Jacinta C. Conrad

Bacteria Use Type IV Pili to Walk Upright and Detach from Surfaces

A searchable database of images allows detailed analysis of bacterial motility. Bacterial biofilms are structured multicellular communities involved in a broad range of infections. Knowing how free-swimming bacteria adapt their motility mechanisms near surfaces is crucial for understanding the transition between planktonic and biofilm phenotypes. By translating microscopy movies into searchable databases of bacterial behavior, we identified fundamental type IV pili–driven mechanisms for Pseudomonas aeruginosa surface motility involved in distinct foraging strategies. Bacteria stood upright and “walked” with trajectories optimized for two-dimensional surface exploration. Vertical orientation facilitated surface detachment and could influence biofilm morphology.

Flagella and Pili-Mediated Near-Surface Single-Cell Motility Mechanisms in P. aeruginosa

Bacterial biofilms are structured multicellular communities that are responsible for a broad range of infections. Knowing how free-swimming bacteria adapt their motility mechanisms near a surface is crucial for understanding the transition from the planktonic to the biofilm phenotype. By translating microscopy movies into searchable databases of bacterial behavior and developing image-based search engines, we were able to identify fundamental appendage-specific mechanisms for the surface motility of Pseudomonas aeruginosa. Type IV pili mediate two surface motility mechanisms: horizontally oriented crawling, by which the bacterium moves lengthwise with high directional persistence, and vertically oriented walking, by which the bacterium moves with low directional persistence and high instantaneous velocity, allowing it to rapidly explore microenvironments. The flagellum mediates two additional motility mechanisms: near-surface swimming and surface-anchored spinning, which often precedes detachment from a surface. Flagella and pili interact cooperatively in a launch sequence whereby bacteria change orientation from horizontal to vertical and then detach. Vertical orientation facilitates detachment from surfaces and thereby influences biofilm morphology.

Glasslike arrest in spinodal decomposition as a route to colloidal gelation

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.

Diffusive Dynamics of Nanoparticles in Arrays of Nanoposts

The diffusive dynamics of dilute dispersions of nanoparticles of diameter 200-400 nm were studied in microfabricated arrays of nanoposts using differential dynamic microscopy and single particle tracking. Posts of diameter 500 nm and height 10 μm were spaced by 1.2-10 μm on a square lattice. As the spacing between posts was decreased, the dynamics of the nanoparticles slowed. Moreover, the dynamics at all length scales were best represented by a stretched exponential rather than a simple exponential. Both the relative diffusivity and the stretching exponent decreased linearly with increased confinement and, equivalently, with decreased void volume. The slowing of the overall diffusive dynamics and the broadening distribution of nanoparticle displacements with increased confinement are consistent with the onset of dynamic heterogeneity and the approach to vitrification.

Physics of bacterial near-surface motility using flagella and type IV pili: implications for biofilm formation

We review physically-motivated studies of bacterial near-surface motility driven by flagella and type IV pili (TfP) in the context of biofilm formation. We describe the motility mechanisms that individual bacteria deploying flagella and TfP use to move on and near surfaces, and discuss how the interactions of motility appendages with fluid and surfaces promote motility, attachment and dispersal of bacteria on surfaces prior to biofilm formation.

Arrested fluid-fluid phase separation in depletion systems: Implications of the characteristic length on gel formation and rheology

We investigate the structural, dynamical, and rheological properties of colloid-polymer mixtures in a volume fraction range of ϕ=0.15–0.35. Our systems are density-matched, residual charges are screened, and the polymer-colloid size ratio is ∼0.37. For these systems, the transition to kinetically arrested states, including disconnected clusters and gels, coincides with the fluid-fluid phase separation boundary. Structural investigations reveal that the characteristic length, L, of the networks is a strong function of the quench depth: for shallow quenches, L is significantly larger than that obtained for deep quenches. By contrast, L is for a given quench depth almost independent of ϕ; this indicates that the strand thickness increases with ϕ. The strand thickness determines the linear rheology: the final relaxation time exhibits a strong dependence on ϕ, whereas the high frequency modulus does not. We present a simple model based on estimates of the strand breaking time and shear modulus that semiquantit...

Structure of colloidal gels during microchannel flow.

We investigate the structure and flow behavior of colloidal gels in microchannels using confocal microscopy. Silica particles are first coated with a cationic polyelectrolyte and then flocculated by the addition of an anionic polyelectrolyte. In the quiescent state, the suspension is an isotropic and homogeneous gel. Under shear flow, the suspension contains dense clusters that yield at intercluster boundaries, resulting in network breakup at high shear rates. These structural changes coincide with a transition from pluglike flow at low pressures to fluidlike flow at high pressures.

Evaporative lithographic patterning of binary colloidal films

Evaporative lithography offers a promising new route for patterning a broad array of soft materials. In this approach, a mask is placed above a drying film to create regions of free and hindered evaporation, which drive fluid convection and entrained particles to regions of highest evaporative flux. We show that binary colloidal films exhibit remarkable pattern formation when subjected to a periodic evaporative landscape during drying.

Transport and Dispersion of Nanoparticles in Periodic Nanopost Arrays

Nanoparticles transported through highly confined porous media exhibit faster breakthrough than small molecule tracers. Despite important technological applications in advanced materials, human health, energy, and environment, the microscale mechanisms leading to early breakthrough have not been identified. Here, we measure dispersion of nanoparticles at the single-particle scale in regular arrays of nanoposts and show that for highly confined flows of dilute suspensions of nanoparticles the longitudinal and transverse velocities exhibit distinct scaling behaviors. The distributions of transverse particle velocities become narrower and more non-Gaussian when the particles are strongly confined. As a result, the transverse dispersion of highly confined nanoparticles at low Péclet numbers is significantly less important than longitudinal dispersion, leading to early breakthrough. This finding suggests a fundamental mechanism by which to control dispersion and thereby improve efficacy of nanoparticles applied for advanced polymer nanocomposites, drug delivery, hydrocarbon production, and environmental remediation.

Regulation of directional cell migration by membrane-induced actin bundling.

Summary During embryonic development and in metastatic cancers, cells detach from the epithelium and migrate with persistent directionality. Directional cell migration is also crucial for the regeneration and maintenance of the epithelium and impaired directional migration is linked to chronic inflammatory diseases. Despite its significance, the mechanisms controlling epithelial cell migration remain poorly understood. Villin is an epithelial-cell-specific actin modifying protein that regulates epithelial cell plasticity and motility. In motile cells villin is associated with the highly branched and the unbranched actin filaments of lamellipodia and filopodia, respectively. In this study we demonstrate for the first time that villin regulates directionally persistent epithelial cell migration. Functional characterization of wild-type and mutant villin proteins revealed that the ability of villin to self-associate and bundle actin as well as its direct interaction with phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] regulates villin-induced filopodial assembly and directional cell migration. Our findings suggest that convergence of different signaling cascades could spatially restrict villin activity to areas of high PtdIns(4,5)P2 and F-actin concentration to assemble filopodia. Furthermore, our data reveal the ability of villin to undergo actin- and PtdIns(4,5)P2-induced self-association, which may be particularly suited to coalesce and reorganize actin bundles within the filopodia.