Intracellular transport of large cargoes, such as organelles, vesicles or large proteins, is a complex dynamical process that involves the interplay of ATP-consuming molecular motors, cytoskeleton filaments and the viscoelastic cytoplasm. The displacements of particles or probes in the cell cytoplasm as a function of time are characterized by different (anomalous) diffusion regimes. We investigate here the motion of pigment organelles (melanosomes) driven by myosin-V motors in Xenopus laevis melanocytes using a high spatio-temporal resolution tracking technique. By analyzing the mean square displacement (MSD) of the obtained trajectories as a function of the time lag, we show that the melanosomes display a transition between subdiffusive to superdiffusive behavior. A stochastic theoretical model is introduced to generalize the interpretation of our data. Starting from a generalized Langevin equation that explicitly considers the collective action of the molecular motors we derive an analytical expression for the MSD as a function of the time lag, which also takes into account the experimental noise. By fitting our model to the experimental data we were able to discriminate the exponents that characterize the passive and active contributions to melanosome dynamics. The model also estimates the "global" motor forces correctly. In this sense, our model gives a quantitative description of active transport in living cells with a reduced number of parameters.
Bistability, excitability, and self-pulsing regimes in an InP-based two-dimensional (2D) photonic crystal nanocavity with quantum wells as an active medium are investigated. A resonant cw beam is evanescently coupled into the cavity through a tapered microfiber. In such conditions, we show that the cavity exhibits class II excitability, which arises from the competition between a fast electronic nonlinear effect, given by carrier-induced refractive index change, and slow thermal dynamics. Multiple perturbation-pulse experiments allow us to measure the refractory time ("dead time" between two excitable pulses) of the excitable nanocavity system.
Bias (dc electric field) dependence of the dielectric constant of ferroelectric ceramics of the Pb(Zr,Ti)O3 type has been investigated. Measurements were carried out at various temperatures in the ferroelectric phase and at several frequencies between 1 kHz and 1 MHz. The bias characteristics of the dielectric constant show a temperature-dependent asymmetry in the electric field direction. A model in which the coercive fields of the 180° and 90° domains are distributed around some average value is considered. The total dielectric constant of the material is calculated by considering the contributions of each type of domain, taking into account the difference between the differential and incremental dielectric constants. This model explains the dependence of dielectric constant on electric field. A comparison of the calculated and experimental results is presented.
Human inducible pluripotent stem cells (hiPSCs) hold a large potential for disease modeling. hiPSC-derived human astrocyte and neuronal cultures permit investigations of neural signaling pathways with subcellular resolution. Combinatorial cultures, and three-dimensional (3-D) embryonic bodies (EBs) enlarge the scope of investigations to multi-cellular phenomena. The highest level of complexity, brain organoids that—in many aspects—recapitulate anatomical and functional features of the developing brain permit the study of developmental and morphological aspects of human disease. An ideal microscope for 3-D tissue imaging at these different scales would combine features from both confocal laser-scanning and light-sheet microscopes: a micrometric optical sectioning capacity and sub-micrometric spatial resolution, a large field of view and high frame rate, and a low degree of invasiveness, i.e., ideally, a better photon efficiency than that of a confocal microscope. In the present work, we describe such an instrument that uses planar two-photon (2P) excitation. Its particularity is that—unlike two- or three-lens light-sheet microscopes—it uses a single, low-magnification, high-numerical aperture objective for the generation and scanning of a virtual light sheet. The microscope builds on a modified Nipkow-Petráň spinning-disk scheme for achieving wide-field excitation. However, unlike the Yokogawa design that uses a tandem disk, our concept combines micro lenses, dichroic mirrors and detection pinholes on a single disk. This new design, advantageous for 2P excitation, circumvents problems arising with the tandem disk from the large wavelength difference between the infrared excitation light and visible fluorescence. 2P fluorescence excited by the light sheet is collected with the same objective and imaged onto a fast sCMOS camera. We demonstrate 3-D imaging of TO-PRO3-stained EBs and of brain organoids, uncleared and after rapid partial transparisation with triethanolamine formamide (RTF) and we compare the performance of our instrument to that of a confocal laser-scanning microscope (CLSM) having a similar numerical aperture. Our large-field 2P-spinning disk microscope permits one order of magnitude faster imaging, affords less photobleaching and permits better depth penetration than a confocal microscope with similar spatial resolution.
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