Multimodal wide-field two-photon excitation imaging: characterization of the technique for in vivo applications

We report fast, non-scanning, wide-field two-photon fluorescence excitation with spectral and lifetime detection for in vivo biomedical applications. We determined the optical characteristics of the technique, developed a Gaussian flat-field correction method to reduce artifacts resulting from non-uniform excitation such that contrast is enhanced, and showed that it can be used for ex vivo and in vivo cellular-level imaging. Two applications were demonstrated: (i) ex vivo measurements of beta-amyloid plaques in retinas of transgenic mice, and (ii) in vivo imaging of sulfonated gallium(III) corroles injected into tumors. We demonstrate that wide-field two photon fluorescence excitation with flat-field correction provides more penetration depth as well as better contrast and axial resolution than the corresponding one-photon wide field excitation for the same dye. Importantly, when this technique is used together with spectral and fluorescence lifetime detection modules, it offers improved discrimination between fluorescence from molecules of interest and autofluorescence, with higher sensitivity and specificity for in vivo applications

Correlated electron emission in laser-induced nonsequence double ionization of Helium

In this paper, we have investigated the correlated electron emission of the nonsequence double ionization (NSDI) in an intense linearly polarized field. The theoretical model we employed is the semiclassical rescattering model, the model atom we used is the helium. We find a significant correlation between magnitude and direction of the momentum of two emission electrons, and give a good explanation for this striking phenomenon by observing the classical collisional trajectories. We argue that this correlation phenomenon is universal in NSDI process, as revealed by the recent experiment on the argon.Comment: 4 pages, 3 figures, accepted for publication in Phys. Rev.

Shot noise limited characterization of femtosecond light pulses

Probing the evolution of physical systems at the femto- or attosecond timescale with light requires accurate characterization of ultrashort optical pulses. The time profiles of such pulses are usually retrieved by methods utilizing optical nonlinearities, which require significant signal powers and operate in a limited spectral range\cite{Trebino_Review_of_Scientific_Instruments97,Walmsley_Review_09}. We present a linear self-referencing characterization technique based on time domain localization of the pulse spectral components, operated in the single-photon regime. Accurate timing of the spectral slices is achieved with standard single photon detectors, rendering the technique applicable in any spectral range from near infrared to deep UV. Using detection electronics with about $70$ ps response, we retrieve the temporal profile of a picowatt pulse train with $\sim10$ fs resolution, setting a new scale of sensitivity in ultrashort pulse characterization.Comment: Supplementary information contained in raw dat

Functional Analysis of the SPA Gene Family in Arabidopsis thaliana

Ambient light conditions affect development throughout the plant life cycle, including seed germination, seedling development and the induction of flowering. In the model plant Arabidopsis, the COP1-SPA ubiquitin ligase complex plays a central role in suppressing light signaling in darkness. The COP1-SPA complex targets positively acting factors like HY5, a protein necessary for normal seedling development in the light, several photoreceptors and the flowering time regulator CONSTANS for degradation via the 26S proteasome. Therefore, one of the major functions of the light signal transduction pathways is the inactivation of the COP1-SPA complex. While COP1 is a single copy gene, the SPA proteins are encoded by four different loci (SPA1-SPA4). All SPA proteins have redundant, but also distinct functions in regulating plant development. SPA1 and SPA2 are the key regulators that suppress photomorphogenesis in dark-grown seedlings. Over-stimulation in light-grown seedlings is primarily prevented by SPA1, and to a minor extent, also by SPA3 and SPA4. SPA2, in contrast has only negligible function in the light. SPA1 is sufficient for repressing flowering under non-inductive short-day conditions. Here, I show that distinct functions of the SPA genes partially correlate with their distinct gene expression patterns. RNA gel blot-analysis revealed that the expression of SPA1, SPA3 and SPA4, but not that of SPA2, is positively influenced by light of different wavelengths. All main photoreceptors contribute to the up-regulation of these SPA transcripts, implying that photoreceptors initiate a negative feedback regulation, which might protect plants from over-stimulation by light. GUS reporter gene experiments show that SPA genes exhibit somewhat distinct tissue-specific expression patterns, which might be important for tissue specific regulation of COP1-SPA targets. However, differences in SPA gene expression cannot account for all distinct SPA gene functions. Promoter-swap experiments with SPA1, SPA2 and SPA4 show that all SPA proteins are potent repressors in dark-grown seedlings. SPA1 and SPA4 also act as repressor in the light. SPA2, however, can never act as a repressor in the light, not even when it is expressed from the strong light-induced SPA1 promoter. These results show that SPA proteins themselves feature properties that determine characteristic SPA protein functions. All SPA proteins feature a characteristic domain structure with a C-terminal WD-repeat, a central coiled-coil domain and a less well-conserved N-terminus that includes a kinase-like motif. The WD-repeat domain and the coiled-coil domain are essential for formation of the COP1-SPA complex as well as interactions with various ubiquitination targets. In contrast, the function of the N-terminal domain is unknown. Aiming to determine the importance of the N-terminal domain of SPA1, I conducted a structure-function analysis. While the N-terminal domain of SPA1 is dispensable for SPA1 function in the seedling stage, this domain is required for SPA1-mediated repression of flowering in non-inductive short-day conditions. These results indicate, that the SPA1 N-terminal domain can full-fill an essential function

Strong field double ionization of H2 : Insights from nonlinear dynamics

The uncorrelated (``sequential'') and correlated (``nonsequential'') double ionization of the H2 molecule in strong laser pulses is investigated using the tools of nonlinear dynamics. We focus on the phase-space dynamics of this system, specifically by finding the dynamical structures that regulate these ionization processes. The emerging picture complements the recollision scenario by clarifying the distinct roles played by the recolliding and core electrons. Our analysis leads to verifiable predictions of the intensities where qualitative changes in ionization occur. We also show how these findings depend on the internuclear distance

Correlation dynamics between electrons and ions in the fragmentation of D2_2 molecules by short laser pulses

We studied the recollision dynamics between the electrons and D$_2^+$ ions following the tunneling ionization of D$_2$ molecules in an intense short pulse laser field. The returning electron collisionally excites the D$_2^+$ ion to excited electronic states from there D$_2^+$ can dissociate or be further ionized by the laser field, resulting in D$^+$ + D or D$^+$ + D$^+$, respectively. We modeled the fragmentation dynamics and calculated the resulting kinetic energy spectrum of D$^+$ to compare with recent experiments. Since the recollision time is locked to the tunneling ionization time which occurs only within fraction of an optical cycle, the peaks in the D$^+$ kinetic energy spectra provides a measure of the time when the recollision occurs. This collision dynamics forms the basis of the molecular clock where the clock can be read with attosecond precision, as first proposed by Corkum and coworkers. By analyzing each of the elementary processes leading to the fragmentation quantitatively, we identified how the molecular clock is to be read from the measured kinetic energy spectra of D$^+$ and what laser parameters be used in order to measure the clock more accurately.Comment: 13 pages with 14 figure

A two-dimensional, two-electron model atom in a laser pulse: exact treatment, single active electron-analysis, time-dependent density functional theory, classical calculations, and non-sequential ionization

Owing to its numerical simplicity, a two-dimensional two-electron model atom, with each electron moving in one direction, is an ideal system to study non-perturbatively a fully correlated atom exposed to a laser field. Frequently made assumptions, such as the ``single active electron''- approach and calculational approximations, e.g. time dependent density functional theory or (semi-) classical techniques, can be tested. In this paper we examine the multiphoton short pulse-regime. We observe ``non-sequential'' ionization, i.e.\ double ionization at lower field strengths as expected from a sequential, single active electron-point of view. Since we find non-sequential ionization also in purely classical simulations, we are able to clarify the mechanism behind this effect in terms of single particle trajectories. PACS Number(s): 32.80.RmComment: 10 pages, 16 figures (gzipped postscript), see also http://www.physik.tu-darmstadt.de/tqe

Collinear helium under periodic driving: stabilization of the asymmetric stretch orbit

The collinear eZe configuration of helium, with the electrons on opposite sides of the nucleus, is studied in the presence of an external electromagnetic (laser or microwave) field. We show that the classically unstable "asymmetric stretch" orbit, on which doubly excited intrashell states of helium with maximum interelectronic angle are anchored, can be stabilized by means of a resonant driving where the frequency of the electromagnetic field equals the frequency of Kepler-like oscillations along the orbit. A static magnetic field, oriented parallel to the oscillating electric field of the driving, can be used to enforce the stability of the configuration with respect to deviations from collinearity. Quantum Floquet calculations within a collinear model of the driven two-electron atom reveal the existence of nondispersive wave packets localized on the stabilized asymmetric stretch orbit, for double excitations corresponding to principal quantum numbers of the order of N > 10.Comment: 13 pages, 12 figure

Atomic excitation during recollision-free ultrafast multi-electron tunnel ionization

Modern intense ultrafast pulsed lasers generate an electric field of sufficient strength to permit tunnel ionization of the valence electrons in atoms. This process is usually treated as a rapid succession of isolated events, in which the states of the remaining electrons are neglected. Such electronic interactions are predicted to be weak, the exception being recollision excitation and ionization caused by linearly-polarized radiation. In contrast, it has recently been suggested that intense field ionization may be accompanied by a two-stage `shake-up' reaction. Here we report a unique combination of experimental techniques that enables us to accurately measure the tunnel ionization probability for argon exposed to 50 femtosecond laser pulses. Most significantly for the current study, this measurement is independent of the optical focal geometry, equivalent to a homogenous electric field. Furthermore, circularly-polarized radiation negates recollision. The present measurements indicate that tunnel ionization results in simultaneous excitation of one or more remaining electrons through shake-up. From an atomic physics standpoint, it may be possible to induce ionization from specific states, and will influence the development of coherent attosecond XUV radiation sources. Such pulses have vital scientific and economic potential in areas such as high-resolution imaging of in-vivo cells and nanoscale XUV lithography.Comment: 17 pages, 4 figures, original format as accepted by Nature Physic