Single Molecule Characterization of UV-Activated Antibodies on Gold by Atomic Force Microscopy.

The interaction between proteins and solid surfaces can influence their conformation and therefore also their activity and affinity. These interactions are highly specific for the respective combination of proteins and solids. Consequently, it is desirable to investigate the conformation of proteins on technical surfaces, ideally at single molecule level, and to correlate the results with their activity. This is in particular true for biosensors where the conformation-dependent target affinity of an immobilized receptor determines the sensitivity of the sensor. Here, we investigate for the first time the immobilization and orientation of antibodies (Abs) photoactivated by a photonic immobilization technique (PIT), which has previously demonstrated to enhance binding capabilities of antibody receptors. The photoactivated immunoglobulins are immobilized on ultrasmooth template stripped gold films and investigated by atomic force microscopy (AFM) at the level of individual molecules. The observed protein orientations are compared with results of nonactivated antibodies adsorbed on similar gold films and mica reference samples. We find that the behavior of Abs is similar for mica and gold when the protein are not treated (physisorption), whereas smaller contact area and larger heights are measured when Abs are treated (PIT). This is explained by assuming that the activated antibodies tend to be more upright compared with nonirradiated ones, thereby providing a better exposure of the binding sites. This finding matches the observed enhancement of Abs binding efficiency when PIT is used to functionalize gold surface of QCM-based biosensors.

Extension of high harmonic spectroscopy in molecules by a 1300 nm laser field.

The emerging techniques of molecular spectroscopy by high order harmonic generation have hitherto been conducted only with Ti:Sapphire lasers which are restricted to molecules with high ionization potentials. In order to gain information on the molecular structure, a broad enough range of harmonics is required. This implies using high laser intensities which would saturate the ionization of most molecular systems of interest, e.g. organic molecules. Using a laser at 1300 nm, we are able to extend the technique to molecules with relatively low ionization potentials (approximately 11 eV), observing wide harmonic spectra reaching up to 60 eV. This energy range improves spatial resolution of the high harmonic spectroscopy to the point where interference minima in harmonic spectra of N(2)O and C(2)H(2) can be observed.

Generation of high energy, 30 fs pulses at 527 nm by hollow-fiber compression technique.

The compression of 300-fs-long, chirp-free laser pulses at 527 nm down to 30 fs is reported. The laser pulses, originated from a frequency-doubled, mode-locked Nd:glass laser, were compressed by a 0.7-m-long, 150-microm-bore-diameter, argon-filled hollow fiber, and a pair of SF10 prisms with a final energy of 160 microJ. These are the shortest, high energy pulses ever produced by direct pulse compression at the central wavelength of 527 nm. The spectral broadening of the pulses propagating inside the hollow fiber was experimentally examined for various filling-gas pressures and input pulse energies. The spectral width of the pulses was broadened up to 25 nm, and 27 nm for argon- and krypton-filled hollow fiber, respectively, at a gas pressure lower than 2 bar. The physical limitations of the hollow-fiber pulse compression technique applied in the visible range are also studied.

Hollow-fiber compression of visible, 200 fs laser pulses to 40 fs pulse duration.

We demonstrate the use of a very simple, compact, and versatile method, based on the hollow-fiber compression technique, to shorten the temporal length of visible laser pulses of 100-300 fs to pulse durations shorter than approximately 50 fs. In particular, 200 fs, frequency-doubled, Nd:glass laser pulses (527 nm) were spectrally broadened to final bandwidths as large as 25 nm by nonlinear propagation through an Ar-filled hollow fiber. A compact, dispersive, prism-pair compressor was then used to produce as short as 40 fs, 150 microJ pulses. A very satisfactory agreement between numerical simulations and measurements is found.

Probing orbital structure of polyatomic molecules by high-order harmonic generation.

The effects of electronic structure and symmetry are observed in laser driven high-order harmonic generation for laser aligned conjugated polyatomic molecular systems. The dependence of the harmonic yield on the angle between the molecular axis and the polarization of the driving laser field is seen to contain the fingerprint of the highest occupied molecular orbitals in acetylene and allene, a good quantitative agreement with calculations employing the strong field approximation was found. These measurements support the extension of the recently proposed molecular orbital imaging techniques beyond simple diatomic molecules to larger molecular systems.

Isolated single-cycle attosecond pulses.

We generated single-cycle isolated attosecond pulses around approximately 36 electron volts using phase-stabilized 5-femtosecond driving pulses with a modulated polarization state. Using a complete temporal characterization technique, we demonstrated the compression of the generated pulses for as low as 130 attoseconds, corresponding to less than 1.2 optical cycles. Numerical simulations of the generation process show that the carrier-envelope phase of the attosecond pulses is stable. The availability of single-cycle isolated attosecond pulses opens the way to a new regime in ultrafast physics, in which the strong-field electron dynamics in atoms and molecules is driven by the electric field of the attosecond pulses rather than by their intensity profile.

Controlling two-center interference in molecular high harmonic generation.

We experimentally investigate the process of intramolecular quantum interference in high-order harmonic generation in impulsively aligned CO2 molecules. The recombination interference effect is clearly seen through the order dependence of the harmonic yield in an aligned sample. The experimental results can be well modeled assuming that the effective de Broglie wavelength of the returning electron wave is not significantly altered by the Coulomb field of the molecular ion. We demonstrate that such interference effects can be effectively controlled by changing the ellipticity of the driving laser field.

Role of the intramolecular phase in high-harmonic generation.

We study numerically the generation of high-order harmonics by two-center molecules for arbitrary angles between the molecular axis and the laser polarization axis. For fixed angle, the harmonic spectrum exhibits a minimum at a frequency which is independent of the laser parameters. The amplitude of each harmonic is strongly angle dependent, and a pronounced minimum is found at the same angle where a sudden jump in the harmonic phase occurs. By calculating the spatial dependence of the harmonic amplitudes and phases, we are able to explain these effects in terms of interfering contributions from various regions within the molecule.

Analysis of the receiver response in lidar measurements.

We report on the calculation of the effective telescope area in lidar applications by a ray-tracing approach. This method allows one to consider the true experimental working conditions and hence to obtain accurate values of the effective telescope area as a function of the height. This in turn allows the retrieval of the signal from the ranges where the overlap function is not constant (e.g., lower ranges), thus increasing the useful range interval. Moreover, we show that the spherical mirrors are more appropriate than the parabolic ones for most of the lidar measurements, although a particular alignment procedure, such as the one we describe, must be used.