24156074

  15. Studies on wide-field-of-view multiphoton imaging using the flexible clinical multiphoton tomograph MPTflex

    NASA Astrophysics Data System (ADS)

    Weinigel, Martin; Breunig, Hans Georg; Fischer, Peter; Kellner-Hfer, Marcel; Bckle, Rainer; Knig, Karsten

    2012-03-01

    Multiphoton imaging systems are capable of high-resolution 3-D image acquisition of deep tissue. A first commercially available CE-certified biomedical system for subcelluar resolution of human skin has been launched by JenLab company with the DermaInspectR in 2002. The demand for more flexibility caused the development of the MPTflexR, which provides an increased flexibility and accessibility especially for clinical and cosmetic examinations. However the high resolution of clinical multiphoton tomographs are adherent with a small field-of-view (FOV) of about 360360?m2. Especially time-consuming is the relocation of areas of interest (AOI) like lesions, sweat glands or hair shafts during a multiphoton examination. This limitation can be be overcome by macroscopic large-area (wide-field-ofview) multiphoton tomography, which is tested first within this work.

  16. High resolution multiphoton spectroscopy by a tunable free-electron-laser light.

    PubMed

    itnik, M; Miheli?, A; Bu?ar, K; Kav?i?, M; Rubensson, J-E; Svanquist, M; Sderstrm, J; Feifel, R; Sthe, C; Ovcharenko, Y; Lyamayev, V; Mazza, T; Meyer, M; Simon, M; Journel, L; Lning, J; Plekan, O; Coreno, M; Devetta, M; Di Fraia, M; Finetti, P; Richter, R; Grazioli, C; Prince, K C; Callegari, C

    2014-11-01

    Seeded free electron lasers theoretically have the intensity, tunability, and resolution required for multiphoton spectroscopy of atomic and molecular species. Using the seeded free electron laser FERMI and a novel detection scheme, we have revealed the two-photon excitation spectra of dipole-forbidden doubly excited states in helium. The spectral profiles of the lowest (-1,0)(+1) (1)S(e) and (0,1)(0) (1)D(e) resonances display energy shifts in the meV range that depend on the pulse intensity. The results are explained by an effective two-level model based on calculated Rabi frequencies and decay rates. PMID:25415905

  17. ALMA DETECTION OF THE VIBRATIONALLY EXCITED HCN J = 4-3 EMISSION LINE IN THE AGN-HOSTING LUMINOUS INFRARED GALAXY IRAS 205514250

    SciTech Connect

    Imanishi, Masatoshi; Nakanishi, Kouichiro

    2013-10-01

    We present results from our ALMA Cycle 0 observations, at the frequencies around the HCN, HCO{sup +}, and HNC J = 4-3 transition lines, of the luminous infrared galaxy IRAS 205514250 at z = 0.043, which is known to host an energetically important obscured active galactic nucleus (AGN). In addition to the targeted HCN, HCO{sup +}, and HNC J = 4-3 emission lines, two additional strong emission lines are seen, which we attribute to H{sub 2}S and CH{sub 3}CN(+CCH). The HCN-to-HCO{sup +} J = 4-3 flux ratio (?0.7) is higher than in the other starburst-dominated galaxy (?0.2) observed in our ALMA Cycle 0 program. We tentatively (?5?) detected the vibrationally excited (v {sub 2} = 1) HCN J = 4-3 (l = 1f) emission line, which is important for testing an infrared radiative pumping scenario for HCN. This is the second detection of this molecular transition in external galaxies. The most likely reason for this detection is not only the high flux of this emission line, but also the small molecular line widths observed in this galaxy, suggesting that vibrational excitation of HCN may be relatively common in AGN-hosting galaxies.

  18. Infrared phonon anomaly and magnetic excitations in single-crystal Cu3Bi(SeO3)2O2Cl

    NASA Astrophysics Data System (ADS)

    Miller, K. H.; Stephens, P. W.; Martin, C.; Constable, E.; Lewis, R. A.; Berger, H.; Carr, G. L.; Tanner, D. B.

    2012-11-01

    Infrared reflection and transmission as a function of temperature have been measured on single crystals of Cu3Bi(SeO3)2O2Cl. The complex dielectric function and optical properties along all three principal axes of the orthorhombic cell were obtained via Kramers-Kronig analysis and by fits to a Drude-Lorentz model. Below 115 K, 16 additional modes [8(E?)+6(E?b?)+2(E??)] appear in the phonon spectra; however, powder x-ray diffraction measurements do not detect a new structure at 85 K. Potential explanations for the new phonon modes are discussed. Transmission in the far infrared as a function of temperature has revealed magnetic excitations originating below the magnetic ordering temperature (Tc24 K). The origin of the excitations in the magnetically ordered state will be discussed in terms of their response to different polarizations of incident light, behavior in externally applied magnetic fields, and the anisotropic magnetic properties of Cu3Bi(SeO3)2O2Cl as determined by dc susceptibility measurements.

  19. Infrared study on room-temperature atomic layer deposition of HfO{sub 2} using tetrakis(ethylmethylamino)hafnium and remote plasma-excited oxidizing agents

    SciTech Connect

    Kanomata, Kensaku; Ohba, Hisashi; Pungboon Pansila, P.; Ahmmad, Bashir; Kubota, Shigeru; Hirahara, Kazuhiro; Hirose, Fumihiko

    2015-01-01

    Room-temperature atomic layer deposition (ALD) of HfO{sub 2} was examined using tetrakis (ethylmethylamino)hafnium (TEMAH) and remote plasma-excited water and oxygen. A growth rate of 0.26 nm/cycle at room temperature was achieved, and the TEMAH adsorption and its oxidization on HfO{sub 2} were investigated by multiple internal reflection infrared absorption spectroscopy. It was observed that saturated adsorption of TEMAH occurs at exposures of ∼1 × 10{sup 5} L (1 L = 1 × 10{sup −6} Torr s) at room temperature, and the use of remote plasma-excited water and oxygen vapor is effective in oxidizing the TEMAH molecules on the HfO{sub 2} surface, to produce OH sites. The infrared study suggested that Hf–OH plays a role as an adsorption site for TEMAH. The reaction mechanism of room temperature HfO{sub 2} ALD is discussed in this paper.

  20. In vivo multiphoton imaging of mitochondrial structure and function during acute kidney injury

    PubMed Central

    Hall, Andrew M.; Rhodes, George J.; Sandoval, Ruben M.; Corridon, Peter R.; Molitoris, Bruce A.

    2014-01-01

    Mitochondrial dysfunction has been implicated in the pathogenesis of acute kidney injury due to ischemia and toxic drugs. Methods for imaging mitochondrial function in cells using confocal microscopy are well established; more recently, it was shown that these techniques can be utilized in ex vivo kidney tissue using multiphoton microscopy. We extended this approach in vivo and found that kidney mitochondrial structure and function can be imaged in anesthetized rodents using multiphoton excitation of endogenous and exogenous fluorophores. Mitochondrial nicotinamide adenine dinucleotide increased markedly in rat kidneys in response to ischemia. Following intravenous injection, the mitochondrial membrane potentialdependent dye TMRM was taken up by proximal tubules; in response to ischemia, the membrane potential dissipated rapidly and mitochondria became shortened and fragmented in proximal tubules. In contrast, the mitochondrial membrane potential and structure were better maintained in distal tubules. Changes in mitochondrial structure, nicotinamide adenine dinucleotide, and membrane potential were found in the proximal, but not distal, tubules after gentamicin exposure. These changes were sporadic, highly variable among animals, and were preceded by changes in non-mitochondrial structures. Thus, real-time changes in mitochondrial structure and function can be imaged in rodent kidneys in vivo using multiphoton excitation of endogenous and exogenous fluorophores in response to ischemiareperfusion injury or drug toxicity. PMID:22992467

  1. Characterization of human normal and cancerous gastric submucosa based on multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Zhong, Jiazhao; Chen, G.; Liu, Y. C.; Zhuo, S. M.; Chen, J. X.; Yan, J.

    2011-11-01

    Gastric cancer is one of the most frequent cancers in the world; almost two-thirds of gastric cancer cases and deaths occur in less developed regions. The initial diagnosis of gastric cancer often is delayed because up to 80 percent of patients are asymptomatic during the early stages of stomach cancer. So the ability to perform real-time in vivo histological diagnosis for early gastric cancer at the cellular level during ongoing endoscopy is a long-standing goal of endoscopists. In this paper, using multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG), MPM images of human normal and cancerous gastric submucosa were obtained at excitation wavelength of 800 nm. The features such as the appearance of abnormal cells and the large loss of collagen in cancerous gastric submucosa were extracted to be as significant indicators to distinguish cancerous submucosa from normal submucosa. With the implementation of multiphoton microscopy concept in endoscopy applications, multiphoton endoscopy might realize in vivo histological diagnosis goal of endoscopists.

  2. Characterization of human normal and cancerous gastric submucosa based on multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Zhong, Jiazhao; Chen, G.; Liu, Y. C.; Zhuo, S. M.; Chen, J. X.; Yan, J.

    2012-03-01

    Gastric cancer is one of the most frequent cancers in the world; almost two-thirds of gastric cancer cases and deaths occur in less developed regions. The initial diagnosis of gastric cancer often is delayed because up to 80 percent of patients are asymptomatic during the early stages of stomach cancer. So the ability to perform real-time in vivo histological diagnosis for early gastric cancer at the cellular level during ongoing endoscopy is a long-standing goal of endoscopists. In this paper, using multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG), MPM images of human normal and cancerous gastric submucosa were obtained at excitation wavelength of 800 nm. The features such as the appearance of abnormal cells and the large loss of collagen in cancerous gastric submucosa were extracted to be as significant indicators to distinguish cancerous submucosa from normal submucosa. With the implementation of multiphoton microscopy concept in endoscopy applications, multiphoton endoscopy might realize in vivo histological diagnosis goal of endoscopists.

  3. Identification of normal and cancerous human colorectal muscularis propria by multiphoton microscopy in different sections

    NASA Astrophysics Data System (ADS)

    Zhou, Yi; Chen, Zhifen; Kang, Deyong; li, Lianhuang; Zhuo, Shuangmu; Zhu, Xiaoqin; Guan, Guoxian; Chen, Jianxin

    2016-01-01

    Multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) as a potential diagnostic tool is attractive. MPM can effectively provide information about morphological and biochemical changes in biological tissues at the molecular level. In this paper, we attempt to identify normal and cancerous human colorectal muscularis propria by multiphoton microscopy in different sections (both in transverse and longitudinal sections). The results show that MPM can display different microstructure changes in the transverse and longitudinal sections of colorectal muscularis propria. MPM also can quantitatively describe the alteration of collagen content between normal and cancerous muscle layers. These are important pathological findings that MPM images can bring more detailed complementary information about tissue architecture and cell morphology through observing the transverse and longitudinal sections of colorectal muscularis propria. This work demonstrates that MPM can be better for identifying the microstructural characteristics of normal and cancerous human colorectal muscularis propria in different sections.

  4. Label-free discrimination of normal and pulmonary cancer tissues using multiphoton fluorescence ratiometric microscopy

    NASA Astrophysics Data System (ADS)

    Wang, Chun-Chin; Wu, Ruei-Jr; Lin, Sung-Jan; Chen, Yang-Fang; Dong, Chen-Yuan

    2010-07-01

    We performed multiphoton excited autofluorescence and second harmonic generation microscopy for the distinction of normal, lung adenocarcinoma (LAC), and squamous cell carcinoma (SCC) specimens. In addition to morphological distinction, we derived quantitative metrics of cellular redox ratios for cancer discrimination. Specifically, the redox ratios of paired normal/SCC and normal/LAC specimens were found to be 0.530.05/0.410.06 and 0.560.02/0.350.06, respectively. The lower redox ratios in cancer specimens, indicating an increase in metabolic activity. These results show that the combination of morphological multiphoton imaging along with redox ratio indices can be used for the discrimination of normal and pulmonary cancer tissues.

  5. Exploring the molecular chemistry and excitation in obscured luminous infrared galaxies. An ALMA mm-wave spectral scan of NGC 4418

    NASA Astrophysics Data System (ADS)

    Costagliola, F.; Sakamoto, K.; Muller, S.; Martín, S.; Aalto, S.; Harada, N.; van der Werf, P.; Viti, S.; Garcia-Burillo, S.; Spaans, M.

    2015-10-01

    Context. Extragalactic observations allow the study of molecular chemistry and excitation under physical conditions which may differ greatly from those found in the Milky Way. The compact, obscured nuclei (CON) of luminous infrared galaxies (LIRG) combine large molecular columns with intense infrared (IR), ultra-violet (UV), and X- radiation and represent ideal laboratories for the study of the chemistry of the interstellar medium (ISM) under extreme conditions. Aims: Our aim was to obtain for the first time a multi-band spectral scan of a LIRG, and to derive molecular abundances and excitation to be compared to other Galactic and extragalactic environments. Methods: We obtained an ALMA Cycle 0 spectral scan of the dusty LIRG NGC 4418, spanning a total of 70.7 GHz in bands 3, 6, and 7. We use a combined local thermal equilibrium (LTE) and non-LTE (NLTE) fit of the spectrum in order to identify the molecular species and to derive column densities and excitation temperatures. We derive molecular abundances and compare them with other Galactic and extragalactic sources by means of a principal component analysis. Results: We detect 317 emission lines from a total of 45 molecular species, including 15 isotopic substitutions and 6 vibrationally excited variants. Our LTE/NLTE fit find kinetic temperatures from 20 to 350 K, and densities between 105 and 107 cm-3. The spectrum is dominated by vibrationally excited HC3N, HCN, and HNC, with vibrational temperatures from 300 to 450 K. We find that the chemistry of NCG 4418 is characterized by high abundances of HC3N, SiO, H2S, and c-HCCCH but a low CH3OH abundance. A principal component analysis shows that NGC 4418 and Arp 220 share very similar molecular abundances and excitation, which clearly set them apart from other Galactic and extragalactic environments. Conclusions: Our spectral scan confirms that the chemical complexity in the nucleus of NGC 4418 is one of the highest ever observed outside our Galaxy. The similar molecular abundances observed toward NCG 4418 and Arp 220 are consistent with a hot gas-phase chemistry, with the relative abundances of SiO and CH3OH being regulated by shocks and X-ray driven dissociation. The bright emission from vibrationally excited species confirms the presence of a compact IR source, with an effective diameter smaller than 5 pc and brightness temperatures higher than 350 K. The molecular abundances and the vibrationally excited spectrum are consistent with a young AGN/starburst system. We suggest that NGC 4418 may be a template for a new kind of chemistry and excitation, typical of CON. Because of the narrow line widths and bright molecular emission, NGC 4418 is the ideal target for further studies of the chemistry in CONs. The spectrum in Fig. 1 is only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/582/A91Appendices are available in electronic form at http://www.aanda.org

  6. Near infrared excited micro-Raman spectra of 4:1 methanol-ethanol mixture and ruby fluorescence at high pressure

    NASA Astrophysics Data System (ADS)

    Wang, X. B.; Shen, Z. X.; Tang, S. H.; Kuok, M. H.

    1999-06-01

    Near infrared (NIR) lasers, as a new excitation source for Raman spectroscopy, has shown its unique advantages and is being increasingly used for some special samples, such as those emitting strong fluorescence in the visible region. This article focuses on some issues related to high-pressure micro-Raman spectroscopy using NIR excitation source. The Raman spectra of 4:1 methanol-ethanol mixture (4:1 M-E) show a linear variation in both Raman shifts and linewidths under pressure up to 18 GPa. This result is useful in distinguishing Raman scattering of samples from that of the alcohol mixture, an extensively used pressure-transmitting medium. The R1 fluorescence in the red region induced by two-photon absorption of the NIR laser is strong enough to be used as pressure scale. The frequency and line width of the R1 lines are very sensitive to pressure change and the glass transition of the pressure medium. Our results manifest that it is reliable and convenient to use NIR induced two-photon excited fluorescence of ruby for both pressure calibration and distribution of pressure in the 4:1 M-E pressure transmitting medium.

  7. Studies of atmospheric molecules by multiphoton spectroscopy. Progress report, July 15, 1989--October, 1991

    SciTech Connect

    Johnson, P.M.

    1991-10-01

    Carbon dioxide presents a great challenge to spectroscopy because of its propensity toward dissociation in all of its excited states. Multiphoton ionization spectroscopy is usually not applicable to the study of dissociating molecules because the dissociation competes effectively with ionization, resulting in no signal. We reasoned, however, that with high enough laser fluence, ionization could compete with dissociation in the longer lived states, exposing them for study from the continuous spectral background resulting from rapidly dissociating states. We describe the various spectroscopic and photophysical effects found through the multiphoton ionization and multiphoton photoelectron spectra. A recently developed variant of threshold ionization spectroscopy, usually called ZEKE, has shown a great deal of usefulness in providing the same information as traditional photoelectron spectroscopy but with higher resolution and much better signal-to-noise when using standard laboratory lasers. Threshold ionization techniques locate the states of an ion by scanning a light source across the ionization continuum of a neutral and somehow detecting when electrons are produced with no kinetic energy. We chose to develop our capabilities in threshold ionization spectroscopy using aromatic molecules because of their importance and because their electronic structure allows a pump-probe type of excitation scheme which avoids the use of vacuum ultraviolet laser beams. Among aromatics, the azines are noted for their small S{sub 1}-T{sub 1} energy gap which give them unique and interesting photophysical properties. We have continued our work on the multiphoton spectrum of metastable nitrogen produced by an electric discharge in supersonic beam. We have been able to assign more of the lines and simulated their rotational structure but many peaks remain unassigned.

  8. Facile and high spatial resolution ratio-metric luminescence thermal mapping in microfluidics by near infrared excited upconversion nanoparticles

    NASA Astrophysics Data System (ADS)

    Wang, Yu; Cao, Wenbin; Li, Shunbo; Wen, Weijia

    2016-02-01

    A local area temperature monitor is important for precise control of chemical and biological processes in microfluidics. In this work, we developed a facile method to realize micron spatial resolution of temperature mapping in a microfluidic channel quickly and cost effectively. Based on the temperature dependent fluorescence emission of NaYF4:Yb3+, Er3+ upconversion nanoparticles (UCNPs) under near-infrared irradiation, ratio-metric imaging of UCNPs doped polydimethylsiloxane can map detailed temperature distribution in the channel. Unlike some reported strategies that utilize temperature sensitive organic dye (such as Rhodamine) to achieve thermal sensing, our method is highly chemically inert and physically stable without any performance degradation in long term operation. Moreover, this method can be easily scaled up or down, since the spatial and temperature resolution is determined by an optical imaging system. Our method supplied a simple and efficient solution for temperature mapping on a heterogeneous surface where usage of an infrared thermal camera was limited.

  9. Molecular engineering of intensely near-infrared absorbing excited states in highly conjugated oligo(porphinato)zinc-(polypyridyl)metal(II) supermolecules.

    PubMed

    Duncan, Timothy V; Ishizuka, Tomoya; Therien, Michael J

    2007-08-01

    A new series of chromophores, MPZn(n), which combine ethyne-bridged bis(terpyridyl)metal(II)-(porphinato)zinc(II) (MPZ(n)) and oligomeric, ethyne-bridged (porphinato)zinc(II) (PZn(n)) architectures, have been synthesized and characterized, along with a series of derivatives bearing pyrrolidinyl electron-releasing groups on the ancillary terpyridine units (Pyr(m)MPZn(n)). Cyclic voltammetric studies, as well as NMR, electronic absorption, fluorescence, and femtosecond pump-probe transient absorption spectroscopies, have been employed to study the ground- and excited-state properties of these unusual chromophores. All of these species possess intensely absorbing excited states having large spectral bandwidth that penetrate deep in the near-infrared (NIR) energy regime. Electronic structural variation of the molecular framework shows that the excited-state absorption maximum can be extensively modulated [lambdamax(T(1) --> T(n))] (880 nm < lambdamax < 1126 nm), while concomitantly maintaining impressively large T(1) --> T(n) absorption manifold spectral bandwidth (full width at half-maximum, fwhm, approximately 2000-2500 cm(-1)). Furthermore, these studies enable correlation of supermolecular electronic structure with the magnitude of the excited-state lifetime (tau(es)) and demonstrate that this parameter can be modulated over 4 orders of magnitude ( approximately 1 ns < tau(es) < 45 micros). Terpyridyl pyrrolidinyl substituents can be utilized to destabilize terpyridyl ligand pi(*) energy levels and diminish the E1/2 (M3+/2+) value of the bis(terpyridyl)metal(II) center: such perturbations determine the relative energies of the PZn(n)-derived 1pi-pi(*) and bis(terpyridyl)metal(II) charge-transfer states and establish whether the T(1)-state wave functions of MPZn(n) and PyrmMPZn(n) species manifest the extensive electronic delocalization and charge-separated (CS) features characteristic of long-lived triplet states that absorb strongly in the NIR. PMID:17629267

  10. Two-photon imaging with longer wavelength excitation in intact Arabidopsis tissues.

    PubMed

    Mizuta, Yoko; Kurihara, Daisuke; Higashiyama, Tetsuya

    2015-09-01

    In vivo imaging of living organisms is an important tool to investigate biological phenomena. Two-photon excitation microscopy (2PEM) is a laser-scanning microscopy that provides noninvasive, deep imaging in living organisms based on the principle of multiphoton excitation. However, application of 2PEM to plant tissues has not been fully developed, as plant-specific autofluorescence, optically dense tissues, and multiple light-scattering structures diminish the clarity of imaging. In this study, the advantages of 2PEM were identified for deep imaging of living and intact Arabidopsis thaliana tissues. When compared to single-photon imaging, near-infrared 2PEM, especially at 1000 nm, reduced chloroplast autofluorescence; autofluorescence also decreased in leaves, roots, pistils, and pollen grains. For clear and deep imaging, longer excitation wavelengths using the orange fluorescent proteins (FPs) TagRFP and tdTomato gave better results than with other colors. 2PEM at 980 nm also provided multicolor imaging by simultaneous excitation, and the combination of suitable FPs and excitation wavelengths allowed deep imaging of intact cells in root tips and pistils. Our results demonstrated the importance of choosing both suitable FPs and excitation wavelengths for clear two-photon imaging. Further advances in in vivo analysis using 2PEM will facilitate more extensive studies in the plant biological sciences. PMID:25588923

  11. Imaging tissue engineering scaffolds using multiphoton microscopy.

    PubMed

    Sun, Yen; Tan, Hsin-Yuan; Lin, Sung-Jan; Lee, Hsuan-Shu; Lin, Tzu-Yu; Jee, Shiou-Hwa; Young, Tai-Horng; Lo, Wen; Chen, Wei-Liang; Dong, Chen-Yuan

    2008-02-01

    In this study, we combined two-photon autofluorescence and second harmonic generation imaging to investigate the three-dimensional microstructure and nonlinear optical properties of tissue engineering scaffolds. We focused on five different types of scaffold materials commonly used in tissue engineering, including: open-cell polylactic acid, polyglycolic acid, collagen composite scaffold, collagraft bone graft matrix strip, and nylon. By the use of multiphoton microscopy and a motorized stage, we obtained high resolution, spectrally resolved structural information of the scaffolds over large areas or in three-dimensions. Our results show that the nonlinear optical properties of the scaffolds will enable us to spectrally and morphologically distinguish the different types of scaffold materials investigated. We envision multiphoton microscopy to be a useful technique in tissue engineering applications in understanding the interplay between cultured cells and the scaffold materials. PMID:17943985

  12. Multiphoton adiabatic passage for atom optics applications

    SciTech Connect

    Demeter, Gabor; Djotyan, Gagik P.

    2009-04-15

    We study the force exerted on two-level atoms by short, counterpropagating laser pulses. When the counterpropagating pulses overlap each other partially, multiphoton adiabatic processes are possible in several configurations, which amplify the force exerted on the atoms. We investigate the practical usefulness of such multiphoton adiabatic transitions for the manipulation of the atoms' mechanical state. In particular, we compare the efficiency of a pair of constant frequency, oppositely detuned laser pulses and that of a pair of frequency-chirped pulses. We also consider the case of prolonged exposure to a sequence of laser pulses for a duration that is comparable to or much larger than the spontaneous lifetime of the atoms. We use numerical methods to calculate the reduction of the force and the heating of the atomic ensemble when spontaneous emission cannot be neglected during the interaction. In addition, we derive simple approximate formulas for the force and the heating, and compare them to the numerical results.

  13. Fundamental studies of molecular multiphoton ionization

    SciTech Connect

    Miller, J.C.; Compton, R.N.

    1984-04-01

    For several years the authors have performed fundamental studies of molecular multiphoton ionization (MPI). We will present a potpourri of techniques and results chosen to illustrate the interesting complexities of molecular MPI. Techniques used include time-of-flight mass spectroscopy, photoelectron spectroscopy, supersonic expansion cooling of molecular beams, harmonic generation, two-color laser MPI, and polarization spectroscopy. Whenever possible the relevance of these results to resonance ionization spectroscopy schemes will be delineated. 23 references, 10 figures.

  14. Low-frequency theory of multiphoton ionization

    NASA Astrophysics Data System (ADS)

    Abranyos, Yonatan; Mittleman, Marvin H.

    1990-10-01

    We evaluate the ionization rate from a recent (adiabatic) theory of multiphoton ionization for the case of a recent experiment: a circularly polarized CO2 laser at an intensity of about 1014 W/cm2. For S states we obtain a rate 104-105 times larger than the Keldysh-Faisal-Reiss theory. For P states we obtain a rate about 109 times larger. We also make a prediction about the angular distribution from S states.

  15. Video-rate resonant scanning multiphoton microscopy

    PubMed Central

    Kirkpatrick, Nathaniel D.; Chung, Euiheon; Cook, Daniel C.; Han, Xiaoxing; Gruionu, Gabriel; Liao, Shan; Munn, Lance L.; Padera, Timothy P.; Fukumura, Dai; Jain, Rakesh K.

    2013-01-01

    The abnormal tumor microenvironment fuels tumor progression, metastasis, immune suppression, and treatment resistance. Over last several decades, developments in and applications of intravital microscopy have provided unprecedented insights into the dynamics of the tumor microenvironment. In particular, intravital multiphoton microscopy has revealed the abnormal structure and function of tumor-associated blood and lymphatic vessels, the role of aberrant tumor matrix in drug delivery, invasion and metastasis of tumor cells, the dynamics of immune cell trafficking to and within tumors, and gene expression in tumors. However, traditional multiphoton microscopy suffers from inherently slow imaging ratesonly a few frames per second, thus unable to capture more rapid events such as blood flow, lymphatic flow, and cell movement within vessels. Here, we report the development and implementation of a video-rate multiphoton microscope (VR-MPLSM) based on resonant galvanometer mirror scanning that is capable of recording at 30 frames per second and acquiring intravital multispectral images. We show that the design of the system can be readily implemented and is adaptable to various experimental models. As examples, we demonstrate the utility of the system to directly measure flow within tumors, capture metastatic cancer cells moving within the brain vasculature and cells in lymphatic vessels, and image acute responses to changes in a vascular network. VR-MPLSM thus has the potential to further advance intravital imaging and provide new insight into the biology of the tumor microenvironment. PMID:24353926

  16. The 3.1 ?m Infrared Spectra of Vibrationally Excited C_3 in a Supersonic Plasma Jet

    NASA Astrophysics Data System (ADS)

    Zhao, Dongfeng; Doney, Kirstin D.; Linnartz, Harold

    2014-06-01

    The linear triatomic carbon (C_3), one of the most important molecules that have been identified in both dense and diffuse interstellar environments, has attracted great interest to astronomers and astrochemists. It is also of fundamental interest as it serves as a benchmark system for quantum chemistry. In this presentation, we report the high-resolution infrared spectra of C_3 in the 3.1 ?m region. The C_3 molecules are produced in a supersonic pulsed planar plasma by discharging a propyne/helium/argon gas mixture. Continuous-wave cavity ringdown spectroscopy is used to record the infrared absorption spectra of C_3. In total, eighteen vibrational bands are observed in the 3110 - 3290 wn range, and sixteen of them are reported for the first time. It is found that, the vibrational temperatures for the two CC stretch modes of C_3 are up to 8000 K in our plasma source, allowing to experimentally determine the ro-vibrational levels of C_3 to the 10 000 wn region. Accurate spectroscopic parameters are obtained from the detailed analysis of our spectra. The molecular data reported here are used to test the very recent theoretical work beyond the 'gold standard' P. Botschiwina, private communication.} for a comprehensive understanding of the ground-state potential energy surface of C_3.

  17. Discrimination of zeolites and beryllium containing silicates using portable Raman spectroscometric equipment with near-infrared excitation

    NASA Astrophysics Data System (ADS)

    Jehlička, J.; Vandenabeele, P.; Edwards, H. G. M.

    2012-02-01

    In this paper Raman spectra were obtained for a series of zeolites (thomsonite, stilbite, natrolite) and beryllium containing silicates (beryl, chrysoberyl, euclase, phenacite, bavenite, milarite) using a portable Raman specrometer with a 785 nm laser excitation to show the possibility to apply this setting for unambiguous detection and discrimination of these silicate minerals. Obtained spectra contain the most intense Raman bands at the same positions ±2-4 cm -1 as reported in the literature. The use of these bands permits the unambiguous identification of these phases. Data show the possibility to discriminate individual species of similar whitish color and aspect. Measurements showed an excellent correspondence of Raman bands obtained using the portable system and a laboratory Raman microspectrometer (with the same excitation laser wavelenght). However, for several minerals of these groups (chrysoberyl, bertrandite, chiavennite) Raman spectra were not of sufficient quality to permit unambiguous identification. The reasons are discussed. Raman spectrum of chiavennite CaMnBe 2Si 5O 13(OH) 2·2(H 2O) - a transformation product occurring together with bavenite on the surface of beryl crystals was obtained for the first time using the laboratory Raman spectrometer.

  18. Discrimination of zeolites and beryllium containing silicates using portable Raman spectroscometric equipment with near-infrared excitation.

    PubMed

    Jehlička, J; Vandenabeele, P; Edwards, H G M

    2012-02-01

    In this paper Raman spectra were obtained for a series of zeolites (thomsonite, stilbite, natrolite) and beryllium containing silicates (beryl, chrysoberyl, euclase, phenacite, bavenite, milarite) using a portable Raman specrometer with a 785 nm laser excitation to show the possibility to apply this setting for unambiguous detection and discrimination of these silicate minerals. Obtained spectra contain the most intense Raman bands at the same positions ±2-4 cm(-1) as reported in the literature. The use of these bands permits the unambiguous identification of these phases. Data show the possibility to discriminate individual species of similar whitish color and aspect. Measurements showed an excellent correspondence of Raman bands obtained using the portable system and a laboratory Raman microspectrometer (with the same excitation laser wavelenght). However, for several minerals of these groups (chrysoberyl, bertrandite, chiavennite) Raman spectra were not of sufficient quality to permit unambiguous identification. The reasons are discussed. Raman spectrum of chiavennite CaMnBe(2)Si(5)O(13)(OH)(2)·2(H(2)O) - a transformation product occurring together with bavenite on the surface of beryl crystals was obtained for the first time using the laboratory Raman spectrometer. PMID:22099060

  19. Multiphoton Imaging of Upconverting Lanthanide Nanoparticles in Three Dimensional Models of Cancer

    PubMed Central

    Gainer, Christian F.; Romanowski, Marek

    2013-01-01

    While upconverting lanthanide nanoparticles have numerous advantages over other exogenous contrast agents used in scanned multiphoton imaging, their long luminescence lifetimes cause images collected with non-descanned detection to be greatly blurred. We demonstrate herein the use of Richardson-Lucy deconvolution to deblur luminescence images obtained via multiphoton scanning microscopy. Images were taken of three dimensional models of colon and ovarian cancer following incubation with NaYF4:Yb,Er nanoparticles functionalized with an antibody for EGFR and folic acid respectively. Following deconvolution, images had a lateral resolution on par with the optimal performance of the imaging system used, ~1.2 μm, and an axial resolution of ~5 μm. Due to the relatively high multiphoton excitation efficiency of these nanoparticles, it is possible to follow binding of individual particles in tissue. In addition, their extreme photostability allows for prolonged imaging without significant loss in luminescence signal. With these advantageous properties in mind, we also discuss the potential application of upconverting lanthanide nanoparticles for tracking of specific, cancer relevant receptors in tissue. PMID:24353385

  20. Multiphoton Imaging of Upconverting Lanthanide Nanoparticles in Three Dimensional Models of Cancer.

    PubMed

    Gainer, Christian F; Romanowski, Marek

    2013-02-22

    While upconverting lanthanide nanoparticles have numerous advantages over other exogenous contrast agents used in scanned multiphoton imaging, their long luminescence lifetimes cause images collected with non-descanned detection to be greatly blurred. We demonstrate herein the use of Richardson-Lucy deconvolution to deblur luminescence images obtained via multiphoton scanning microscopy. Images were taken of three dimensional models of colon and ovarian cancer following incubation with NaYF4:Yb,Er nanoparticles functionalized with an antibody for EGFR and folic acid respectively. Following deconvolution, images had a lateral resolution on par with the optimal performance of the imaging system used, ~1.2 μm, and an axial resolution of ~5 μm. Due to the relatively high multiphoton excitation efficiency of these nanoparticles, it is possible to follow binding of individual particles in tissue. In addition, their extreme photostability allows for prolonged imaging without significant loss in luminescence signal. With these advantageous properties in mind, we also discuss the potential application of upconverting lanthanide nanoparticles for tracking of specific, cancer relevant receptors in tissue. PMID:24353385

  1. Multiphoton imaging of upconverting lanthanide nanoparticles in three dimensional models of cancer

    NASA Astrophysics Data System (ADS)

    Gainer, Christian F.; Romanowski, Marek

    2013-02-01

    While upconverting lanthanide nanoparticles have numerous advantages over other exogenous contrast agents used in scanned multiphoton imaging, their long luminescence lifetimes cause images collected with non-descanned detection to be greatly blurred. We demonstrate herein the use of Richardson-Lucy deconvolution to deblur luminescence images obtained via multiphoton scanning microscopy. Images were taken of three dimensional models of colon and ovarian cancer following incubation with NaYF4:Yb,Er nanoparticles functionalized with an antibody for EGFR and folic acid respectively. Following deconvolution, images had a lateral resolution on par with the optimal performance of the imaging system used, ~1.2 μm, and an axial resolution of ~5 μm. Due to the relatively high multiphoton excitation efficiency of these nanoparticles, it is possible to follow binding of individual particles in tissue. In addition, their extreme photostability allows for prolonged imaging without significant loss in luminescence signal. With these advantageous properties in mind, we also discuss the potential application of upconverting lanthanide nanoparticles for tracking of specific, cancer relevant receptors in tissue.

  2. Clinical optical coherence tomography combined with multiphoton tomography for evaluation of several skin disorders

    NASA Astrophysics Data System (ADS)

    König, Karsten; Speicher, Marco; Bückle, Rainer; Reckfort, Julia; McKenzie, Gordon; Welzel, Julia; Koehler, Martin J.; Elsner, Peter; Kaatz, Martin

    2010-02-01

    The first clinical trial of optical coherence tomography (OCT) combined with multiphoton tomography (MPT) and dermoscopy is reported. State-of-the-art (i) OCT systems for dermatology (e.g. multibeam swept source OCT), (ii) the femtosecond laser multiphoton tomograph DermaInspectTM, and (iii) digital dermoscopes were applied to 47 patients with a diversity of skin diseases and disorders such as skin cancer, psoriasis, hemangioma, connective tissue diseases, pigmented lesions, and autoimmune bullous skin diseases. Dermoscopy, also called 'epiluminescent microscopy', provides two-dimensional color images of the skin surface. OCT imaging is based on the detection of optical reflections within the tissue measured interferometrically whereas nonlinear excitation of endogenous fluorophores and the second harmonic generation are the bases of MPT images. OCT cross sectional "wide field" image provides a typical field of view of 5 x 2 mm2 and offers fast information on the depth and the volume of the investigated lesion. In comparison, multiphoton tomography presents 0.36 x 0.36 mm2 horizontal or diagonal sections of the region of interest within seconds with submicron resolution and down to a tissue depth of 200 μm. The combination of OCT and MPT provides a synergistic optical imaging modality for early detection of skin cancer and other skin diseases.

  3. Ultralow-threshold multiphoton-pumped lasing from colloidal nanoplatelets in solution

    NASA Astrophysics Data System (ADS)

    Li, Mingjie; Zhi, Min; Zhu, Hai; Wu, Wen-Ya; Xu, Qing-Hua; Jhon, Mark Hyunpong; Chan, Yinthai

    2015-09-01

    Although multiphoton-pumped lasing from a solution of chromophores is important in the emerging fields of nonlinear optofluidics and bio-photonics, conventionally used organic dyes are often rendered unsuitable because of relatively small multiphoton absorption cross-sections and low photostability. Here, we demonstrate highly photostable, ultralow-threshold multiphoton-pumped biexcitonic lasing from a solution of colloidal CdSe/CdS nanoplatelets within a cuvette-based Fabry-Pérot optical resonator. We find that colloidal nanoplatelets surprisingly exhibit an optimal lateral size that minimizes lasing threshold. These nanoplatelets possess very large gain cross-sections of 7.3 × 10-14 cm2 and ultralow lasing thresholds of 1.2 and 4.3 mJ cm-2 under two-photon (λexc=800 nm) and three-photon (λexc=1.3 μm) excitation, respectively. The highly polarized emission from the nanoplatelet laser shows no significant photodegradation over 107 laser shots. These findings constitute a more comprehensive understanding of the utility of colloidal semiconductor nanoparticles as the gain medium in high-performance frequency-upconversion liquid lasers.

  4. Intrinsic indicator of photodamage during label-free multiphoton microscopy of cells and tissues.

    PubMed

    Galli, Roberta; Uckermann, Ortrud; Andresen, Elisabeth F; Geiger, Kathrin D; Koch, Edmund; Schackert, Gabriele; Steiner, Gerald; Kirsch, Matthias

    2014-01-01

    Multiphoton imaging has evolved as an indispensable tool in cell biology and holds prospects for clinical applications. When addressing endogenous signals such as coherent anti-Stokes Raman scattering (CARS) or second harmonic generation, it requires intense laser irradiation that may cause photodamage. We report that increasing endogenous fluorescence signal upon multiphoton imaging constitutes a marker of photodamage. The effect was studied on mouse brain in vivo and ex vivo, on ex vivo human brain tissue samples, as well as on glioblastoma cells in vitro, demonstrating that this phenomenon is common to a variety of different systems, both ex vivo and in vivo. CARS microscopy and vibrational spectroscopy were used to analyze the photodamage. The development of a standard easy-to-use model that employs rehydrated cryosections allowed the characterization of the irradiation-induced fluorescence and related it to nonlinear photodamage. In conclusion, the monitoring of endogenous two-photon excited fluorescence during label-free multiphoton microscopy enables to estimate damage thresholds ex vivo as well as detect photodamage during in vivo experiments. PMID:25343251

  5. In vivo multiphoton imaging of collagen remodeling after microablative fractional rejuvenation

    NASA Astrophysics Data System (ADS)

    Cicchi, Riccardo; Kapsokalyvas, Dimitrios; Troiano, Michela; Campolmi, Piero; Morini, Cristiano; Cosci, Alessandro; Massi, Daniela; Lotti, Torello; Pavone, Francesco S.

    2011-03-01

    The potential of multiphoton microscopy in providing in-vivo early diagnosis of skin lesions has already been demonstrated, while its capability in therapy follow-up has not been deeply explored so far. Two-photon excited fluorescence and second-harmonic generation microscopy were used in combination to follow-up collagen remodeling after laser micro-ablative rejuvenation. Treated regions of volunteers were imaged with multiphoton microscopy before and after treatment, and we found a strong age-dependence of the treatment effectiveness. In particular, the photorejuvenating effect was negligible in young subjects (< 30 years), whereas a significant production of new collagen was observed in aged subjects (> 70 years). Quantification of the amount of newly produced collagen and its organization were performed by means of visual examination of two-photon images. The obtained results demonstrate the performance of laser fractional micro-ablative rejuvenation without the need of an invasive biopsy as well as the wide applicability range of applications for multiphoton microscopy in clinical dermatology.

  6. Ultralow-threshold multiphoton-pumped lasing from colloidal nanoplatelets in solution

    PubMed Central

    Li, Mingjie; Zhi, Min; Zhu, Hai; Wu, Wen-Ya; Xu, Qing-Hua; Jhon, Mark Hyunpong; Chan, Yinthai

    2015-01-01

    Although multiphoton-pumped lasing from a solution of chromophores is important in the emerging fields of nonlinear optofluidics and bio-photonics, conventionally used organic dyes are often rendered unsuitable because of relatively small multiphoton absorption cross-sections and low photostability. Here, we demonstrate highly photostable, ultralow-threshold multiphoton-pumped biexcitonic lasing from a solution of colloidal CdSe/CdS nanoplatelets within a cuvette-based FabryProt optical resonator. We find that colloidal nanoplatelets surprisingly exhibit an optimal lateral size that minimizes lasing threshold. These nanoplatelets possess very large gain cross-sections of 7.3 10?14?cm2 and ultralow lasing thresholds of 1.2 and 4.3?mJ?cm?2 under two-photon (?exc=800?nm) and three-photon (?exc=1.3??m) excitation, respectively. The highly polarized emission from the nanoplatelet laser shows no significant photodegradation over 107 laser shots. These findings constitute a more comprehensive understanding of the utility of colloidal semiconductor nanoparticles as the gain medium in high-performance frequency-upconversion liquid lasers. PMID:26419950

  7. Ultralow-threshold multiphoton-pumped lasing from colloidal nanoplatelets in solution.

    PubMed

    Li, Mingjie; Zhi, Min; Zhu, Hai; Wu, Wen-Ya; Xu, Qing-Hua; Jhon, Mark Hyunpong; Chan, Yinthai

    2015-01-01

    Although multiphoton-pumped lasing from a solution of chromophores is important in the emerging fields of nonlinear optofluidics and bio-photonics, conventionally used organic dyes are often rendered unsuitable because of relatively small multiphoton absorption cross-sections and low photostability. Here, we demonstrate highly photostable, ultralow-threshold multiphoton-pumped biexcitonic lasing from a solution of colloidal CdSe/CdS nanoplatelets within a cuvette-based Fabry-Pérot optical resonator. We find that colloidal nanoplatelets surprisingly exhibit an optimal lateral size that minimizes lasing threshold. These nanoplatelets possess very large gain cross-sections of 7.3 × 10(-14) cm(2) and ultralow lasing thresholds of 1.2 and 4.3 mJ cm(-2) under two-photon (λexc=800 nm) and three-photon (λexc=1.3 μm) excitation, respectively. The highly polarized emission from the nanoplatelet laser shows no significant photodegradation over 10(7) laser shots. These findings constitute a more comprehensive understanding of the utility of colloidal semiconductor nanoparticles as the gain medium in high-performance frequency-upconversion liquid lasers. PMID:26419950

  8. Bioinspired near-infrared-excited sensing platform for in vitro antioxidant capacity assay based on upconversion nanoparticles and a dopamine-melanin hybrid system.

    PubMed

    Wang, Dong; Chen, Chuan; Ke, Xuebin; Kang, Ning; Shen, Yuqing; Liu, Yongliang; Zhou, Xi; Wang, Hongjun; Chen, Changqing; Ren, Lei

    2015-02-11

    A novel core-shell structure based on upconversion fluorescent nanoparticles (UCNPs) and dopamine-melanin has been developed for evaluation of the antioxidant capacity of biological fluids. In this approach, dopamine-melanin nanoshells facilely formed on the surface of UCNPs act as ultraefficient quenchers for upconversion fluorescence, contributing to a photoinduced electron-transfer mechanism. This spontaneous oxidative polymerization of the dopamine-induced quenching effect could be effectively prevented by the presence of various antioxidants (typically biothiols, ascorbic acid (Vitamin C), and Trolox). The chemical response of the UCNPs@dopamine-melanin hybrid system exhibited great selectivity and sensitivity toward antioxidants relative to other compounds at 100-fold higher concentration. A satisfactory correlation was established between the ratio of the "anti-quenching" fluorescence intensity and the concentration of antioxidants. Besides the response of the upconversion fluorescence signal, a specific evaluation process for antioxidants could be visualized by the color change from colorless to dark gray accompanied by the spontaneous oxidation of dopamine. The near-infrared (NIR)-excited UCNP-based antioxidant capacity assay platform was further used to evaluate the antioxidant capacity of cell extracts and human plasma, and satisfactory sensitivity, repeatability, and recovery rate were obtained. This approach features easy preparation, fluorescence/visual dual mode detection, high specificity to antioxidants, and enhanced sensitivity with NIR excitation, showing great potential for screening and quantitative evaluation of antioxidants in biological systems. PMID:25604145

  9. Luminescence quenching of conductive Si nanocrystals via “Linkage emission”: Hopping-like propagation of infrared-excited Auger electrons

    SciTech Connect

    Ishii, Masashi; Crowe, Iain F.; Halsall, Matthew P.; Hamilton, Bruce; Knights, Andrew P.; Gwilliam, Russell M.

    2014-08-14

    Phosphorus (P) is an n-type dopant for conductive silicon nanocrystals (Si-nc's), the electrical activation of which may be monitored through a non-radiative Auger recombination process that quenches the Si-nc luminescence. We investigated this quenching mechanism through electrical measurements of Si-nc's. Infrared-excited Auger electron emission as the non-radiative process was directly probed and the dynamics of the process are determined from a frequency response analysis. To explain the dynamics, we propose a model in which Auger electrons with a low kinetic energy establish a local inter-nanocrystal conductance and the repetition of this local conductance results in a constant photocurrent (“linkage emission”). This emission becomes significant by electron filling in the Si-nc's owing to the electrical activation of P, which is consistent with observed luminescence quenching behavior. We found that the IR photo-excited emission is distinct from the thermally induced hopping conduction and show that confined, rather than trapped, charges are the source of the Auger electrons. Thus, the process consumes both confined charges and the recombination energy for Auger emission, which explains the luminescence quenching mechanism of Si-nc:P.

  10. Simulated infrared emission spectra of highly excited polyatomic molecules: a detailed model of the PAH-UIR hypothesis

    NASA Technical Reports Server (NTRS)

    Cook, D. J.; Saykally, R. J.

    1998-01-01

    A detailed description of the polycyclic aromatic hydrocarbon (PAH)/unidentified infrared band (UIR) mechanism is presented in which experimental spectral bandshape functions are used to simulate IR emission spectra for individual molecules. These spectra are additively superimposed to produce a conglomerate spectrum representative of a family of PAH molecules. Ab initio vibrational frequencies and intensities for nine PAHs (neutral and cationic) as large as ovalene are used in conjunction with measured bandshape and temperature-dependent redshift data to simulate the UIR bands. The calculated spectra of cations provide a closer match to the UIRs than do those of the neutrals. However, the PAH cations used in the simulations fail to reproduce the details of the UIR emission spectra. The discrepancies are potentially alleviated if both larger PAHs and a greater number of PAHs were included in the simulation.

  11. Infrared spectroscopy and intramolecular vibrational relaxation of c-C 6F 12 excited above the dissociation threshold

    NASA Astrophysics Data System (ADS)

    Bagratashvili, V. N.; Burimov, V. N.; Ionov, S. I.; Sviridov, A. P.; Stuchebrukhov, A. A.; Turovetz, I. M.

    1987-05-01

    The IR spectrum of c-C 6F 12 at a vibrational energy of twice the dissociation threshold was investigated. Absorption of cw CO 2 laser radiation was measured at various frequencies. Our experimental conditions were chosen such that during absorption measurements all vibrational degrees of freedom were in equilibrium, the molecular rotation being at room temperature. The Boltzmann vibrational distribution allowed computer simulations of the spectrum to be made to determine the homogeneous contribution. The homogeneous half-width of the spectrum is ?=130.5 cm -1 and the homogeneous spectrum of c-C 6F 12 at E= 60000 cm -1 is non-Lorentzian. We attribute this to the influence of higher-order anharmonicities on the relaxation from the excited mode ( v27) to other modes in the molecule.

  12. Multiphoton dissociation of H2S by intense x-ray pulses from the Linac Coherent Light Source FEL

    NASA Astrophysics Data System (ADS)

    Murphy, Brendan; Fang, Li; Hoener, Matthias; Kukk, Edwin; Kanter, Elliot; Bozek, John; Berrah, Nora

    2011-05-01

    We will report on sequential multiphoton ionization of H2S by intense pulses of x-rays from the LCLS free electron laser. Following L-shell excitation, fragmentation of the molecule is observed by Auger electron, photoelectron, and ion time-of-flight spectroscopy. Intensity dependent features in these spectra shed light on the dynamics of the dissociation process. This work is funded by the DOE-SC-BES, Chemical Sciences, Geosciences and Biosciences Division.

  13. Waveguide-integrated single- and multi-photon detection at telecom wavelengths using superconducting nanowires

    SciTech Connect

    Ferrari, Simone; Kahl, Oliver; Kovalyuk, Vadim; Goltsman, Gregory N.; Korneev, Alexander; Pernice, Wolfram H. P.

    2015-04-13

    We investigate single- and multi-photon detection regimes of superconducting nanowire detectors embedded in silicon nitride nanophotonic circuits. At near-infrared wavelengths, simultaneous detection of up to three photons is observed for 120?nm wide nanowires biased far from the critical current, while narrow nanowires below 100?nm provide efficient single photon detection. A theoretical model is proposed to determine the different detection regimes and to calculate the corresponding internal quantum efficiency. The predicted saturation of the internal quantum efficiency in the single photon regime agrees well with plateau behavior observed at high bias currents.

  14. Waveguide-integrated single- and multi-photon detection at telecom wavelengths using superconducting nanowires

    NASA Astrophysics Data System (ADS)

    Ferrari, Simone; Kahl, Oliver; Kovalyuk, Vadim; Goltsman, Gregory N.; Korneev, Alexander; Pernice, Wolfram H. P.

    2015-04-01

    We investigate single- and multi-photon detection regimes of superconducting nanowire detectors embedded in silicon nitride nanophotonic circuits. At near-infrared wavelengths, simultaneous detection of up to three photons is observed for 120 nm wide nanowires biased far from the critical current, while narrow nanowires below 100 nm provide efficient single photon detection. A theoretical model is proposed to determine the different detection regimes and to calculate the corresponding internal quantum efficiency. The predicted saturation of the internal quantum efficiency in the single photon regime agrees well with plateau behavior observed at high bias currents.

  15. Effects of multiphoton absorption on parametric comb generation in silicon microresonators.

    PubMed

    Lau, Ryan K W; Lamont, Michael R E; Okawachi, Yoshitomo; Gaeta, Alexander L

    2015-06-15

    We investigate theoretically the parametric frequency comb generation in silicon microresonators at telecom and mid-infrared (MIR) wavelengths in the presence of multiphoton absorption and free-carrier effects using a modified Lugiato-Lefever model. We show that parametric oscillation may occur at MIR wavelengths, provided that the free-carrier lifetime is sufficiently short or the optical pump power is sufficiently low, but is inhibited at telecom wavelengths. In addition, we propose an etchless, air-clad silicon microresonator that enables an octave-spanning frequency comb in a completely passive device. PMID:26076260

  16. Collision dynamics of methyl radicals and highly vibrationally excited molecules using crossed molecular beams

    SciTech Connect

    Chu, P.M.Y.

    1991-10-01

    The vibrational to translational (V{yields}T) energy transfer in collisions between large highly vibrationally excited polyatomics and rare gases was investigated by time-of-flight techniques. Two different methods, UV excitation followed by intemal conversion and infrared multiphoton excitation (IRMPE), were used to form vibrationally excited molecular beams of hexafluorobenzene and sulfur hexafluoride, respectively. The product translational energy was found to be independent of the vibrational excitation. These results indicate that the probability distribution function for V{yields}T energy transfer is peaked at zero. The collisional relaxation of large polyatomic molecules with rare gases most likely occurs through a rotationally mediated process. Photodissociation of nitrobenzene in a molecular beam was studied at 266 nm. Two primary dissociation channels were identified including simple bond rupture to produce nitrogen dioxide and phenyl radical and isomerization to form nitric oxide and phenoxy radical. The time-of-flight spectra indicate that simple bond rupture and isomerization occurs via two different mechanisms. Secondary dissociation of the phenoxy radicals to carbon monoxide and cyclopentadienyl radicals was observed as well as secondary photodissociation of phenyl radical to give H atom and benzyne. A supersonic methyl radical beam source is developed. The beam source configuration and conditions were optimized for CH{sub 3} production from the thermal decomposition of azomethane. Elastic scattering of methyl radical and neon was used to differentiate between the methyl radicals and the residual azomethane in the molecular beam.

  17. Photon absorption in step-wise multi-photon activation fluorescence (SMPAF) of Sepia melanin

    NASA Astrophysics Data System (ADS)

    Lai, Zhenhua; Kerimo, Josef; DiMarzio, Charles

    2013-02-01

    Previous research has shown that melanin goes through a step-wise three-photon absorption process when the fluorescence is activated with high laser intensity. We have conducted further research using even higher laser intensity for the activation, and have shown the possibility of observing power dependence other than third-order. This article discusses the possible energy states of Sepia melanin by studying the power dependence curves of the step-wise multi-photon activated fluorescence signal. Three different excitation channels are activated. Possible reasons causing the three channels are discussed.

  18. Identification of dirty necrosis in colorectal carcinoma based on multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Li, Lianhuang; Jiang, Weizhong; Yang, Yinghong; Chen, Zhifen; Feng, Changyin; Li, Hongsheng; Guan, Guoxian; Chen, Jianxin

    2014-06-01

    Dirty necrosis within glandular lumina is often considered as a characteristic of colorectal carcinomas (CRCs) that is a diagnostically useful feature of CRCs with DNA microsatellite instability (MSI). Multiphoton microscopy (MPM), which is based on the second-harmonic generation and two-photon excited fluorescence signals, was used to identify dirty necrosis. Our results demonstrated that MPM has the ability to exhibit the microstructure of dirty necrosis and the signal intensity as well as an emission spectrum that can help to differentiate dirty necrosis from cancer cells. These findings indicate that MPM may be helpful in distinguishing MSI colorectal carcinoma via the identification of dirty necrosis.

  19. Resonant multiphoton ionization spectra of molecules and molecular fragments. Technical report, October 1987--September 1988

    SciTech Connect

    1997-07-01

    The objective of our research under this contract is to carry out studies of resonant enhanced multiphoton ionization (REMPI) processes in molecules. In the (n + 1) - REMPI process of interest an atom or molecule in a specific initial state absorbs n photons making a transition to an intermediate state from which it is subsequently ionized by absorption of an additional photon. A remarkable feature of resonant enhanced multiphoton ionization is that the narrow bandwidth radiation of lasers makes it possible (i) to select a specific rovibrational level in the initial state of a molecule or fragment, (ii) to resonantly pump this level up to a selected rotational-vibrational level of an excited electronic state, and (iii) to subsequently photoionize the state that has been resonantly excited. The extreme state-selectivity and sensitivity make REMPI both a tool with several practical applications and an important technique for probing the photoionization dynamics of vibrationally and electronically excited states. Some significant applications of this technique include its use for state-specific detection of species and diagnostics in combustion and chemical etching media and plasmas, (ii) for state-specific generation of molecular ions for use in ion-molecule reaction studies, and (iii) as a probe of photofragmentation and gas-surface scattering including alignment and orientation effects in these processes.

  20. Optimal spectral filtering in soliton self-frequency shift for deep-tissue multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Wang, Ke; Qiu, Ping

    2015-05-01

    Tunable optical solitons generated by soliton self-frequency shift (SSFS) have become valuable tools for multiphoton microscopy (MPM). Recent progress in MPM using 1700 nm excitation enabled visualizing subcortical structures in mouse brain in vivo for the first time. Such an excitation source can be readily obtained by SSFS in a large effective-mode-area photonic crystal rod with a 1550-nm fiber femtosecond laser. A longpass filter was typically used to isolate the soliton from the residual in order to avoid excessive energy deposit on the sample, which ultimately leads to optical damage. However, since the soliton was not cleanly separated from the residual, the criterion for choosing the optimal filtering wavelength is lacking. Here, we propose maximizing the ratio between the multiphoton signal and the n'th power of the excitation pulse energy as a criterion for optimal spectral filtering in SSFS when the soliton shows dramatic overlapping with the residual. This optimization is based on the most efficient signal generation and entirely depends on physical quantities that can be easily measured experimentally. Its application to MPM may reduce tissue damage, while maintaining high signal levels for efficient deep penetration.

  1. Picosecond ultraviolet multiphoton laser photolysis and transient absorption spectroscopy of liquid benzenes

    SciTech Connect

    Miyasaka, H.; Masuhara, H.; Mataga, N.

    1985-04-25

    Ultraviolet multiphoton excitation with a picosecond laser pulse at the wavelength of no appreciable ground-state absorption has been used to excite benzene and alkylbenzenes homogeneously in the neat liquid state and to investigate their photophysical and photochemical primary processes by picosecond time-resolved transient absorption spectral measurements. The excimer formation processes in neat liquid benzenes excited by two-photon absorption of the 355-nm picosecond pulse from a Nd/sup 3 -/:YAG laser have been confirmed to proceed via ionization and recombination processes, and the lifetime of the ionized (ion pair) state has been determined to be 10-20 ps in neat benzene. 26 references, 4 figures, 4 tables.

  2. Time-resolved two-color photoacoustic and multiphoton ionization spectroscopy of aniline

    SciTech Connect

    Moll, D.J.; Parker, G.R. Jr.; Kuppermann, A.

    1984-05-15

    The multiphoton ionization and photoacoustic signals from aniline have been detected as a function of time delay between two laser pulses of different wavelength. The first pulse excited the S/sub 0/ ..-->.. S/sub 1/ 0--0 transition. The second pulse either excited S/sub 1/ to a higher singlet state, or a triplet state produced by intersystem crossing to a higher triplet state, depending on the time delay between pulses. Both ionization and dissociation processes are observed. By varying the conditions of excitation it appears that a given amount of energy can be selectively channeled almost exclusively into either ionization or dissociation pathways. The results are explained using a simple Franck--Condon factor model.

  3. Adiabatic theory of nonresonant multiphoton ionization

    NASA Astrophysics Data System (ADS)

    Mittleman, Marvin H.

    1989-07-01

    The Keldysh-Faisal-Reiss theory of nonresonant multiphoton ionization seems to give reasonably good agreement with experiments for production rates in the above-threshold-ionization peaks. The theory has some conceptual difficulties, since physically equivalent Hamiltonians give radically different results when treated by this method. An alternate form of the S matrix is used here in which approximation must be made upon the exact wave function evolving from the initial state rather than the time-reversed form used in the Keldysh-Reiss treatment. The exact initial state is approximated by an adiabatic form which results in an S matrix that eliminates the conceptual difficulties described above.

  4. Kinetic studies following state-selective laser excitation. Progress report, May 1, 1991--April 29, 1992

    SciTech Connect

    Keto, J.W.

    1992-01-01

    We have made measurements of state-to-state deactivation cross sections and radiative lifetimes for Xe*(6p,6p{prime},7p) and Kr*(5p) states in xenon and krypton buffer gases. These results are relevant to kinetic models and both excimer lasers and the infrared xenon laser; and they are a significant improvement in the precision of the known radiative lifetimes. This type of experiment can now be compared with recent calculations of state-to-state collisional relaxation in rare-gases by Hickman, Huestis, and Saxon. We have also made significant progress in the study of the electronic spectra of small molecules of the rare gases. Spectra have been obtained for Xe{sub 2}, Xe{sub 3}, Xe{sub 4}, and larger clusters. As guidance for the larger clusters of the rare gases we have obtained the first multiphoton spectra for excitons in condensed xenon. In collaboration with research on the multiphoton spectra of the rare gases, we have continued experiments using synchrotron radiation in collaboration with the University of Hamburg. In experiments there we have observed excitation and fluorescence spectra for single xenon atoms at the surface, within the second layer, and within the bulk of large argon clusters.

  5. Achieving molecular selectivity in imaging using multiphoton Raman spectroscopy techniques

    SciTech Connect

    Holtom, Gary R. ); Thrall, Brian D. ); Chin, Beek Yoke ); Wiley, H Steven ); Colson, Steven D. )

    2000-12-01

    In the case of most imaging methods, contrast is generated either by physical properties of the sample (Differential Image Contrast, Phase Contrast), or by fluorescent labels that are localized to a particular protein or organelle. Standard Raman and infrared methods for obtaining images are based upon the intrinsic vibrational properties of molecules, and thus obviate the need for attached flurophores. Unfortunately, they have significant limitations for live-cell imaging. However, an active Raman method, called Coherent Anti-Stokes Raman Scattering (CARS), is well suited for microscopy, and provides a new means for imaging specific molecules. Vibrational imaging techniques, such as CARS, avoid problems associated with photobleaching and photo-induced toxicity often associated with the use of fluorescent labels with live cells. Because the laser configuration needed to implement CARS technology is similar to that used in other multiphoton microscopy methods, such as two -photon fluorescence and harmonic generation, it is possible to combine imaging modalities, thus generating simultaneous CARS and fluorescence images. A particularly powerful aspect of CARS microscopy is its ability to selectively image deuterated compounds, thus allowing the visualization of molecules, such as lipids, that are chemically indistinguishable from the native species.

  6. Spectroscopic analysis of skin intrinsic signals for multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Pena, Ana-Maria; Strupler, Mathias; Boulesteix, Thierry; Senni, Karim; Godeau, Gaston; Beaurepaire, Emmanuel; Schanne-Klein, Marie-Claire

    2006-02-01

    We recorded multiphoton images of human skin biopsies using endogenous sources of nonlinear optical signals. We detected simultaneously two-photon excited fluorescence (2PEF) from intrinsic fluorophores and second harmonic generation (SHG) from collagen. We observed SHG from fibrillar collagens in the dermis, whereas no SHG was detectable from the non fibrillar type IV collagen in the basal laminae. We compared these distinct behaviours of collagens I and IV in SHG microscopy to polarization-resolved surface SHG experiments on thin films of collagens I and IV molecules. We observed similar signals for both types of molecular films, except for the chiroptical contributions which are present only for collagen I and enhance the signal typically by a factor of 2. We concluded that SHG microscopy is a sensitive probe of the micrometer-scale structural organization of collagen in biological tissues. In order to elucidate the origin of the endogenous fluorescence signals, we recorded 2PEF spectra at various positions in the skin biopsies, and compared these data to in vitro spectroscopic analysis. In particular, we studied the keratin fluorescence and determined its 2PEF action cross section. We observed a good agreement between 2PEF spectra recorded in the keratinized upper layers of the epidermis and in a solution of purified keratin. Finally, to illustrate the capabilities of this technique, we recorded 2PEF/SHG images of skin biopsies obtained from patients of various ages.

  7. The multiphoton ionization of uranium hexafluoride. Revision 1

    SciTech Connect

    Armstrong, D.P.

    1992-05-01

    Multiphoton ionization (MPI) time-of-flight mass spectroscopy and photoelectron spectroscopy studies of UF{sub 6} have been conducted using focused light from the Nd:YAG laser fundamental ({lambda}=1064 nm) and its harmonics ({lambda}=532, 355, or 266 nm), as well as other wavelengths provided by a tunable dye laser. The MPI mass spectra are dominated by the singly and multiply charged uranium ions rather than by the UF{sub x}{sup +} fragment ions even at the lowest laser power densities at which signal could be detected. The laser power dependence of U{sup n+} ions signals indicates that saturation can occur for many of the steps required for their ionization. In general, the doubly-charged uranium ion (U{sup 2+}) intensity is much greater than that of the singly-charged uranium ion (U{sup +}). For the case of the tunable dye laser experiments, the U{sup n+} (n = 1- 4) wavelength dependence is relatively unstructured and does not show observable resonance enhancement at known atomic uranium excitation wavelengths. The dominance of the U{sup 2+} ion and the absence or very small intensities of UF{sub x}{sup +} fragments, along with the unsaturated wavelength dependence, indicate that mechanisms may exist other than ionization of bare U atoms after the stepwise photodissociation of F atoms from the parent molecule.

  8. Insights on the excited state electronic structures of ruthenium(II) polypyridine complexes obtained by step-scan Fourier transform infrared absorption difference time-resolved spectroscopy

    SciTech Connect

    Chen, Pingyun; Palmer, R.A.; Omberg, K.M.

    1997-03-12

    Application of time-resolved infrared spectroscopy has had an important impact on transition metal photochemistry. The emphasis has been on metal carbonyl and metal cyano complexes because the oscillator strengths of {anti v}(CO) and {anti v}(CN) are high, and tunable lasers are available in the relevant spectral region. Until recently, time-resolved infrared spectroscopy using Fourier transform interferometry has been limited to a time resolution of {ge}5 {mu}s. However, application of step-scan FT-IR has greatly expanded the time window. It is now possible to acquire spectra with high resolution and sensitivity on a time scale of tens of nanoseconds over the entire mid-IR region. In this communication, the authors report the application of step-scan FT-IR absorbance difference time-resolved spectroscopy ({sup 2}FT-IR {Delta}A TRS) with spectra acquired on the 10 ns time scale to the study of electronic structure in the metal-to-ligand charge transfer (MLCT) excited states of two related complexes of ruthenium(II) containing only the ligands 2,2{prime}-bipyridine (bpy), 4-(carboxyethyl)-4{prime}-methyl-2,2{prime}-bipyridine (4-COOEt-4{prime}-CH{sub 3}bpy) and 4,4{prime}-(dicarboxyethyl)-2,2{prime}-bipyridine (4,4{prime}-(COOEt){sub 2}bpy). Comparison of the relative vibrational energies of the MLCT states leads to specific and significant conclusions regarding the distribution of electron density in these states.

  9. In vivo non-invasive multiphoton tomography of human skin

    NASA Astrophysics Data System (ADS)

    König, Karsten; Riemann, Iris; Ehlers, Alexander; Le Harzic, Ronan

    2005-10-01

    High resolution non-invasive 3D imaging devices are required to detect pathogenic microorganisms such as Anthrax spores, bacteria, viruses, fungi and chemical agents entering biological tissues such as the epidermis. Due to the low light penetration depth and the biodamage potential, ultraviolet light sources can not be employed to realize intratissue imaging of bio- and chemohazards. We report on the novel near infrared laser technology multiphoton tomography and the high resolution 4D imaging tool DermaInspect for non-invasive detection of intratissue agents and their influence on cellular metabolism based on multiphoton autofluorescence imaging (MAI) and second harmonic generation (SHG). Femtosecond laser pulses in the spectral range of 750 nm to 850 nm have been used to image in vivo human skin with subcellular spatial and picosecond temporal resolution. The non-linear induced autofluorescence of both, skin tissues and microorganisms, originates mainly from naturally endogenous fluorophores/protein structures like NAD(P)H, flavins, keratin, collagen, elastin, porphyrins and melanin. Bacteria emit in the blue/green spectral range due to NAD(P)H and flavoproteins and, in certain cases, in the red spectral range due to the biosynthesis of Zn-porphyrins, coproporphyrin and protoporphyrin. Collagen and exogenous non-centrosymmetric molecules can be detected by SHG signals. The system DermaInspect consists of a wavelength-tunable compact 80/90 MHz Ti:sapphire laser, a scan module with galvo scan mirrors, piezo-driven objective, fast photon detector and time-resolved single photon counting unit. It can be used to perform optical sectioning and 3D autofluorescence lifetime imaging (τ-mapping) with 1 μm spatial resolution and 270 ps temporal resolution. The parameter fluorescence lifetime depends on the type of fluorophore and its microenvironment and can be used to distinguish bio- and chemohazards from cellular background and to gain information for pathogen identification. The novel in vivo non-invasive imaging system offers the possibility to detect and to localize CB agents in tissues and to gain information on their impact on respiratory chain activity, cell division and metabolism. The system DermaInspect can also be used to detect food and water contamination.

  10. Multiphoton ionization in circularly polarized standing waves

    NASA Astrophysics Data System (ADS)

    Guo, Dong-Sheng; Drake, G. W. F.

    1992-05-01

    We derive the wave functions for an electron moving in a quantized circularly polarized standing-wave radiation field. By applying these solutions as the final states, we derive multiphoton ionization transition-rate formulas according to Keldysh-Faisal-Reiss (KFR) theory. Numerical calculations and theoretical analysis show that in the case of two light beams with the same angular momentum, KFR theory does not predict the peak splittings in the photoelectron angular distributions that were observed in an experiment by Bucksbaum, Schumacher, and Bashkansky (BSB) [Phys. Rev. Lett. 61, 1162 (1988)]. A transition-rate formula from a scattering theory developed for the case of single-mode multiphoton ionization by Guo, berg, and Crasemann [Phys. Rev. A 40, 4997 (1989)] is extended to the standing-wave case. The theory predicts the peak splitting observed by BSB. Numerical results of the splitting angle from the theory show good agreement with experimental measurements. We also prove that the sine function of the half splitting angle is inversely proportional to the square root of the photoelectron kinetic energy E, i.e., sin(?/2-?max)~1/E1/2, where ?max is the maximum scattering angle smaller than ?/2. Some predictions are made for future experimental observations, such as quantum effects associated with ponderomotive-energy decay.

  11. Multiphoton ionization of large water clusters

    SciTech Connect

    Apicella, B.; Li, X.; Passaro, M.; Spinelli, N.; Wang, X.

    2014-05-28

    Water clusters are multimers of water molecules held together by hydrogen bonds. In the present work, multiphoton ionization in the UV range coupled with time of flight mass spectrometry has been applied to water clusters with up to 160 molecules in order to obtain information on the electronic states of clusters of different sizes up to dimensions that can approximate the bulk phase. The dependence of ion intensities of water clusters and their metastable fragments produced by laser ionization at 355 nm on laser power density indicates a (3+1)-photon resonance-enhanced multiphoton ionization process. It also explains the large increase of ionization efficiency at 355 nm compared to that at 266 nm. Indeed, it was found, by applying both nanosecond and picosecond laser ionization with the two different UV wavelengths, that no water cluster sequences after n = 9 could be observed at 266 nm, whereas water clusters up to m/z 2000 Th in reflectron mode and m/z 3000 Th in linear mode were detected at 355 nm. The agreement between our findings on clusters of water, especially true in the range with n > 10, and reported data for liquid water supports the hypothesis that clusters above a critical dimension can approximate the liquid phase. It should thus be possible to study clusters just above 10 water molecules, for getting information on the bulk phase structure.

  12. Multiphoton ionization of large water clusters.

    PubMed

    Apicella, B; Li, X; Passaro, M; Spinelli, N; Wang, X

    2014-05-28

    Water clusters are multimers of water molecules held together by hydrogen bonds. In the present work, multiphoton ionization in the UV range coupled with time of flight mass spectrometry has been applied to water clusters with up to 160 molecules in order to obtain information on the electronic states of clusters of different sizes up to dimensions that can approximate the bulk phase. The dependence of ion intensities of water clusters and their metastable fragments produced by laser ionization at 355 nm on laser power density indicates a (3+1)-photon resonance-enhanced multiphoton ionization process. It also explains the large increase of ionization efficiency at 355 nm compared to that at 266 nm. Indeed, it was found, by applying both nanosecond and picosecond laser ionization with the two different UV wavelengths, that no water cluster sequences after n = 9 could be observed at 266 nm, whereas water clusters up to m/z 2000 Th in reflectron mode and m/z 3000 Th in linear mode were detected at 355 nm. The agreement between our findings on clusters of water, especially true in the range with n > 10, and reported data for liquid water supports the hypothesis that clusters above a critical dimension can approximate the liquid phase. It should thus be possible to study clusters just above 10 water molecules, for getting information on the bulk phase structure. PMID:24880286

  13. Multiphoton absorption of HF laser photons by molecules containing a hydroxyl group

    NASA Astrophysics Data System (ADS)

    McAlpine, Robert D.; Evans, D. K.; McClusky, F. K.

    1980-08-01

    Multiphoton absorption (MPA) of HF laser radiation has been studied, as a function of pressure (15 Pa to 1.3 kPa) and fluence (2 mJ/cm2 to 75 J/cm2) for the series: water, methanol, methan-d3-ol, ethanol, and 2,2,2-trifluoroethanol. As the group attached to the -OH is made more complex, the quasicontinuum occurs after fewer excitation steps, and under ''collisionless'' conditions, the same degree of multiphoton excitation is found to require a lower fluence. For water, at pressures between 73 Pa and 1.3 kPa, the cross sections are considerably lower than those for the other molecules, and MPA requires fluences in excess of 75 J/cm2. The remaining molecules divide into two groups, the ''small'' molecules (CH3OH and CD3OH) and the ''large'' molecules (C2H5OH and CF3CH2OH). For the small molecules at low pressures, the cross sections decrease with increasing fluence, an effect which is thought to be due to anharmonic bottlenecking. As pressure increases, the fluence dependence of the cross sections disappear. For the large molecules, anharmonic bottlenecking appears to be reduced, due to the greater density of states, and cross sections increase with increasing fluence according to the empirical form: ?(E, P)=K'Eb'Pa (where P is pressure, E is fluence and b', a, and K' are constants). The facility of HF laser-induced collisionless multiphoton dissociation of the -OH containing molecules is discussed in light of these results.

  14. Infrared-active optical phonons and magnetic excitations in the hexagonal manganites RMnO3 (R =Ho, Er, Tm, Yb, and Lu)

    NASA Astrophysics Data System (ADS)

    Basistyy, R.; Stanislavchuk, T. N.; Sirenko, A. A.; Litvinchuk, A. P.; Kotelyanskii, M.; Carr, G. L.; Lee, N.; Wang, X.; Cheong, S.-W.

    2014-07-01

    Optical properties of hexagonal multiferroic oxides RMnO3, where R=Ho, Er, Tm, Yb, and Lu, have been studied in the far-infrared spectral range between 100 and 2000 cm-1 and temperatures between 1.5 and 300 K by means of several experimental techniques: Mueller matrix spectroscopic ellipsometry, rotating analyzer ellipsometry, and optical transmission spectroscopy. Spectra of the optical phonons are described in terms of the temperature dependencies of their frequency, damping, and oscillator strength. For all studies, oxide materials' clear signatures of the spin-phonon interaction have been found below the temperature of the antiferromagnetic phase transition TN due to magnetic ordering of Mn3+ spins. A decrease of the ionic radius for R3+ ions between Ho3+ and Lu3+ in the corresponding RMnO3 compounds resulted in systematic variation of the frequency for several optical phonons. A magnetic excitation at 190 cm-1 was observed at low temperatures below TN and interpreted as resulting from two-magnon absorption.

  15. Nonperturbative multiphoton processes and electron-positron pair production

    SciTech Connect

    Hatsagortsyan, K. Z.; Mueller, C.; Keitel, C. H.

    2006-04-07

    Various regimes of pair production in laser fields are analyzed. Particularly, the question of the observability of pair production in a nonperturbative multiphoton regime is discussed. A simple heuristic method is employed which gives order-of-magnitude estimates for probabilities of multiphoton processes and allows to describe its main features. The method is initially probed upon the known process of pair production in a Coulomb and a strong laser field. Then it is applied to the nonperturbative multiphoton regime of the pair production process in a standing laser wave.

  16. Infrared photodissociation of a water molecule from a flexible molecule-H{sub 2}O complex: Rates and conformational product yields following XH stretch excitation

    SciTech Connect

    Clarkson, Jasper R.; Herbert, John M.; Zwier, Timothy S.

    2007-04-07

    Infrared-ultraviolet hole-burning and hole-filling spectroscopies have been used to study IR-induced dissociation of the tryptamine{center_dot}H{sub 2}O and tryptamine{center_dot}D{sub 2}O complexes. Upon complexation of a single water molecule, the seven conformational isomers of tryptamine collapse to a single structure that retains the same ethylamine side chain conformation present in the most highly populated conformer of tryptamine monomer. Infrared excitation of the tryptamine{center_dot}H{sub 2}O complex was carried out using a series of infrared absorptions spanning the range of 2470-3715 cm{sup -1}. The authors have determined the conformational product yield over this range and the dissociation rate near threshold, where it is slow enough to be measured by our methods. The observed threshold for dissociation occurred at 2872 cm{sup -1} in tryptamine{center_dot}H{sub 2}O and at 2869 cm{sup -1} in tryptamine{center_dot}D{sub 2}O, with no dissociation occurring on the time scale of the experiment ({approx}2 {mu}s) at 2745 cm{sup -1}. The dissociation time constants varied from {approx}200 ns for the 2869 cm{sup -1} band of tryptamine{center_dot}D{sub 2}O to {approx}25 ns for the 2872 cm{sup -1} band of tryptamine{center_dot}H{sub 2}O. This large isotope dependence is associated with a zero-point energy effect that increases the binding energy of the deuterated complex by {approx}190 cm{sup -1}, thereby reducing the excess energy available at the same excitation energy. At all higher energies, the dissociation lifetime was shorter than the pulse duration of our lasers (8 ns). At all wavelengths, the observed products in the presence of collisions are dominated by conformers A and B of tryptamine monomer, with small contributions from the other minor conformers. In addition, right at threshold (2869 cm{sup -1}), tryptamine{center_dot}D{sub 2}O dissociates exclusively to conformer A in the absence of collisions with helium, while both A and B conformational products are observed in the presence of collisions with helium. Using resolution-of-identity approximation to second-order Moeller-Plesset binding energies extrapolated to the complete basis set limit and harmonic vibrational frequencies and transition states calculated at the density functional limit B3LYP/6-31+G{sup *} level of theory, Rice-Ramsperger-Kassel-Marcus (RRKM) predictions for the dissociation, isomerization, and water shuttling rates as a function of energy are made. At threshold, the experimental dissociation rate is almost 10{sup 3} faster than RRKM predictions. Reasons for this apparent non-RRKM behavior will be discussed.

  17. Minimally invasive multiphoton and harmonic generation imaging of extracellular matrix structures in lung airway and related diseases.

    PubMed

    Abraham, Thomas; Hirota, Jeremy A; Wadsworth, Samuel; Knight, Darryl A

    2011-10-01

    Multiphoton microscopy has become a powerful imaging method for minimally invasive evaluation of extracellular matrix (ECM) and cellular structures deep within tissues in their native environments. This technology, which uses ultra-short femto-second laser pulses as the excitation source, is efficient in multiphoton excitation fluorescence (MPEF) of endogenously fluorescent macromolecular systems and induction of highly specific second harmonic generation (SHG) signals from non-centrosymmetric macromolecules such as fibrillar collagens. Both these signals can be captured simultaneously to provide spatially resolved 3D structural organization of ECM as well as cellular morphologies in lung or airway tissue with spectral specificity and sensitivity. These imaging modalities are minimally invasive since structures deep within tissues can be visualized without the need for tissue fixation and/or sectioning. Much of the traditional histological and chemical procedures associated with conventional microscopy methods, which may alter native structure of lung tissue samples, can be circumvented to generate more accurate 3D morphological and fine structural information. In addition to outlining basic principles associated with MPEF and SHG microscopy methods, this review reports potential uses of these high resolution imaging modalities in lung structural imaging. We place special emphasis on imaging 3D structural features of airways, visualizing and quantifying ECM remodeling associated with mouse asthma model as well as the potential uses for multiphoton microscopy in in vitro airway applications. PMID:21497667

  18. Multiphoton microscopy using intrinsic signals for pharmacological studies in unstained cardiac and vascular tissue

    NASA Astrophysics Data System (ADS)

    Beaurepaire, Emmanuel; Boulesteix, Thierry; Pena, Ana-Maria; Pages, Nicole; Senni, Karim; Godeau, Gaston; Sauviat, Martin-Pierre; Schanne-Klein, Marie-Claire

    2005-03-01

    We report two novel applications of multiphoton microscopy for pharmacological studies of unstained cardiovascular tissue. First, we show that second harmonic generation (SHG) microscopy of unstained cardiac myocytes can be used to determine the sarcomere length with sub-resolution accuracy, owing to the remarkable contrast of the SHG signal originating from myosin filaments. A measurement precision of 20 nm is achieved, taking the sample variability into account. We used this technique to measure sarcomere contracture in the presence of saxitoxin, and results were in agreement with mechanical measurements of atrial tissue contracture. Second, we characterized multiphoton microscopy of intact unlabeled arteries. We performed simultaneous detection of two-photon-excited fluorescence (2PEF) from elastin laminae and SHG from collagen fibers upon 860 nm excitation. Combined 2PEF/SHG images provide a highly specific, micron scale description of the architecture of these two major components of the vessel wall. We used this methodology to study the effects of lindane (a pesticide) on the artery wall structure and evidenced structural alteration of the vessel morphology.

  19. Multiphoton microscopy and microspectroscopy for diagnostics of inflammatory and neoplastic lung

    NASA Astrophysics Data System (ADS)

    Pavlova, Ina; Hume, Kelly R.; Yazinski, Stephanie A.; Flanders, James; Southard, Teresa L.; Weiss, Robert S.; Webb, Watt W.

    2012-03-01

    Limitations of current medical procedures for detecting early lung cancers inspire the need for new diagnostic imaging modalities for the direct microscopic visualization of lung nodules. Multiphoton microscopy (MPM) provides for subcellular resolution imaging of intrinsic fluorescence from unprocessed tissue with minimal optical attenuation and photodamage. We demonstrate that MPM detects morphological and spectral features of lung tissue and differentiates between normal, inflammatory and neoplastic lung. Ex vivo MPM imaging of intrinsic two-photon excited fluorescence was performed on mouse and canine neoplastic, inflammatory and tumor-free lung sites. Results showed that MPM detected microanatomical differences between tumor-free and neoplastic lung tissue similar to standard histopathology but without the need for tissue processing. Furthermore, inflammatory sites displayed a distinct red-shifted fluorescence compared to neoplasms in both mouse and canine lung, and adenocarcinomas displayed a less pronounced fluorescence emission in the 500 to 550 nm region compared to adenomas in mouse models of lung cancer. These spectral distinctions were also confirmed by two-photon excited fluorescence microspectroscopy. We demonstrate the feasibility of applying MPM imaging of intrinsic fluorescence for the differentiation of lung neoplasms, inflammatory and tumor-free lung, which motivates the application of multiphoton endoscopy for the in situ imaging of lung nodules.

  20. Molecular photoelectron angular distribution rotations in multi-photon resonant ionization of H{sub 2}{sup +} by circularly polarized ultraviolet laser pulses

    SciTech Connect

    Yuan, Kai-Jun Chelkowski, Szczepan; Bandrauk, André D.

    2015-04-14

    We study effects of pulse durations on molecular photoelectron angular distributions (MPADs) in ultrafast circular polarization ultraviolet resonant ionization processes. Simulations performed on aligned H{sub 2}{sup +} by numerically solving time dependent Schrödinger equations show rotations of MPADs with respect to the molecular symmetry axes. It is found that in multi-photon resonant ionization processes, rotation angles are sensitive to pulse durations, which we attribute to the coherent resonant excitation between the ground state and the intermediate excited electronic state induced by Rabi oscillations. Multi-photon nonresonant and single photon ionization processes are simulated and compared which exhibit a constant rotation angle. An asymmetry parameter is introduced to describe the pulse duration sensitivity by perturbation theory models. Influence of pulse frequency detunings on MPADs is also investigated where oscillations of rotations are absent at long pulse durations due to nonresonance excitation.

  1. Rotational averaging of multiphoton absorption cross sections

    SciTech Connect

    Friese, Daniel H. Beerepoot, Maarten T. P.; Ruud, Kenneth

    2014-11-28

    Rotational averaging of tensors is a crucial step in the calculation of molecular properties in isotropic media. We present a scheme for the rotational averaging of multiphoton absorption cross sections. We extend existing literature on rotational averaging to even-rank tensors of arbitrary order and derive equations that require only the number of photons as input. In particular, we derive the first explicit expressions for the rotational average of five-, six-, and seven-photon absorption cross sections. This work is one of the required steps in making the calculation of these higher-order absorption properties possible. The results can be applied to any even-rank tensor provided linearly polarized light is used.

  2. Multiphoton microscopy of cleared mouse organs

    NASA Astrophysics Data System (ADS)

    Parra, Sonia G.; Chia, Thomas H.; Zinter, Joseph P.; Levene, Michael J.

    2010-05-01

    Typical imaging depths with multiphoton microscopy (MPM) are limited to less than 300 ?m in many tissues due to light scattering. Optical clearing significantly reduces light scattering by replacing water in the organ tissue with a fluid having a similar index of refraction to that of proteins. We demonstrate MPM of intact, fixed, cleared mouse organs with penetration depths and fields of view in excess of 2 mm. MPM enables the creation of large 3-D data sets with flexibility in pixel format and ready access to intrinsic fluorescence and second-harmonic generation. We present high-resolution images and 3-D image stacks of the brain, small intestine, large intestine, kidney, lung, and testicle with image sizes as large as 40964096 pixels.

  3. REVIEW ARTICLE Multiphoton polymerization of hybrid materials

    NASA Astrophysics Data System (ADS)

    Farsari, Maria; Vamvakaki, Maria; Chichkov, Boris N.

    2010-12-01

    Multiphoton polymerization has been developed as a direct laser writing technique for the preparation of complex 3D structures with resolution beyond the diffraction limit of light. The combination of two or more hybrid materials with different functionalities in the same system has allowed the preparation of structures with advanced properties and functions. Furthermore, the surface functionalization of the 3D structures opens new avenues for their applications in a variety of nanobiotechnological fields. This paper describes the principles of 2PP and the experimental set-up used for 3D structure fabrication. It also gives an overview of the materials that have been employed in 2PP so far and depicts the perspectives of this technique in the development of new active components.

  4. Single and Multiphoton Fluorescence Recovery after Photobleaching

    PubMed Central

    Sullivan, Kelley D.; Majewska, Ania K.

    2015-01-01

    Fluorescence recovery after photobleaching (FRAP) is a microscopy technique for measuring the kinetics of fluorescently labeled molecules, and can be applied both in vitro and in vivo for two-and three-dimensional systems. This chapter discusses the three basic FRAP methods: traditional FRAP, multi-photon FRAP (MPFRAP), and FRAP with spatial Fourier analysis (SFA-FRAP). Each discussion is accompanied by a description of the appropriate mathematical analysis appropriate for situations in which the recovery kinetics are dictated by free diffusion. In some experiments, the recovery kinetics are dictated by the boundary conditions of the system, and FRAP is then used to quantify the connectivity of various compartments. Since the appropriate mathematical analysis is independent of the bleaching method, the analysis of compartmental connectivity is discussed last, in a separate section. PMID:25561627

  5. Multiphoton imaging of biological samples during freezing and heating

    NASA Astrophysics Data System (ADS)

    Breunig, H. G.; Uchugonova, A.; König, K.

    2014-02-01

    We applied multiphoton microscopic imaging to observe freezing and heating effects in plant- and animal cell samples. The experimental setups consisted of a multiphoton imaging system and a heating and cooling stage which allows for precise temperature control from liquid nitrogen temperature (-196°C 77 K) up to +600°C (873 K) with heating/freezing rates between 0.01 K/min and 150 K/min. Two multiphoton imaging systems were used: a system based on a modified optical microscope and a flexible mobile system. To illustrate the imaging capabilities, plant leafs as well as animal cells were microscopically imaged in vivo during freezing based on autofluorescence lifetime and intensity of intrinsic molecules. The measurements illustrate the usefulness of multiphoton imaging to investigate freezing effects on animal and plant cells.

  6. Multiphoton microscopy system with a compact fiber-based femtosecond-pulse laser and handheld probe.

    PubMed

    Liu, Gangjun; Kieu, Khanh; Wise, Frank W; Chen, Zhongping

    2011-01-01

    We report on the development of a compact multiphoton microscopy (MPM) system that integrates a compact and robust fiber laser with a miniature probe. The all normal dispersion fiber femtosecond laser has a central wavelength of 1.06 ?m, pulse width of 125 fs and average power of more than 1 W. A double cladding photonic crystal fiber was used to deliver the excitation beam and to collect the two-photon signal. The hand-held probe included galvanometer-based mirror scanners, relay lenses and a focusing lens. The packaged probe had a diameter of 16 mm. Second harmonic generation (SHG) images and two-photon excited fluorescence (TPEF) images of biological tissues were demonstrated using the system. PMID:20635426

  7. Photophysical characterization of sickle cell disease hemoglobin by multi-photon microscopy

    PubMed Central

    Vigil, Genevieve D.; Howard, Scott S.

    2015-01-01

    The photophysical properties of human sickle cell disease (SCD) Hemoglobin (Hb) is characterized by multi-photon microscopy (MPM). The intrinsic two-photon excited fluorescence (TPEF) signal associated with extracted hemoglobin was investigated and the solidified SCD variant (HbS) was found to demonstrate broad emission peaking around 510 nm when excited at 800 nm. MPM is used to dynamically induce and image HbS gelling by photolysis of deoxygenated HbS. For comparison, photolysis conditions were applied to a healthy variant of human hemoglobin (HbA) and found to remain in solution not forming fibers. The use of this signal to study the mechanism of HbS polymerization associated with the sickling of SCD erythrocytes is discussed. PMID:26504657

  8. Generation of High-Order Squeezing in Multiphoton Micromaser

    NASA Technical Reports Server (NTRS)

    Li, Fu-Li; Huang, Qing

    1996-01-01

    The generation of steady state higher-order squeezing in the sense of Hong and Mandel and also of Hillery in a multiphoton micromaser is studied. The results show that the cotangent state which is generated by the coherent trapping scheme in a multiphoton micromaser can exhibit not only second-order squeezing but also fourth-order and squared field amplitude squeezings. The influence of the cavity loss on the squeezings is investigated.

  9. Visualizing laser-skin interaction in vivo by multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Tsai, Tsung-Hua; Jee, Shiou-Hwa; Chan, Jung-Yi; Lee, Jin-Ning; Lee, Woan-Ruoh; Dong, Chen-Yuan; Lin, Sung-Jan

    2009-03-01

    Recently, multiphoton microscopy has gained much popularity as a noninvasive imaging modality in biomedical research. We evaluate the potential of multiphoton microscopy for monitoring laser-skin reaction in vivo. Nude mouse skin is irradiated with an erbium:YAG laser at various fluences and immediately imaged by a multiphoton microscope. The alterations of cutaneous nonlinear optical properties including multiphoton autofluorescence and second-harmonic generation associated with laser irradiation are evaluated morphologically and quantitatively. Our results show that an erbium:YAG laser at a low fluence can selectively disrupt the stratum corneum, and this alteration may account for the penetration enhancing effect of laser-assisted transcutaneous drug delivery. At a higher fluence, the zone of tissue ablation as well as the disruption of the surrounding stratum corneum, keratinocytes, and dermal extracellular matrix can be better characterized by multiphoton microscopy as compared with conventional histology. Furthermore, the degree of collagen damage in the residual thermal zone can be quantified by second-harmonic generation signals, which have significant difference between control skin, skin irradiated with a 1.5-, 8-, and 16-J/cm2 erbium:YAG laser (P<0.05). We show that multiphoton microscopy can be a useful noninvasive imaging modality for monitoring laser-skin reaction in vivo.

  10. New photocalorimetric references for UV excitation

    NASA Astrophysics Data System (ADS)

    Murgida, D. H.; Erra-Balsells, R.; Bilmes, G. M.

    1996-02-01

    A set of calorimetric references for excitation at ? = 266 nm in acetonitrile solutions is proposed. Theophylline and several purinic compounds (purine, 6-methylpurine, 6-methoxypurine, 6-chloropurine, and 6-aminopurine) were investigated using pulsed laser optoacoustic spectroscopy with resolution times between 150 ns and 1 ?s and compared with 2-hydroxybenzophenone. At room temperature, all these compounds show no fluorescence, good stability and no photochemical processes as photodegradation or multiphotonic effects below fluences of 1000 J/m 2.

  11. Distinguishing between benign and malignant melanocytic nevi by in vivo multiphoton microscopy

    PubMed Central

    Balu, Mihaela; Kelly, Kristen M.; Zachary, Christopher B.; Harris, Ronald M.; Krasieva, Tatiana B.; Knig, Karsten; Durkin, Anthony J.; Tromberg, Bruce J.

    2014-01-01

    Monitoring of atypical nevi is an important step in early detection of melanoma, a clinical imperative in preventing the disease progression. Current standard diagnosis is based on biopsy and histopathological examination, a method that is invasive and highly dependent upon the physicians experience. In this work, we employed a clinical multiphoton microscope to image in vivo and non-invasively melanocytic nevi at three different stages: common nevi without dysplastic changes, dysplastic nevi with structural and architectural atypia, and melanoma. We analyzed multiphoton microscopy (MPM) images corresponding to 15 lesions (5 in each group) both qualitatively and quantitatively. For the qualitative analysis, we identified the morphological features characteristic of each group. MPM images corresponding to dysplastic nevi and melanoma were compared with standard histopathology in order to determine correlations between tissue constituents and morphology and to evaluate whether standard histopathology criteria can be identified in the MPM images. Prominent qualitative correlations included the morphology of epidermal keratinocytes, the appearance of nests of nevus cells surrounded by collagen fibers, and the structure of the epidermal-dermal junction. For the quantitative analysis, we defined a numerical multiphoton melanoma index (MMI) based on 3D in vivo image analysis that scores signals derived from two-photon excited fluorescence, second harmonic generation, and melanocyte morphology features on a continuous 9-point scale. Indices corresponding to common nevi (01), dysplastic nevi (14) and melanoma (58) were significantly different (p<0.05), suggesting the potential of the method to distinguish between melanocytic nevi in vivo. PMID:24686168

  12. Excitation of the low lying vibrational levels of H2O by O(3P) as measured on Spacelab 2

    NASA Astrophysics Data System (ADS)

    Meyerott, R. E.; Swenson, G. R.; Schweitzer, E. L.; Koch, D. G.

    1994-09-01

    The data from the infrared telescope (IRT), which was flown on space shuttle Challenger Spacelab 2 mission (July 1985), were originally reported by Koch et al. (1987) as originating from near orbital emissions, primarily H2O. In this study, analysis of this data was extended to determine the collisional cross sections for the excitation of the low lying vibrational levels of H2O, present in the orbiter cloud, by atmospheric O(3P). The evaluation of the contribution to the measured signal from solar excitation and ram O excitation of outgassing H2O permits the determination of the H2O column density and the excitation cross section of the (101) level at an O(3P) velocity of approximately 7.75 km/s. Contributions to the radiation in the 1.7-3.0 micron band by transitions from the (100), (001), and multiquantum excited levels are discussed. The findings of the study are (1) the IRT data for the 4.5-9.5 micron and the nighttime data for the 1.7-3.0 micron sensors are consistent with being explained by collision excitation of H2O by O(3P), (2) diurnal variations of 4.5-9.5 micron intensities follow the model predicted O density for a full orbit, (3) daytime increases in the H2O cloud density were not evident, (4) the cross sections for the collisional excitation process are derived and compared to values computated by Johnson (1986) and Redmon et al. (1986), (5) theoretical investigation suggests greater than 60% of the radiation from H2O is a result of multiphoton emission resulting from collisional multiquanta excitation, and (6) the large daytime increase in the 1.7-3.0 micron intensity data suggests that O(+) may likely be instrumental in producing excited H2O(+) through charge exchange.

  13. Particle modeling of microplasma generated by resonance enhanced multiphoton ionization

    NASA Astrophysics Data System (ADS)

    Tholeti, Siva Sashank

    Resonance-enhanced multiphoton ionization (REMPI) is a technique applied to the spectroscopy of atoms. The REMPI technique typically involves a resonant single or multiple photon absorption to an electronically excited intermediate state followed by another photon which ionizes the atom. Rayleigh scattering of REMPI plasma has given rise to a non-intrusive, time accurate measurement of electron formation and loss, which lead to many applications viz. trace species detection and micro-plasma diagnostics. It is very important to quantify the expansion process and the evolution of energy of electrons and ions. The operation scale of this process is in microns and non continuum nature of the process lead to the use of PIC/MCC scheme to compu- tationally model REMPI technique. This work attempts to understand and analyze the processes taking place during the expansion of REMPI plasma computationally using the PIC/MCC scheme. One dimensional and two dimensional approximations are considered to analyze the REMPI plasma expansion in Argon gas generated by a laser with a focal shape of a prolate ellipsoid. The expansion of the plasma is found to be very sensitive to the initial velocity distribution of the electrons. REMPI plasma expansion is shown to be ambipolar in nature, with the radial expansion more predominant than axial expansion, hence requiring the 2D model. Electron energy distribution functions(EEDFs) are found at various radial locations along with the corresponding mean energies. The deviation of the EEDFs from that of equilibrium Maxwell-Boltzmann energy distribution is presented both qualitatively and quanti- tatively, indicating the predominant processes at various instances in time.

  14. Dynamics of C-Br bond dissociation in methyl 2-bromopropionate at 235 nm: A resonance-enhanced multiphoton ionization study

    NASA Astrophysics Data System (ADS)

    Saha, Ankur; Kumar, Awadhesh; Naik, Prakash D.

    2016-01-01

    The dynamics of the C-Br bond dissociation on UV excitation of methyl 2-bromopropionate mainly to the 1(n?*) state, repulsive in the C-Br bond, has been investigated, employing resonance-enhanced multiphoton ionization. Both the ground state and spin-orbits excited bromine atoms were detected, with the former being the major channel. Bromine fragments show bimodal translational energy distributions, with slow and fast (major) bromine atoms arising mainly from the ground and excited electronic states, respectively. The measured recoil anisotropy suggests isotropic angular distributions of bromine atoms. Molecular orbital calculations reveal an important role of avoided curve crossing on C-Br bond dissociation dynamics.

  15. High-resolution multiphoton optical tomography of tissues: an in vitro and in vivo study

    NASA Astrophysics Data System (ADS)

    Riemann, Iris; Schenke-Layland, Katja; Ehlers, Alexander; Dimitrow, Enrico; Kaatz, Martin; Elsner, Peter; Martin, Sven; Knig, Karsten

    2006-03-01

    Multiphoton optical tomography based on NIR (near-infrared) femtosecond laser pulses provides non-invasive optical sectioning of skin with high spatial intracellular resolution and high tissue penetration. The imaging system DermaInspect was used to perform this technology in clinical studies in vivo on patients with suspicious melanoma. Pigmented cell clusters based on non-linear luminescence were clearly distinguished from non-pigmented cells in the epidermis using the autofluorescence of endogenous fluorophores like NAD(P)H, flavins, keratin, elastin, collagen and melanin. Some of the investigated tissues showed differences in the structure of the epidermal layers and the presence of dendritic cells compared to normal skin. Multiphoton laser microscopy was used to visualize extracellular matrix (ECM) structures of native and tissueengineered heart valves. The quality of the resulting 3-D images allowed an exact differentiation between collagenous and elastic fibers. The analysis of heart valve tissues of patients with cardiomyopathy revealed a dramatic loss of its capability to generate SH (second harmonic), indicating a structural deformation of the collagenous fibers, which was virtually impossible to obtain by routine histological or immunohistological staining. These results indicate that NIR femtosecond laser scanning systems can be employed as novel non-invasive optical technology for 3-D resolved ECM component imaging and in vitro and in vivo tissue diagnosis.

  16. Photon-momentum transfer in multiphoton ionization and in time-resolved holography with photoelectrons

    NASA Astrophysics Data System (ADS)

    Chelkowski, Szczepan; Bandrauk, Andr D.; Corkum, Paul B.

    2015-11-01

    In most models and theoretical calculations describing multiphoton ionization by infrared light, the dipole approximation is used. This is equivalent to setting the very small photon momentum to zero. Using numerical solutions of the two-dimensional (2-D) time-dependent Schrdinger equation for one electron (H-like) systems, we show that, for linear polarization, the radiation pressure on photoelectrons is very sensitive to the details of the ionization mechanism. The directly ionized photoelectrons, those that never recollide with the parent ion, are driven in the direction of the laser photon momentum, whereas a fraction of slower photoelectrons are pushed in the opposite direction, leading to the counterintuitive shifts observed in recent experiments [Phys. Rev. Lett. 113, 243001 (2014), 10.1103/PhysRevLett.113.243001]. This complex response is due to the interplay between the Lorentz force and the Coulomb attraction from the ion. On average, however, the photoelectron momentum is in the direction of the photon momentum as in the case of circular polarization. The influence of the photon momentum is shown to be discernible in the holographic patterns of time-resolved atomic and molecular holography with photoelectrons, thus suggesting a new research subject in multiphoton ionization.

  17. Stepwise multiphoton activation fluorescence reveals a new method of melanin detection

    NASA Astrophysics Data System (ADS)

    Lai, Zhenhua; Kerimo, Josef; Mega, Yair; DiMarzio, Charles A.

    2013-06-01

    The stepwise multiphoton activated fluorescence (SMPAF) of melanin, activated by a continuous-wave mode near infrared (NIR) laser, reveals a broad spectrum extending from the visible spectra to the NIR and has potential application for a low-cost, reliable method of detecting melanin. SMPAF images of melanin in mouse hair and skin are compared with conventional multiphoton fluorescence microscopy and confocal reflectance microscopy (CRM). By combining CRM with SMPAF, we can locate melanin reliably. However, we have the added benefit of eliminating background interference from other components inside mouse hair and skin. The melanin SMPAF signal from the mouse hair is a mixture of a two-photon process and a third-order process. The melanin SMPAF emission spectrum is activated by a 1505.9-nm laser light, and the resulting spectrum has a peak at 960 nm. The discovery of the emission peak may lead to a more energy-efficient method of background-free melanin detection with less photo-bleaching.

  18. Insights on proximity effect and multiphoton induced luminescence from gold nanospheres in far field optical microscopy

    NASA Astrophysics Data System (ADS)

    Borglin, Johan; Guldbrand, Stina; Evenbratt, Hanne; Kirejev, Vladimir; Grnbeck, Henrik; Ericson, Marica B.

    2015-12-01

    Gold nanoparticles can be visualized in far-field multiphoton laser-scanning microscopy (MPM) based on the phenomena of multiphoton induced luminescence (MIL). This is of interest for biomedical applications, e.g., for cancer diagnostics, as MPM allows for working in the near-infrared (NIR) optical window of tissue. It is well known that the aggregation of particles causes a redshift of the plasmon resonance, but its implications for MIL applying far-field MPM should be further exploited. Here, we explore MIL from 10 nm gold nanospheres that are chemically deposited on glass substrates in controlled coverage gradients using MPM operating in NIR range. The substrates enable studies of MIL as a function of inter-particle distance and clustering. It was shown that MIL was only detected from areas on the substrates where the particle spacing was less than one particle diameter, or where the particles have aggregated. The results are interpreted in the context that the underlying physical phenomenon of MIL is a sequential two-photon absorption process, where the first event is driven by the plasmon resonance. It is evident that gold nanospheres in this size range have to be closely spaced or clustered to exhibit detectable MIL using far-field MPM operating in the NIR region.

  19. InVivo Multiphoton NADH Fluorescence Reveals Depth-Dependent Keratinocyte Metabolism in Human Skin

    PubMed Central

    Balu, Mihaela; Mazhar, Amaan; Hayakawa, CaroleK.; Mittal, Richa; Krasieva, TatianaB.; Knig, Karsten; Venugopalan, Vasan; Tromberg, BruceJ.

    2013-01-01

    We employ a clinical multiphoton microscope to monitor invivo and noninvasively the changes in reduced nicotinamide adenine dinucleotide (NADH) fluorescence of human epidermal cells during arterial occlusion. We correlate these results with measurements of tissue oxy- and deoxyhemoglobin concentration during oxygen deprivation using spatial frequency domain imaging. During arterial occlusion, a decrease in oxyhemoglobin corresponds to an increase in NADH fluorescence in the basal epidermal cells, implying a reduction in basal cell oxidative phosphorylation. The ischemia-induced oxygen deprivation is associated with a strong increase in NADH fluorescence of keratinocytes in layers close to the stratum basale, whereas keratinocytes from epidermal layers closer to the skin surface are not affected. Spatial frequency domain imaging optical property measurements, combined with a multilayer Monte Carlo-based radiative transport model of multiphoton microscopy signal collection in skin, establish that localized tissue optical property changes during occlusion do not impact the observed NADH signal increase. This outcome supports the hypothesis that the vascular contribution to the basal layer oxygen supply is significant and these cells engage in oxidative metabolism. Keratinocytes in the more superficial stratum granulosum are either supplied by atmospheric oxygen or are functionally anaerobic. Based on combined hemodynamic and two-photon excited fluorescence data, the oxygen consumption rate in the stratum basale is estimated to be ?0.035 ?moles/106 cells/h. PMID:23332078

  20. Multiphoton microscopic imaging of fibrotic focus in invasive ductal carcinoma of the breast

    NASA Astrophysics Data System (ADS)

    Chen, Sijia; Nie, Yuting; Lian, Yuane; Wu, Yan; Fu, Fangmeng; Wang, Chuan; Zhuo, Shuangmu; Chen, Jianxin

    2014-11-01

    During the proliferation of breast cancer, the desmoplastic can evoke a fibrosis response by invading healthy tissue. Fibrotic focus (FF) in invasive ductal carcinoma (IDC) of the breast had been reported to be associated with significantly poorer survival rate than IDC without FF. As an important prognosis indicator, it's difficult to obtain the exact fibrotic information from traditional detection method such as mammography. Multiphoton imaging based on two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) has been recently employed for microscopic examination of unstained tissue. In this study, multiphoton microscopy (MPM) was used to image the fibrotic focus in invasive ductal carcinoma tissue. The morphology and distribution of collagen in fibrotic focus can be demonstrated by the SHG signal. Variation of collagen between IDC with and without FF will be examined and further characterized, which may be greatly related to the metastasis of breast cancer. Our result suggested that the MPM can be efficient in identifying and locating the fibrotic focus in IDC. Combining with the pathology analysis and other detecting methods, MPM owns potential in becoming an advanced histological tool for detecting the fibrotic focus in IDC and collecting prognosis information, which may guide the subsequent surgery option and therapy procedure for patients.

  1. Deep-tissue multiphoton fluorescence lifetime microscopy for intravital imaging of protein-protein interactions

    NASA Astrophysics Data System (ADS)

    Fruhwirth, G. O.; Matthews, D. R.; Brock, A.; Keppler, M.; Vojnovic, B.; Ng, T.; Ameer-Beg, S.

    2009-02-01

    Fluorescent lifetime imaging microscopy (FLIM) has proven to be a valuable tool in beating the Rayleigh criterion for light microscopy by measuring Frster resonance energy transfer (FRET) between two fluorophores. Applying multiphoton FLIM, we previously showed in a human breast cancer cell line that recycling of a membrane receptorgreen fluorescent protein fusion is enhanced concomitantly with the formation of a receptor:protein kinase C ? complex in the endosomal compartment. We have extended this established technique to probe direct protein-protein interactions also in vivo. Therefore, we used various expressible fluorescent tags fused to membrane receptor molecules in order to generate stable two-colour breast carcinoma cell lines via controlled retroviral infection. We used these cell lines for establishing a xenograft tumour model in immune-compromised Nude mice. Using this animal model in conjunction with scanning Ti:Sapphire laser-based two-photon excitation, we established deep-tissue multiphoton FLIM in vivo. For the first time, this novel technique enables us to directly assess donor fluorescence lifetime changes in vivo and we show the application of this method for intravital imaging of direct protein-protein interactions.

  2. Chronic imaging of amyloid plaques in the live mouse brain using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Bacskai, Brian J.; Kajdasz, Stephen T.; Christie, R. H.; Zipfel, Warren R.; Williams, Rebecca M.; Kasischke, Karl A.; Webb, Watt W.; Hyman, B. T.

    2001-04-01

    Transgenic mice expressing the human Amyloid Precursor Protein (APP) develop amyloid plaques as they age. These plaques resemble those found in the human disease. Multiphoton laser scanning microscopy combined with a novel surgical approach was used to measure amyloid plaque dynamics chronically in the cortex of living transgenic mice. Thioflavine S (thioS) was used as a fluorescent marker of amyloid deposits. Multiphoton excitation allowed visualization of amyloid plaques up to 200 micrometers deep into the brain. The surgical site could be imaged repeatedly without overt damage to the tissue, and individual plaques within this volume could be reliably identified over periods of several days to several months. On average, plaque sizes remained constant over time, supporting a model of rapid deposition, followed by relative stability. Alternative reporters for in vivo histology include thiazine red, and FITC-labeled amyloid-(Beta) peptide. We also present examples of multi-color imaging using Hoechst dyes and FITC-labeled tomato lectin. These approaches allow us to observe cell nuclei or microglia simultaneously with amyloid-(Beta) deposits in vivo. Chronic imaging of a variety of reporters in these transgenic mice should provide insight into the dynamics of amyloid-(Beta) activity in the brain.

  3. Setup and use of a two-laser multiphoton microscope for multichannel intravital fluorescence imaging

    PubMed Central

    Entenberg, David; Wyckoff, Jeffrey; Gligorijevic, Bojana; Roussos, Evanthia T; Verkhusha, Vladislav V; Pollard, Jeffrey W; Condeelis, John

    2014-01-01

    Characterizing biological mechanisms dependent upon the interaction of many cell types in vivo requires both multiphoton microscope systems capable of expanding the number and types of fluorophores that can be imaged simultaneously while removing the wavelength and tunability restrictions of existing systems, and enhanced software for extracting critical cellular parameters from voluminous 4D data sets. We present a procedure for constructing a two-laser multiphoton microscope that extends the wavelength range of excitation light, expands the number of simultaneously usable fluorophores and markedly increases signal to noise via over-clocking of detection. We also utilize a custom-written software plug-in that simplifies the quantitative tracking and analysis of 4D intravital image data. We begin by describing the optics, hardware, electronics and software required, and finally the use of the plug-in for analysis. We demonstrate the use of the setup and plug-in by presenting data collected via intravital imaging of a mouse model of breast cancer. The procedure may be completed in ~24 h. PMID:21959234

  4. Label-free monitoring of colorectal adenoma-carcinoma sequence based on multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Chen, J. X.; Li, H. S.; Chen, Z. F.; Feng, C. Y.; Yang, Y. H.; Jiang, W. Z.; Guan, G. X.; Zhu, X. Q.; Zhuo, S. M.; Xu, J.

    2014-06-01

    The monitoring and evaluation of colorectal adenoma-carcinoma sequence during endoscopy are important for endoscopic resection of precursor lesions to disrupt the adenoma-carcinoma sequence and halt progression to invasive neoplastic disease. In this study, multiphoton microscopy (MPM) was used to identify different stages during the development of colorectal adenocarcinoma including adenoma with low-grade and high-grade dysplasia, and adenocarcinoma invading the submucosa. It was found that by combining two-photon excited fluorescence (TPEF) imaging and second harmonic generation (SHG) imaging, MPM can reveal the morphological changes of the epithelial cells and glands, identify the invasive position and depth of atypical glands and quantitatively describe the change of the cellular nucleus and the nuclear-to-cytoplasmic ratio during the stepwise progression of colorectal adenocarcinoma. These are important pathological findings for pathologists when diagnosing colorectal lesions. With the advancement of a compact and flexible multiphoton endoscope for in vivo imaging and clinical applications, MPM has the potential to provide immediate histological diagnosis for the monitoring and evaluation of the colorectal adenoma-carcinoma sequence during endoscopy.

  5. Combined multiphoton microscopy and optical coherence tomography using a 12-fs broadband source

    NASA Astrophysics Data System (ADS)

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

    2006-03-01

    A 12-fs broadband (100-nm) source is used to combine multiphoton microscopy (MPM) and optical coherence tomography (OCT) in a single platform. An ultrafast Ti:sapphire laser simultaneously provides short pulses necessary for efficient MPM excitation and the broad bandwidth required for high-resolution OCT. Using 0.3-m microspheres and a 63, 0.95 numerical aperture objective, we demonstrate that MPM and OCT channels are coregistered with lateral resolution of approximately 0.5 m and axial resolution of approximately 1.5 m. Preliminary studies of a 3-D organotypic epithelial tissue model show that multiphoton images of fluorescence and second harmonic signals are derived from cellular and extracellular matrix structures, respectively, while OCT images are generated from scattering interfaces due to tissue variations in refractive index. The combined MPM/OCT microscope is capable of providing simultaneous functional and structural information from cells and extracellular matrix and is potentially a powerful tool for studying biological processes in thick tissues.

  6. Time- and spectral-resolved multiphoton imaging of fresh bladder biopsies

    NASA Astrophysics Data System (ADS)

    Cicchi, Riccardo; Crisci, Alfonso; Nesi, Gabriella; Cosci, Alessandro; Giancane, Saverio; Carini, Marco; Pavone, Francesco S.

    2009-07-01

    Human tissues intrinsically contains many fluorophores, as such NADH, elastin, collagen, and flavins, that can be excited and imaged using multiphoton microscopy, up to 150 microns depth. In this work we used combined two photon intrinsic fluorescence (TPE), second harmonic generation microscopy (SHG), fluorescence lifetime imaging microscopy (FLIM), and multispectral two photon emission detection (MTPE) to investigate different kinds of human ex-vivo fresh biopsies of bladder. Morphological and spectroscopic analyses allowed to characterize both healthy and pathological tissue samples in a good agreement with common routine histology. In particular, we examined tissue samples from bladder normal mucosa, and bladder carcinoma in-situ (CIS), finding both morphological and spectroscopic differences. From the morphological point of view, cancer cells appeared more elongated with respect to corresponding normal cells; they also exhibited a different nucleus to cytoplasm ratio. From the spectroscopic point of view, we found differences between the two tissue types in both spectral emission and fluorescence lifetime distribution. Even if further analysis, as well as a more significant statistics on a large number of samples would be helpful to discriminate between low and high grade cancer, our method is a promising tool to be used as diagnostic confirmation of histological results, as well as a diagnostic tool in a multiphoton endoscope or cystoscope to be used in in-vivo imaging applications.

  7. Soliton dynamics in the multiphoton plasma regime

    PubMed Central

    Husko, Chad A.; Combrié, Sylvain; Colman, Pierre; Zheng, Jiangjun; De Rossi, Alfredo; Wong, Chee Wei

    2013-01-01

    Solitary waves have consistently captured the imagination of scientists, ranging from fundamental breakthroughs in spectroscopy and metrology enabled by supercontinuum light, to gap solitons for dispersionless slow-light, and discrete spatial solitons in lattices, amongst others. Recent progress in strong-field atomic physics include impressive demonstrations of attosecond pulses and high-harmonic generation via photoionization of free-electrons in gases at extreme intensities of 1014 W/cm2. Here we report the first phase-resolved observations of femtosecond optical solitons in a semiconductor microchip, with multiphoton ionization at picojoule energies and 1010 W/cm2 intensities. The dramatic nonlinearity leads to picojoule observations of free-electron-induced blue-shift at 1016 cm−3 carrier densities and self-chirped femtosecond soliton acceleration. Furthermore, we evidence the time-gated dynamics of soliton splitting on-chip, and the suppression of soliton recurrence due to fast free-electron dynamics. These observations in the highly dispersive slow-light media reveal a rich set of physics governing ultralow-power nonlinear photon-plasma dynamics.

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

    SciTech Connect

    Yoshimi, Kazuyoshi; Koshino, Kazuki

    2010-09-15

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

  9. New perspectives in laser analytics: Resonance-enhanced multiphoton ionization in a Paul ion trap combined with a time-of-flight mass spectrometer

    NASA Astrophysics Data System (ADS)

    Bisling, Peter; Heger, Hans Jrg; Michaelis, Walfried; Weitkamp, Claus; Zobel, Harald

    1995-04-01

    A new laser analytical device has been developed that is based on resonance-enhanced multiphoton ionization in the very center of a radio-frequency quadrupole ion trap. Applications in speciation anlaysis of biological and enviromental samples and in materials science will all benefit from laser-optical selectivity in the resonance excitation process, combined with mass-spectropic sensivity which is further enhanced by the ion accumulation and storage capability.

  10. Far-infrared spectroscopy of CH 3OD in highly excited torsional states and the atlas of the Fourier transform spectra in the range 200-350 cm -1

    NASA Astrophysics Data System (ADS)

    Mukhopadhyay, Indranath; Mellau, Georg Chr; Klee, Stefan

    2000-10-01

    The high resolution Fourier transform far-infrared (FIR) spectrum of the torsion rotation band of CH 3OD has been analyzed for the highly excited torsion states ( n?2) in the vibrational ground state. The spectrum shows splitting of the lines due to strong torsional-rotational-vibrational interactions in the molecule. Assignments were possible for rotational sub-bands in the torsional state as high as n=4 and for K values up to 8 and J values of up to 30 in most cases, for all the symmetry species. For the third excited torsional state n=3 assignments were possible to K=10. The data were analyzed with the help of the energy expansion model, which has been proven very successful in methanol. The state dependent expansion parameters are presented. These molecular parameters were able to reproduce the observed wavenumbers almost to within experimental accuracy of 0.0002 cm -1 for clean unblended lines. These expansion coefficients should prove valuable in the calculation of precise energy values for excited torsional states up to n=4, which is way above the torsional barrier. The detailed high-resolution spectral atlas of CH 3OD has been presented in the range 200-350 cm -1. This atlas is an extension of our earlier atlas in the range 20-205 cm -1. The availability of this atlas in the journal will be very valuable for spectroscopists and astrophysicists seeking information in the infrared (IR) region in the laboratory and in outer space.

  11. Resonance enhanced multiphoton ionization spectra of molecules and molecular fragments. Annual progress report, March 1992--February 1993

    SciTech Connect

    1993-12-31

    In this report, the author will review the progress made in his studies of ion rotational distributions resulting from resonance enhanced multiphoton ionization of excited electronic states and from single-photon ionization of ground electronic states of jet-cooled molecules by coherent VUV and XUV radiation. To do so he will select a few examples from his studies which serve to highlight his progress and to identify the background and significance of the specific spectral features and systems he has chosen to study.

  12. Control of resonance enhanced multi-photon ionization photoelectron spectroscopy by phase-shaped femtosecond laser pulse

    SciTech Connect

    Zhang Shian; Lu Chenhui; Jia Tianqing; Sun Zhenrong; Qiu Jianrong

    2012-11-07

    In this paper, we theoretically demonstrate that the (2+1+1) resonance enhanced multi-photon ionization photoelectron spectroscopy in sodium atom can be effectively controlled by shaping femtosecond laser pulse with a {pi} phase step modulation in weak laser field, involving its total photoelectron energy, maximal photoelectron intensity, and spectroscopic bandwidth. Our results show that the total photoelectron energy can be suppressed but not enhanced, the maximal photoelectron intensity can be enhanced and also suppressed, and the photoelectron spectroscopy can be tremendously narrowed. These theoretical results can provide a feasible scheme to achieve the high-resolution photoelectron spectroscopy and study the excited state structure in atomic and molecular systems.

  13. Photofragmentation, state interaction, and energetics of Rydberg and ion-pair states: Resonance enhanced multiphoton ionization of HI

    SciTech Connect

    Hróðmarsson, Helgi Rafn; Wang, Huasheng; Kvaran, Ágúst

    2014-06-28

    Mass resolved resonance enhanced multiphoton ionization data for hydrogen iodide (HI), for two-photon resonance excitation to Rydberg and ion-pair states in the 69 600–72 400 cm{sup −1} region were recorded and analyzed. Spectral perturbations due to homogeneous and heterogeneous interactions between Rydberg and ion-pair states, showing as deformations in line-positions, line-intensities, and line-widths, were focused on. Parameters relevant to photodissociation processes, state interaction strengths and spectroscopic parameters for deperturbed states were derived. Overall interaction and dynamical schemes to describe the observations are proposed.

  14. Selective enhancement of resonant multiphoton ionization with strong laser fields

    NASA Astrophysics Data System (ADS)

    Li, Min; Zhang, Peng; Luo, Siqiang; Zhou, Yueming; Zhang, Qingbin; Lan, Pengfei; Lu, Peixiang

    2015-12-01

    High-resolution photoelectron momentum distributions of Xe atoms ionized by 800-nm linearly polarized laser fields have been traced at intensities from 1.1 1013 to 3.5 1013W /c m2 using velocity-map imaging techniques. At certain laser intensities, the momentum spectrum exhibits a distinct double-ring structure for low-order above-threshold ionization, which appears to be absent at lower or higher laser intensities. By investigating the intensity-resolved photoelectron energy spectrum, we find that this double-ring structure originates from resonant multiphoton ionization involving multiple Rydberg states of atoms. Varying the laser intensity, we can selectively enhance the resonant multiphoton ionization through certain atomic Rydberg states. The photoelectron angular distributions of multiphoton resonance are also investigated for the low-order above-threshold ionization.

  15. Evaluation of multiphoton effects in down-conversion

    SciTech Connect

    Yoshimi, Kazuyoshi; Koshino, Kazuki

    2010-04-15

    Multiphoton effects in down-conversion are investigated based on the full-quantum multimode formalism by considering a three-level system as a prototype nonlinear system. We analytically derive the three-photon output wave function for two input photons, where one of the two input photons is down-converted and the other one is not. Using this output wave function, we calculate the down-conversion probability, the purity, and the fidelity to evaluate the entanglement between a down-converted photon pair and a non-down-converted photon. It is shown that the saturation effect occurs by multiphoton input and that it affects both the down-conversion probability and the quantum correlation between the down-converted photon pair and the non-down-converted photon. We also reveal the necessary conditions for multiphoton effects to be strong.

  16. Vibrational resonance enhanced broadband multiphoton absorption in a triphenylamine derivative

    SciTech Connect

    Lu Changgui; Cui Yiping; Huang Wei; Yun Binfeng; Wang Zhuyuan; Hu Guohua; Cui Jing; Lu Zhifeng; Qian Ying

    2007-09-17

    Multiphoton absorption of 2,5-bis[4-(2-N,N-diphenylaminostyryl)phenyl]-1,3,4-oxadiazole was experimentally studied by using femtosecond laser pulses. This material demonstrates a very broad multiphoton absorption band of around 300 nm width with two peaks of 1250 and 1475 nm. The first peak results from the three-photon absorption process while the second is attributed to the vibrational resonance enhanced four-photon absorption process. Combination of these two processes provides a much broader multiphoton absorption band. In this letter, the analytical solution to nonlinear transmission of a three-photon absorption process is also given when the incident beam has a Gaussian transverse spatial profile.

  17. Development of a compact supersonic jet/multiphoton ionization/time-of-flight mass spectrometer for the on-site analysis of dioxin, part II: Application to chlorobenzene and dibenzofuran.

    PubMed

    Matsumoto, Junichi; Nakano, Bunji; Imasaka, Totaro

    2003-03-01

    A compact supersonic jet/multiphoton ionization/mass spectrometer (SSJ/MPI/MS) equipped with a single pump system was developed for use in the analysis of dioxin. A mass spectrum and multiphoton ionization spectrum were obtained for chlorobenzene and dibenzofuran using an optical parametric oscillator (OPO) for excitation and subsequent ionization. The detection limit for chlorobenzene (S/N = 3) was 0.3 ppb, which corresponds to an absolute amount of 0.1 pg. The rotational temperature was 35 K. This slightly high temperature can be attributed to the long nozzle throat used in the instrument. PMID:12675344

  18. Multiphoton Coherent Manipulation in Large Spin Qubits

    NASA Astrophysics Data System (ADS)

    Chiorescu, Irinel

    2009-03-01

    Manipulation of quantum information allows certain algorithms to be performed at unparalleled speeds. Photons are an ideal choice to manipulate qubits as they interact with quantum systems in predictable ways. They are a versatile tool for manipulating, reading/coupling qubits and for encoding/transferring quantum information over long distances. Spin-based qubits have well known behavior under photon driving and can be potentially operated up to room temperature. When diluted enough to avoid uncontrolled spin-spin interactions, a variety of spin qubits show long coherence times, e.g. the nitrogen vacancies in pure diamonds (1,2), nitrogen atoms trapped in a C60 cage (3), Ho3+ and Cr5+ ions (4,5) and molecular magnets (6,7). We have used large spin Mn2+ ions (S=5/2) to realize a six level system that can be operated by means of single as well as multi-photon coherent Rabi oscillations (8). This spin system has a very small anisotropy whose effect can be tuned in-situ to turn the system into a multi-level harmonic system. This offer new ways of manipulating, reading and resetting a spin qubit. Decoherence effects are strongly reduced by the quasi-isotropic electron interaction with the crystal field and with the 55Mn nuclear spins. [0pt] 1. R. Hanson et al., Science 320, 352 (2008). [0pt] 2. M.V. Gurudev Dutt et al., Science 316, 1312 (2007). [0pt] 3. G.W. Morley et al., Phys. Rev. Lett. 98, 220501 (2007). [0pt] 4. S. Bertaina et al., Nat. Nanotech. 2, 39 (2007). [0pt] 5. S. Nellutla et al., Phys. Rev. Lett. 99, 137601 (2007). [0pt] 6. A. Ardavan et al., Phys. Rev. Lett. 98, 057201 (2007). [0pt] 7. S. Bertaina et al., Nature 453, 203,(2008). [0pt] 8. S. Bertaina et al., submitted.

  19. Highly efficient avalanche multiphoton luminescence from coupled Au nanowires in the visible region.

    PubMed

    Wang, Qu-Quan; Han, Jun-Bo; Guo, Dong-Lai; Xiao, Si; Han, Yi-Bo; Gong, Hong-Mei; Zou, Xian-Wu

    2007-03-01

    We demonstrate highly efficient avalanche multiphoton luminescence (MPL) from ordered-arrayed gold nanowires (NWs) with low time-average excitation intensity, Iexc (5.0-9.1 kW/cm2). The intensity of avalanche MPL, IMPL, is about 10(4) times larger than that of three-photon luminescence, the slope partial differential log IMPL/ partial differential log Iexc of avalanche MPL reaches as high as 18.3, and the corresponding polarization dependence of IMPL has a form of cos50 phip. The emission dynamics of avalanche MPL and three-photon luminescence are also studied comparatively. These observations indicate that the highly efficient avalanche MPL is attributed to the giant enhancement and coupling of longitudinal surface plasmon resonance of ordered-arrayed gold NWs. PMID:17311463

  20. Three dimensional multiphoton imaging of fresh and whole mount developing mouse mammary glands

    PubMed Central

    2013-01-01

    Background The applications of multiphoton microscopy for deep tissue imaging in basic and clinical research are ever increasing, supplementing confocal imaging of the surface layers of cells in tissue. However, imaging living tissue is made difficult by the light scattering properties of the tissue, and this is extraordinarily apparent in the mouse mammary gland which contains a stroma filled with fat cells surrounding the ductal epithelium. Whole mount mammary glands stained with Carmine Alum are easily archived for later reference and readily viewed using bright field microscopy to observe branching architecture of the ductal network. Here, we report on the advantages of multiphoton imaging of whole mount mammary glands. Chief among them is that optical sectioning of the terminal end bud (TEB) and ductal epithelium allows the appreciation of abnormalities in structure that are very difficult to ascertain using either bright field imaging of the stained gland or the conventional approach of hematoxylin and eosin staining of fixed and paraffin-embedded sections. A second advantage is the detail afforded by second harmonic generation (SHG) in which collagen fiber orientation and abundance can be observed. Methods GFP-mouse mammary glands were imaged live or after whole mount preparation using a Zeiss LSM510/META/NLO multiphoton microscope with the purpose of obtaining high resolution images with 3D content, and evaluating any structural alterations induced by whole mount preparation. We describe a simple means for using a commercial confocal/ multiphoton microscope equipped with a Ti-Sapphire laser to simultaneously image Carmine Alum fluorescence and collagen fiber networks by SHG with laser excitation set to 860nm. Identical terminal end buds (TEBs) were compared before and after fixation, staining, and whole mount preparation and structure of collagen networks and TEB morphologies were determined. Flexibility in excitation and emission filters was explored using the META detector for spectral emission scanning. Backward scattered or reflected SHG (SHG-B) was detected using a conventional confocal detector with maximum aperture and forward scattered or transmitted SHG (SHG-F) detected using a non-descanned detector. Results We show here that the developing mammary gland is encased in a thin but dense layer of collagen fibers. Sparse collagen layers are also interspersed between stromal layers of fat cells surrounding TEBs. At the margins, TEBs approach the outer collagen layer but do not penetrate it. Abnormal mammary glands from an HAI-1 transgenic FVB mouse model were found to contain TEBs with abnormal pockets of cells forming extra lumens and zones of continuous lateral bud formation interspersed with sparse collagen fibers. Parameters influencing live imaging and imaging of fixed unstained and Carmine Alum stained whole mounts were evaluated. Artifacts induced by light scattering of GFP and Carmine Alum signals from epithelial cells were identified in live tissue as primarily due to fat cells and in whole mount tissue as due to dense Carmine Alum staining of epithelium. Carmine Alum autofluorescence was detected at excitation wavelengths from 750 to 950nm with a peak of emission at 623nm (~602-656nm). Images of Carmine Alum fluorescence differed dramatically at emission wavelengths of 565615nm versus 650710nm. In the latter, a mostly epithelial (nuclear) visualization of Carmine Alum predominates. Autofluorescence with a peak emission of 495nm was derived from the fixed and processed tissue itself as it was present in the unstained whole mount. Contribution of autofluorescence to the image decreases with increasing laser excitation wavelengths. SHG-B versus SHG-F signals revealed collagen fibers and could be found within single fibers, or in different fibers within the same layer. These differences presumably reflected different states of collagen fiber maturation. Loss of SHG signals from layer to layer could be ascribed to artifacts rendered by light scat

  1. Five-Photon Absorption and Selective Enhancement of Multiphoton Absorption Processes

    PubMed Central

    2015-01-01

    We study one-, two-, three-, four-, and five-photon absorption of three centrosymmetric molecules using density functional theory. These calculations are the first ab initio calculations of five-photon absorption. Even- and odd-order absorption processes show different trends in the absorption cross sections. The behavior of all even- and odd-photon absorption properties shows a semiquantitative similarity, which can be explained using few-state models. This analysis shows that odd-photon absorption processes are largely determined by the one-photon absorption strength, whereas all even-photon absorption strengths are largely dominated by the two-photon absorption strength, in both cases modulated by powers of the polarizability of the final excited state. We demonstrate how to selectively enhance a specific multiphoton absorption process. PMID:26120588

  2. Multiphoton fluorescence spectra and lifetimes of biliverdins and their protein-associated complex

    NASA Astrophysics Data System (ADS)

    Huang, Chin-Jie; Wu, Cheng-Ham; Liu, Tzu-Ming

    2012-03-01

    To investigate whether endogenous biliverdins can serve as a fluorescence metabolic marker in cancer diagnosis, we measured their multiphoton fluorescence spectra and lifetimes with femtosecond Cr:forsterite laser. Excited at 1230nm, the two-photon fluorescence of biliverdins peaks around 670nm. The corresponding lifetime (<100ps) was much shorter than those of porphyrins (~10ns), which is another commonly present metabolites in living cells. Further mixing biliverdins with proteins like fetal bovine serum (FBS), biliverdins reductase A (BVRA), or heme oxygenase-1 (HO-1), the yields of red autofluorescences didn't change a lot, but the corresponding lifetimes with HO-1 and BSA were lengthened to 200~300ps. This indicates that biliverdin can have an association with these proteins and change its lifetime. These spectral and temporal characteristics of fluorescence make biliverdin a potential marker fluorophore for hyperspectral diagnosis on the heme catabolism in human cells or tissues.

  3. Hypericin-Mediated Destruction of Collagen Fibers Revealed by Multiphoton Microscopy

    NASA Astrophysics Data System (ADS)

    Hovhannisyan, Ararat Zh.; Hovhannisyan, Vladimir A.; Dong, Chen-Yuan

    Collagen is the major component of the extracellular matrix in skin, tendon, cartilage, cornea, bone, etc., and as a main structural protein is the key determinant of mechanical and functional properties of tissues and organs. Proper balance between synthesis and degradation of collagen fibers is critical for maintaining normal physiologic function; therefore, the modification of collagen fibers in a controlled manner is of high importance for biomedicine. In this work, using second harmonic generation (SHG) and two-photon excited auto-fluorescence (TPEF) microscopy, we revealed that hypericin, a natural pigment extracted from plant, induced structural modification of collagen based tissues. Dynamics of the process was monitored by time-lapse multiphoton imaging. It was demonstrated that hypericin-mediated process was considerably irreversible and has a potential to be used for destroying of abnormal tissues and treatment of some diseases.

  4. Marginal characteristics of skin scarred dermis quantitatively extracted from multiphoton microscopic imaging

    NASA Astrophysics Data System (ADS)

    Zhu, Xiaoqin; Zhuo, Shuangmu; Zheng, Liqin; Jiang, Xingshan; Chen, Jianxin; Lin, Bifang

    2010-11-01

    Multiphoton microscopy based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) was applied to examine the marginal regions at dermis of normal, atrophic and keloid scars. High-contrast, high-resolution image showed an obvious boundary at scar margin and different morphological patterns of collagen or elastin on the two sides. Since the degree of the morphological alteration between the two sides of boundary at scar margin was varied among different types of scars, alteration degree of SHG-to-TPEF index was defined as a quantitative indicator for discrimination. It will help to determine the most appropriate clinical treatment strategy for different types of scars and potentially monitor therapy in vivo.

  5. Mapping femtosecond pulse front distortion and group velocity dispersion in multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Tullis, I. D.; Ameer-Beg, S. M.; Barber, P. R.; Rankov, V.; Vojnovic, B.

    2006-02-01

    Group velocity dispersion (GVD) and pulse front distortion of ultrashort pulses are of critical importance in efficient multiphoton excitation microscopy. Since measurement of the pulse front distortion due to a lens is not trivial we have developed an imaging interferometric cross-correlator which allows us to measure temporal delays and pulse-widths across the spatial profile of the beam. The instrument consists of a modified Michelson interferometer with a reference arm containing a voice-coil delay stage and an arm which contains the optics under test. The pulse replicas are recombined and incident on a 22×22 lenslet array. The beamlets are focused in a 0.5 mm thick BBO crystal (cut for Type I second harmonic generation), filtered to remove the IR component of the beam and imaged using a 500 fps camera. The GVD and pulse front distortion are extracted from the temporal stack of beamlet images to produce a low resolution spatio-temporal map.

  6. On-line screening of airborne PAH contamination by simultaneous multiphoton ionization and laser induced fluorescence

    SciTech Connect

    Gridin, V.V.; Inoue, Takanori; Ogawa, Teiichiro; Schechter, I.

    2000-04-01

    In this preliminary study, the feasibility of on-line monitoring of sub-micron polycyclic aromatic hydrocarbon (PAH) aerosols by a combination of laser-induced-fluorescence (LIF) and laser multiphoton ionization (LMPI) techniques was investigated. For this purpose, an aerosol contaminated nitrogen gas was slowly bubbled through a quartz measurement chamber filled with hexane. Continuous PAH monitoring was maintained over two hours of the resultant concentration process. Simultaneous LIF and LMPI excitation was achieved by a pulsed dye laser, operated at 283 nm. For this wavelength, pyrene and naphthalene aerosols were used as target PAH materials. The resultant fluorescence light was collected by an optical fiber, while the LMPI signals were acquired by a pair of stainless steel electrodes immersed in the solution, allowing an automated readout of the photoionization current. The results indicate that PAH sub-micron aerosols, at an air concentration of 1 mg/m{sup 3}, can be monitored in this way.

  7. Multiphoton microscopy using frequency-doubled compact femtosecond erbium-doped fiber laser

    NASA Astrophysics Data System (ADS)

    Huang, Lin; Chong, Shau Poh; Mills, Arthur; Jones, David; Tang, Shuo

    2014-02-01

    We report on the development of a compact multiphoton microscopy (MPM) system based on a frequency-doubled, femtosecond erbium-doped fiber laser source at 1.58 μm. By use of periodically poled MgO:LiNbO3, frequency-doubled pulses at 790 nm with average power of 75 mW and pulse width of 130 fs are applied as the excitation source. The fiber laser is optimized for its parameters along with the dispersive properties of the delivery fiber such that the MPM signal is maximized at the sample location. Micro-electro-mechanical system (MEMS) scanner, miniature objective, and multimode fiber are further used to make the MPM system compact. MPM images are obtained from unstained biological samples. The MPM system with a compact, portable, low-cost fiber laser has a great potential to transform the bench-top MPM system to a portable system for in vivo MPM imaging.

  8. Effect of multiphoton ionization on performance of crystalline lens.

    PubMed

    Gupta, Pradeep Kumar; Singh, Ram Kishor; Strickland, D; Campbell, M C W; Sharma, R P

    2014-12-15

    This Letter presents a model for propagation of a laser pulse in a human crystalline lens. The model contains a transverse beam diffraction effect, laser-induced optical breakdown for the creation of plasma via a multiphoton ionization process, and the gradient index (GRIN) structure. Plasma introduces the nonlinearity in the crystalline lens which affects the propagation of the beam. The multiphoton ionization process generates plasma that changes the refractive index and hence leads to the defocusing of the laser beam. The Letter also points out the relevance of the present investigation to cavitation bubble formation for restoring the elasticity of the eyes. PMID:25502994

  9. Multiphoton autofluorescence spectral analysis for fungus imaging and identification

    NASA Astrophysics Data System (ADS)

    Lin, Sung-Jan; Tan, Hsin-Yuan; Kuo, Chien-Jui; Wu, Ruei-Jr; Wang, Shiou-Han; Chen, Wei-Liang; Jee, Shiou-Hwa; Dong, Chen-Yuan

    2009-07-01

    We performed multiphoton imaging on fungi of medical significance. Fungal hyphae and spores of Aspergillus flavus, Micosporum gypseum, Micosoprum canis, Trichophyton rubrum, and Trichophyton tonsurans were found to be strongly autofluorescent but generate less prominent second harmonic signal. The cell wall and septum of fungal hyphae can be easily identified by autofluorescence imaging. We found that fungi of various species have distinct autofluorescence characteristics. Our result shows that the combination of multiphoton imaging and spectral analysis can be used to visualize and identify fungal species. This approach may be developed into an effective diagnostic tool for fungal identification.

  10. Nonlinear optical imaging characteristics of colonic adenocarcinoma using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Liu, Nenrong; Chen, Rong; Li, Hongsheng; Chen, Jianxin

    2012-12-01

    Multiphoton microscopy (MPM), a noninvasive optical method with high resolution and high sensitivity, can obtain detailed microstructures of biotissues at submolecular level. In this study, MPM is used to image microstructure varieties of human colonic mucosa and submucosa with adenocarcinoma. Some parameters, such as gland configuration, SHG/TPEF intensity ratio, and collagen orientation and so on, should serve the indicators of early colorectal cancer. The exploratory results show that it's potential for the development of multiphoton mini-endoscopy in real-time early diagnosis of colorectal cancer.

  11. Infrared-active quadruple contrast FePt nanoparticles for multiple scale molecular imaging.

    PubMed

    Chou, Shang-Wei; Liu, Chien-Liang; Liu, Tzu-Ming; Shen, Yu-Fang; Kuo, Lun-Chang; Wu, Cheng-Ham; Hsieh, Tsung-Yuan; Wu, Pei-Chun; Tsai, Ming-Rung; Yang, Che-Chang; Chang, Kai-Yao; Lu, Meng-Hua; Li, Pai-Chi; Chen, Shi-Ping; Wang, Yu-Hsin; Lu, Chen-Wen; Chen, Yi-An; Huang, Chih-Chia; Wang, Churng-Ren Chris; Hsiao, Jong-Kai; Li, Meng-Lin; Chou, Pi-Tai

    2016-04-01

    A single nanomaterial with multiple imaging contrasts and functions is highly desired for multiscale theragnosis. Herein, we demonstrate single 1-1.9 μm infrared-active FePt alloy nanoparticles (FePt NPs) offering unprecedented four-contrast-in-one molecular imaging - computed tomography (CT), magnetic resonance imaging (MRI), photoacoustic (PA) imaging, and high-order multiphoton luminescence (HOMPL) microscopy. The PA response of FePt NPs outperforms that of infrared-active gold nanorods by 3- to 5.6-fold under identical excitation fluence and particle concentrations. HOMPL (680 nm) of an isolated FePt NP renders spatial full-width-at-half-maximum values of 432 nm and 300 nm beyond the optical diffraction limit for 1230-nm and 920-nm excitation, respectively. The in vivo targeting function was successfully visualized using HOMPL, PA imaging, CT, and MRI, thereby validating FePt as a single nanomaterial system covering up to four types (Optical/PA/CT/MRI) of molecular imaging contrast, ranging from the microscopic level to whole-body scale investigation. PMID:26854391

  12. Fine tunable red-green upconversion luminescence from glass ceramic containing 5%Er{sup 3+}:NaYF{sub 4} nanocrystals under excitation of two near infrared femtosecond lasers

    SciTech Connect

    Shang, Xiaoying; Cheng, Wenjing; Zhou, Kan; Ma, Jing; Feng, Donghai; Zhang, Shian; Sun, Zhenrong; Jia, Tianqing; Chen, Ping; Qiu, Jianrong

    2014-08-14

    In this paper, we report fine tunable red-green upconversion luminescence of glass ceramic containing 5%Er{sup 3+}: NaYF{sub 4} nanocrystals excited simultaneously by two near infrared femtosecond lasers. When the glass ceramic was irradiated by 800?nm femtosecond laser, weak red emission centered at 670?nm was detected. Bright red light was observed when the fs laser wavelength was tuned to 1490?nm. However, when excited by the two fs lasers simultaneously, the sample emitted bright green light centered at 550?nm, while the red light kept the same intensity. The dependences of the red and the green light intensities on the two pump lasers are much different, which enables us to manipulate the color emission by adjusting the two pump laser intensities, respectively. We present a theoretical model of Er{sup 3+} ions interacting with two fs laser fields, and explain well the experimental results.

  13. Imaging photoelectron circular dichroism of chiral molecules by femtosecond multiphoton coincidence detection

    NASA Astrophysics Data System (ADS)

    Lehmann, C. Stefan; Ram, N. Bhargava; Powis, Ivan; Janssen, Maurice H. M.

    2013-12-01

    Here, we provide a detailed account of novel experiments employing electron-ion coincidence imaging to discriminate chiral molecules. The full three-dimensional angular scattering distribution of electrons is measured after photoexcitation with either left or right circular polarized light. The experiment is performed using a simplified photoelectron-photoion coincidence imaging setup employing only a single particle imaging detector. Results are reported applying this technique to enantiomers of the chiral molecule camphor after three-photon ionization by circularly polarized femtosecond laser pulses at 400 nm and 380 nm. The electron-ion coincidence imaging provides the photoelectron spectrum of mass-selected ions that are observed in the time-of-flight mass spectra. The coincident photoelectron spectra of the parent camphor ion and the various fragment ions are the same, so it can be concluded that fragmentation of camphor happens after ionization. We discuss the forward-backward asymmetry in the photoelectron angular distribution which is expressed in Legendre polynomials with moments up to order six. Furthermore, we present a method, similar to one-photon electron circular dichroism, to quantify the strength of the chiral electron asymmetry in a single parameter. The circular dichroism in the photoelectron angular distribution of camphor is measured to be 8% at 400 nm. The electron circular dichroism using femtosecond multiphoton excitation is of opposite sign and about 60% larger than the electron dichroism observed before in near-threshold one-photon ionization with synchrotron excitation. We interpret our multiphoton ionization as being resonant at the two-photon level with the 3s and 3p Rydberg states of camphor. Theoretical calculations are presented that model the photoelectron angular distribution from a prealigned camphor molecule using density functional theory and continuum multiple scattering X alpha photoelectron scattering calculations. Qualitative agreement is observed between the experimental results and the theoretical calculations of the Legendre moments representing the angular distribution for the two enantiomers. The electron-ion coincidence technique using multiphoton ionization opens new directions in table-top analytical mass-spectrometric applications of mixtures of chiral molecules.

  14. Imaging photoelectron circular dichroism of chiral molecules by femtosecond multiphoton coincidence detection

    SciTech Connect

    Lehmann, C. Stefan; Ram, N. Bhargava; Janssen, Maurice H. M.; Powis, Ivan

    2013-12-21

    Here, we provide a detailed account of novel experiments employing electron-ion coincidence imaging to discriminate chiral molecules. The full three-dimensional angular scattering distribution of electrons is measured after photoexcitation with either left or right circular polarized light. The experiment is performed using a simplified photoelectron-photoion coincidence imaging setup employing only a single particle imaging detector. Results are reported applying this technique to enantiomers of the chiral molecule camphor after three-photon ionization by circularly polarized femtosecond laser pulses at 400 nm and 380 nm. The electron-ion coincidence imaging provides the photoelectron spectrum of mass-selected ions that are observed in the time-of-flight mass spectra. The coincident photoelectron spectra of the parent camphor ion and the various fragment ions are the same, so it can be concluded that fragmentation of camphor happens after ionization. We discuss the forward-backward asymmetry in the photoelectron angular distribution which is expressed in Legendre polynomials with moments up to order six. Furthermore, we present a method, similar to one-photon electron circular dichroism, to quantify the strength of the chiral electron asymmetry in a single parameter. The circular dichroism in the photoelectron angular distribution of camphor is measured to be 8% at 400 nm. The electron circular dichroism using femtosecond multiphoton excitation is of opposite sign and about 60% larger than the electron dichroism observed before in near-threshold one-photon ionization with synchrotron excitation. We interpret our multiphoton ionization as being resonant at the two-photon level with the 3s and 3p Rydberg states of camphor. Theoretical calculations are presented that model the photoelectron angular distribution from a prealigned camphor molecule using density functional theory and continuum multiple scattering X alpha photoelectron scattering calculations. Qualitative agreement is observed between the experimental results and the theoretical calculations of the Legendre moments representing the angular distribution for the two enantiomers. The electron-ion coincidence technique using multiphoton ionization opens new directions in table-top analytical mass-spectrometric applications of mixtures of chiral molecules.

  15. Imaging photoelectron circular dichroism of chiral molecules by femtosecond multiphoton coincidence detection.

    PubMed

    Lehmann, C Stefan; Ram, N Bhargava; Powis, Ivan; Janssen, Maurice H M

    2013-12-21

    Here, we provide a detailed account of novel experiments employing electron-ion coincidence imaging to discriminate chiral molecules. The full three-dimensional angular scattering distribution of electrons is measured after photoexcitation with either left or right circular polarized light. The experiment is performed using a simplified photoelectron-photoion coincidence imaging setup employing only a single particle imaging detector. Results are reported applying this technique to enantiomers of the chiral molecule camphor after three-photon ionization by circularly polarized femtosecond laser pulses at 400 nm and 380 nm. The electron-ion coincidence imaging provides the photoelectron spectrum of mass-selected ions that are observed in the time-of-flight mass spectra. The coincident photoelectron spectra of the parent camphor ion and the various fragment ions are the same, so it can be concluded that fragmentation of camphor happens after ionization. We discuss the forward-backward asymmetry in the photoelectron angular distribution which is expressed in Legendre polynomials with moments up to order six. Furthermore, we present a method, similar to one-photon electron circular dichroism, to quantify the strength of the chiral electron asymmetry in a single parameter. The circular dichroism in the photoelectron angular distribution of camphor is measured to be 8% at 400 nm. The electron circular dichroism using femtosecond multiphoton excitation is of opposite sign and about 60% larger than the electron dichroism observed before in near-threshold one-photon ionization with synchrotron excitation. We interpret our multiphoton ionization as being resonant at the two-photon level with the 3s and 3p Rydberg states of camphor. Theoretical calculations are presented that model the photoelectron angular distribution from a prealigned camphor molecule using density functional theory and continuum multiple scattering X alpha photoelectron scattering calculations. Qualitative agreement is observed between the experimental results and the theoretical calculations of the Legendre moments representing the angular distribution for the two enantiomers. The electron-ion coincidence technique using multiphoton ionization opens new directions in table-top analytical mass-spectrometric applications of mixtures of chiral molecules. PMID:24359367

  16. Coherent beam control through inhomogeneous media in multi-photon microscopy

    NASA Astrophysics Data System (ADS)

    Paudel, Hari Prasad

    Multi-photon fluorescence microscopy has become a primary tool for high-resolution deep tissue imaging because of its sensitivity to ballistic excitation photons in comparison to scattered excitation photons. The imaging depth of multi-photon microscopes in tissue imaging is limited primarily by background fluorescence that is generated by scattered light due to the random fluctuations in refractive index inside the media, and by reduced intensity in the ballistic focal volume due to aberrations within the tissue and at its interface. We built two multi-photon adaptive optics (AO) correction systems, one for combating scattering and aberration problems, and another for compensating interface aberrations. For scattering correction a MEMS segmented deformable mirror (SDM) was inserted at a plane conjugate to the objective back-pupil plane. The SDM can pre-compensate for light scattering by coherent combination of the scattered light to make an apparent focus even at a depths where negligible ballistic light remains (i.e. ballistic limit). This problem was approached by investigating the spatial and temporal focusing characteristics of a broad-band light source through strongly scattering media. A new model was developed for coherent focus enhancement through or inside the strongly media based on the initial speckle contrast. A layer of fluorescent beads under a mouse skull was imaged using an iterative coherent beam control method in the prototype two-photon microscope to demonstrate the technique. We also adapted an AO correction system to an existing in three-photon microscope in a collaborator lab at Cornell University. In the second AO correction approach a continuous deformable mirror (CDM) is placed at a plane conjugate to the plane of an interface aberration. We demonstrated that this "Conjugate AO" technique yields a large field-of-view (FOV) advantage in comparison to Pupil AO. Further, we showed that the extended FOV in conjugate AO is maintained over a relatively large axial misalignment of the conjugate planes of the CDM and the aberrating interface. This dissertation advances the field of microscopy by providing new models and techniques for imaging deeply within strongly scattering tissue, and by describing new adaptive optics approaches to extending imaging FOV due to sample aberrations.

  17. Intense Visible and Near-Infrared Upconversion Photoluminescence in Colloidal LiYF4:Er3+ Nanocrystals under Excitation at 1490 nm

    PubMed Central

    Chen, Guanying; Ohulchanskyy, Tymish Y.; Kachynski, Aliaksandr; gren, Hans; Prasad, Paras N.

    2012-01-01

    We report intense upconversion photoluminescence (PL) in colloidal LiYF4:Er3+ nanocrystals under excitation with telecom-wavelength at 1490 nm. The intensities of two- and three-photon anti-Stokes upconversion PL bands are higher than or comparable to that of the Stokes emission under excitation with low power density in the range of 5120 W/cm2. The quantum yield of the upconversion PL was measured to be as high as ~1.20.1%, which is almost 4 times higher than the highest upconversion PL quantum yield reported up to date for lanthanide-doped nanocrystals in 100 nm sized hexagonal NaYF4:Yb3+20%, Er3+2% using excitation at ~980 nm. Power dependence study revealed that the intensities of all PL bands have linear dependence on the excitation power density, which was explained by saturation effects in the intermediate energy states. PMID:21557587

  18. A multiphoton microscope platform for imaging the mouse eye

    PubMed Central

    Masihzadeh, Omid; Lei, Tim C.; Ammar, David A.; Kahook, Malik Y.

    2012-01-01

    Purpose To demonstrate the ability of multiphoton microscopy to obtain full three-dimensional high-resolution images of the intact mouse eye anterior chamber without need for enucleation. Methods A custom multiphoton microscope was constructed and optimized for deep tissue imaging. Simultaneous two-photon autofluorescence (2PAF) and second harmonic generation (SHG) imaging were performed. A mouse holder and stereotaxic platform were designed to access different parts of the eye for imaging. A reservoir for keeping the eye moist was used during imaging sessions. Results Non-invasive multiphoton images deep inside the anterior chamber of the mouse eye were obtained without the need for enucleation. The iris, corneal epithelium and endothelium, trabecular meshwork region and conjunctiva were visualized by the 2PAF and SHG signals. Identification of the anatomy was achieved by the intrinsic properties of the native tissue without any exogenous labeling. Images as deep as 600 microns into the eye were clearly demonstrated. Full three-dimensional image reconstructions of the entire anterior chamber were performed and analyzed using custom software. Conclusions Multiphoton imaging is a highly promising tool for ophthalmic research. We have demonstrated the ability to image the entire anterior chamber of the mouse eye in its native state. These results provide a foundation for future in vivo studies of the eye. PMID:22815637

  19. Advances in time-dependent methods for multiphoton processes

    SciTech Connect

    Kulander, K.C.; Schafer, K.J.; Krause, J.L.

    1990-09-01

    This paper discusses recent theoretical results on above threshold ionization harmonic generation and high-frequency, high intensity suppression of ionization. These studies of multiphoton processes in atoms and molecules for short, intense pulsed optical lasers have been carried out using techniques which involve the explicit solution of the time-dependent Schroedinger equation. 43 refs., 5 figs.

  20. Resonance enhanced multiphoton ionization spectra of molecules and molecular fragments. Annual technical report

    SciTech Connect

    1997-07-01

    Resonance Enhanced Multiphoton Ionization (REMPI) utilizes pulsed laser radiation to prepare a molecule in an excited state via absorption of one or more photons and to subsequently ionize that state before it can decay. A remarkable feature of REMPI, and one that is very basic to many of its applications and uses, is that the very narrow bandwidth of the {open_quotes}pump{close_quotes} laser makes it possible to select a specific vibrational and rotational level in the initial state and to prepare the excited state of interest in a single vibrational and rotational level. Thus, by suitable choice of the photon pump transition, it is possible to selectively ionize a species of interest without ionizing any other species that might be present. This feature makes REMPI one of the most powerful tools for ultrasensitive detection of species. With REMPI it is also possible to study the photoionization dynamics of a single rotational level of an excited electronic state. Such state-resolved studies can certainly be expected to provide significant insight into the underlying dynamics of molecular photoionization.

  1. Optical depletion mechanism of upconverting luminescence and its potential for multi-photon STED-like microscopy.

    PubMed

    Wu, Ruitao; Zhan, Qiuqiang; Liu, Haichun; Wen, Xuanyuan; Wang, Baoju; He, Sailing

    2015-12-14

    Simulated emission depletion (STED) microscopy is very powerful, but still suffers from small tissue penetration depth, photobleaching of fluorescent probes and complicated imaging systems. Here, we propose an optical luminescence depletion mechanism employing upconverting nanoparticles (UCNPs) and explore its potential for multi-photon STED-like microscopy. With the addition of Yb3+ ions in NaYF4:Er3+ UCNPs, the two-photon green emission of Er3+ under 795-nm excitation was successfully depleted by 1140-nm laser through the synergetic effect of the excited state absorption and the interionic energy transfer. This STED-like depletion mechanism was systematically investigated using steady-state rate equations, evidenced by the surprising emerging of 478-nm emission. The green emission depletion efficiency was about 30%, limited by the current laser source. Our work indicates that NaYF4:Yb3+/Er3+ UCNPs will be potential probes for multi-photon super-resolution microscopy with many advantages, including long-wavelength-induced large penetration, non-photobleaching and non-photoblinking properties, cost-effective and simplified imaging systems. PMID:26699029

  2. Origin and effect of high-order dispersion in ultrashort pulse multiphoton microscopy in the 10 fs regime.

    PubMed

    Wang, Weichao; Liu, Yujia; Xi, Peng; Ren, Qiushi

    2010-12-10

    Short pulses can induce high nonlinear excitation, and thus they should be favorable for use in multiphoton microscopy. However, the large spectral dispersion can easily destroy the advantages of the ultrashort pulse if there is no compensation. The group delay dispersion (GDD), third-order dispersion, and their effects on the intensity and bandwidth of second-harmonic generation (SHG) signal were analyzed. We found that the prism pair used for compensating the GDD of the two-photon microscope actually introduces significant negative high-order dispersion (HOD), which dramatically narrowed down the two-photon absorption probability for ultrashort pulses. We also investigated the SHG signal after GDD and HOD compensation for different pulse durations. Without HOD compensation, the SHG efficiency dropped significantly for a pulse duration below 20 fs. We experimentally compared the SHG and two-photon excited fluorescence (TPEF) signal intensity for 11 fs versus 50 fs pulses, a pulse duration close to that commonly used in conventional multiphoton microscopy. The result suggested that after adaptive phase compensation, the 11fs pulse can yield a 3.2- to 6.0-fold TPEF intensity and a 5.1-fold SHG intensity, compared to 50 fs pulses. PMID:21151226

  3. Excitation with Effective Subcycle Laser Pulses

    NASA Astrophysics Data System (ADS)

    Marceau, C.; Gingras, G.; Witzel, B.

    2013-11-01

    We have used laser pulses with a temporally shaped polarization to demonstrate the multiphoton excitation of the xenon 5g state within a subcycle of a laser pulse. Our polarization gated laser pulses are composed of circularly polarized sections at the leading and trailing edges of the pulse and of an experimentally defined linearly polarized central part. Only the linear part (the gate) of the pulse can excite neutral xenon in the 5g state. The transition cannot be driven with circularly polarized light because the number of photons needed would cause a violation of selection rules for the change of the magnetic quantum number. We show that the linearly polarized central part can be reduced to a subcycle pulse. This allows us to study excitation with an effective pulse as short as 2.3 fs at 800 nm. Electron imaging spectroscopy has been used to visualize the presence of excited states as a function of the pulse duration of the gate.

  4. Multi-photon ionization of atoms in intense short-wavelength radiation fields

    NASA Astrophysics Data System (ADS)

    Meyer, Michael

    2015-05-01

    The unprecedented characteristics of XUV and X-ray Free Electron Lasers (FELs) have stimulated numerous investigations focusing on the detailed understanding of fundamental photon-matter interactions in atoms and molecules. In particular, the high intensities (up to 106 W/cm2) giving rise to non-linear phenomena in the short wavelength regime. The basic phenomenology involves the production of highly charged ions via electron emission to which both sequential and direct multi-photon absorption processes contribute. The detailed investigation of the role and relative weight of these processes under different conditions (wavelength, pulse duration, intensity) is the key element for a comprehensive understanding of the ionization dynamics. Here the results of recent investigations are presented, performed at the FELs in Hamburg (FLASH) and Trieste (FERMI) on atomic systems with electronic structures of increasing complexity (Ar, Ne and Xe). Mainly, electron spectroscopy is used to obtain quantitative information about the relevance of various multi-photon ionization processes. For the case of Ar, a variety of processes including above threshold ionization (ATI) from 3p and 3s valence shells, direct 2p two-photon ionization and resonant 2p-4p two-photon excitations were observed and their role was quantitatively determined comparing the experimental ionization yields to ab-initio calculations of the cross sections for the multi-photon processes. Using Ar as a benchmark to prove the reliability of the combined experimental and theoretical approach, the more complex and intriguing case of Xe was studied. Especially, the analysis of the two-photon ATI from the Xe 4d shell reveals new insight into the character of the 4d giant resonance, which was unresolved in the linear one-photon regime. Finally, the influence of intense XUV radiation to the relaxation dynamics of the Ne 2s-3p resonance was investigated by angle-resolved electron spectroscopy, especially be observing the intensity dependent variation of the angular distribution patterns for the sequential ionization process.

  5. Insights on the CN B 2Σ+ + Ar potential from ultraviolet fluorescence excitation and infrared depletion studies of the CN-Ar complex

    NASA Astrophysics Data System (ADS)

    O'Donnell, Bridget A.; Beames, Joseph M.; Lester, Marsha I.

    2012-06-01

    UV laser-induced fluorescence and IR-UV fluorescence depletion studies have been used to characterize the intermolecular levels of the CN-Ar complex in the excited state correlating with CN B 2Σ+ + Ar. Additional CN-Ar features are identified to lower wavenumber than reported previously. Fluorescence depletion spectra are recorded to confirm that these CN-Ar features and other higher energy features in the B-X spectrum originate from a common ground state level. The UV depletion is induced by IR excitation of CN-Ar from the ground state zero-point level to a hindered internal rotor state (nK = 11) in the CN overtone region. The lowest energy feature in the B-X spectrum at 25 714.1 cm-1 is assigned as a transition to the zero-point level of the B state and also yields its binding energy, D0 = 186(2) cm-1, which is in excellent accord with theoretical predictions. The next feature approximately 40 cm-1 higher is attributed to overlapping transitions to intermolecular levels with bend ({v}_bK = 1^1) or stretch (vs = 1) excitation. Yet higher features (previously reported) are also assigned, based on their transition type and wavenumber, which are consistent with the intermolecular energy level pattern computed theoretically. Finally, the intensity profile of the lowest energy features in the B-X spectrum reflects the predicted change in the CN (B 2Σ+, X 2Σ+) + Ar potentials upon electronic excitation from a weakly anisotropic potential about the linear N≡C-Ar configuration in the ground state to a more strongly bound linear C≡N-Ar structure in the excited B electronic state.

  6. Novel techniques with multiphoton microscopy: Deep-brain imaging with microprisms, neurometabolism of epilepsy, and counterfeit paper money detection

    NASA Astrophysics Data System (ADS)

    Chia, Thomas H.

    Multiphoton microscopy is a laser-scanning fluorescence imaging method with extraordinary potential. We describe three innovative multiphoton microscopy techniques across various disciplines. Traditional in vivo fluorescence microscopy of the mammalian brain has a limited penetration depth (<400 microm). We present a method of imaging 1 mm deep into mouse neocortex by using a glass microprism to relay the excitation and emission light. This technique enables simultaneous imaging of multiple cortical layers, including layer V, at an angle typical of slice preparations. At high-magnification imaging using an objective with 1-mm of coverglass correction, resolution was sufficient to resolve dendritic spines on layer V GFP neurons. Functional imaging of blood flow at various neocortical depths is also presented, allowing for quantification of red blood cell flux and velocity. Multiphoton fluorescence lifetime imaging (FLIM) of NADH reveals information on neurometabolism. NADH, an intrinsic fluorescent molecule and ubiquitous metabolic coenzyme, has a lifetime dependent on enzymatic binding. A novel NADH FLIM algorithm is presented that produces images showing spatially distinct NADH fluorescence lifetimes in mammalian brain slices. This program provides advantages over traditional FLIM processing of multi-component lifetime data. We applied this technique to a GFP-GFAP pilocarpine mouse model of temporal lobe epilepsy. Results indicated significant changes in the neurometabolism of astrocytes and neuropil in the cell and dendritic layers of the hippocampus when compared to control tissue. Data obtained with NADH FLIM were subsequently interpreted based on the abnormal activity reported in epileptic tissue. Genuine U.S. Federal Reserve Notes have a consistent, two-component intrinsic fluorescence lifetime. This allows for detection of counterfeit paper money because of its significant differences in fluorescence lifetime when compared to genuine paper money. We used scanning multiphoton laser excitation to sample a 4 mm2 region from 54 genuine Reserve Notes. Three types of counterfeit samples were tested. Four out of the nine counterfeit samples fit to a one-component decay. Five out of nine counterfeit samples fit to a two-component model, but are identified as counterfeit due to significant deviations in the longer lifetime component compared to genuine bills.

  7. Coherent Control of Multiphoton Transitions in the Gas and Condensed Phases with Shaped Ultrashort Pulses

    SciTech Connect

    Marcos Dantus

    2008-09-23

    Controlling laser-molecule interactions has become an integral part of developing devices and applications in spectroscopy, microscopy, optical switching, micromachining and photochemistry. Coherent control of multiphoton transitions could bring a significant improvement of these methods. In microscopy, multi-photon transitions are used to activate different contrast agents and suppress background fluorescence; coherent control could generate selective probe excitation. In photochemistry, different dissociative states are accessed through two, three, or more photon transitions; coherent control could be used to select the reaction pathway and therefore the yield-specific products. For micromachining and processing a wide variety of materials, femtosecond lasers are now used routinely. Understanding the interactions between the intense femtosecond pulse and the material could lead to technologically important advances. Pulse shaping could then be used to optimize the desired outcome. The scope of our research program is to develop robust and efficient strategies to control nonlinear laser-matter interactions using ultrashort shaped pulses in gas and condensed phases. Our systematic research has led to significant developments in a number of areas relevant to the AMO Physics group at DOE, among them: generation of ultrashort phase shaped pulses, coherent control and manipulation of quantum mechanical states in gas and condensed phases, behavior of isolated molecules under intense laser fields, behavior of condensed phase matter under intense laser field and implications on micromachining with ultrashort pulses, coherent control of nanoparticles their surface plasmon waves and their nonlinear optical behavior, and observation of coherent Coulomb explosion processes at 10^16 W/cm^2. In all, the research has resulted in 36 publications (five journal covers) and nine invention disclosures, five of which have continued on to patenting

  8. Yb(3+)-doped GeS(2)-Ga2S(3)-CsCl glass with broad and adjustable absorption/excitation band for near-infrared luminescence.

    PubMed

    Fan, Bo; Xue, Bai; Zhang, Xianghua; Adam, Jean-Luc; Fan, Xianping

    2013-07-01

    The luminescent property of Yb(3+) ions in GeS(2)-Ga(2)S(3)-CsCl glasses with different CsCl contents has been studied. All the samples demonstrate a broad excitation band in the UV or/and visible range, depending on the composition, which is attributed to the charge transfer of the Yb(3+)-S(2-)/Cl(-) couple. The width of the excitation/absorption band can be as large as 150 nm. Moreover, with the increase of CsCl content, the peak position of the band can be continuously adjusted from 458 to 380 nm, due to the increase of the local average electronegativity around Yb(3+) ions. The broad and adjustable excitation band makes the Yb(3+)doped GeS(2)-Ga(2)S(3)-CsCl glass interesting for modifying the solar spectrum by absorbing strongly in the UV/blue region for emission around 1 ?m. This kind of material is the key to adapting the solar spectrum to the response of silicon photovoltaic solar cells. PMID:23811902

  9. Hybrid label-free multiphoton and optoacoustic microscopy (MPOM)

    NASA Astrophysics Data System (ADS)

    Soliman, Dominik; Tserevelakis, George J.; Omar, Murad; Ntziachristos, Vasilis

    2015-07-01

    Many biological applications require a simultaneous observation of different anatomical features. However, unless potentially harmful staining of the specimens is employed, individual microscopy techniques do generally not provide multi-contrast capabilities. We present a hybrid microscope integrating optoacoustic microscopy and multiphoton microscopy, including second-harmonic generation, into a single device. This combined multiphoton and optoacoustic microscope (MPOM) offers visualization of a broad range of structures by employing different contrast mechanisms and at the same time enables pure label-free imaging of biological systems. We investigate the relative performance of the two microscopy modalities and demonstrate their multi-contrast abilities through the label-free imaging of a zebrafish larva ex vivo, simultaneously visualizing muscles and pigments. This hybrid microscopy application bears great potential for developmental biology studies, enabling more comprehensive information to be obtained from biological specimens without the necessity of staining.

  10. The role of resonances in strong-field multiphoton processes

    SciTech Connect

    Perry, M.D.; Kulander, K.C.

    1990-10-01

    Resonantly-enhanced multiphoton ionization (REMPI) has been the subject of extensive experimental and theoretical study since the invention of the laser. Until recently, the overwhelming majority of REMPI research have been conducted at intensities less than 10{sup 12} W/cm{sup 2}. At these intensities, the strength of the applied field remains less than one percent of the atomic Coulomb field experienced by the outer electrons in a typical noble gas atom. In this regime, treatment of the applied field as a weak perturbation on the atomic system yields excellent agreement with experiment. Here, we investigate the role of resonances in multiphoton ionization at much higher intensities, specifically, we examine the behavior and influence of resonances as the strength of the applied field becomes a significant fraction of the atomic field. 33 refs., 7 figs., 2 tabs.

  11. Multichannel Multiphoton Imaging of Metal Oxides Nanoparticles in Biological System

    SciTech Connect

    Zheng, Yuangang; Holtom, Gary R.; Colson, Steve D.; Periasamy, A. and So, S.T.C.

    2004-09-15

    Near-IR ultrafast pulse laser and confocal microscope are combined to create a multiphoton multichannel non-linear imaging technique, which allows in situ 3-D characterization of nonfluorescent nanoparticles in biological systems. We observed intense CARS signals generated from various metal oxides due to their high third-order nonlinear susceptibilities (x(3)), which do not depend on the vibrational resonance but on the electronic resonance. We show that fine and ultrafine particles of metal oxides in alveolar macrophage cells may be imaged in vitro using CARS and multiphoton fluorescence microscopy with highest optical resolution for extended periods without photobleaching effects. The advantage of the epidetection over the forward detection for imaging sub-micron particles has been investigated.

  12. Characteristics of subgingival calculus detection by multiphoton fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Tung, Oi-Hong; Lee, Shyh-Yuan; Lai, Yu-Lin; Chen, How-Foo

    2011-06-01

    Subgingival calculus has been recognized as a major cause of periodontitis, which is one of the main chronic infectious diseases of oral cavities and a principal cause of tooth loss in humans. Bacteria deposited in subgingival calculus or plaque cause gingival inflammation, function deterioration, and then periodontitis. However, subgingival calculus within the periodontal pocket is a complicated and potentially delicate structure to be detected with current dental armamentaria, namely dental x-rays and dental probes. Consequently, complete removal of subgingival calculus remains a challenge to periodontal therapies. In this study, the detection of subgingival calculus employing a multiphoton autofluorescence imaging method was characterized in comparison with a one-photon confocal fluorescence imaging technique. Feasibility of such a system was studied based on fluorescence response of gingiva, healthy teeth, and calculus with and without gingiva covered. The multiphoton fluorescence technology perceived the tissue-covered subgingival calculus that cannot be observed by the one-photon confocal fluorescence method.

  13. Multi-photon intracellular sodium imaging combined with UV-mediated focal uncaging of glutamate in CA1 pyramidal neurons.

    PubMed

    Kleinhans, Christian; Kafitz, Karl W; Rose, Christine R

    2014-01-01

    Multi-photon fluorescence microscopy has enabled the analysis of morphological and physiological parameters of brain cells in the intact tissue with high spatial and temporal resolution. Combined with electrophysiology, it is widely used to study activity-related calcium signals in small subcellular compartments such as dendrites and dendritic spines. In addition to calcium transients, synaptic activity also induces postsynaptic sodium signals, the properties of which are only marginally understood. Here, we describe a method for combined whole-cell patch-clamp and multi-photon sodium imaging in cellular micro domains of central neurons. Furthermore, we introduce a modified procedure for ultra-violet (UV)-light-induced uncaging of glutamate, which allows reliable and focal activation of glutamate receptors in the tissue. To this end, whole-cell recordings were performed on Cornu Ammonis subdivision 1 (CA1) pyramidal neurons in acute tissue slices of the mouse hippocampus. Neurons were filled with the sodium-sensitive fluorescent dye SBFI through the patch-pipette, and multi-photon excitation of SBFI enabled the visualization of dendrites and adjacent spines. To establish UV-induced focal uncaging, several parameters including light intensity, volume affected by the UV uncaging beam, positioning of the beam as well as concentration of the caged compound were tested and optimized. Our results show that local perfusion with caged glutamate (MNI-Glutamate) and its focal UV-uncaging result in inward currents and sodium transients in dendrites and spines. Time course and amplitude of both inward currents and sodium signals correlate with the duration of the uncaging pulse. Furthermore, our results show that intracellular sodium signals are blocked in the presence of blockers for ionotropic glutamate receptors, demonstrating that they are mediated by sodium influx though this pathway. In summary, our method provides a reliable tool for the investigation of intracellular sodium signals induced by focal receptor activation in intact brain tissue. PMID:25350367

  14. Enhancing collection efficiency in large field of view multiphoton microscopy.

    PubMed

    McMullen, J D; Kwan, A C; Williams, R M; Zipfel, W R

    2011-02-01

    Many multiphoton imaging applications would benefit from a larger field of view; however, large field of views (>mm) require low magnification objectives which have low light collection efficiencies. We demonstrate a light collection system mounted on a low magnification objective that increases fluorescence collection by as much as 20-fold in scattering tissues. This peripheral detector results in an effective numerical aperture of collection >0.8 with a 3-4 mm field of view. PMID:21118215

  15. Vectorizable wave propagation FORTRANcode for calculations of multiphoton dissociation

    NASA Astrophysics Data System (ADS)

    Ting, Julian J.-L.; Yuan, J. M.; Jiang, T.-F.

    1992-06-01

    A vectorizable FORTRAN code for the ETA10 or other vector machines for numerical calculation of the time-dependent Schrdinger equation is presented. Preliminary results for the multiphoton dissociation of nitrogen oxide are shown. The applied electric field was approximated by a classical sinusoidal wave; both sinusoidal and cosinusoidal wave forms were tested, and the internuclear potential was taken to be a Morse function. Dissociation histories of three types were found.

  16. All-optical bidirectional neural interfacing using hybrid multiphoton holographic optogenetic stimulation.

    PubMed

    Paluch-Siegler, Shir; Mayblum, Tom; Dana, Hod; Brosh, Inbar; Gefen, Inna; Shoham, Shy

    2015-07-01

    Our understanding of neural information processing could potentially be advanced by combining flexible three-dimensional (3-D) neuroimaging and stimulation. Recent developments in optogenetics suggest that neurophotonic approaches are in principle highly suited for noncontact stimulation of network activity patterns. In particular, two-photon holographic optical neural stimulation (2P-HONS) has emerged as a leading approach for multisite 3-D excitation, and combining it with temporal focusing (TF) further enables axially confined yet spatially extended light patterns. Here, we study key steps toward bidirectional cell-targeted 3-D interfacing by introducing and testing a hybrid new 2P-TF-HONS stimulation path for accurate parallel optogenetic excitation into a recently developed hybrid multiphoton 3-D imaging system. The system is shown to allow targeted all-optical probing of in vitro cortical networks expressing channelrhodopsin-2 using a regeneratively amplified femtosecond laser source tuned to 905nm. These developments further advance a prospective new tool for studying and achieving distributed control over 3-D neuronal circuits both in vitro and in vivo. PMID:26217673

  17. Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

    PubMed Central

    Cha, Jae Won; Yew, Elijah Y. S.; Kim, Daekeun; Subramanian, Jaichandar; Nedivi, Elly; So, Peter T. C.

    2015-01-01

    Multifocal multiphoton microscopy (MMM) improves imaging speed over a point scanning approach by parallelizing the excitation process. Early versions of MMM relied on imaging detectors to record emission signals from multiple foci simultaneously. For many turbid biological specimens, the scattering of emission photons results in blurred images and degrades the signal-to-noise ratio (SNR). We have recently demonstrated that a multianode photomultiplier tube (MAPMT) placed in a descanned configuration can effectively collect scattered emission photons from each focus into their corresponding anodes significantly improving image SNR for highly scattering specimens. Unfortunately, a descanned MMM has a longer detection path resulting in substantial emission photon loss. Optical design constraints in a descanned geometry further results in significant optical aberrations especially for large field-of-view (FOV), high NA objectives. Here, we introduce a non-descanned MMM based on MAPMT that substantially overcomes most of these drawbacks. We show that we improve signal efficiency up to fourfold with limited image SNR degradation due to scattered emission photons. The excitation foci can also be spaced wider to cover the full FOV of the objective with minimal aberrations. The performance of this system is demonstrated by imaging interneuron morphological structures deep in the brains of living mice. PMID:25874160

  18. Non-descanned multifocal multiphoton microscopy with a multianode photomultiplier tube

    NASA Astrophysics Data System (ADS)

    Cha, Jae Won; Yew, Elijah Y. S.; Kim, Daekeun; Subramanian, Jaichandar; Nedivi, Elly; So, Peter T. C.

    2015-08-01

    Multifocal multiphoton microscopy (MMM) improves imaging speed over a point scanning approach by parallelizing the excitation process. Early versions of MMM relied on imaging detectors to record emission signals from multiple foci simultaneously. For many turbid biological specimens, the scattering of emission photons results in blurred images and degrades the signal-to-noise ratio (SNR). We have recently demonstrated that a multianode photomultiplier tube (MAPMT) placed in a descanned configuration can effectively collect scattered emission photons from each focus into their corresponding anodes significantly improving image SNR for highly scattering specimens. Unfortunately, a descanned MMM has a longer detection path resulting in substantial emission photon loss. Optical design constraints in a descanned geometry further results in significant optical aberrations especially for large field-of-view (FOV), high NA objectives. Here, we introduce a non-descanned MMM based on MAPMT that substantially overcomes most of these drawbacks. We show that we improve signal efficiency up to fourfold with limited image SNR degradation due to scattered emission photons. The excitation foci can also be spaced wider to cover the full FOV of the objective with minimal aberrations. The performance of this system is demonstrated by imaging interneuron morphological structures deep in the brains of living mice.

  19. Multiphoton ionization mass spectrometry of nitrated polycyclic aromatic hydrocarbons.

    PubMed

    Tang, Yuanyuan; Imasaka, Tomoko; Yamamoto, Shigekazu; Imasaka, Totaro

    2015-08-01

    In order to suppress the fragmentation and improve the sensitivity for determination of nitrated polycyclic aromatic hydrocarbons (NPAHs), the mechanism of multiphoton ionization was studied for the following representative NPAHs, 9-nitroanthracene, 3-nitrofluoranthene, and 1-nitropyrene. The analytes were extracted from the PM2.5 on the sampling filter ultrasonically, and were measured using gas chromatography/multiphoton ionization/time-of-flight mass spectrometry with a femtosecond tunable laser in the range from 267 to 405 nm. As a result, a molecular ion was observed as the major ion and fragmentation was suppressed at wavelengths longer than 345 nm. Furthermore, the detection limit measured at 345 nm was measured to be the subpicogram level. The organic compounds were extracted from a 2.19 mg sample of particulate matter 2.5 (PM2.5), and the extract was subjected to multiphoton ionization mass spectrometry after gas chromatograph separation. The background signals were drastically suppressed at 345 nm, and the target NPAHs, including 9-nitroanthracene and 1-nitropyrene, were detected, and their concentrations were determined to be 5 and 3 pg/m(3), respectively. PMID:26048831

  20. Mid-J CO shock tracing observations of infrared dark clouds. II. Low-J CO constraints on excitation, depletion, and kinematics

    NASA Astrophysics Data System (ADS)

    Pon, A.; Johnstone, D.; Caselli, P.; Fontani, F.; Palau, A.; Butler, M. J.; Kaufman, M.; Jiménez-Serra, I.; Tan, J. C.

    2016-03-01

    Infrared dark clouds are kinematically complex molecular structures in the interstellar medium that can host sites of massive star formation. We present maps measuring 4 square arcminutes of the 12CO, 13CO, and C18O J = 3 to 2 lines from selected locations within the C and F (G028.37+00.07 and G034.43+00.24) infrared dark clouds (IRDCs), as well as single pointing observations of the 13CO and C18O J = 2 to 1 lines towards three cores within these clouds. We derive CO gas temperatures throughout the maps and find that CO is significantly frozen out within these IRDCs. We find that the CO depletion tends to be the highest near column density peaks with maximum depletion factors between 5 and 9 in IRDC F and between 16 and 31 in IRDC C. We also detect multiple velocity components and complex kinematic structure in both IRDCs. Therefore, the kinematics of IRDCs seem to point to dynamically evolving structures yielding dense cores with considerable depletion factors. Based on observations carried out with the JCMT and IRAM 30 m Telescopes. IRAM is supported by INSU/CNRS (France), MPG (Germany) and IGN (Spain).IRAM CO observations are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/587/A96

  1. Study of the infrared emission from the O I-smcaps (3d TD) and (4s VS) states produced by electron impact excitation of O2

    SciTech Connect

    Erdman, P.W.; Zipf, E.C.

    1987-09-15

    We have measured the absolute emission cross section value for the O I(3p TP--3d TD, lambda = 11 287 A) and the (3p VP--4s VS, lambda = 11 299 A) multiplets excited by electron impact on O2. The former cross section has a value of 3.05 x 10 Z cmS +- 15% at 100 eV, while the latter's value is 4.14 x 10 Z cmS +- 15%. The quintet emission at lambda = 11 299 A is the only deexcitation channel for the 4s VS state, and, therefore, represents the total excitation cross section for that state. However, the lambda = 11 287 A branch from the O I(3d TD) state only constitutes --25% of the total 3d TD cross section, with the major branch from this state being the transition to the ground state observed at lambda = 1027 A. The branching ratio obtained in this study is in good agreement with the theoretical calculation of Pradhan and Saraph (omega-tilde/sub 11//sub //sub 287/ = 0.28), but our absolute sigma(lambda = 11 287 A) value is substantially smaller than the result reported recently by Schulman et al.

  2. Subcycle dynamics of high harmonic generation in valence-shell and virtual states of Ar atoms excited by attosecond pulses and driven by near-infrared laser fields: A self-interaction-free TDDFT theoretical approach

    NASA Astrophysics Data System (ADS)

    Heslar, John; Telnov, Dmitry A.; Chu, Shih-I.

    2015-05-01

    In the framework of the self-interaction-free time-dependent density functional theory (TDDFT), we have performed an ab initio all-electron study of subcycle structure, dynamics, and spectra of high harmonic generation (HHG) processes of Ar atoms in the presence of extreme ultraviolet (XUV) attosecond pulses and near-infrared (NIR) laser fields. The TDDFT equations are solved accurately and efficiently via the time-dependent generalized pseudospectral (TDGPS) method. We focus on the subcycle (with respect to NIR field) temporal behavior of the level shift of the excited energy levels and related dynamics of harmonic photon emission. We observe and identify the subcycle shifts in the harmonic emission spectrum as a function of the time delay between the XUV and NIR pulses. We present and analyze the harmonic emission spectra from 3snp0, 3p0ns, 3p1nd1,3p1np1, 3p0nd0, 3p0np0, and 3p0ns excited states and the 3p04p0-virtual state as functions of the time delay. In addition, we explore the subcycle a.c. Stark shift phenomenon in NIR fields and its influence on the harmonic emission process. Our analysis reveals several novel features of the subcycle HHG dynamics and spectra as well as temporal energy level shift. This work was partially supported by DOE.

  3. A study of the infrared emission from the O I (3d 3D0) and (4s 5S0) states produced by electron impact excitation of O2

    NASA Technical Reports Server (NTRS)

    Erdman, P. W.; Zipf, E. C.

    1987-01-01

    The absolute emission cross section values for the O I(3p 3P-3d 3D0, lambda = 11,287 A) and the (3p 5P-4s 5S0, lambda = 11,299 A) multiplets excited by electron impact on O2 have been measured. These infrared oxygen emission features appear prominently in auroral spectra and were observed in electron bombardment of O2. The former cross section has a value of 3.05 x 10 to the -19th + or - 15 percent at 100 eV, while the latter's value is 4.14 x 10 to the -19th sq cm + or - 15 percent. The quintet emission at lambda = 11,299 A is the only deexcitation channel for the 4s 5S0 state and thus represents the total excitation cross section for that state. However, the lambda = 11,287 A branch from the O I (3d 3D0) state only constitutes about 25 percent of the total 3d 3D0 cross section, with the major branch from this state being the transition to the ground state observed at lambda = 1027 A. The branching ratio is in good agreement with the theoretical calculation of Padhan and Saraph (1977).

  4. Quenching nitrogen-vacancy center photoluminescence with an infrared pulsed laser

    NASA Astrophysics Data System (ADS)

    Lai, N. D.; Faklaris, O.; Zheng, D.; Jacques, V.; Chang, H.-C.; Roch, J.-F.; Treussart, F.

    2013-03-01

    Diamond nanocrystals containing nitrogen-vacancy (NV) color centers have been used in recent years as fluorescent probes for near-field and cellular imaging. In this work, we report that an infrared (IR) pulsed excitation beam can quench the photoluminescence of a NV color center in a diamond nanocrystal (size <50 nm) with an extinction ratio as high as ?90%. We attribute this effect to the heating of the nanocrystal consecutive to multi-photon absorption by the diamond matrix. This quenching is reversible: the photoluminescence intensity goes back to its original value when the IR laser beam is turned off, with a typical response time of 100 ps, allowing for fast control of NV color center photoluminescence. We used this effect to achieve the sub-diffraction-limited imaging of fluorescent diamond nanocrystals on a coverglass. For that, as in the ground state depletion super-resolution technique, we combined the green excitation laser beam with the control IR depleting one after shaping its intensity profile in a doughnut form, so that the emission comes only from the sub-wavelength size central part.

  5. Multiphoton microscopy of engineered dermal substitutes: assessment of 3-D collagen matrix remodeling induced by fibroblast contraction.

    PubMed

    Pena, Ana-Maria; Fagot, Dominique; Olive, Christian; Michelet, Jean-Franois; Galey, Jean-Baptiste; Leroy, Frdric; Beaurepaire, Emmanuel; Martin, Jean-Louis; Colonna, Anne; Schanne-Klein, Marie-Claire

    2010-01-01

    Dermal fibroblasts are responsible for the generation of mechanical forces within their surrounding extracellular matrix and can be potentially targeted by anti-aging ingredients. Investigation of the modulation of fibroblast contraction by these ingredients requires the implementation of three-dimensional in situ imaging methodologies. We use multiphoton microscopy to visualize unstained engineered dermal tissue by combining second-harmonic generation that reveals specifically fibrillar collagen and two-photon excited fluorescence from endogenous cellular chromophores. We study the fibroblast-induced reorganization of the collagen matrix and quantitatively evaluate the effect of Y-27632, a RhoA-kinase inhibitor, on dermal substitute contraction. We observe that collagen fibrils rearrange around fibroblasts with increasing density in control samples, whereas collagen fibrils show no remodeling in the samples containing the RhoA-kinase inhibitor. Moreover, we show that the inhibitory effects are reversible. Our study demonstrates the relevance of multiphoton microscopy to visualize three-dimensional remodeling of the extracellular matrix induced by fibroblast contraction or other processes. PMID:21054112

  6. (1+1) Resonance-enhanced multiphoton ionization and photodissociation study of CS2 via the 1B2 state.

    PubMed

    Hu, Zhengfa; Lee, Wei-Bin; Zhang, Xiao-Peng; Wei, Pei-Ying; Lin, King-Chuen

    2008-02-22

    (1+1) Resonance-enhanced multiphoton ionization (REMPI) spectra of CS(2) and molecular dissociation dynamics are investigated using a time-of-flight mass spectrometer equipped with velocity imaging detection. The REMPI spectra via a linear-bent 1Sigma(g)+-->(1)B(2)(1Sigma(u)+) transition are acquired in the wavelength range of 208-217 nm. Each ro-vibrational band profile of the (1)B(2)(1Sigma(u)+) state is deconvoluted to yield the corresponding predissociative lifetime from 0.3 to 3 ps. Upon excitation at 210.25 and 212.54 nm, the resulting images of S(+) and CS(+) fragments are analyzed to give individual translational energy distributions, which are resolved into two components corresponding to the CS+S((3)P) and CS+S((1)D) channels. The product branching ratios of S((3)P)/S((1)D) are evaluated to be 5.7+/-1.0 and 9.6+/-2.5 at 210.25 and 212.54 nm, respectively. Despite the difficulty avoiding the effect of multiphoton absorption, the molecular dissociation channel is verified to prevail over the dissociative ionization channel of CS(2). The anisotropy parameters for the triplet and singlet channels are determined to be approximately 0.8 and 1.1-1.3, respectively, suggesting that the predissociative state should have a bent configuration with a short lifetime. PMID:18236487

  7. Multiphoton and magnetic resonance imaging of Barley embryos: comparing micro-imaging techniques across scale and parameter barriers

    NASA Astrophysics Data System (ADS)

    Stark, Martin; Manz, Bertram; Riemann, Iris; Volke, Frank; Weschke, Winfriede; König, Karsten

    2007-02-01

    Multiphoton stimulated autofluorescence microscopy and Magnetic resonance imaging (MRI) address different molecular properties of the sample and reach to a different length scale. MRI maps density or mobility of nuclei (here: hydrogen), and targets at whole objects from the scale of sub-millimetres to meters. Multiphoton imaging profits from the nonlinear absorption of light in the focus of a femtosecond laser source stimulating the autofluorescence of biomolecules. As this effect relies on a high light intensity the accessible field of view is limited, but the resolution is very high. Studying a plant embryo (barley) we compare the two techniques. At 770 nm excitation the cell walls of the embryo exhibited significant autofluorescence, allowing for a subcellular resolution. While details where imaged with an objective of N.A. 1.3, an overview was generated with a N.A. as low as 0.25. The overview image as well as merged images and tomographical data were used to link the high-resolution optical data with the three-dimensional highresolution MR images. There, images of the proton density were acquired using a standard 3D spin-echo imaging pulse sequence. While the optical high-resolution data provides a field of view restricted to only a small part of the embryo, the MR image contains the whole grain. Bridging the scales it might be possible to trace transport of e.g. nutrients from large structure of the plant to the cellular level.

  8. Multiphoton microscopy of engineered dermal substitutes: assessment of 3-D collagen matrix remodeling induced by fibroblast contraction

    NASA Astrophysics Data System (ADS)

    Pena, Ana-Maria; Fagot, Dominique; Olive, Christian; Michelet, Jean-Franois; Galey, Jean-Baptiste; Leroy, Frdric; Beaurepaire, Emmanuel; Martin, Jean-Louis; Colonna, Anne; Schanne-Klein, Marie-Claire

    2010-09-01

    Dermal fibroblasts are responsible for the generation of mechanical forces within their surrounding extracellular matrix and can be potentially targeted by anti-aging ingredients. Investigation of the modulation of fibroblast contraction by these ingredients requires the implementation of three-dimensional in situ imaging methodologies. We use multiphoton microscopy to visualize unstained engineered dermal tissue by combining second-harmonic generation that reveals specifically fibrillar collagen and two-photon excited fluorescence from endogenous cellular chromophores. We study the fibroblast-induced reorganization of the collagen matrix and quantitatively evaluate the effect of Y-27632, a RhoA-kinase inhibitor, on dermal substitute contraction. We observe that collagen fibrils rearrange around fibroblasts with increasing density in control samples, whereas collagen fibrils show no remodeling in the samples containing the RhoA-kinase inhibitor. Moreover, we show that the inhibitory effects are reversible. Our study demonstrates the relevance of multiphoton microscopy to visualize three-dimensional remodeling of the extracellular matrix induced by fibroblast contraction or other processes.

  9. The near-infrared structure and spectra of the bipolar nebulae M2-9 and AFGL 2688: The role of ultraviolet pumping and shocks in molecular hydrogen excitation

    NASA Technical Reports Server (NTRS)

    Hora, Joseph L.; Latter, William B.

    1994-01-01

    High-resolution near-infrared images and moderate resolution spectra were obtained of the bipolar nebulae M2-9 and AFGL 2688. The ability to spatially and spectrally resolve the various components of the nebulae has proved to be important in determining their physical structure and characteristics. In M2-9, the lobes are found to have a double-shell structure. The inner shell is dominated by emission from hydrogen recombination lines, and the outer shell is primarily emission from H2 lines in teh 2-2.5 micron region. Analysis of H2 line ratios indicates that the H2 emission is radiatively excited. A well-resolved photodissociation region is observed in the lobes. The spectrum of the central source is dominated by H recombination lines and a strong continuum rising toward longer wavelengths consistent with a T = 795 K blackbody. Also present are lines of He I and Fe II. In contrast, the N knot and E lobe of M2-9 show little continuum emission. The N knot spectrum consists of lines of (Fe II) and hydrogen recombination lines. In AGFL 2688, the emission from the bright lobes is mainly continuum reflected from the central star. Several molecular features from C2 and CN are present. In the extreme end of the N lobe and in the E equatorial region, the emission is dominated by lines of H2 in the 2-2.5 region. The observed H2 line ratios indicate that the emission is collisionally excited, with an excitation temperature T(sub ex) approixmately = 1600 +/- 100 K.

  10. Further observations of rotationally excited far infrared OH16 and OH18 emission in Orion-KL: Tighter constraints on the nature of the emitting region

    NASA Technical Reports Server (NTRS)

    Melnick, G. J.; Stacey, G. J.; Genzel, R.; Lugten, J. B.; Poglitsch, A.

    1989-01-01

    The Orion-KL region, within 1 arc minute, is observed. The rotational cross ladder (53.351 microns) and rotational ground state (120.1719 microns) transitions are studied. It is shown that these lines exhibit a P-Cygni profile and unambiguously show that the OH gas is expanding out from the central BN-KL infrared cluster. The OH-16 rotational ground state transition (119.234 microns) is velocity resolved and it is found that its intrinsic full width at half maximum is 75 km/s. The line fluxes and line profiles are modeled and it is shown that no single temperature and density component can reproduce the data. Rather, the best fit to the data requires emission from three main components of the gas: post shocked gas with the profiles of temperature, density, and OH abundance; a high density component to the cool post shocked region; and the plateau region.

  11. Silver nanoparticle based surface enhanced Raman scattering spectroscopy of diabetic and normal rat pancreatic tissue under near-infrared laser excitation

    NASA Astrophysics Data System (ADS)

    Huang, H.; Shi, H.; Feng, S.; Lin, J.; Chen, W.; Huang, Z.; Li, Y.; Yu, Y.; Lin, D.; Xu, Q.; Chen, R.

    2013-04-01

    This paper presents the use of high spatial resolution silver nanoparticle based near-infrared surface enhanced Raman scattering (SERS) from rat pancreatic tissue to obtain biochrmical information about the tissue. A high quality SERS signal from a mixture of pancreatic tissues and silver nanoparticles can be obtained within 10 s using a Renishaw micro-Raman system. Prominent SERS bands of pancreatic tissue were assigned to known molecular vibrations, such as the vibrations of DNA bases, RNA bases, proteins and lipids. Different tissue structures of diabetic and normal rat pancreatic tissues have characteristic features in SERS spectra. This exploratory study demonstrated great potential for using SERS imaging to distinguish diabetic and normal pancreatic tissues on frozen sections without using dye labeling of functionalized binding sites.

  12. Cryogenic exciter

    DOEpatents

    Bray, James William; Garces, Luis Jose

    2012-03-13

    The disclosed technology is a cryogenic static exciter. The cryogenic static exciter is connected to a synchronous electric machine that has a field winding. The synchronous electric machine is cooled via a refrigerator or cryogen like liquid nitrogen. The static exciter is in communication with the field winding and is operating at ambient temperature. The static exciter receives cooling from a refrigerator or cryogen source, which may also service the synchronous machine, to selected areas of the static exciter and the cooling selectively reduces the operating temperature of the selected areas of the static exciter.

  13. Comparing the dynamical effects of symmetric and antisymmetric stretch excitation of methane in the Cl+CH4 reaction

    NASA Astrophysics Data System (ADS)

    Bechtel, Hans A.; Camden, Jon P.; Brown, Davida J. Ankeny; Zare, Richard N.

    2004-03-01

    The effects of two nearly isoenergetic C-H stretching motions on the gas-phase reaction of atomic chlorine with methane are examined. First, a 1:4:9 mixture of Cl2, CH4, and He is coexpanded into a vacuum chamber. Then, either the antisymmetric stretch (ν3=3019 cm-1) of CH4 is prepared by direct infrared absorption or the infrared-inactive symmetric stretch (ν1=2917 cm-1) of CH4 is prepared by stimulated Raman pumping. Photolysis of Cl2 at 355 nm generates fast Cl atoms that initiate the reaction with a collision energy of 1290±175 cm-1 (0.16±0.02 eV). Finally, the nascent HCl or CH3 products are detected state-specifically via resonance enhanced multiphoton ionization and separated by mass in a time-of-flight spectrometer. We find that the rovibrational distributions and state-selected differential cross sections of the HCl and CH3 products from the two vibrationally excited reactions are nearly indistinguishable. Although Yoon et al. [J. Chem. Phys. 119, 9568 (2003)] report that the reactivities of these two different types of vibrational excitation are quite different, the present results indicate that the reactions of symmetric-stretch excited or antisymmetric-stretch excited methane with atomic chlorine follow closely related product pathways. Approximately 37% of the reaction products are formed in HCl(v=1,J) states with little rotational excitation. At low J states these products are sharply forward scattered, but become almost equally forward and backward scattered at higher J states. The remaining reaction products are formed in HCl(v=0,J) and have more rotational excitation. The HCl(v=0,J) products are predominantly back and side scattered. Measurements of the CH3 products indicate production of a non-negligible amount of umbrella bend excited methyl radicals primarily in coincidence with the HCl(v=0,J) products. The data are consistent with a model in which the impact parameter governs the scattering dynamics.

  14. Near-infrared excited state dynamics of melanins: the effects of iron content, photo-damage, chemical oxidation, and aggregate size.

    PubMed

    Simpson, Mary Jane; Wilson, Jesse W; Robles, Francisco E; Dall, Christopher P; Glass, Keely; Simon, John D; Warren, Warren S

    2014-02-13

    Ultrafast pump-probe measurements can discriminate the two forms of melanin found in biological tissue (eumelanin and pheomelanin), which may be useful for diagnosing and grading melanoma. However, recent work has shown that bound iron content changes eumelanin's pump-probe response, making it more similar to that of pheomelanin. Here we record the pump-probe response of these melanins at a wider range of wavelengths than previous work and show that with shorter pump wavelengths the response crosses over from being dominated by ground-state bleaching to being dominated by excited-state absorption. The crossover wavelength is different for each type of melanin. In our analysis, we found that the mechanism by which iron modifies eumelanin's pump-probe response cannot be attributed to Raman resonances or differences in melanin aggregation and is more likely caused by iron acting to broaden the unit spectra of individual chromophores in the heterogeneous melanin aggregate. We analyze the dependence on optical intensity, finding that iron-loaded eumelanin undergoes irreversible changes to the pump-probe response after intense laser exposure. Simultaneously acquired fluorescence data suggest that the previously reported "activation" of eumelanin fluorescence may be caused in part by the dissociation of metal ions or the selective degradation of iron-containing melanin. PMID:24446774

  15. Near-Infrared Excited State Dynamics of Melanins: The Effects of Iron Content, Photo-Damage, Chemical Oxidation, and Aggregate Size

    PubMed Central

    2015-01-01

    Ultrafast pumpprobe measurements can discriminate the two forms of melanin found in biological tissue (eumelanin and pheomelanin), which may be useful for diagnosing and grading melanoma. However, recent work has shown that bound iron content changes eumelanins pumpprobe response, making it more similar to that of pheomelanin. Here we record the pumpprobe response of these melanins at a wider range of wavelengths than previous work and show that with shorter pump wavelengths the response crosses over from being dominated by ground-state bleaching to being dominated by excited-state absorption. The crossover wavelength is different for each type of melanin. In our analysis, we found that the mechanism by which iron modifies eumelanins pumpprobe response cannot be attributed to Raman resonances or differences in melanin aggregation and is more likely caused by iron acting to broaden the unit spectra of individual chromophores in the heterogeneous melanin aggregate. We analyze the dependence on optical intensity, finding that iron-loaded eumelanin undergoes irreversible changes to the pumpprobe response after intense laser exposure. Simultaneously acquired fluorescence data suggest that the previously reported activation of eumelanin fluorescence may be caused in part by the dissociation of metal ions or the selective degradation of iron-containing melanin. PMID:24446774

  16. Multiphoton ionization of ions, neutrals, and clusters. Final report

    SciTech Connect

    Wessel, J.

    1995-12-28

    A multiyear research program investigating molecular detection methods based on multiphoton spectroscopy has been completed under DOE sponsorship. A number of new laser-based spectroscopic methods were developed and applied to a variety of aromatic hydrocarbons, including monomer and cluster species. The objectives of sensitivities approaching single molecule detection combined with high selectivity were achieved. This report references the status of the field at the beginning of this work and summarizes the significant progress during the period from 1987 onward. Detailed scientific findings from the studies are presented in the published literature referenced throughout this report.

  17. Exploration of multiphoton entangled states by using weak nonlinearities.

    PubMed

    He, Ying-Qiu; Ding, Dong; Yan, Feng-Li; Gao, Ting

    2016-01-01

    We propose a fruitful scheme for exploring multiphoton entangled states based on linear optics and weak nonlinearities. Compared with the previous schemes the present method is more feasible because there are only small phase shifts instead of a series of related functions of photon numbers in the process of interaction with Kerr nonlinearities. In the absence of decoherence we analyze the error probabilities induced by homodyne measurement and show that the maximal error probability can be made small enough even when the number of photons is large. This implies that the present scheme is quite tractable and it is possible to produce entangled states involving a large number of photons. PMID:26751044

  18. Quantum Radiation Reaction Effects in Multiphoton Compton Scattering

    SciTech Connect

    Di Piazza, A.; Hatsagortsyan, K. Z.; Keitel, C. H.

    2010-11-26

    Radiation reaction effects in the interaction of an electron and a strong laser field are investigated in the realm of quantum electrodynamics. We identify the quantum radiation reaction with the multiple photon recoils experienced by the laser-driven electron due to consecutive incoherent photon emissions. After determining a quantum radiation dominated regime, we demonstrate how in this regime quantum signatures of the radiation reaction strongly affect multiphoton Compton scattering spectra and that they could be measurable in principle with presently available laser technology.

  19. Coulomb effects in multiphoton above-threshold ionization

    NASA Astrophysics Data System (ADS)

    Kami?ski, J. Z.; Jaro?, A.; Ehlotzky, F.

    1996-03-01

    Multiphoton above-threshold ionization (ATI) is considered for hydrogen in the framework of the Keldysh-Faisal-Reiss (KFR) models but treating the Coulomb effects of the residual ions on the ionized electrons more completely than in previous work. The spectrum of the ATI peaks is evaluated for linearly polarized laser light, and it is shown that with the appropriate inclusion of the Coulomb interaction more hot electrons are predicted than by the original KFR theories. Moreover, the angular distributions of the ATI electrons are evaluated and are shown to have sidelobes.

  20. Coulomb effects in multiphoton above-threshold ionization

    SciTech Connect

    Kaminski, J.Z.; Jaron, A.; Ehlotzky, F.

    1996-03-01

    Multiphoton above-threshold ionization (ATI) is considered for hydrogen in the framework of the Keldysh-Faisal-Reiss (KFR) models but treating the Coulomb effects of the residual ions on the ionized electrons more completely than in previous work. The spectrum of the ATI peaks is evaluated for linearly polarized laser light, and it is shown that with the appropriate inclusion of the Coulomb interaction more hot electrons are predicted than by the original KFR theories. Moreover, the angular distributions of the ATI electrons are evaluated and are shown to have sidelobes. {copyright} {ital 1996 The American Physical Society.}

  1. Atomic force microscopy based, multiphoton, photoelectron emission imaging

    SciTech Connect

    Spanakis, E.; Chimmalgi, A.; Stratakis, E.; Grigoropoulos, C. P.; Fotakis, C.; Tzanetakis, P.

    2006-07-03

    Images of photoelectron emission from metallic surfaces were obtained with a modified atomic force microscope operating in air. Illumination of the samples was achieved in the near field of a metal-coated microcantilever tip, placed in the beam of a femtosecond pulsed laser that is incident at a grazing angle with respect to the sample surface. Photoelectron currents were measured through the tip with a prototype amplifier. The power law dependence of average photocurrent on light intensity is compatible with multiphoton photoelectric effect and the work function of the metal covering a particular area on the two-metal patterned samples used.

  2. Use of adaptive optics for improved multiphoton imaging

    NASA Astrophysics Data System (ADS)

    Girkin, John M.; Marsh, Paul N.

    2004-06-01

    We report on the integration of active optical elements in a multiphoton microscope to improve the resolution and overall image quality when imaging deeply into biological samples. Optical models were generated of sample systems and these have been compared with the performance of the complete imaging system. The active elements used were commercially available flexible membrane mirrors controlled by custom, home written, software. Significant improvements in image quality have been demonstrated using a range of optimisation routines based on the analysis of the images produced by the system, rather than with a wavefront sensor. A three-fold increase in the resolution 100?m into the sample was achieved.

  3. Exploration of multiphoton entangled states by using weak nonlinearities

    PubMed Central

    He, Ying-Qiu; Ding, Dong; Yan, Feng-Li; Gao, Ting

    2016-01-01

    We propose a fruitful scheme for exploring multiphoton entangled states based on linear optics and weak nonlinearities. Compared with the previous schemes the present method is more feasible because there are only small phase shifts instead of a series of related functions of photon numbers in the process of interaction with Kerr nonlinearities. In the absence of decoherence we analyze the error probabilities induced by homodyne measurement and show that the maximal error probability can be made small enough even when the number of photons is large. This implies that the present scheme is quite tractable and it is possible to produce entangled states involving a large number of photons. PMID:26751044

  4. Dynamic Multiphoton Microscopy: Focusing Light on Acute Kidney Injury

    PubMed Central

    Molitoris, Bruce A.

    2014-01-01

    Acute kidney injury (AKI) is a major global health problem; much research has been conducted on AKI, and numerous agents have shown benefit in animal studies, but none have translated into treatments. There is, therefore, a pressing unmet need to increase knowledge of the pathophysiology of AKI. Multiphoton microscopy (MPM) provides a tool to non-invasively visualize dynamic events in real time and at high resolution in rodent kidneys, and in this article we review its application to study novel mechanisms and treatments in different forms of AKI. PMID:25180263

  5. Multifocal multiphoton microscopy based on a spatial light modulator

    PubMed Central

    Shao, Y.; Qin, W.; Liu, H.; Peng, X.; Niu, H.

    2013-01-01

    We present a new multifocal multiphoton microscope that employs a programmable spatial light modulator to generate dynamic multifocus arrays which can be rapidly scanned by changing the incident angle of the laser beam using a pair of galvo scanners. Using this microscope, we can rapidly select the number and the spatial density of focal points in a multifocus array, as well as the locations and shapes of arrays according to the features of the areas of interest in the field of view without any change to the hardware. PMID:23894222

  6. Microstructure imaging of human rectal mucosa using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Liu, N. R.; Chen, G.; Chen, J. X.; Yan, J.; Zhuo, S. M.; Zheng, L. Q.; Jiang, X. S.

    2011-01-01

    Multiphoton microscopy (MPM) has high resolution and sensitivity. In this study, MPM was used to image microstructure of human rectal mucosa. The morphology and distribution of the main components in mucosa layer, absorptive cells and goblet cells in the epithelium, abundant intestinal glands in the lamina propria and smooth muscle fibers in the muscularis mucosa were clearly monitored. The variations of these components were tightly relevant to the pathology in gastrointestine system, especially early rectal cancer. The obtained images will be helpful for the diagnosis of early colorectal cancer.

  7. Multiphoton microscopic imaging of histological sections without hematoxylin and eosin staining differentiates carcinoma in situ lesion from normal oesophagus

    NASA Astrophysics Data System (ADS)

    Chen, Jianxin; Xu, Jian; Kang, Deyong; Xu, Meifang; Zhuo, Shuangmu; Zhu, Xiaoqin; Jiang, Xingshan

    2013-10-01

    Multiphoton microscopy (MPM) has become a powerful, important tool for tissues imaging at the molecular level. In this paper, this technique was extended to histological investigations, differentiating carcinoma in situ (CIS) lesion from normal oesophagus by imaging histological sections without hematoxylin and eosin (H&E) staining. The results show that the histology procedures of dehydration, paraffin embedding, and de-paraffinizing highlighted two photon excited fluorescence of cytoplasm and nucleolus of epithelial cell and collagen in stroma. MPM has the ability to identify the characteristics of CIS lesion including changes of squamous cells and full epithelium, identification of basement membrane, especially prominent nucleolus. The studies described here show that MPM has the potential for future retrospective studies of tumor staging by employing on histological section specimens without H&E staining.

  8. Resonance enhanced multiphoton ionization spectra of molecules and molecular fragments. Annual technical report, March 1991--February 1992

    SciTech Connect

    1993-12-31

    Resonance Enhanced Multiphoton Ionization (REMPI) utilizes pulsed laser radiation to prepare a molecule in an excited state via absorption of one or more photons and to subsequently ionize that state before it can decay. The overall objective of this effort is to carry out theoretical studies of these REMPI processes in molecules and molecular fragments which are designed to provide a robust analysis and prediction of key spectral features of interest in several experimental studies and applications of this technique. A specific and very important objective of the effort is to predict the vibrational and rotational ion distributions which result from REMPI of representative molecules and to understand the underlying mechanisms that give use to these ion distributions. The author highlights progress made during this period.

  9. In vivo three-dimensional optical coherence tomography and multiphoton microscopy in a mouse model of ovarian neoplasia

    NASA Astrophysics Data System (ADS)

    Watson, Jennifer M.; Marion, Samuel L.; Rice, Photini Faith; Bentley, David L.; Besselsen, David; Utzinger, Urs; Hoyer, Patricia B.; Barton, Jennifer K.

    2013-03-01

    Our goal is to use optical coherence tomography (OCT) and multiphoton microscopy (MPM) to detect early tumor development in a mouse model of ovarian neoplasia. We hope to use information regarding early tumor development to create a diagnostic test for high-risk patients. In this study we collect in vivo images using OCT, second harmonic generation and two-photon excited fluorescence from non-vinylcyclohexene diepoxide (VCD)-dosed and VCD-dosed mice. VCD causes follicular apoptosis (simulating menopause) and leads to tumor development. Using OCT and MPM we visualized the ovarian microstructure and were able to see differences between non-VCD-dosed and VCD-dosed animals. This leads us to believe that OCT and MPM may be useful for detecting changes due to early tumor development.

  10. Effect of laser spectral bandwidth on coherent control of resonance-enhanced multiphoton-ionization photoelectron spectroscopy

    SciTech Connect

    Xu, Shuwu; School of Science, Nantong University, Nantong 226007 ; Ding, Jingxin Lu, Chenhui; Jia, Tianqing; Zhang, Shian Sun, Zhenrong

    2014-02-28

    The high-resolution (2 + 1) resonance-enhanced multiphoton-ionization photoelectron spectroscopy (REMPI-PS) can be obtained by measuring the photoelectron intensity at a given kinetic energy and scanning the single ? phase step position. In this paper, we further demonstrate that the high-resolution (2 + 1) REMPI-PS cannot be achieved at any measured position of the kinetic energy by this measurement method, which is affected by the laser spectral bandwidth. We propose a double ? phase step modulation to eliminate the effect of the laser spectral bandwidth, and show the advantage of the double ? phase step modulation on achieving the high-resolution (2 + 1) REMPI-PS by considering the contributions involving on- and near-resonant three-photon excitation pathways.

  11. Label-free imaging and quantitative chemical analysis of Alzheimer's disease brain samples with multimodal multiphoton nonlinear optical microspectroscopy

    NASA Astrophysics Data System (ADS)

    Lee, Jang Hyuk; Kim, Dae Hwan; Song, Woo Keun; Oh, Myoung-Kyu; Ko, Do-Kyeong

    2015-05-01

    We developed multimodal multiphoton microspectroscopy using a small-diameter probe with gradient-index lenses and applied it to unstained Alzheimer's disease (AD) brain samples. Our system maintained the image quality and spatial resolution of images obtained using an objective lens of similar numerical aperture. Multicolor images of AD brain samples were obtained simultaneously by integrating two-photon excited fluorescence and second-harmonic generation on a coherent anti-Stokes Raman scattering (CARS) microendoscope platform. Measurements of two hippocampal regions, the cornus ammonis-1 and dentate gyrus, revealed more lipids, amyloid fibers, and collagen in the AD samples than in the normal samples. Normal and AD brains were clearly distinguished by a large spectral difference and quantitative analysis of the CH mode using CARS microendoscope spectroscopy. We expect this system to be an important diagnosis tool in AD research.

  12. Signal enhancement in multiphoton imaging by the use of coated glass substrates

    PubMed Central

    Lee, Sheng-Lin; Guo, Han-Wen; Chen, Yang-Fan; Dong, Chen-Yuan

    2015-01-01

    In nonlinear optical imaging of biological specimens, more than half of the generated luminescence signal is lost, when signal collection is performed in the epi-illuminated geometry. In this study, we enhanced the collected luminescence signal by the use of alternating multiply-coated layers of tantalum pentoxide (Ta2O5) and silicon dioxide (SiO2) on standard microscope cover glasses that has high transmission in the near-infrared wavelength region and high reflection of the visible, luminescence signal. Our coating is biocompatible, allows visual examination of the specimens and optimize collection of the luminescence signal. We demonstrated this approach on a number of specimens including sulforhodamine solution, fluorescence microspheres, and labeled 3T3 cells. In all cases, the use of coated cover glass enhanced signal, optimally by a factor of about 2. Image analysis of labeled 3T3 cells also shows signal enhancement did not contribute to additional photobleaching. Our results show that properly designed coated cover glass can enhance detected signal in multiphoton microscopy and result in improved image quality. PMID:26417521

  13. Signal enhancement in multiphoton imaging by the use of coated glass substrates.

    PubMed

    Lee, Sheng-Lin; Guo, Han-Wen; Chen, Yang-Fan; Dong, Chen-Yuan

    2015-09-01

    In nonlinear optical imaging of biological specimens, more than half of the generated luminescence signal is lost, when signal collection is performed in the epi-illuminated geometry. In this study, we enhanced the collected luminescence signal by the use of alternating multiply-coated layers of tantalum pentoxide (Ta2O5) and silicon dioxide (SiO2) on standard microscope cover glasses that has high transmission in the near-infrared wavelength region and high reflection of the visible, luminescence signal. Our coating is biocompatible, allows visual examination of the specimens and optimize collection of the luminescence signal. We demonstrated this approach on a number of specimens including sulforhodamine solution, fluorescence microspheres, and labeled 3T3 cells. In all cases, the use of coated cover glass enhanced signal, optimally by a factor of about 2. Image analysis of labeled 3T3 cells also shows signal enhancement did not contribute to additional photobleaching. Our results show that properly designed coated cover glass can enhance detected signal in multiphoton microscopy and result in improved image quality. PMID:26417521

  14. Cooper minima in electron spectra after multiphoton above-threshold ionization

    NASA Astrophysics Data System (ADS)

    Telnov, Dmitry A.; Chu, Shih-I.

    2015-05-01

    We have performed calculations of electron momentum and energy distributions after multiphoton above-threshold ionization (ATI) for several one-electron quantum systems (H, He+, H2+,and HeH2+) in intense laser fields. We use the carrier wavelengths in the near-infrared band (730 to 800 nm) and the peak intensities 5 1013 to 1 1014 W/cm2. For some initial states of the systems under consideration, the spectra exhibit minima in the low-energy region (3 to 7 eV), which resemble the famous Cooper minima in one-photon ionization processes. The minima are well pronounced for the initial states with the electronic orbitals that have nodal surfaces, such as 2 s state of He+, 1?u state of H2+,and 2 ? state of HeH2+. Such minima are not observed for the initial ground electronic states, as well as for initial 2 p state of He+, which possess nodeless orbitals. The effect is essentially non-perturbative; the positions of the minima depend on the intensity and frequency of the laser field. Nonetheless, it seems the nodal structure of the initial electronic orbital plays a crucial role in shaping these minima in the ATI electron spectra. This work is partially supported by DOE.

  15. Digital deconvolution filter derived from linear discriminant analysis and application for multiphoton fluorescence microscopy.

    PubMed

    Sullivan, Shane Z; Schmitt, Paul D; Muir, Ryan D; DeWalt, Emma L; Simpson, Garth J

    2014-04-01

    A digital filter derived from linear discriminant analysis (LDA) is developed for recovering impulse responses in photon counting from a high speed photodetector (rise time of ~1 ns) and applied to remove ringing distortions from impedance mismatch in multiphoton fluorescence microscopy. Training of the digital filter was achieved by defining temporally coincident and noncoincident transients and identifying the projection within filter-space that best separated the two classes. Once trained, data analysis by digital filtering can be performed quickly. Assessment of the reliability of the approach was performed through comparisons of simulated voltage transients, in which the ground truth results were known a priori. The LDA filter was also found to recover deconvolved impulses for single photon counting from highly distorted ringing waveforms from an impedance mismatched photomultiplier tube. The LDA filter was successful in removing these ringing distortions from two-photon excited fluorescence micrographs and through data simulations was found to extend the dynamic range of photon counting by approximately 3 orders of magnitude through minimization of detector paralysis. PMID:24559143

  16. Ultrasensitive standoff chemical sensing based on nonlinear multi-photon laser wave-mixing spectroscopy

    NASA Astrophysics Data System (ADS)

    Gregerson, Marc; Hetu, Marcel; Iwabuchi, Manna; Jimenez, Jorge; Warren, Ashley; Tong, William G.

    2012-10-01

    Nonlinear multi-photon laser wave mixing is presented as an ultrasensitive optical detection method for chem/bio agents in thin films and gas- and liquid-phase samples. Laser wave mixing is an unusually sensitive optical absorption-based detection method that offers significant inherent advantages including excellent sensitivity, small sample requirements, short optical path lengths, high spatial resolution, high spectral resolution and standoff remote detection capability. Wave mixing can detect trace amounts of chemicals even when using micrometer-thin samples, and hence, it can be conveniently interfaced to fibers, microarrays, microfluidic systems, lab-on-a-chip, capillary electrophoresis and other capillary- or fiber-based chemical separation systems. The wave-mixing signal is generated instantaneously as the two input laser beams intersect inside the analyte of interest. Laser excitation wavelengths can be tuned to detect multiple chemicals in their native form since wave mixing can detect both fluorescing and non-fluorescing samples at parts-pertrillion or better detection sensitivity levels. The wave-mixing signal is a laser-like coherent beam, and hence, it allows reliable and effective remote sensing of chemicals. Sensitive wave-mixing detectors offer many potential applications including sensitive detection of biomarkers, early detection of diseases, sensitive monitoring of environmental samples, and reliable detection of hazardous chem/bio agents with a standoff detection capability.

  17. Clinical studies of pigmented lesions in human skin by using a multiphoton tomograph

    NASA Astrophysics Data System (ADS)

    Balu, Mihaela; Kelly, Kristen M.; Zachary, Christopher B.; Harris, Ronald M.; Krasieva, Tatiana B.; Knig, Karsten; Tromberg, Bruce J.

    2013-02-01

    In vivo imaging of pigmented lesions in human skin was performed with a clinical multiphoton microscopy (MPM)-based tomograph (MPTflex, JenLab, Germany). Two-photon excited fluorescence was used for visualizing endogenous fluorophores such as NADH/FAD, keratin, melanin in the epidermal cells and elastin fibers in the dermis. Collagen fibers were imaged by second harmonic generation. Our study involved in vivo imaging of benign melanocytic nevi, atypical nevi and melanoma. The goal of this preliminary study was to identify in vivo the characteristic features and their frequency in pigmented lesions at different stages (benign, atypical and malignant) and to evaluate the ability of in vivo MPM to distinguish atypical nevi from melanoma. Comparison with histopathology was performed for the biopsied lesions. Benign melanocytic nevi were characterized by the presence of nevus cell nests at the epidermal-dermal junction. In atypical nevi, features such as lentiginous hyperplasia, acanthosis and architectural disorder were imaged. Cytological atypia was present in all the melanoma lesions imaged, showing the strongest correlation with malignancy. The MPM images demonstrated very good correlation with corresponding histological images, suggesting that MPM could be a promising tool for in vivo non-invasive pigmented lesion diagnosis, particularly distinguishing atypical nevi from melanoma.

  18. Monitoring chemically enhanced transdermal delivery of zinc oxide nanoparticles by using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Lo, Wen; Hsu, Chih-Ting; Kuo, Tsung-Rong; Wu, Chung-Long; Chiang, Shu-Jen; Lin, Sung-Jan; Chen, Shean-Jen; Chen, Chia-Chun; Dong, Chen-Yuan

    2010-02-01

    Zinc oxide nanoparticles (ZnO NPs) are commonly used in sunscreens to reduce the risk of skin cancer by blocking ultraviolet radiation. ZnO NPs absorption through the transdermal route may not cause high health risk as inhalation or ingestion. However, in practical usage of sunscreens and cosmetics, ZnO NPs are topically applied to a large area of skin with long periods hence the potential absorption amount of ZnO NPs is still need to be concerned. Therefore, if the ZnO NPs are able the pass the barrier of normal skin, the pathways of transdermal delivery and the factors of enhancements become important issues. In this work, multiphoton microscopy provides us a non-invasive visualization of ZnO NPs in skin. Moreover, we quantitatively analyzed the enhancement of oleic acid and ethanol. Due to the fact that photoluminance of ZnO NPs spectrally overlaps autofluorence from skin stratum corneum (SC) and high turbidity of both ZnO NPs and SC, it is difficult to resolve the distribution of ZnO NPs in skin by using fluorescence microscopy. In this work, the second harmonic generation (SHG) signals from ZnO NPs which double the frequency of excitation source to characterize the delivery pathways and penetration depth in skin. Moreover, we quantitatively compare the ZnO NPs delivery efficiency in normal skin and in skins with three chemically enhancing conditions: ethanol, oleic acid and the combination of ethanol and oleic acid.

  19. Identification of non-neoplastic and neoplastic gastric polyps using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Jiang, Shanghai; Kang, Deyong; Xu, Meifang; Zhu, Xiaoqin; Zhuo, Shuangmu; Chen, Jianxin

    2012-12-01

    Gastric polyps can be broadly defined as luminal lesions projecting above the plane of the mucosal surface. They are generally divided into non-neoplastic and neoplastic polyps. Accurate diagnosis of neoplastic polyps is important because of their well-known relationship with gastric cancer. Multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) is one of the most important recent inventions in biological imaging. In this study, we used MPM to image the microstructure of gastric polyps, including fundic gland polyps, hyperplastic polyps, inflammatory fibroid polyps and adenomas, then compared with gold-standard hematoxylin- eosin(H-E)-stained histopathology. MPM images showed that different gastric polyps have different gland architecture and cell morphology. Dilated, elongated or branch-like hyperplastic polyps are arranged by columnar epithelial cells. Inflammatory fibroid polyps are composed of small, thin-walled blood vessels surrounded by short spindle cells. Fundic glands polyps are lined by parietal cells and chief cells, admixed with normal glands. Gastric adenomas are generally composed of tubules or villi of dysplastic epithelium, which usually show some degree of intestinal-type differentiation toward absorptive cells, goblet cells, endocrine cells. Our results demonstrated that MPM can be used to identify non- neoplastic and neoplastic gastric polyps without the need of any staining procedure.

  20. Optical tweezers and multiphoton microscopies integrated photonic tool for mechanical and biochemical cell processes studies

    NASA Astrophysics Data System (ADS)

    de Thomaz, A. A.; Faustino, W. M.; Fontes, A.; Fernandes, H. P.; Barjas-Castro, M. d. L.; Metze, K.; Giorgio, S.; Barbosa, L. C.; Cesar, C. L.

    2007-09-01

    The research in biomedical photonics is clearly evolving in the direction of the understanding of biological processes at the cell level. The spatial resolution to accomplish this task practically requires photonics tools. However, an integration of different photonic tools and a multimodal and functional approach will be necessary to access the mechanical and biochemical cell processes. This way we can observe mechanicaly triggered biochemical events or biochemicaly triggered mechanical events, or even observe simultaneously mechanical and biochemical events triggered by other means, e.g. electricaly. One great advantage of the photonic tools is its easiness for integration. Therefore, we developed such integrated tool by incorporating single and double Optical Tweezers with Confocal Single and Multiphoton Microscopies. This system can perform 2-photon excited fluorescence and Second Harmonic Generation microscopies together with optical manipulations. It also can acquire Fluorescence and SHG spectra of specific spots. Force, elasticity and viscosity measurements of stretched membranes can be followed by real time confocal microscopies. Also opticaly trapped living protozoas, such as leishmania amazonensis. Integration with CARS microscopy is under way. We will show several examples of the use of such integrated instrument and its potential to observe mechanical and biochemical processes at cell level.

  1. Scaling up multiphoton neural scanning: the SSA algorithm.

    PubMed

    Schuck, Renaud; Annecchino, Luca A; Schultz, Simon R

    2014-01-01

    In order to reverse-engineer the information processing capabilities of the cortical circuit, we need to densely sample neural circuit; it may be necessary to sample the activity of thousands of neurons simultaneously. Frame scanning techniques do not scale well in this regard, due to the time "wasted" scanning extracellular space. For scanners in which inertia can be neglected, path length minimization strategies enable large populations to be imaged at relatively high sampling rates. However, in a standard multiphoton microscope, the scanners responsible for beam deflection are inertial, indicating that an optimal solution should take rotor and mirror momentum into account. We therefore characterized the galvanometric scanners of a commercial multiphoton microscope, in order to develop and validate a MATLAB model of microscope scanning dynamics. We tested the model by simulating scan paths across pseudo-randomly positioned neuronal populations of differing neuronal density and field of view. This model motivated the development of a novel scanning algorithm, Adaptive Spiral Scanning (SSA), in which the radius of a circular trajectory is constantly updated such that it follows a spiral trajectory scanning all the cells. Due to the kinematic efficiency of near-circular trajectories, this algorithm achieves higher sampling rates than shortest path approaches, while retaining a relatively efficient coverage fraction in comparison to raster or resonance based frame-scanning approaches. PMID:25570582

  2. Rigid and high NA multiphoton fluorescence GRIN-endoscopes

    NASA Astrophysics Data System (ADS)

    Schenkl, Selma; Ehlers, Alexander; Le Harzic, Ronan; Stark, Martin; Riemann, Iris; Messerschmidt, Bernhard; Kaatz, Martin; König, Karsten

    2007-07-01

    Multiphoton autofluorescence imaging offers minimal-invasive examination of cells without the need of staining and complicated confocal detection systems. Therefore, it is especially interesting for non-invasive clinical diagnostics. To extend this sophisticated technique from superficial regions to deep lying cell layers, internal body parts and specimens difficult of access, the bulky optics need to be reduced in diameter. This is done by tiny GRIN-optics, based on a radial gradient in the reflective index. Of especial interest for multi-photon applications is the newly developed GRIN-lens assembly with increased numerical aperture. High resolution images of plant tissue, hair and cells show the improved image quality,compared to classical GRIN-lenses. The rigid GRIN-endoscopes are already applied in wound healing studies. Here, the GRIN-lenses with diameters smaller than 3 mm enter small skin depressions. They reproduce the focus of a conventional laser scanning tomograph tens of mm apart in the specimen under study. We present first clinical measurements of elastin and SHG of collagen of in-vivo human skin of venous ulcers (ulcer curis).

  3. The effect of radial polarization in multiphoton lithography

    NASA Astrophysics Data System (ADS)

    Lin, Le; Zheng, Mei-Ling; Dong, Xian-Zi; Duan, Xuan-Ming; Zhao, Zhen-Sheng

    2015-10-01

    Considering the axially symmetric polarization and intensity distribution, radially polarized (RP) laser beam has comparatively higher axial component of electric field and smaller size of focal spot compared to linearly polarized (LP) laser. In this study, the effect of radial polarization on multiphoton fabrication has been studied, and polymer spots and lines are chosen as the study objects of 2D micro/nano structures of multiphoton lithography. These structures were fabricated with IP-L, a commercial negative photoresist, by RP fs-pulse laser beam which was tightly focused by an objective lens with high numerical aperture. Multiple experimental conditions, such as fabrication power, exposure time and scanning velocity, were verified in order to observe the structural variation of these polymer structures. On the basis of measurement from images of the scanning electron microscope, the transverse and longitudinal sizes of polymer spots and lines could be analyzed, and the relationship between the aspect ratio (AR) and the above experimental conditions could be acquired. The statistical results agree with our predictions that the RP laser beam can significantly reduce the AR, and the AR in RP laser fabrication has little correlation with conditions besides fabrication power, such as exposure time and scanning velocity.

  4. Using adaptive optics for deep in-vivo multiphoton FLIM

    NASA Astrophysics Data System (ADS)

    Poland, Simon; Fruhwirth, Gilbert; Ng, Tony; Ameer-beg, Simon

    2011-03-01

    Multiphoton microscopy (MPM) is a high resolution (sub-?m) 3D optical imaging technique that has seen widespread use for microscopy at moderate depth within biological tissue (~1 mm). MPM combined with Fluorescence lifetime imaging (FLIM) and Fluorescent resonant energy transfer (FRET) provides the ability to image protein-protein interactions. When applied in-vivo at depth, it will be a key component to identifying and evaluating drug interaction in tumours. Unfortunately as one images more deeply into biological tissue, depth is restricted due to the specimen induced aberrations, which result in deterioration in both the image quality and resolution. Adaptive optics (AO), a technique first developed for astronomy, has been shown to be successful in overcoming problems associated with imaging in depth in confocal, multiphoton, CARS and SHG microscopy. The principle relies on shaping the wavefront with a wavefront modulator to compensate for the distortions introduced by the biological tissue sample. The success of such a technique relies on being able to correctly determine the mirror shape required In this paper we will discuss the development a dedicated MPM FLIM-FRET microscope incorporating an AO for use in-vivo applications. Using a deformable membrane mirror as a wavefront modulator, a strategy for implementation will be discussed.

  5. Non-invasive multiphoton imaging of extracellular matrix structures.

    PubMed

    Schenke-Layland, Katja

    2008-12-01

    Multiphoton microscopy has become a powerful method for the artifact-free, nondestructive evaluation of deep-tissue cells and extracellular matrix (ECM) structures in their native environment. By interacting with highly non-centrosymmetric molecular assemblies such as fibrillar collagen, the non-linear process called second harmonic generation (SHG) has also proven to be an important diagnostic tool for the visualization of ECM compartments in situ with submicron resolution without the need for tissue processing. This review reports on applications of multiphoton-induced autofluorescence and SHG microscopy to identify collagen and elastic fiber orientation in native, tissue-engineered and processed, as well as healthy and diseased, tissues and organs. SHG signal profiling was used to quantify ECM damage in various cardiovascular and exocrine tissues, as well as cartilage. These novel imaging modalities open the general possibility of high-resolution in situ and more important in vivo imaging of ECM structures, cells and intracellular organelles in living intact tissues. PMID:19343671

  6. Multiphoton imaging: a view to understanding sulfur mustard lesions

    NASA Astrophysics Data System (ADS)

    Werrlein, Robert J. S.; Madren-Whalley, Janna S.

    2003-07-01

    It is well known that topical exposure to sulfur mustard (SM) produces persistent, incapacitating blisters of the skin. However, the primary lesions effecting epidermal-dermal separation and disabling of mechanisms for cutaneous repair remain uncertain. Immunofluorescent staining plus multiphoton imaging of human epidermal tissues and keratinocytes exposed to SM (400 μM x 5 min)have revealed that SM disrupts adhesion-complex molecules which are also disrupted by epidermolysis bullosa-type blistering diseases of the skin. Images of keratin-14 showed early, progressive, postexposure collapse of the K5/K14 cytoskeleton that resulted in ventral displacement of the nuclei beneath its collapsing filaments. This effectively corrupted the dynamic filament assemblies that link basal-cell nuclei to the extracellular matrix via α6β4-integrin and laminin-5. At 1 h postexposure, there was disruption in the surface organization of α6β4 integrins, associated displacement of laminin-5 anchoring sites and a concomitant loss of functional asymmetry. Accordingly, our multiphoton images are providing compelling evidence that SM induces prevesicating lesions that disrupt the receptor-ligand organization and cytoskeletal systems required for maintaining dermal-epidermal attachment, signal transduction, and polarized mobility.

  7. Mg in electromagnetic fields: Theoretical partial multiphoton cross sections

    SciTech Connect

    Nikolopoulos, L.A.A.

    2005-03-01

    We present ab initio calculations of multiphoton ionization cross sections--up to four--in atomic magnesium. We have followed a configuration interaction approach with the basis set constructed in terms of L{sup 2} integrable B-spline polynomials. The multiphoton ionization cross sections are given for a range of photon energies where the ion is left in its ground state. For the two-photon ionization process we calculate the cross section as a function of the photon energy and compare with known theoretical results. We also provide the corresponding angular asymmetry parameters which determine the angular distribution of the ionized electron. We have extended the energy range of reported theoretical three-photon ionization cross section while we present the four-photon cross section in the region between the 3s and 3p ionization thresholds. In this region the 3p{sup 2} {sup 1}S autoionizing state is identified through the four-photon absorption process and the related four-photon Fano q parameter is obtained.

  8. Nanoscale spatial induction of ultraviolet photoproducts in cellular DNA by three-photon near-infrared absorption

    PubMed Central

    Meldrum, Rosalind A; Botchway, Stanley W; Wharton, Christopher W; Hirst, Graeme J

    2003-01-01

    The high-resolution spatial induction of ultraviolet (UV) photoproducts in mammalian cellular DNA is a goal of many scientists who study UV damage and repair. Here we describe how UV photoproducts can be induced in cellular DNA within nanometre dimensions by near-diffraction-limited 750 nm infrared laser radiation. The use of multiphoton excitation to induce highly localized DNA damage in an individual cell nucleus or mitochondrion will provide much greater resolution for studies of DNA repair dynamics and intracellular localization as well as intracellular signalling processes and cellcell communication. The technique offers an advantage over the masking method for localized irradiation of cells, as the laser radiation can specifically target a single cell and subnuclear structures such as nucleoli, nuclear membranes or any structure that can be labelled and visualized by a fluorescent tag. It also increases the time resolution with which migration of DNA repair proteins to damage sites can be monitored. We define the characteristics of localized DNA damage induction by near-infrared radiation and suggest how it may be used for new biological investigations. PMID:14618160

  9. Photolytic production of C/sup 2/H: collisional quenching of A tilde /sup 2/II. -->. X tilde /sup 2/Sigma/sup +/ infrared emission and the removal of excited C/sub 2/H

    SciTech Connect

    Shokoohi, F.; Watson, T.A.; Reisler, H.; Kong, F.; Renlund, A.M.; Wittig, C.

    1986-10-23

    The authors report the observation of time-resolved C/sub 2/H A tilde /sup 2/II ..-->.. X tilde /sup 2/Sigma/sup +/ infrared emission (1-5 ..mu..m) following the 193-nm photolyses of C/sub 2/H/sub 2/ and C/sub 2/HBr. Quenching of this emission by numerous collision partners (M) under pseudo-first-order conditions leads to large bimolecular rate coefficients (e.g. > 10/sup -11/ cm/sup 3/ molecule/sup -1/ s/sup -1/, except when M is a rare gas or N/sub 2/). Although such rate coefficients can be assigned to the quenching of fluorescence, they do not represent state-to-state processes, since quenching is due to an intricate combination of reactive, radiative, and energy-transfer processes. In separate experiments, rate coefficients are determined by monitoring the time-resolved CH A/sup 2/..delta.. ..-->.. X/sup 2/II chemiluminescence which is produced directly by the reaction of C/sub 2/H with O/sub 2/, and the C/sub 2/H species responsible for the CH emissions is identified as electronically and/or vibrationally excited C/sub 2/H. The above results are in agreement with recent molecular beam experiments that show that nascent C/sub 2/H contains considerably internal energy following the 193-nm photolysis of C/sub 2/H/sub 2/.

  10. Fringe-free, Background-free, Collinear Third Harmonic Generation FROG Measurements for Multiphoton Microscopy

    SciTech Connect

    Chadwick, R; Spahr, E; Squier, J A; Durfee, C G; Walker, B C; Fittinghoff, D N

    2006-07-21

    Collinear pulse measurement tools useful at the full numerical aperture (NA) of multiphoton microscope objectives are a necessity for a quantitative characterization of the femtosecond pulses focused by these systems. In this letter, we demonstrate a simple new technique, for characterizing the pulse at the focus in a multiphoton microscope. This technique, a background-free, fringe-free, form of frequency-resolved optical gating, uses the third harmonic signal generated from a glass coverslip. Here it is used to characterize 100 fs pulses (typical values for a multiphoton microscope) at the focus of a 0.65 NA objective.

  11. Multiphoton fluorescence and second harmonic generation microscopy of different skin states

    NASA Astrophysics Data System (ADS)

    Lin, Sung-Jan; Lo, Wen; Sun, Yen; Jee, Shiou-Hwa; Dong, Chen-Yuan

    2005-04-01

    In recent years, non-linear imaging techniques such as multiphoton fluorescence and harmonic generation microscopy have been successfully applied to dermatological imaging. Confocal-like image quality, enhanced depth penetration, and non-linear spectral signatures are among the main advantages of this family of techniques. In this presentation, we will focus on the applications of multiphoton microscopy to skin specimens in different physiological states. Images of normal and diseased tissue specimens will be presented and spectrally characterized. Our work has potential applications in developing multiphoton microscopy into a clinically applicable diagnostic tool.

  12. Multiphoton imaging with a novel compact diode-pumped Ti:sapphire oscillator.

    PubMed

    Knig, Karsten; Andersen, Peter; Le, Tuan; Breunig, Hans Georg

    2015-12-01

    Multiphoton laser scanning microscopy commonly relies on bulky and expensive femtosecond lasers. We integrated a novel minimal-footprint Ti:sapphire oscillator, pumped by a frequency-doubled distributed Bragg reflector tapered diode laser, into a clinical multiphoton tomograph and evaluated its imaging capability using different biological samples, i.e. cell monolayers, corneal tissue, and human skin. With the novel laser, the realization of very compact Ti:sapphire-based systems for high-quality multiphoton imaging at a significantly size and weight compared to current systems will become possible. Microsc. Res. Tech. 78:1154-1158, 2015. 2015 Wiley Periodicals, Inc. PMID:26534831

  13. Modulation of the pupil function of microscope objective lens for multifocal multi-photon microscopy using a spatial light modulator

    NASA Astrophysics Data System (ADS)

    Matsumoto, Naoya; Okazaki, Shigetoshi; Takamoto, Hisayoshi; Inoue, Takashi; Terakawa, Susumu

    2014-02-01

    We propose a method for high precision modulation of the pupil function of a microscope objective lens to improve the performance of multifocal multi-photon microscopy (MMM). To modulate the pupil function, we adopt a spatial light modulator (SLM) and place it at the conjugate position of the objective lens. The SLM can generate an arbitrary number of spots to excite the multiple fluorescence spots (MFS) at the desired positions and intensities by applying an appropriate computer-generated hologram (CGH). This flexibility allows us to control the MFS according to the photobleaching level of a fluorescent protein and phototoxicity of a specimen. However, when a large number of excitation spots are generated, the intensity distribution of the MFS is significantly different from the one originally designed due to misalignment of the optical setup and characteristics of the SLM. As a result, the image of a specimen obtained using laser scanning for the MFS has block noise segments because the SLM could not generate a uniform MFS. To improve the intensity distribution of the MFS, we adaptively redesigned the CGH based on the observed MFS. We experimentally demonstrate an improvement in the uniformity of a 10 × 10 MFS grid using a dye solution. The simplicity of the proposed method will allow it to be applied for calibration of MMM before observing living tissue. After the MMM calibration, we performed laser scanning with two-photon excitation to observe a real specimen without detecting block noise segments.

  14. Vacuum ultraviolet and near-infrared excited luminescence properties of Ca{sub 3}(PO{sub 4}){sub 2}:RE{sup 3+}, Na{sup +} (RE=Tb, Yb, Er, Tm, and Ho)

    SciTech Connect

    Zhang Jia; Wang Yuhua; Guo Linna; Zhang Feng; Wen Yan; Liu Bitao; Huang Yan

    2011-08-15

    Tb{sup 3+}, Yb{sup 3+}, Tm{sup 3+}, Er{sup 3+}, and Ho{sup 3+} doped Ca{sub 3}(PO{sub 4}){sub 2} were synthesized by solid-state reaction, and their luminescence properties were studied by spectra techniques. Tb{sup 3+}-doped samples can exhibit intense green emission under VUV excitation, and the brightness for the optimal Tb{sup 3+} content is comparable with that of the commercial Zn{sub 2}SiO{sub 4}:Mn{sup 2+} green phosphor. Under near-infrared laser excitation, the upconversion luminescence spectra of Yb{sup 3+}, Tm{sup 3+}, Er{sup 3+}, and Ho{sup 3+} doped samples demonstrate that the red, green, and blue tricolored fluorescence could be obtained by codoping Yb{sup 3+}-Ho{sup 3+}, Yb{sup 3+}-Er{sup 3+}, and Yb{sup 3+}-Tm{sup 3+} in Ca{sub 3}(PO{sub 4}){sub 2}, respectively. Good white upconversion emission with CIE chromaticity coordinates (0.358, 0.362) is achieved by quadri-doping Yb{sup 3+}-Tm{sup 3+}-Er{sup 3+}-Ho{sup 3+} in Ca{sub 3}(PO{sub 4}){sub 2}, in which the cross-relaxation process between Er{sup 3+} and Tm{sup 3+}, producing the {sup 1}D{sub 2}-{sup 3}F{sub 4} transition of Tm{sup 3+}, is found. The upconversion mechanisms are elucidated through the laser power dependence of the upconverted emissions and the energy level diagrams. - Graphical abstract: The CPO:0.25Tb{sup 3+}, 0.25Na{sup +} exhibits a comparable brightness to the commercial Zn{sub 2}SiO{sub 4}:Mn{sup 2+} upon 147 nm excitation. Good white light color is achieved in CPO:Yb{sup 3+}-Tm{sup 3+}-Er{sup 3+}-Ho{sup 3+} under 980 nm excitation. Highlights: > Ca{sub 3}(PO{sub 4}){sub 2}:Tb{sup 3+},Na{sup +} exhibits a comparable brightness with commercial Zn{sub 2}SiO{sub 4}:Mn{sup 2+}. > Red, green and blue colors are achieved in Yb{sup 3+}, Ho{sup 3+}, Er{sup 3+}, Tm{sup 3+} doped Ca{sub 3}(PO{sub 4}){sub 2}. > Good white emission is obtained in Yb{sup 3+}-Ho{sup 3+}-Er{sup 3+}-Tm{sup 3+} quadri-doped Ca{sub 3}(PO{sub 4}){sub 2}. > Ca{sub 3}(PO{sub 4}){sub 2}:Tb{sup 3+}, Yb{sup 3+}, Ho{sup 3+}, Er{sup 3+}, Tm{sup 3+} could be potential phosphors.

  15. Multiphoton intravital microscopy setup to visualize the mouse mammary gland

    NASA Astrophysics Data System (ADS)

    Adur, Javier; Herrera Torres, Ana M.; Masedunskas, Andrius; Baratti, Mariana O.; de Thomaz, Andre A.; Pelegati, Vitor B.; Carvalho, Hernandes F.; Cesar, Carlos L.

    2013-06-01

    Recently, light microscopy-based techniques have been extended to live mammalian models leading to the development of a new imaging approach called intravital microscopy (IVM). Although IVM has been introduced at the beginning of the last century, its major advancements have occurred in the last twenty years with the development of non-linear microscopy that has enabled performing deep tissue imaging. IVM has been utilized to address many biological questions in basic research and is now a fundamental tool that provide information on tissues such as morphology, cellular architecture, and metabolic status. IVM has become an indispensable tool in numerous areas. This study presents and describes the practical aspects of IVM necessary to visualize epithelial cells of live mouse mammary gland with multiphoton techniques.

  16. Watching stem cells at work with a flexible multiphoton tomograph

    NASA Astrophysics Data System (ADS)

    Uchugonova, Aisada; Hoffmann, Robert; Weinigel, Martin; König, Karsten

    2012-03-01

    There is a high demand for non-invasive imaging techniques that allow observation of stem cells in their native environment without significant input on cell metabolism, reproduction, and behavior. Easy accessible hair follicle pluripotent stem cells in the bulge area and dermal papilla are potential sources for stem cell based therapy. It has been shown that these cells are able to generate hair, non-follicle skin cells, nerves, vessels, smooth muscles etc. and may participate in wound healing processes. We report on the finding of nestin-GFP expressing stem cells in their native niche in the bulge of the hair follicle of living mice by using high-resolution in-vivo multiphoton tomography. The 3D imaging with submicron resolution was based on two-photon induced fluorescence and second harmonic generation (SHG) of collagen. Migrating stem cells from the bulge to their microenvironment have been detected inside the skin during optical deep tissue sectioning.

  17. High-Resolution Multiphoton Imaging of Tumors In Vivo

    PubMed Central

    Wyckoff, Jeffrey; Gligorijevic, Bojana; Entenberg, David; Segall, Jeffrey; Condeelis, John

    2014-01-01

    Analysis of the individual steps in metastasis is crucial if insights at the molecular level are to be linked to the cell biology of cancer. A technical hurdle to achieving the analysis of the individual steps of metastasis is the fact that, at the gross level, tumors are heterogeneous in both animal models and patients. Human primary tumors show extensive variation in all properties ranging from growth and morphology of the tumor through tumor-cell density in the blood and formation and growth of metastases. Methods capable of the direct visualization and analysis of tumor-cell behavior at single-cell resolution in vivo have become crucial in advancing the understanding of mechanisms of metastasis, the definition of microenvironment, and the markers related to both. This article discusses the use of high-resolution multiphoton imaging of tumors (specifically breast tumors in mice) in vivo. PMID:21969629

  18. Multiphoton quantum Rabi oscillations in ultrastrong cavity QED

    NASA Astrophysics Data System (ADS)

    Garziano, Luigi; Stassi, Roberto; Macr?, Vincenzo; Kockum, Anton Frisk; Savasta, Salvatore; Nori, Franco

    2015-12-01

    When an atom is strongly coupled to a cavity, the two systems can exchange a single photon through a coherent Rabi oscillation. This process enables precise quantum-state engineering and manipulation of atoms and photons in a cavity, which play a central role in quantum information and measurement. Recently, a new regime of cavity QED was reached experimentally where the strength of the interaction between light and artificial atoms (qubits) becomes comparable to the atomic transition frequency or the resonance frequency of the cavity mode. Here we show that this regime can strongly modify the concept of vacuum Rabi oscillations, enabling multiphoton exchanges between the qubit and the resonator. We find that experimental state-of-the-art circuit-QED systems can undergo two- and three-photon vacuum Rabi oscillations. These anomalous Rabi oscillations can be exploited for the realization of efficient Fock-state sources of light and complex entangled states of qubits.

  19. Live-animal imaging of renal function by multiphoton microscopy.

    PubMed

    Dunn, Kenneth W; Sutton, Timothy A; Sandoval, Ruben M

    2007-07-01

    Intravital microscopy, microscopy of living animals, is a powerful research technique that combines the resolution and sensitivity found in microscopic studies of cultured cells with the relevance and systemic influences of cells in the context of the intact animal. The power of intravital microscopy has recently been extended with the development of multiphoton fluorescence microscopy systems capable of collecting optical sections from deep within the kidney at subcellular resolution, supporting high-resolution characterizations of the structure and function of glomeruli, tubules, and vasculature in the living kidney. Fluorescent probes are administered to an anesthetized, surgically prepared animal, followed by image acquisition for up to 3 hr. Images are transferred via a high-speed network to specialized computer systems for digital image analysis. This general approach can be used with different combinations of fluorescent probes to evaluate processes such as glomerular permeability, proximal tubule endocytosis, microvascular flow, vascular permeability, mitochondrial function, and cellular apoptosis/necrosis. PMID:18770850

  20. Multiphoton Microscopy and Interaction of Intense Light Pulses with Polymers

    NASA Astrophysics Data System (ADS)

    Guay, Jean-Michel

    2011-07-01

    The nanoscale manipulation of soft-matter, such as biological tissues, in its native environment has promising applications in medicine to correct for defects (eg. eye cataracts) or to destroy malignant regions (eg. cancerous tumours). To achieve this we need the ability to first image and then do precise ablation with sub-micron resolution with the same setup. For this purpose, we designed and built a multiphoton microscope and tested it on goldfish gills and bovine cells. We then studied light-matter interaction on a hard polymer (PMMA) because the nature of ablation of soft-matter in its native environment is complex and not well understood. Ablation and modification thresholds for successive laser shots were obtained. The ablation craters revealed 3D nanostructures and polarization dependent orientation. The interaction also induced localized porosity in PMMA that can be controlled.