Label-free 3D visualization of cellular and tissue structures in intact muscle with second and third harmonic generation microscopy.

Second and Third Harmonic Generation (SHG and THG) microscopy is based on optical effects which are induced by specific inherent physical properties of a specimen. As a multi-photon laser scanning approach which is not based on fluorescence it combines the advantages of a label-free technique with restriction of signal generation to the focal plane, thus allowing high resolution 3D reconstruction of image volumes without out-of-focus background several hundred micrometers deep into the tissue. While in mammalian soft tissues SHG is mostly restricted to collagen fibers and striated muscle myosin, THG is induced at a large variety of structures, since it is generated at interfaces such as refraction index changes within the focal volume of the excitation laser. Besides, colorants such as hemoglobin can cause resonance enhancement, leading to intense THG signals. We applied SHG and THG microscopy to murine (Mus musculus) muscles, an established model system for physiological research, to investigate their potential for label-free tissue imaging. In addition to collagen fibers and muscle fiber substructure, THG allowed us to visualize blood vessel walls and erythrocytes as well as white blood cells adhering to vessel walls, residing in or moving through the extravascular tissue. Moreover peripheral nerve fibers could be clearly identified. Structure down to the nuclear chromatin distribution was visualized in 3D and with more detail than obtainable by bright field microscopy. To our knowledge, most of these objects have not been visualized previously by THG or any label-free 3D approach. THG allows label-free microscopy with inherent optical sectioning and therefore may offer similar improvements compared to bright field microscopy as does confocal laser scanning microscopy compared to conventional fluorescence microscopy

Designing Game Based Learning – a Participatory Approach

Game Based Learning seems to be an interesting new possibility of teaching and learning, but the effort spent on designing games and the possible positive outcomes have to be weighed carefully. The following paper describes the development process and the conceptual design of a simulation game on sustainability for teenagers. The design process is participatory in nature. Members of the future group of learners are involved in the design process at every stage. This involvement is especially important to overcome the contradiction between the goal of the game as such and the pedagogical goal of the designers

Selective activation as an explanation fro hindsight bias

In hindsight, people often claim to have known more in foresight than they actually did. For example, the confidence for one of several possible outcomes is larger when it is known that this particular outcome occurred. A widespread explanation of hindsight bias assumes that the feedback serves as an anchor. How precisely this anchor takes effect and why it leads to a bias towards the anchor value has not been satisfactorily answered yet. One possible mechanism to explain hindsight bias assumes that the encoding of the feedback leads to a selective activation of the item-specific knowledge base. As a result, specific information units are strengthened and are thus more likely to be recalled when a person tries to reconstruct his or her original judgment. We tested the effect of selective activation in two hindsight experiments. The results showed a clear hindsight bias in that the recalled confidence ratings were distorted towards the feedback. Moreover, the consequences of selective activation were evident in that more information favoring the feedback was recalled

Spectral Energy Distributions of Gamma Ray Bursts Energized by External Shocks

Sari, Piran, and Narayan have derived analytic formulas to model the spectra from gamma-ray burst blast waves that are energized by sweeping up material from the surrounding medium. We extend these expressions to apply to general radiative regimes and to include the effects of synchrotron self-absorption. Electron energy losses due to the synchrotron self-Compton process are also treated in a very approximate way. The calculated spectra are compared with detailed numerical simulation results. We find that the spectral and temporal breaks from the detailed numerical simulation are much smoother than the analytic formulas imply, and that the discrepancies between the analytic and numerical results are greatest near the breaks and endpoints of the synchrotron spectra. The expressions are most accurate (within a factor of ~ 3) in the optical/X-ray regime during the afterglow phase, and are more accurate when epsilon_e, the fraction of swept-up particle energy that is transferred to the electrons, is <~ 0.1. The analytic results provide at best order-of-magnitude accuracy in the self-absorbed radio/infrared regime, and give poor fits to the self-Compton spectra due to complications from Klein-Nishina effects and photon-photon opacity.Comment: 16 pages, 7 figures, ApJ, in press, 537, July 1, 2000. Minor changes in response to referee report, corrected figure

Visualization of Endothelial Actin Cytoskeleton in the Mouse Retina

Angiogenesis requires coordinated changes in cell shape of endothelial cells (ECs), orchestrated by the actin cytoskeleton. The mechanisms that regulate this rearrangement in vivo are poorly understood - largely because of the difficulty to visualize filamentous actin (F-actin) structures with sufficient resolution. Here, we use transgenic mice expressing Lifeact-EGFP to visualize F-actin in ECs. We show that in the retina, Lifeact-EGFP expression is largely restricted to ECs allowing detailed visualization of F-actin in ECs in situ. Lifeact-EGFP labels actin associated with cell-cell junctions, apical and basal membranes and highlights actin-based structures such as filopodia and stress fiber-like cytoplasmic bundles. We also show that in the skin and the skeletal muscle, Lifeact-EGFP is highly expressed in vascular mural cells (vMCs), enabling vMC imaging. In summary, our results indicate that the Lifeact-EGFP transgenic mouse in combination with the postnatal retinal angiogenic model constitutes an excellent system for vascular cell biology research. Our approach is ideally suited to address structural and mechanistic details of angiogenic processes, such as endothelial tip cell migration and fusion, EC polarization or lumen formation

A hadronic synchrotron mirror model for the "orphan" TeV flare in 1ES 1959+650

Very-high-energy gamma-ray flares of TeV blazars are generally accompanied by simultaneous flaring activity in X-rays. The recent observations by the Whipple collaboration of an ``orphan'' TeV flare of 1ES 1959+650 (without simultaneous X-ray flare) is very hard to reconcile with the standard leptonic SSC model which is routinely very successfully employed to explain the SED and spectral variability of TeV blazars. In this paper, an alternative scenario is suggested in which the ``orphan'' TeV flare may originate from relativistic protons, interacting with an external photon field supplied by electron-synchrotron radiation reflected off a dilute reflector. While the external photons will be virtually ``invisible'' to the co-moving ultrarelativistic electrons in the jet due to Klein-Nishina effects, their Doppler boosted energy is high enough to excite the $\Delta$ resonance from relativistic protons with Lorentz factors of ~ 10^3 - 10^4. This model is capable of explaining the ``orphan'' TeV flare of 1ES 1959+650 with plausible parameters, thus constraining the number and characteristic energy of relativistic protons in the jet of this blazar.Comment: Accepted for publication in ApJ. 14 preprint pages, including 2 .eps figure

Magnetic Field Amplification and Flat Spectrum Radio Quasars

We perform time-dependent, spatially-resolved simulations of blazar emission to evaluate several flaring scenarios related to magnetic-field amplification and enhanced particle acceleration. The code explicitly accounts for light-travel-time effects and is applied to flares observed in the flat spectrum radio quasar (FSRQ) PKS 0208-512, which show optical/{\gamma}-ray correlation at some times, but orphan optical flares at other times. Changes in both the magnetic field and the particle acceleration efficiency are explored as causes of flares. Generally, external Compton emission appears to describe the available data better than a synchrotron self-Compton scenario, and in particular orphan optical flares are difficult to produce in the SSC framework. X-ray soft-excesses, {\gamma}-ray spectral hardening, and the detections at very high energies of certain FSRQs during flares find natural explanations in the EC scenario with particle acceleration change. Likewise, optical flares with/without {\gamma}-ray counterparts can be explained by different allocations of energy between the magnetization and particle acceleration, which may be related to the orientation of the magnetic field relative to the jet flow. We also calculate the degree of linear polarization and polarization angle as a function of time for a jet with helical magnetic field. Tightening of the magnetic helix immediately downstream of the jet perturbations, where flares occur, can be sufficient to explain the increases in the degree of polarization and a rotation by >= 180 degree of the observed polarization angle, if light-travel-time effects are properly considered.Comment: 12 pages, 9 figures. Accepted for publication in MNRA

Strong amplitude-phase coupling in submonolayer quantum dots

This article may be downloaded for personal use only. Any other use requires prior permission of the author and AIP Publishing. This article appeared in Appl. Phys. Lett. 109, 201102 (2016) and may be found at https://doi.org/10.1063/1.4967833.Submonolayer quantum dots promise to combine the beneficial features of zero- and two-dimensional carrier confinement. To explore their potential with respect to all-optical signal processing, we investigate the amplitude-phase coupling (α-parameter) in semiconductor optical amplifiers based on InAs/GaAs submonolayer quantum dots in ultrafast pump-probe experiments. Lateral coupling provides an efficient carrier reservoir and gives rise to a large α-parameter. Combined with a high modal gain and an ultrafast gain recovery, this makes the submonolayer quantum dots an attractive gain medium for nonlinear optical signal processing

Observation of Long-Lived Muonic Hydrogen in the 2S State

The kinetic energy distribution of ground state muonic hydrogen atoms mu-p(1S) is determined from time-of-flight spectra measured at 4, 16, and 64 hPa H2 room-temperature gas. A 0.9 keV-component is discovered and attributed to radiationless deexcitation of long-lived mu-p(2S) atoms in collisions with H2 molecules. The analysis reveals a relative population of about 1%, and a pressure-dependent lifetime (e.g. (30.4 +21.4 -9.7) ns at 64 hPa) of the long-lived mu-p(2S) population, equivalent to a 2S-quench rate in mu-p(2S) + H2 collisions of (4.4 +2.1 -1.8) 10^11 s^-1 at liquid hydrogen density.Comment: 4 pages, 2 figures, accepted for publication in Physical Review Letter