Polymer Patterning by Laser-Induced Multipoint Initiation of Frontal Polymerization.

Frontal polymerization (FP) is an approach for thermosetting plastics at a lower energy cost than an autoclave. The potential to generate simultaneous propagation of multiple polymerization fronts has been discussed as an exciting possibility. However, FP initiated at more than two points simultaneously has not been demonstrated. Multipoint initiation could enable both large-scale material fabrication and unique pattern generation. Here, the authors present laser-patterned photothermal heating as a method for simultaneous initiation of FP at multiple locations in a 2-D sample. Carbon black particles are mixed into liquid resin (dicyclopentadiene) to enhance absorption of light from a Ti:sapphire laser (800 nm) focused on a sample. The laser is time-shared by rapid steering among initiation points, generating polymerization using up to seven simultaneous points of initiation. This process results in the formation of both symmetric and asymmetric seam patterns resulting from the collision of fronts. The authors also present and validate a theoretical framework for predicting the seam patterns formed by front collisions. This framework allows the design of novel patterns via an inverse solution for determining the initiation points required to form a desired pattern. Future applications of this approach could enable rapid, energy-efficient manufacturing of novel composite-like patterned materials.

Interparticle hydrogen bonding can elicit shear jamming in dense suspensions.

Dense suspensions of hard particles in a liquid can exhibit strikingly counter-intuitive behaviour, such as discontinuous shear thickening (DST)1-7 and reversible shear jamming (SJ) into a state where flow is arrested and the suspension is solid-like8-12. A stress-activated crossover from hydrodynamic interactions to frictional particle contacts is key for these behaviours2-4,6,7,9,13. However, in experiments, many suspensions show only DST, not SJ. Here we show that particle surface chemistry plays a central role in creating conditions that make SJ readily observable. We find the system's ability to form interparticle hydrogen bonds when sheared into contact elicits SJ. We demonstrate this with charge-stabilized polymer microspheres and non-spherical cornstarch particles, controlling hydrogen bond formation with solvents. The propensity for SJ is quantified by tensile tests12 and linked to an enhanced friction by atomic force microscopy. Our results extend the fundamental understanding of the SJ mechanism and open avenues for designing strongly non-Newtonian fluids.

A simple approach to spectrally resolved fluorescence and bright field microscopy over select regions of interest.

A standard wide field inverted microscope was converted to a spatially selective spectrally resolved microscope through the addition of a polarizing beam splitter, a pair of polarizers, an amplitude-mode liquid crystal-spatial light modulator, and a USB spectrometer. The instrument is capable of simultaneously imaging and acquiring spectra over user defined regions of interest. The microscope can also be operated in a bright-field mode to acquire absorption spectra of micron scale objects. The utility of the instrument is demonstrated on three different samples. First, the instrument is used to resolve three differently labeled fluorescent beads in vitro. Second, the instrument is used to recover time dependent bleaching dynamics that have distinct spectral changes in the cyanobacteria, Synechococcus leopoliensis UTEX 625. Lastly, the technique is used to acquire the absorption spectra of CH3NH3PbBr3 perovskites and measure differences between nanocrystal films and micron scale crystals.

Intracellular transport of insulin granules is a subordinated random walk.

We quantitatively analyzed particle tracking data on insulin granules expressing fluorescent fusion proteins in MIN6 cells to better understand the motions contributing to intracellular transport and, more generally, the means for characterizing systems far from equilibrium. Care was taken to ensure that the statistics reflected intrinsic features of the individual granules rather than details of the measurement and overall cell state. We find anomalous diffusion. Interpreting such data conventionally requires assuming that a process is either ergodic with particles working against fluctuating obstacles (fractional brownian motion) or nonergodic with a broad distribution of dwell times for traps (continuous-time random walk). However, we find that statistical tests based on these two models give conflicting results. We resolve this issue by introducing a subordinated scheme in which particles in cages with random dwell times undergo correlated motions owing to interactions with a fluctuating environment. We relate this picture to the underlying microtubule structure by imaging in the presence of vinblastine. Our results provide a simple physical picture for how diverse pools of insulin granules and, in turn, biphasic secretion could arise.

Controlling the position and orientation of single silver nanowires on a surface using structured optical fields.

We demonstrate controlled trapping and manipulation of single silver (Ag) nanowires in two dimensions at a surface using structured light fields generated with a spatial light modulator. The Ag nanowires are attracted toward the regions of maximal optical intensity along the surface when the trapping laser light is linearly polarized and are repelled toward the minima of optical intensity when the light is circularly polarized. For linearly polarized light, stably trapped nanowires are oriented perpendicular to the polarization direction due to a torque induced by an asymmetrical response of the nanowire to the electric field. The attractive interactions with linearly polarized trapping laser light, which is at 800 nm for all measurements, enable stable trapping and translation of Ag nanowires in the antinodes of optical gratings and in zero-order Bessel beams. Trapped nanowires can be positioned and oriented on a transparent dielectric substrate, making possible the nonmechanical assembly of plasmonic nanostructures for particular functions.

Three-dimensional optical trapping and manipulation of single silver nanowires.

We report the first experimental realization of all-optical trapping and manipulation of plasmonic nanowires in three dimensions. The optical beam used for trapping is the Fourier transform of a linearly polarized Bessel beam (termed FT-Bessel). The extended depth of focus of this beam enables the use of a retroreflection geometry to cancel radiation pressure in the beam propagation direction, making it possible to trap highly scattering and absorbing silver nanowires. Individual silver nanowires with lengths of several micrometers can be positioned by the trapping beam with a precision better than 100 nm and are oriented by the polarization of the trapping light with a precision of approximately 1°. Multiple nanowires can be trapped simultaneously in spatially separated maxima of the trapping field. Since trapping in the interferometric FT-Bessel potential is robust in bulk solution and near surfaces, it will enable the controlled assembly of metal nanowires into plasmonic nanostructures.

Isopeptide bonds of the major pilin protein BcpA influence pilus structure and bundle formation on the surface of Bacillus cereus.

Bacillus cereus strains elaborate pili on their surface using a mechanism of sortase-mediated cross-linking of major and minor pilus components. Here we used a combination of electron microscopy and atomic force microscopy to visualize these structures. Pili occur as single, double or higher order assemblies of filaments formed from monomers of the major pilin, BcpA, capped by the minor pilin, BcpB. Previous studies demonstrated that within assembled pili, four domains of BcpA - CNA(1), CNA(2), XNA and CNA(3) - each acquire intramolecular lysine-asparagine isopeptide bonds formed via catalytic glutamic acid or aspartic acid residues. Here we showed that mutants unable to form the intramolecular isopeptide bonds in the CNA(2) or CNA(3) domains retain the ability to form pilus bundles. A mutant lacking the CNA(1) isopeptide bond assembled deformed pilin subunits that failed to associate as bundles. X-ray crystallography revealed that the BcpA variant Asp(312) Ala, lacking an aspartyl catalyst, did not generate the isopeptide bond within the jelly-roll structure of XNA. The Asp(312) Ala mutant was also unable to form bundles and promoted the assembly of deformed pili. Thus, structural integrity of the CNA(1) and XNA domains are determinants for the association of pili into higher order bundle structures and determine native pilus structure.

Protein-protein docking and analysis reveal that two homologous bacterial adenylyl cyclase toxins interact with calmodulin differently.

Calmodulin (CaM), a eukaryotic calcium sensor that regulates diverse biological activities, consists of N- and C-terminal globular domains (N-CaM and C-CaM, respectively). CaM serves as the activator of CyaA, a 188-kDa adenylyl cyclase toxin secreted by Bordetella pertussis, which is the etiologic agent for whooping cough. Upon insertion of the N-terminal adenylyl cyclase domain (ACD) of CyaA to its targeted eukaryotic cells, CaM binds to this domain tightly ( approximately 200 pm affinity). This interaction activates the adenylyl cyclase activity of CyaA, leading to a rise in intracellular cAMP levels to disrupt normal cellular signaling. We recently solved the structure of CyaA-ACD in complex with C-CaM to elucidate the mechanism of catalytic activation. However, the structure of the interface between N-CaM and CyaA, the formation of which contributes a 400-fold increase of binding affinity between CyaA and CaM, remains elusive. Here, we used site-directed mutations and molecular dynamic simulations to generate several working models of CaM-bound CyaA-ACD. The validity of these models was evaluated by disulfide bond cross-linking, point mutations, and fluorescence resonance energy transfer experiments. Our study reveals that a beta-hairpin region (amino acids 259-273) of CyaA-ACD likely makes contacts with the second calcium binding motif of the extended CaM. This mode of interaction differs from the interaction of N-CaM with anthrax edema factor, which binds N-CaM via its helical domain. Thus, two structurally conserved, bacterial adenylyl cyclase toxins have evolved to utilize distinct binding surfaces and modes of activation in their interaction with CaM, a highly conserved eukaryotic signaling protein.

Stochastic scanning multiphoton multifocal microscopy.

Multiparticle tracking with scanning confocal and multiphoton fluorescence imaging is increasingly important for elucidating biological function, as in the transport of intracellular cargo-carrying vesicles. We demonstrate a simple rapid-sampling stochastic scanning multifocal multiphoton microscopy (SS-MMM) fluorescence imaging technique that enables multiparticle tracking without specialized hardware at rates 1,000 times greater than conventional single point raster scanning. Stochastic scanning of a diffractive optic generated 10x10 hexagonal array of foci with a white noise driven galvanometer yields a scan pattern that is random yet space-filling. SS-MMM creates a more uniformly sampled image with fewer spatio-temporal artifacts than obtained by conventional or multibeam raster scanning. SS-MMM is verified by simulation and experimentally demonstrated by tracking microsphere diffusion in solution.

Generation of optical vector beams with a diffractive optical element interferometer.

We present a novel approach to generating radially and azimuthally polarized vector beams that utilize an interferometer constructed from two identical diffractive optical elements. The measured polarization properties of four vector beam states and their phase relationships are in good agreement with theoretical expectations. This interferometer is passively phase stable and robust, making it suitable for linear and nonlinear optical (superresolution) microscopy.

Correlating second harmonic optical responses of single Ag nanoparticles with morphology.

Femtosecond laser excited second harmonic (SH) activity from single Ag nanoparticles is reported. A correlation of SH single-particle measurements with high-resolution imaging of particle morphology by TEM was achieved by creating position markers on an optical and electron transparent substrate (Si3N4 thin film, approximately 100 nm). We compared the SH activity of single Ag nanoparticles (nanospheres versus nanorods) and cluster structures (composed of two or multiple particles, e.g., dimers and trimers). The direct correlation of single-particle structures and SH activity, spectral and power dependence, strongly suggests one-photon resonant driven nonlinear oscillator response mechanism.