Spatial and temporal pulse shaping for lateral and depth resolved two-photon excited fluorescence contrast

We report combined temporal and spatial laser pulse shaping to perform lateral and depth dependent two-photon excited fluorescence of dyes. For generating the specific spatially and temporally phase tailored pulses a temporal pulse shaper and a subsequent spatial pulse shaper are employed. Simultaneous spatial and temporal shaping is presented for two-photon excited fluorescence by applying temporal third order phase functions on spatially different light field components. Moreover, the prospects of spatial shaping are demonstrated by applying various lateral two-photon fluorescence pattern. In particular, a depth dependent excitation of different dyes is performed which leads to a high axially resolved fluorescence contrast. The introduced spatial and temporal shaping technique provides new perspectives for biophotonic imaging applications

Contrast improvement by using tailored laser pulses to circumvent undesired excitations

We report on fluorescence contrast improvement by using phase, amplitude, and polarization shaped laser pulses. The measurements were conducted by applying phase functions at different spectral amplitudes for excitations of dyes and agree very well with calculations. In particular, undesired one-photon excitations are circumvented with phase and amplitude tailored pulses for two-photon transition. This is realized by cutting out the laser spectrum at the wavelength of the one-photon process while utilizing an antisymmetric phase function that allows for constructive interference of the remaining outer spectral contributions for two-photon absorption. Moreover, polarization enhanced contrast between dyes is demonstrated where the two-photon dye is predominantly excited in one polarization direction and simultaneously the one-photon dye in the other polarization direction. The presented methods of shaping ultrashort laser pulses have a high potential for imaging applications

Combined temporal and spatial laser pulse shaping for two-photon excited fluorescence contrast improvement

We report on combined simultaneous temporal and spatial laser pulse shaping by utilizing light polarization properties. Thereto, a setup comprising a temporal pulse shaper, a waveplate, and a spatial shaper was developed and characterized by comparison with simulations. This enables to simultaneously shape one polarization component temporally and spatially while the perpendicular polarization component is modified temporally. The spatially and temporally modulated light fields were recorded and visualized by suitable contour plots, which was particularly demonstrated for cylindrically symmetric pulse profiles. Moreover, temporally and spatially shaped pulses were applied for two-photon excited fluorescence of dyes. These measurements were conducted by scanning third order phase functions for specific spatial pulse components which yields an enhanced contrast difference between fluorescing dyes. The presented temporal and spatial shaping method of ultrashort laser pulses has a high potential for biophotonic applications

Fluorescence contrast improvement by polarization shaped laser pulses for autofluorescent biomolecules

We present contrast enhancement for the autofluorescing coenzymes flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide (NADH) in glycerol using phase and polarization shaped laser pulses after the transmission through a kagome fiber. Thereto, we report a way to calculate the optimal light modulator incident polarization angle, which in general differs from the horizontal. Combining phase and polarization shaping, we can selectively excite FAD in one polarization and simultaneously NADH in the other polarization direction by third order phase functions. Due to high anisotropy, the contrast of the fluorescence depends on the polarization direction. The effect of the fiber on the phase is precompensated in order to obtain the desired phase function after the fiber. Since the relative amounts of NADH and FAD give information about cellular metabolic activity which in turn helps understand disease processes, the method promises high biophotonic potential

Modifications of filament spectra by shaped octave-spanning laser pulses

In this paper we examine the spectral changes in a white light laser filament due to different pulse shapes generated by a pulse-shaping setup. We particularly explore how the properties of the filament spectra can be controlled by parametrically tailored white light pulses. The experiments are carried out in a gas cell with up to 9 bars of argon. Plasma generation and self-phase modulation strongly affect the pulse in the spectral and temporal domains. By exploiting these effects we show that the pulse spectrum can be modified in a desired way by either using second-order parametric chirp functions to shift the filament spectrum to higher or lower wavelengths, or by optimizing pulse shapes with a genetic algorithm to generate more complex filament spectra. This paper is an example of the application of complex, parametrically shaped white light pulses

Localized helium excitations in 4He_N-benzene clusters

We compute ground and excited state properties of small helium clusters 4He_N containing a single benzene impurity molecule. Ground-state structures and energies are obtained for N=1,2,3,14 from importance-sampled, rigid-body diffusion Monte Carlo (DMC). Excited state energies due to helium vibrational motion near the molecule surface are evaluated using the projection operator, imaginary time spectral evolution (POITSE) method. We find excitation energies of up to ~23 K above the ground state. These states all possess vibrational character of helium atoms in a highly anisotropic potential due to the aromatic molecule, and can be categorized in terms of localized and collective vibrational modes. These results appear to provide precursors for a transition from localized to collective helium excitations at molecular nanosubstrates of increasing size. We discuss the implications of these results for analysis of anomalous spectral features in recent spectroscopic studies of large aromatic molecules in helium clusters.Comment: 15 pages, 5 figures, submitted to Phys. Rev.

Atomically resolved phase transition of fullerene cations solvated in helium droplets

Helium has a unique phase diagram and below 25 bar it does not form a solid even at the lowest temperatures. Electrostriction leads to the formation of a solid layer of helium around charged impurities at much lower pressures in liquid and superfluid helium. These so-called ‘Atkins snowballs’ have been investigated for several simple ions. Here we form HenC60+ complexes with n exceeding 100 via electron ionization of helium nanodroplets doped with C60. Photofragmentation of these complexes is measured by merging a tunable narrow- bandwidth laser beam with the ions. A switch from red- to blueshift of the absorption frequency of HenC60+ on addition of He atoms at n=32 is associated with a phase transition in the attached helium layer from solid to partly liquid (melting of the Atkins snowball). Elaborate molecular dynamics simulations using a realistic force field and including quantum effects support this interpretation

Extracellular Charge Adsorption Influences Intracellular Electrochemical Homeostasis in Amphibian Skeletal Muscle

The membrane potential measured by intracellular electrodes, Em, is the sum of the transmembrane potential difference (E1) between inner and outer cell membrane surfaces and a smaller potential difference (E2) between a volume containing fixed charges on or near the outer membrane surface and the bulk extracellular space. This study investigates the influence of E2 upon transmembrane ion fluxes, and hence cellular electrochemical homeostasis, using an integrative approach that combines computational and experimental methods. First, analytic equations were developed to calculate the influence of charges constrained within a three-dimensional glycocalyceal matrix enveloping the cell membrane outer surface upon local electrical potentials and ion concentrations. Electron microscopy confirmed predictions of these equations that extracellular charge adsorption influences glycocalyceal volume. Second, the novel analytic glycocalyx formulation was incorporated into the charge-difference cellular model of Fraser and Huang to simulate the influence of extracellular fixed charges upon intracellular ionic homeostasis. Experimental measurements of Em supported the resulting predictions that an increased magnitude of extracellular fixed charge increases net transmembrane ionic leak currents, resulting in either a compensatory increase in Na+/K+-ATPase activity, or, in cells with reduced Na+/K+-ATPase activity, a partial dissipation of transmembrane ionic gradients and depolarization of Em