Effect of curvature on the diffusion of colloidal bananas.

Anisotropic colloidal particles exhibit complex dynamics which play a crucial role in their functionality, transport, and phase behavior. In this Letter, we investigate the two-dimensional diffusion of smoothly curved colloidal rods-also known as colloidal bananas-as a function of their opening angle α. We measure the translational and rotational diffusion coefficients of the particles with opening angles ranging from 0^{∘} (straight rods) to nearly 360^{∘}(closed rings). In particular, we find that the anisotropic diffusion of the particles varies nonmonotonically with their opening angle and that the axis of fastest diffusion switches from the long to the short axis of the particles when α>180^{∘}. We also find that the rotational diffusion coefficient of nearly closed rings is approximately an order of magnitude higher than that of straight rods of the same length. Finally, we show that the experimental results are consistent with slender body theory, indicating that the dynamical behavior of the particles arises primarily from their local drag anisotropy. These results highlight the impact of curvature on the Brownian motion of elongated colloidal particles, which must be taken into account when seeking to understand the behavior of curved colloidal particles.

Bonded straight and helical flagellar filaments form ultra-low-density glasses.

We study how the three-dimensional shape of rigid filaments determines the microscopic dynamics and macroscopic rheology of entangled semidilute Brownian suspensions. To control the filament shape we use bacterial flagella, which are microns-long helical or straight filaments assembled from flagellin monomers. We compare the dynamics of straight rods, helical filaments, and shape-diblock copolymers composed of seamlessly joined straight and helical segments. Caged by their neighbors, straight rods preferentially diffuse along their long axis, but exhibit significantly suppressed rotational diffusion. Entangled helical filaments escape their confining tube by corkscrewing through the dense obstacles created by other filaments. By comparison, the adjoining segments of the rod-helix shape-diblocks suppress both the translation and the corkscrewing dynamics. Consequently, the shape-diblock filaments become permanently jammed at exceedingly low densities. We also measure the rheological properties of semidilute suspensions and relate their mechanical properties to the microscopic dynamics of constituent filaments. In particular, rheology shows that an entangled suspension of shape rod-helix copolymers forms a low-density glass whose elastic modulus can be estimated by accounting for how shear deformations reduce the entropic degrees of freedom of constrained filaments. Our results demonstrate that the three-dimensional shape of rigid filaments can be used to design rheological properties of semidilute fibrous suspensions.

Structure of propagating high-stress fronts in a shear-thickening suspension.

We report direct measurements of spatially resolved stress at the boundary of a shear-thickening cornstarch suspension revealing persistent regions of high local stress propagating in the flow direction at the speed of the top boundary. The persistence of these propagating fronts enables precise measurements of their structure, including the profile of boundary stress measured by boundary stress microscopy (BSM) and the nonaffine velocity of particles at the bottom boundary of the suspension measured by particle image velocimetry (PIV). In addition, we directly measure the relative flow between the particle phase and the suspending fluid (fluid migration) and find the migration is highly localized to the fronts and changes direction across the front, indicating that the fronts are composed of a localized region of high dilatant pressure and low particle concentration. The magnitude of the flow indicates that the pore pressure difference driving the fluid migration is comparable to the critical shear stress for the onset of shear thickening. The propagating fronts fully account for the increase in viscosity with applied stress reported by the rheometer and are consistent with the existence of a stable jammed region in contact with one boundary of the system that generates a propagating network of percolated frictional contacts spanning the gap between the rheometer plates and producing strong localized dilatant pressure.

Synthesis of Rough Colloidal SU-8 Rods and Bananas via Nanoprecipitation.

Surface roughness plays an important role in determining the mechanical properties, wettability, and self-assembly in colloidal systems. In this work, we develop a simple and fast method to produce rough colloidal SU-8 rods, bananas, and spheres, via the nanoprecipitation of SU-8 in water. During this process, SU-8 nanospheres are absorbed onto the surface of the colloidal SU-8 particles and then cross-linked using UV-light. The size of the spherical asperities and the asperity density are controlled by the concentration of SU-8 used during the nanoprecipitation reaction. Fluorescent labeling of the rough SU-8 colloidal particles allows for their confocal imaging, which demonstrates their stability at high packing fractions. With these newly developed rough particles, we provide a colloidal model system that allows for studies addressing the impact of surface roughness on materials composed of anisotropic particles.

Dynamics and memory of boundary stresses in discontinuous shear thickening suspensions during oscillatory shear.

We report direct measurements of spatially resolved surface stresses of a dense suspension during large amplitude oscillatory shear (LAOS) in the discontinuous shear thickening regime using boundary stress microscopy. Consistent with previous studies, bulk rheology shows a dramatic increase in the complex viscosity above a frequency-dependent critical strain. We find that the viscosity increase is coincident with that appearance of large heterogeneous boundary stresses, indicative of the formation of transient solid-like phases (SLPs) on spatial scales large compared to the particle size. The critical strain for the appearance of SLPs is largely determined by the peak oscillatory stress, which depends on the peak shear rate and the frequency-dependent suspension viscosity. The SLPs dissipate and reform on each cycle, with a spatial pattern that is highly variable at low frequencies but remarkably persistent at the highest frequency measured (ω = 10 rad s-1).

Effective aspect ratio of helices in shear flow.

We report the results of simulations of rigid colloidal helices suspended in a shear flow, using dissipative particle dynamics for a coarse-grained representation of the suspending fluid, as well as deterministic trajectories of non-Brownian helices calculated from the resistance tensor derived under the slender-body approximation. The shear flow produces nonuniform rotation of the helices, similarly to other high aspect ratio particles, such that more elongated helices spend more time aligned with the fluid velocity. We introduce a geometric effective aspect ratio calculated directly from the helix geometry and a dynamical effective aspect ratio derived from the trajectories of the particles and find that the two effective aspect ratios are approximately equal over the entire parameter range tested. We also describe observed transient deflections of the helical axis into the vorticity direction that can occur when the helix is rotating through the gradient direction and that depend on the rotation of the helix about its axis.

Role of particle orientational order during shear thickening in suspensions of colloidal rods.

Rheology of dense anisotropic colloidal suspensions often exhibits unsteady flow at constant imposed shear stress and/or shear rate. Using simultaneous high-resolution confocal microscopy and rheology, we find that the temporal behavior arises due to a strong coupling between shear flow and particle orientation. At smaller applied stresses, the orientation of rods fluctuates around the flow direction. A transition to an intermittent disordered state is observed at higher stresses when the angle between the flow and the rod orientation reaches a critical value. This disordered state is associated with transient drop in shear rate and an increase in viscosity. Simultaneous visualization of boundary stresses and orientation shows that the disordered regions lead to heterogeneous stresses and positive normal forces at the boundary, indicating the formation of systems spanning disordered particle contact networks.

Localized stress fluctuations drive shear thickening in dense suspensions.

Dense particulate suspensions exhibit a dramatic increase in average viscosity above a critical, material-dependent shear stress. This thickening changes from continuous to discontinuous as the concentration is increased. Using direct measurements of spatially resolved surface stresses in the continuous thickening regime, we report the existence of clearly defined dynamic localized regions of substantially increased stress that appear intermittently at stresses above the critical stress. With increasing applied stress, these regions occupy an increasing fraction of the system, and the increase accounts quantitatively for the observed shear thickening. The regions represent high-viscosity fluid phases, with a size determined by the distance between the shearing surfaces and a viscosity that is nearly independent of shear rate but that increases rapidly with concentration. Thus, we find that continuous shear thickening arises from increasingly frequent localized discontinuous transitions between distinct fluid phases with widely differing viscosities.

Live Cell Imaging and 3D Analysis of Angiotensin Receptor Type 1a Trafficking in Transfected Human Embryonic Kidney Cells Using Confocal Microscopy.

Live-cell imaging is used to simultaneously capture time-lapse images of angiotensin type 1a receptors (AT1aR) and intracellular compartments in transfected human embryonic kidney-293 (HEK) cells following stimulation with angiotensin II (Ang II). HEK cells are transiently transfected with plasmid DNA containing AT1aR tagged with enhanced green fluorescent protein (EGFP). Lysosomes are identified with a red fluorescent dye. Live-cell images are captured on a laser scanning confocal microscope after Ang II stimulation and analyzed by software in three dimensions (3D, voxels) over time. Live-cell imaging enables investigations into receptor trafficking and avoids confounds associated with fixation, and in particular, the loss or artefactual displacement of EGFP-tagged membrane receptors. Thus, as individual cells are tracked through time, the subcellular localization of receptors can be imaged and measured. Images must be acquired sufficiently rapidly to capture rapid vesicle movement. Yet, at faster imaging speeds, the number of photons collected is reduced. Compromises must also be made in the selection of imaging parameters like voxel size in order to gain imaging speed. Significant applications of live-cell imaging are to study protein trafficking, migration, proliferation, cell cycle, apoptosis, autophagy and protein-protein interaction and dynamics, to name but a few.

Torsional stiffness determines aggregate structure in sheared colloidal rod suspensions.

We report the results of simulations of suspensions of sheared rigid rods in the presence of strong attractive inter-particle interactions, using dissipative particle dynamics for a coarse-grained representation of the suspending fluid. We find that the combined effect of the attractive interactions and shear-induced alignment generically produces aggregates of aligned bundles when the contact points between rods are free to rotate. However, the introduction of substantial torsional stiffness to the inter-particle contacts recapitulates the disordered aggregates often observed in suspensions of high aspect ratio particles. We show that the degree of alignment within the aggregates depends on the strength of the torsional stiffness, while the stability of the aggregates and their impact on system viscosity depend on the competition between the attractive interaction and the shear stresses.

Traction force and tension fluctuations in growing axons.

Actively generated mechanical forces play a central role in axon growth and guidance, but the mechanisms that underly force generation and regulation in growing axons remain poorly understood. We report measurements of the dynamics of traction stresses from growth cones of actively advancing axons from postnatal rat DRG neurons. By tracking the movement of the growth cone and analyzing the traction stress field from a reference frame that moves with it, we are able to show that there is a clear and consistent average stress field that underlies the complex spatial stresses present at any one time. The average stress field has strong maxima on the sides of the growth cone, directed inward toward the growth cone neck. This pattern represents a contractile stress contained within the growth cone, and a net force that is balanced by the axon tension. Using high time-resolution measurements of the growth cone traction stresses, we show that the stress field is composed of fluctuating local stress peaks, with a large number peaks that live for a short time, a population of peaks whose lifetime distribution follows an exponential decay, and a small number of very long-lived peaks. We show that the high time-resolution data also reveal that the tension appears to vary randomly over short time scales, roughly consistent with the lifetime of the stress peaks, suggesting that the tension fluctuations originate from stochastic adhesion dynamics.

Stress heterogeneities in sheared type-I collagen networks revealed by Boundary Stress Microscopy.

Disordered fiber networks provide structural support to a wide range of important materials, and the combination of spatial and dynamic complexity may produce large inhomogeneities in mechanical properties, an effect that is largely unexplored experimentally. In this work, we introduce Boundary Stress Microscopy to quantify the non-uniform surface stresses in sheared collagen gels. We find local stresses exceeding average stresses by an order of magnitude, with variations over length scales much larger than the network mesh size. The strain stiffening behavior observed over a wide range of network mesh sizes can be parameterized by a single characteristic strain and associated stress, which describes both the strain stiffening regime and network yielding. The characteristic stress is approximately proportional to network density, but the peak boundary stress at both the characteristic strain and at yielding are remarkably insensitive to concentration.

Shear-driven aggregation of SU-8 microrods in suspension.

A non-Brownian suspension of micron scale rods exhibits reversible shear-driven formation of disordered aggregates resulting in dramatic viscosity enhancement at low shear rates. Aggregate formation is imaged using a combined rheometer and fluorescence microscope. The size and structure of these aggregates are found to be a function of shear rate and concentration, with larger aggregates present at lower shear rates and higher concentrations. Quantitative measurements of the early-stage aggregation process are modeled by collision driven growth of porous structures which suggest that the aggregate density increases with shear rate. This result is combined with a Krieger-Dougherty type constitutive relationship and steady-state viscosity measurements to estimate the intrinsic viscosity of complex structures developed under shear. These results represent a direct, quantitative, experimental demonstration of the association between aggregation and viscosity enhancement for a rod suspension, and demonstrate a way of inferring microscopic geometric properties of a dynamic system through the combination of quantitative imaging and rheology.

The effects of confinement on neuronal growth cone morphology and velocity.

Optimizing growth cone guidance through the use of patterned substrates is important for designing regenerative substrates to aid in recovery from neuronal injury. Using laser ablation, we designed micron-scale patterns capable of confining dissociated mouse cerebellar granule neuron growth cones to channels of different widths ranging from 1.5 to 12 µm. Growth cone dynamics in these channels were observed using time-lapse microscopy. Growth cone area was decreased in channels between 1.5 and 6 µm as compared to that in 12 µm and unpatterned substrates. Growth cone aspect ratio was also affected as narrower channels forced growth cones into a narrow, elongated shape. There was no difference in the overall rate of growth cone advance in uniform channels between 1.5 and 12 µm as compared to growth on unpatterned substrates. The percentage of time growth cones advanced, paused, and retracted was also similar. However, growth cones did respond to changes in confinement: growth cones in narrow lanes rapidly sped up when encountering a wide region and then slowed down as they entered another narrow region. Our results suggest that the rate of neurite extension is not affected by the degree of confinement, but does respond to changes in confinement.

Optical and physicochemical characterization of the luminous mucous secreted by the marine worm Chaetopterus sp.

Bioluminescence of the marine worm Chaetopterus variopedatus was first investigated several decades ago mainly using tissue extract. Light production of the worm, however, originates from a secreted mucus only. Here, we report the optical and physicochemical properties of the luminous mucus. We show that the produced light occurs as a long glow in the blue range (455 nm), which is an unusual color for a shallow benthic invertebrate. We also show that the light originates from a photoprotein whose light production is independent of molecular oxygen yet somewhat related to the physicochemical (rheological) characteristics of the mucus itself. Indeed, the mucus seems to polymerize and become more viscous on exposure to H2O2, which in turn seems to inhibit the light production. Ferrous iron was not associated with any strong stimulatory effect. This is in contrast to past studies on worm tissues showing that the light production is strongly stimulated by H2O2 and ferrous iron. Overall, our results highlight the fact that working on the luminous mucus only (vs. worm tissues) provides the ability to study its chemical properties possibly involved in the fine control of light production-as well as its rheological properties-and identify the possible interactions between these two properties.

Strength in the periphery: growth cone biomechanics and substrate rigidity response in peripheral and central nervous system neurons.

There is now considerable evidence of the importance of mechanical cues in neuronal development and regeneration. Motivated by the difference in the mechanical properties of the tissue environment between the peripheral (PNS) and central (CNS) nervous systems, we compare substrate-stiffness-dependent outgrowth and traction forces from PNS (dorsal root ganglion (DRG)) and CNS (hippocampal) neurons. We show that neurites from DRG neurons display maximal outgrowth on substrates with a Young's modulus of ∼1000 Pa, whereas hippocampal neurite outgrowth is independent of substrate stiffness. Using traction force microscopy, we also find a substantial difference in growth cone traction force generation, with DRG growth cones exerting severalfold larger forces compared with hippocampal growth cones. The traction forces generated by DRG and hippocampal growth cones both increase with increasing stiffness, and DRG growth cones growing on substrates with a Young's modulus of 1000 Pa strengthen considerably after 18-30 h. Finally, we find that retrograde actin flow is almost three times faster in hippocampal growth cones than in DRG. Moreover, the density of paxillin puncta is significantly lower in hippocampal growth cones, suggesting that stronger substrate coupling of the DRG cytoskeleton is responsible for the remarkable difference in traction force generation. These findings reveal a differential adaptation of cytoskeletal dynamics to substrate stiffness in growth cones of different neuronal types, and highlight the potential importance of the mechanical properties of the cellular environment for neuronal navigation during embryonic development and nerve regeneration.

Size-dependent rheology of type-I collagen networks.

We investigate the system size-dependent rheological response of branched type I collagen gels. When subjected to a shear strain, the highly interconnected mesh dynamically reorients, resulting in overall stiffening of the network. When a continuous shear strain is applied to a collagen network, we observe that the local apparent modulus, in the strain-stiffening regime, is strongly dependent on the gel thickness. In addition, we demonstrate that the overall network failure is determined by the ratio of the gel thickness to the mesh size. These findings have broad implications for cell-matrix interactions, the interpretation of rheological tissue data, and the engineering of biomimetic scaffolds.

Event ordering in live-cell imaging determined from temporal cross-correlation asymmetry.

We use the temporal asymmetry of the cross-correlation function to determine the temporal ordering of spatially localized cellular events in live-cell multichannel fluorescence imaging. The analysis is well suited to noisy, stochastic systems where the temporal order may not be apparent in the raw data. The approach is applicable to any biochemical reaction not in chemical equilibrium, including protein complex assembly, sequential enzymatic processes, gene regulation, and other cellular signaling events. As an automated quantitative measure, this approach allows the data to be readily interpreted statistically with minimal subjective biases. We first test the technique using simulations of simple biophysical models with a definite temporal ordering. We then demonstrate the approach by extracting the temporal ordering of three proteins-actin, sorting nexin 9, and clathrin-in the endocytic pathway.