Wide-Field Multi-Parameter FLIM: Long-Term Minimal Invasive Observation of Proteins in Living Cells.

Time-domain Fluorescence Lifetime Imaging Microscopy (FLIM) is a remarkable tool to monitor the dynamics of fluorophore-tagged protein domains inside living cells. We propose a Wide-Field Multi-Parameter FLIM method (WFMP-FLIM) aimed to monitor continuously living cells under minimum light intensity at a given illumination energy dose. A powerful data analysis technique applied to the WFMP-FLIM data sets allows to optimize the estimation accuracy of physical parameters at very low fluorescence signal levels approaching the lower bound theoretical limit. We demonstrate the efficiency of WFMP-FLIM by presenting two independent and relevant long-term experiments in cell biology: 1) FRET analysis of simultaneously recorded donor and acceptor fluorescence in living HeLa cells and 2) tracking of mitochondrial transport combined with fluorescence lifetime analysis in neuronal processes

A Physicsl Model of Electron According to the Basic Structures of Matter Hypothesis

A physical model of the electron is suggested according to the basic structures of matter (BSM) hypothesis. BSM is based on an alternative concept about the physical vacuum, assuming that space contains an underlying grid structure of nodes formed of superdense subelementary particles, which are also involved in the structure of the elementary particles. The proposed grid structure is formed of vibrating nodes that possess quantum features and energy well. It is admitted that this hypothetical structure could account for the missing “dark matter” in the universe. The signature of this dark matter is apparent in the galactic rotational curves and in the relation between masses of the supermassive black hole in the galactic center and the host galaxy. The suggested model of the electron possesses oscillation features with anomalous magnetic moment and embedded signatures of the Compton wavelength and the fine-structure constant. The analysis of the interactions between the oscillating electron and the nodes of the vacuum grid structure allows us to obtain physical meaning for some fundamental constants

Airborne chemical sensing with mobile robots

Airborne chemical sensing with mobile robots has been an active research areasince the beginning of the 1990s. This article presents a review of research work in this field,including gas distribution mapping, trail guidance, and the different subtasks of gas sourcelocalisation. Due to the difficulty of modelling gas distribution in a real world environmentwith currently available simulation techniques, we focus largely on experimental work and donot consider publications that are purely based on simulations

Matter-wave entanglement and teleportation by molecular dissociation and collisions

We propose dissociation of cold diatomic molecules as a source of atom pairs with highly correlated (entangled) positions and momenta, approximating the original quantum state introduced by Einstein, Podolsky and Rosen (EPR) [Phys. Rev. 47, 777 (1935)]. Wavepacket teleportation is shown to be achievable by its collision with one of the EPR correlated atoms and manipulation of the other atom in the pair.Comment: REVTeX, 4 pages, 3 figures. Text reformulated, modified figs. 1 and 2. Accepted by Phys. Rev. Let

Inference in particle tracking experiments by passing messages between images

Methods to extract information from the tracking of mobile objects/particles have broad interest in biological and physical sciences. Techniques based on simple criteria of proximity in time-consecutive snapshots are useful to identify the trajectories of the particles. However, they become problematic as the motility and/or the density of the particles increases due to uncertainties on the trajectories that particles followed during the images' acquisition time. Here, we report an efficient method for learning parameters of the dynamics of the particles from their positions in time-consecutive images. Our algorithm belongs to the class of message-passing algorithms, known in computer science, information theory and statistical physics as Belief Propagation (BP). The algorithm is distributed, thus allowing parallel implementation suitable for computations on multiple machines without significant inter-machine overhead. We test our method on the model example of particle tracking in turbulent flows, which is particularly challenging due to the strong transport that those flows produce. Our numerical experiments show that the BP algorithm compares in quality with exact Markov Chain Monte-Carlo algorithms, yet BP is far superior in speed. We also suggest and analyze a random-distance model that provides theoretical justification for BP accuracy. Methods developed here systematically formulate the problem of particle tracking and provide fast and reliable tools for its extensive range of applications.Comment: 18 pages, 9 figure

Secretory vesicles are preferentially targeted to areas of low molecular SNARE density

Intercellular communication is commonly mediated by the regulated fusion, or exocytosis, of vesicles with the cell surface. SNARE (soluble N-ethymaleimide sensitive factor attachment protein receptor) proteins are the catalytic core of the secretory machinery, driving vesicle and plasma membrane merger. Plasma membrane SNAREs (tSNAREs) are proposed to reside in dense clusters containing many molecules, thus providing a concentrated reservoir to promote membrane fusion. However, biophysical experiments suggest that a small number of SNAREs are sufficient to drive a single fusion event. Here we show, using molecular imaging, that the majority of tSNARE molecules are spatially separated from secretory vesicles. Furthermore, the motilities of the individual tSNAREs are constrained in membrane micro-domains, maintaining a non-random molecular distribution and limiting the maximum number of molecules encountered by secretory vesicles. Together our results provide a new model for the molecular mechanism of regulated exocytosis and demonstrate the exquisite organization of the plasma membrane at the level of individual molecular machines

Inferring Latent States and Refining Force Estimates via Hierarchical Dirichlet Process Modeling in Single Particle Tracking Experiments

Optical microscopy provides rich spatio-temporal information characterizing in vivo molecular motion. However, effective forces and other parameters used to summarize molecular motion change over time in live cells due to latent state changes, e.g., changes induced by dynamic micro-environments, photobleaching, and other heterogeneity inherent in biological processes. This study focuses on techniques for analyzing Single Particle Tracking (SPT) data experiencing abrupt state changes. We demonstrate the approach on GFP tagged chromatids experiencing metaphase in yeast cells and probe the effective forces resulting from dynamic interactions that reflect the sum of a number of physical phenomena. State changes are induced by factors such as microtubule dynamics exerting force through the centromere, thermal polymer fluctuations, etc. Simulations are used to demonstrate the relevance of the approach in more general SPT data analyses. Refined force estimates are obtained by adopting and modifying a nonparametric Bayesian modeling technique, the Hierarchical Dirichlet Process Switching Linear Dynamical System (HDP-SLDS), for SPT applications. The HDP-SLDS method shows promise in systematically identifying dynamical regime changes induced by unobserved state changes when the number of underlying states is unknown in advance (a common problem in SPT applications). We expand on the relevance of the HDP-SLDS approach, review the relevant background of Hierarchical Dirichlet Processes, show how to map discrete time HDP-SLDS models to classic SPT models, and discuss limitations of the approach. In addition, we demonstrate new computational techniques for tuning hyperparameters and for checking the statistical consistency of model assumptions directly against individual experimental trajectories; the techniques circumvent the need for "ground-truth" and subjective information.Comment: 25 pages, 6 figures. Differs only typographically from PLoS One publication available freely as an open-access article at http://journals.plos.org/plosone/article?id=10.1371/journal.pone.013763

Formation of plasma around a small meteoroid: 2. Implications for radar head echo

This paper calculates the spatial distribution of the plasma responsible for radar head echoes by applying the kinetic theory developed in the companion paper. This results in a set of analytic expressions for the plasma density as a function of distance from the meteoroid. It shows that at distances less than a collisional mean free path from the meteoroid surface, the plasma density drops in proportion to 1/R where R is the distance from the meteoroid center; and, at distances much longer than the mean‐free‐path behind the meteoroid, the density diminishes at a rate proportional to 1/R2. The results of this paper should be used for modeling and analysis of radar head echoes.This work was supported by NSF grant AGS-1244842. (AGS-1244842 - NSF

Electrostatic correlations in inhomogeneous charged fluids beyond loop expansion

Electrostatic correlation effects in inhomogeneous symmetric electrolytes are investigated within a previously developed electrostatic self-consistent (SC) theory (R.R. Netz and H. Orland, Eur. Phys.J. E 11, 301 (2003)). To this aim, we introduce two computational approaches that allow to solve the SC equations beyond the loop expansion. Both approaches can handle the case of dielectrically discontinuous boundaries where the one-loop theory is known to fail. By comparing the theoretical results obtained from these schemes with the results of the MC simulations that we ran for ions at neutral single dielectric interfaces as well as with previous MC data for charged interfaces, we first show that the weak coupling (WC) Debye-Huckel (DH) theory remains quantitatively accurate up to the bulk ion density rhob=0.01 M, whereas the SC theory exhibits a good quantitative accuracy up to rhob=0.2 M. Then, we derive from the perturbative SC scheme the one-loop theory of asymmetrically partitioned salt systems around a dielectrically homogeneous charged surface. It is shown that correlation effects originate in these systems from a competition between the salt screening loss at the interface driving the ions to the bulk region, and the interfacial counterion screening excess attracting them towards the surface. In the case of weak surface charges, the interfacial salt screening loss is the dominant effect. As a result, correlations decrease the MF density of both coions and counterions. With increasing surface charge, the surface-attractive counterion screening excess starts to dominate, and correlation effects amplify in this regime the MF density of both type of ions. We also show that at a characteristic value of the electrostatic coupling parameter, electrostatic correlations result in a charge inversion effect