We directly probe the microscopic structure, connectivity, and elasticity of colloidal gels using confocal microscopy. We show that the gel is a random network of one-dimensional chains of particles. By measuring thermal fluctuations, we determine the effective spring constant between pairs of particles as a function of separation; this is in agreement with the theory for fractal chains. Long-range attractions between particles lead to freely rotating bonds, and the gel is stabilized by multiple connections among the chains. By contrast, short-range attractions lead to bonds that resist bending, with dramatically suppressed formation of loops of particles.

Material moduli of the cytoskeleton (CSK) influence a wide range of cell functions(1-3). There is substantial evidence from reconstituted F-actin gels that a regime exists in which the moduli scale with frequency with a universal exponent of 3/4. Such behaviour is entropic in origin and is attributable to fluctuations in semi flexible polymers driven by thermal forces(4-7), but it is not obvious a priori that such entropic effects are responsible for the elasticity of the CSK. Here we demonstrate the existence of such a regime in the living cell, but only at high frequencies. Fast events scaled with frequency in a manner comparable to semi flexible-polymer dynamics, but slow events scaled with a non-universal exponent that was systematically smaller than 3/4 and probably more consistent with a soft-glass regime(8,9). These findings strongly suggest that at smaller timescales elasticity arises from entropic fluctuations of a semi flexible-filament network, whereas on longer timescales slow (soft-glass-like) dynamics of a different origin prevail. The transition between these two regimes occurred on timescales of the order of 0.01 s, thus setting within the slow glassy regime cellular events such as spreading, crawling, contracting, and invading.

We use confocal microscopy to visualize individual particles near the colloidal glass transition. We identify the most slowly-relaxing particles and show that they form spatially correlated clusters that percolate across the sample. In supercooled fluids, the largest cluster spans the system on short time scales but breaks up on longer time scales. In contrast, in glasses, a percolating cluster exists on all accessible time scales. Using molecular dynamics simulation, we show that these clusters make the dominant contribution to the bulk elasticity of the sample.

We study dense colloidal crystals under oscillatory shear using a confocal microscope. At large strains the crystals yield and the suspensions form shear bands. The pure harmonic response exhibited by the suspension rules out the applicability of nonlinear rheology models typically used to describe shear banding in other types of complex fluids. Instead, we show that a model based on the coexistence of linearly responding phases of the colloidal suspension accounts for the observed flows. These results highlight a new use of oscillatory measurements in distinguishing the contribution of linear and nonlinear local rheology to a globally nonlinear material response.

Cytoskeletal microtubules have been proposed to influence cell shape and mechanics based on their ability to resist large-scale compressive forces exerted by the surrounding contractile cytoskeleton. Consistent with this, cytoplasmic microtubules are often highly curved and appear buckled because of compressive loads. However, the results of in vitro studies suggest that microtubules should buckle at much larger length scales, withstanding only exceedingly small compressive forces. This discrepancy calls into question the structural role of microtubules, and highlights our lack of quantitative knowledge of the magnitude of the forces they experience and can withstand in living cells. We show that intracellular microtubules do bear large-scale compressive loads from a variety of physiological forces, but their buckling wavelength is reduced significantly because of mechanical coupling to the surrounding elastic cytoskeleton. We quantitatively explain this behavior, and show that this coupling dramatically increases the compressive forces that microtubules can sustain, suggesting they can make a more significant structural contribution to the mechanical behavior of the cell than previously thought possible.

We demonstrate a high-throughput drop sorter for microfluidic devices that uses dielectrophoretic forces. Microelectrodes underneath a polydimethylsiloxane channel produce forces of more than 10 nN on a water drop in an inert oil, resulting in sorting rates greater than 1.6 kHz. We investigate the dependence of such forces on drop size and flow. Alternate designs with electrodes on either side of a symmetric channel Y junction provide refined control over droplet selection.

The use of microfluidic devices to control drops of water in a carrier oil is a promising means of performing biological and chemical assays. An essential requirement for this is the controlled coalescence of pairs of drops to mix reagents together. We show that this can be accomplished through electrocoalescence of drops synchronized by size-dependent flow in microfluidic channels. Smaller drops move faster due to the Poiseuille flow, allowing pairs of surfactant-stabilized drops to be brought into contact where they are coalesced with an electric field. We apply this method to an enzyme assay to measure enzyme kinetic constants.

Suspensions of carbon black in oil, stabilized with adsorbed polyisobutylene succinimide (PIBSI) dispersant, are commonly used as model systems for investigating the soot-handling characteristics of motor oils. The structure of the carbon-black agglomerates changes dramatically with temperature; this results in a concomitant change in the suspension rheology. Linear and nonlinear rheological experiments indicate a large increase of the interparticle attractions as the temperature is raised. To elucidate the origin of this behavior, we investigate the effect of temperature on the stabilizing effect of the dispersant. Measurements of adsorption isotherms of the dispersant on carbon black indicate that there is little variation of the binding energy with temperature. Intrinsic viscosity measurements of PIBSI dispersants in solution clearly exhibit an inverse dependence of the dispersant chain dimension with temperature. These results suggest that the temperature-dependent changes in the chain conformation of the PIBSI dispersant are primarily responsible for the changes in the dispersion rheology, and we propose a simple model to account for these data.

Cytoplasmic extracts prepared from Xenopus laevis eggs are used for the reconstitution of a wide range of processes in cell biology, and offer a unique environment in which to investigate the role of cytoplasmic mechanics without the complication of preorganized cellular structures. As a step toward understanding the mechanical properties of this system, we have characterized the rheology of crude interphase extracts. At macroscopic length scales, the extract forms a soft viscoelastic solid. Using a conventional mechanical rheometer, we measure the elastic modulus to be in the range of 2 - 10 Pa, and loss modulus in the range of 0.5 - 5 Pa. Using pharmacological and immunological disruption methods, we establish that actin. laments and microtubules cooperate to give mechanical strength, whereas the intermediate. lament cytokeratin does not contribute to viscoelasticity. At microscopic length scales smaller than the average network mesh size, the response is predominantly viscous. We use multiple particle tracking methods to measure the thermal fluctuations of 1 mum embedded tracer particles, and measure the viscosity to be similar to20 mPa-s. We explore the impact of rheology on actin-dependent cytoplasmic contraction, and find that although microtubules modulate contractile forces in vitro, their interactions are not purely mechanical.

Double emulsions are highly structured fluids consisting of emulsion drops that contain smaller droplets inside. Although double emulsions are potentially of commercial value, traditional fabrication by means of two emulsification steps leads to very ill-controlled structuring. Using a microcapillary device, we fabricated double emulsions that contained a single internal droplet in a coreshell geometry. We show that the droplet size can be quantitatively predicted from the flow profiles of the fluids. The double emulsions were used to generate encapsulation structures by manipulating the properties of the fluid that makes up the shell. The high degree of control afforded by this method and the completely separate fluid streams make this a flexible and promising technique.

Minute concentrations of suspended particles can dramatically alter the behavior of a drying droplet. After a period of isotropic shrinkage, similar to droplets of a pure liquid, these droplets suddenly buckle like an elastic shell. While linear elasticity is able to describe the morphology of the buckled droplets, it fails to predict the onset of buckling. Instead, we find that buckling is coincident with a stress-induced fluid to solid transition in a shell of particles at a droplet's surface, occurring when attractive capillary forces overcome stabilizing electrostatic forces between particles.

Colloid-polymer mixtures can undergo spinodal decomposition into colloid-rich and colloid-poor regions. Gelation results when interconnected colloid-rich regions solidify. We show that this occurs when these regions undergo a glass transition, leading to dynamic arrest of the spinodal decomposition. The characteristic length scale of the gel decreases with increasing quench depth, and the nonergodicity parameter exhibits a pronounced dependence on scattering vector. Mode coupling theory gives a good description of the dynamics, provided we use the full static structure as input.

We present a unified framework for understanding the compaction of colloidal gels under their own weight. The dynamics of the collapse are determined by the value of the gravitational stress σ(g), as compared to the yield stress σ(Y) of the network. For σ(g)<σ(Y), gels collapse poroelastically, and their rate of compression decays exponentially in time. For σ(g)>σ(Y), the network eventually yields, leading to rapid settling. In both cases, the rate of collapse is backflow limited, while its overall magnitude is determined by a balance between gravitational stress and network elastic stress.

Colloidal silica gels are shown to stiffen with time, as demonstrated by both dynamic light scattering and bulk rheological measurements. Their elastic moduli increase as a power law with time, independent of particle volume fraction; however, static light scattering indicates that there are no large-scale structural changes. We propose that increases in local elasticity arising from bonding between neighboring colloidal particles can account for the strengthening of the network, while preserving network structure.

Diblock copolymers are known to spontaneously organize into polymer vesicles. Typically, this is achieved through the techniques of film rehydration or electroformation. We present a new method for generating polymer vesicles from double emulsions. We generate precision water-in-oil-in-water double emulsions from the breakup of concentric fluid streams; the hydrophobic fluid is a volatile mixture of organic solvent that contains dissolved diblock copolymers. We collect the double emulsions and slowly evaporate the organic solvent, which ultimately directs the self-assembly of the dissolved diblock copolymers into vesicular structures. Independent control over all three fluid streams enables precision assembly of polymer vesicles and provides for highly efficient encapsulation of active ingredients within the polymerosomes. We also use double emulsions with several internal drops to form new polymerosome structures.

We study the growth and invasion of glioblastoma multiforme (GBM) in three-dimensional collagen I matrices of varying collagen concentration. Phase-contrast microscopy studies of the entire GBM system show that invasiveness at early times is limited by available collagen fibers. At early times, high collagen concentration correlates with more effective invasion. Conversely, high collagen concentration correlates with inhibition in the growth of the central portion of GBM, the multicellular tumor spheroid. Analysis of confocal reflectance images of the collagen matrices quantifies how the collagen matrices differ as a function of concentration. Studying invasion on the length scale of individual invading cells with a combination of confocal and coherent anti-Stokes Raman scattering microscopy reveals that the invasive GBM cells rely heavily on cell-matrix interactions during invasion and remodeling.

While the important role of electrostatic interactions in aqueous colloidal suspensions is widely known and reasonably well-understood, their relevance to nonpolar suspensions remains mysterious. We measure the interaction potentials of colloidal particles in a nonpolar solvent with reverse micelles. We find surprisingly strong electrostatic interactions characterized by surface potentials, vertical bar e zeta vertical bar, from 2.0 to 4.4 k(B)T and screening lengths, kappa(-1), from 0.2 to 1.4 mu m. Interactions depend on the concentration of reverse micelles and the degree of confinement. Furthermore, when the particles are weakly confined, the values of vertical bar e zeta vertical bar and kappa extracted from interaction measurements are consistent with bulk measurements of conductivity and electrophoretic mobility. A simple thermodynamic model, relating the structure of the micelles to the equilibrium ionic strength, is in good agreement with both conductivity and interaction measurements. Since dissociated ions are solubilized by reverse micelles, the entropic incentive to charge a particle surface is qualitatively changed from aqueous systems, and surface entropy plays an important role.

We construct shells with tunable morphology and mechanical response with colloidal particles that self-assemble at the interface of emulsion droplets. Particles self-assemble to minimize the total interfacial energy, spontaneously forming a particle layer that encapsulates the droplets. We stabilize these layers to form solid shells at the droplet interface by aggregating the particles, connecting the particles with adsorbed polymer, or fusing the particles. These techniques reproducibly yield shells with controllable properties such as elastic moduli and breaking forces. To enable diffusive exchange through the particle shells, we transfer them into solvents that are miscible with the encapsulant. We characterize the mechanical properties of the shells by measuring the response to deformation by calibrated microcantilevers.

Calcite crystals often nucleate and grow in solutions of calcium carbonate, and these crystallites can become trapped at the air water interface, where they form unusual structures. The most common is a fractal structure, which can extend over a large fraction of the interface, and whose origin is understood in terms of the aggregation of the particles. Much more rarely, a different and entirely unexpected structure is observed: the particles remain well separated on the interface, forming an ordered phase reminiscent of a two-dimensional colloidal crystal. The structure of the crystallites that form this ordered phase is always observed to be tetrahedral, in contrast to the much more common rhombohedral structure of the crystallites that form the fractal phase. We show that the interparticle interaction potential that leads to this ordered phase is a balance between a long-range attractive interaction and a long-range repulsive interaction. The attraction results from gravity-induced capillary forces, while the repulsion results from a dipole-dipole interaction due to the charged surface of the tetrahedral crystals. The interaction potential is estimated from the thermal motion of the particles, and fits to the theoretically expected values suggest that the effective surface charge on the tetrahedral crystals is sigma similar to 0.01 charges/nm(2).