A modular spring-loaded actuator for mechanical activation of membrane proteins.

How cells respond to mechanical forces by converting them into biological signals underlie crucial cellular processes. Our understanding of mechanotransduction has been hindered by technical barriers, including limitations in our ability to effectively apply low range piconewton forces to specific mechanoreceptors on cell membranes without laborious and repetitive trials. To overcome these challenges we introduce the Nano-winch, a robust, easily assembled, programmable DNA origami-based molecular actuator. The Nano-winch is designed to manipulate multiple mechanoreceptors in parallel by exerting fine-tuned, low- piconewton forces in autonomous and remotely activated modes via adjustable single- and double-stranded DNA linkages, respectively. Nano-winches in autonomous mode can land and operate on the cell surface. Targeting the device to integrin stimulated detectable downstream phosphorylation of focal adhesion kinase, an indication that Nano-winches can be applied to study cellular mechanical processes. Remote activation mode allowed finer extension control and greater force exertion. We united remotely activated Nano-winches with single-channel bilayer experiments to directly observe the opening of a channel by mechanical force in the force responsive gated channel protein, BtuB. This customizable origami provides an instrument-free approach that can be applied to control and explore a diversity of mechanotransduction circuits on living cells.

High-resolution-scanning waveguide microscopy: spatial refractive index and topography quantification.

We report on a high-resolution metal-clad waveguide scanning microscopic method with a diffraction-limited resolution. This microscope can be operated in both TM and TE waveguide modes with radially and azimuthally polarized beams, respectively, and allows both refractive index and topography of dielectric objects to be evaluated at high resolution and sensitivity. We emphasize the performance of this microscopic method from calibrated 3D polymer microstructures with rectangular, disk, and ring shapes.

A tensegrity driven DNA nanopore.

Control of transport across membranes, whether natural or synthetic, is fundamental in many biotechnology applications, including sensing and drug release. Mutations of naturally existing protein channels, such as hemolysin, have been explored in the past. More recently, DNA channels with conductivities in the nanosiemens range have been designed. Regulating transport across DNA channels in response to external stimuli remains an important challenge. Previous designs relied on steric hindrance to control the inner diameter of the channel, which resulted in unstable electric signatures. In this paper we introduce a new design to control electric channel conductance of a DNA nanopore. The tensegrity driven mechanism inhibits the flux of small analytes while keeping a tightly controlled ionic transport modulated by the addition of specific DNA sequences. Current signals are clearly defined, with no sign of gating, opening new perspectives in single molecule DNA sensing.

Wavelet-based decomposition of high resolution surface plasmon microscopy V(Z) curves at visible and near infrared wavelengths.

Surface plasmon resonance is conventionally conducted in the visible range and, during the past decades, it has proved its efficiency in probing molecular scale interactions. Here we elaborate on the first implementation of a high resolution surface plasmon microscope that operates at near infrared (IR) wavelength for the specific purpose of living matter imaging. We analyze the characteristic angular and spatial frequencies of plasmon resonance in visible and near IR lights and how these combined quantities contribute to the V(Z) response of a scanning surface plasmon microscope (SSPM). Using a space-frequency wavelet decomposition, we show that the V(Z) response of the SSPM for red (632.8 nm) and near IR (1550 nm) lights includes the frequential response of plasmon resonance together with additional parasitic frequencies induced by the objective pupil. Because the objective lens pupil profile is often unknown, this space-frequency decomposition turns out to be very useful to decipher the characteristic frequencies of the experimental V(Z) curves. Comparing the visible and near IR light responses of the SSPM, we show that our objective lens, primarily designed for visible light microscopy, is still operating very efficiently in near IR light. Actually, despite their loss in resolution, the SSPM images obtained with near IR light remain contrasted for a wider range of defocus values from negative to positive Z values. We illustrate our theoretical modeling with a preliminary experimental application to blood cell imaging.

Amplitude and phase images of cellular structures with a scanning surface plasmon microscope.

Imaging cellular internal structure at nanometer scale axial resolution with non invasive microscopy techniques has been a major technical challenge since the nineties. We propose here a complement to fluorescence based microscopies with no need of staining the biological samples, based on a Scanning Surface Plasmon Microscope (SSPM). We describe the advantages of this microscope, namely the possibility of both amplitude and phase imaging and, due to evanescent field enhancement by the surface plasmon resonance, a very high resolution in Z scanning (Z being the axis normal to the sample). We show for fibroblast cells (IMR90) that SSPM offers an enhanced detection of index gradient regions, and we conclude it is very well suited to discriminate regions of variable density in biological media such as cell compartments, nucleus, nucleoli and membranes.

Selection rules for the tip-splitting instability.

The local destabilization of a Saffman-Taylor viscous finger occurs by a splitting of its tip and results in the formation of two branches separated by a fjord. The accumulation of such instabilities leads to complex patterns. In this paper we present a detailed analysis of a dynamical model that accounts for the selection of both the width and the orientation of the fjords growing in a wedge of angle theta(0). It is shown that the selection rules have a dynamical origin and are related to the existence of attracting sets that disappear in the absence of surface tension. We also infer the existence of a critical angle theta(c)=60 degrees such that if theta(0)

Exploring the natural conformational changes of the C-terminal domain of calmodulin.

Several experimental results suggest that the Ca2+-loaded C-terminal domain of calmodulin (or some of its mutants) exhibits conformational changes triggered solely by thermal fluctuations. The time scales involved are in the 10(-6)-10(-3) s range. Here we develop a theoretical method to explore this type of motions based on a modified version of molecular dynamics algorithm where the secondary structure motifs are held fixed. In this version, increasing the temperature enhances the sampling of conformations with locally fixed secondary structures. From the temperature dependence of the transition rate between various conformational states, we obtain characteristic times that are consistent with those observed experimentally.

Dense branching morphology in electrodeposition experiments: characterization and mean-field modeling

Dense branching morphologies (DBM) obtained in thin gap electrodeposition cells are characterized by a dense array of branches behind a flat advancing envelope. In this Letter, we show the existence in DBM of a new (porous) phase, qualitatively different from a (compact) metal deposit. The local porosity inside the branches is found to be much more robust than geometric characteristics such as the width or the distance between branches. This fact seems to be unreported in previous modeling of DBM. A mean-field model is proposed that displays overall features observed in the experiments, such as concentration profiles, front velocity, and branched internal structure.

Internal structure of dense electrodeposits

We report experimental investigations of the structure of dense patterns obtained during electrochemical deposition of copper in thin cells. The deposit correlation function reveals the periodic structuration of the patterns but shows that the primary spacing is not steady during the growth and that moreover it is not simply related to the diffusion length. Another measurable quantity is the occupancy ratio of the fingers in the cell. Its variation as a function of the experimental parameters is interpreted from specific properties of electrochemical growth. The results are discussed with respect to the well-known behavior of cellular solidification fronts.

Modeling large-scale dynamics of proteins.

We study a dynamical model for the large-scale motions of bovine pancreatic trypsin inhibitor in vacuum. The model is obtained by projecting Newton equations onto some set of anharmonic modes. We compare the statistics of the so-obtained trajectories with those obtained by standard techniques, and conclude that our dynamical model is able to reproduce fairly well the average properties of the large-scale motions of this protein. Moreover, it allows for time steps one order of magnitude larger than the standard ones.