Session R35: Active Matter: Collective Phenomena in Living Systems III

Phase separation dynamics during \textit{Myxococcus xanthus} fruiting body formation

Many living systems take advantage of collective behavior for group survival. We use the soil-dwelling bacterium \textit{Myxococcus xanthus} as a model to study out-of-equilibrium phase separation during fruiting body formation. \textit{M. xanthus} cells have the ability to glide on solid surfaces and reverse their direction periodically. When starved, \textit{M. xanthus} cells aggregate together and form structures called fruiting bodies, inside of which cells sporulate to survive stressful conditions. We show that at high cell density the formation of fruiting bodies is a phase separation process. From experimental data that combines single-cell tracking, population-scale imaging, mutants, and drug applications, we construct the phase diagram of \textit{M. xanthus} in the space of P\'{e}clet number and cell density. When wild type cells are starved, we find that they actively increase their P\'{e}clet number by modulating gliding speed and reversal frequency which induces a phase separation from a gas-like state to an aggregated fruiting body state.   

Active matter model of Myxococcus xanthus aggregation

\emph{Myxococcus xanthus} is a soil-dwelling bacterium that exhibits several fascinating collective behaviors including streaming, swarming, and generation of fruiting bodies. A striking feature of \emph{M. xanthus} is that it periodically reverses its motility direction. The first stage of fruiting body formation is characterized by the aggregation of cells on a surface into round mesoscopic structures. Experiments have shown that this aggregation relies heavily on regulation of the reversal rate and local mechanical interactions, suggesting motility-induced phase separation may play an important role. We have adapted self-propelled particle models to include cell reversal and motility suppression resulting from sporulation observed in aggregates. Using 2D molecular dynamics simulations, we map the phase behavior in the space of P\'eclet number and local density and examine the kinetics of aggregation for comparison to experiments.   

Thermodynamics of the motility-induced phase separation

Self-propelled particles are known to accumulate in regions of space where their velocity is lowered. In addition, if their velocity diminishes when the local density increases (for example due to crowding effects), a positive feedback loop leads to the now well-established motility-induced phase separation (MIPS) between a dense immotile phase and a dilute motile phase. Understanding the phase equilibrium of MIPS is still a matter of debate. Although, depending on the models used to study the transition, a chemical potential or a pressure can be defined, these quantities do not play their usual thermodynamic role. In particular, the usual common tangent or equal-area constructions fail in these systems. Indeed, we will show that describing the phase equilibrium of MIPS necessitates generalized thermodynamics that include non-equilibrium contributions. This approach allows us to predict correctly the phase diagram of MIPS and to gain insight into the thermodynamics of active systems. It also sheds light on the (in)equivalence of statistical ensembles for these systems, paving the way for more efficient computational studies.   

Epithelial gap closure governed by forces and geometry.

The closure of gaps within epithelia is crucial to maintain the integrity and the homeostasis of the tissue during wound healing or cell extrusion processes. Cells mediate gap closure through either the assembly of multicellular actin-based contractile cables (purse-string contraction) or the protrusive activity of border cells into the gap (cell crawling). I will present experimental data and numerical modeling that show how these mechanisms can mutually interact to promote efficient epithelial gap closure and how mechanical constraints can regulate these mechanisms. I will first present how geometrical constraints dictate mechanisms of epithelial gap closure. We determine the importance of tissue shape during closure and the role of curvature of cell boundaries in this process. An essential difference between the two closure mechanisms is that cell crawling always pulls the edge of the tissue forward (i.e. towards the gap) while purse string pulls the edge forward or backwards depending on the local geometry. Our study demonstrates how the interplay between these two mechanisms is crucial for closing gaps and wounds, which naturally come in arbitrary shapes. Then I will focus on epithelial closure mechanism during cell extrusion. Within confluent cell layers, cellular motions coupled between neighbors are tightly regulated by the packing density of the epithelium inducing drastic changes in the dynamics of these tissues. I will show how cell density and tissue mechanics regulate the extrusion of cells within a confluent epithelial cell sheet, simultaneously measuring collective movements and traction forces. Epithelial packing and collective cell dynamics dictate the modes of cellular extrusions from lamellipodia crawling of the neighboring cells at low densities to coordinated actin-based contractile purse-string mechanism at higher density.   

Self-Driven Jamming of Growing Microbial Populations

When cells grow in confined spaces, they assemble into dense populations that interact both chemically and physically. Although in recent years scientists have uncovered a previously hidden layer of mechanical regulation in mammalian tissues that impacts gene expression and development, little is known about the consequences of mechanical constraints on single-celled microbes. This is largely due to a lack of appropriate culturing techniques and accurate computational models. Using physically explicit computer models that are developed alongside microfluidic experiments, we address two fundamental questions: (1) what structures self-assemble in confined geometries due to the cell growth and division process? and (2) how do those structures and associated stresses feed back on to cell physiology? We find that microbial growth in confinement can lead to jamming, heterogeneous stress fields, and intermittent flow that in turn result in spatially and temporally heterogeneous physiological responses. With computer simulations, we further explore the differences between this 'active' flow that is driven internally by cell growth and 'inactive' flow, such as shear and hopper flow, that is driven externally.   

Cell Size Clues for the Allee Effect in Vegetative Amoeba Suspension Culture

That cells proliferate at higher rates with increasing density helps us appreciate and understand the development of multicellular behavior through the study of dilute cell systems. However, arduous cell counting with a microscope reveals that in the model eukaryote, Dictyostelium discoideum this transition is difficult to ascertain and thereby further explore despite our earlier progress (Phys. Rev. E 77, 041905, (2008)). Here we report preliminary evidence that the slow proliferation phase is well characterized by reduced cell size compared to the wide distribution of cell sizes in the familiar exponential proliferation phase of moderate densities. This observation is enabled by a new system for characterizing cells in stirred suspension cultures. Our technique relies on quickly acquiring magnitude distributions of detected flashes of laser light scattered in situ by cell targets.   

Manipulation of long-term dynamics in a colloidal active matter system using speckle light fields

Particles undergoing a stochastic motion within a disordered medium is a ubiquitous physical and biological phenomena. Examples can be given from organelles performing tasks in the cytoplasm to large animals moving in patchy environment. Here, we use speckle light fields to study the anomalous diffusion in an active matter system consisting of micron-sized silica particles(diameter 5$\mu$m) and motile bacterial cells (E. coli). The speckle light fields are generated by mode mixing inside a multimode optical fiber where a small amount of incident laser power is needed to obtain an effective disordered optical landscape for the purpose of optical manipulation. We experimentally show how complex potentials contribute to the long-term dynamics of the active matter system and observed an enhanced diffusion of particles interacting with the active bacterial bath in the speckle light fields. We showed that this effect can be tuned and controlled by varying the intensity and the statistical properties of the speckle pattern. Potentially, these results could be of interest for many technological applications, such as the manipulation of microparticles inside optically disordered media of biological interest.   

A Density-Independent Flocking Transition in Confluent Tissues

Some of us recently demonstrated a density-independent solid-liquid transition in confluent tissues controlled by cell motility and a cell shape parameter measuring the interplay of cortical tension and cell-cell adhesion. An important insight of this work is that the rigidity and dynamics of cell layers depends sensitively on cell shape. To explore the influence of cell shape on collective states, we have constructed continuum equations that couple a scalar field describing cell-shape anisotropy to cell polarization. The model displays a density independent transition to a polarized state of elongated cells driven by a cellular “shape-index” parameter. We map out the phase diagram using linear stability analysis and numerical solution of the nonlinear hydrodynamic equations. The proposed transition constitutes a density-independent flocking transition.   

Multicellular contractility contributes to the emergence of mesothelioma nodules.

Malignant pleural mesothelioma (MPM) nodules arise from the mesothelial lining of the pleural cavity by a poorly understood mechanism. We demonstrate that macroscopic multicellular aggregates, reminiscent of the MPM nodules found in patients, develop when MPM cell lines are cultured at high cell densities for several weeks. Surprisingly, the nodule-like aggregates do not arise by excessive local cell proliferation, but by myosin II-driven cell contractility. Contractile nodules contain prominent actin cables that can span several cells. Several features of the in vitro MPM nodule development can be explained by a computational model that assumes uniform and steady intercellular contractile forces within a monolayer of cells, and a mechanical load-dependent lifetime of cell-cell contacts. The model behaves as a self-tensioned Maxwell fluid and exhibits an instability that leads to pattern formation. Altogether, our findings suggest that inhibition of the actomyosin system may provide a hitherto not utilized therapeutic approach to affect MPM growth.   

Cilia driven flow networks in the brain

Neurons exchange soluble substances via the cerebrospinal fluid (CSF) that fills the ventricular system. The walls of the ventricular cavities are covered with motile cilia that constantly beat and thereby induce a directional flow. We recently discovered that cilia in the third ventricle generate a complex flow pattern leading to partitioning of the ventricular volume and site-directed transport paths along the walls. Transient and daily recurrent alterations in the cilia beating direction lead to changes in the flow pattern. This has consequences for delivery of CSF components along the near wall flow. The contribution of this cilia-induced flow to overall CSF flow remains to be investigated. The state-of-art lattice Boltzmann method is adapted for studying the CFS flow. The 3D geometry of the third ventricle at high resolution was reconstructed. Simulation of CSF flow without cilia in this geometry confirmed that the previous idea about unidirectional flow does not explain how different components of CSF can be delivered to their various target sites. We study the contribution of the cilia-induced flow pattern to overall CSF flow and identify target areas for site-specific delivery of CSF-constituents with respect to the temporal changes. ~   

Collective motion of motile cilia: from human airways to model systems

Mammalian airways are a fantastic playground of nonlinear phenomena, from the function of individual active filaments, to the emerging collective behaviour, to the rheology of the mucus solution surrounding cilia. We have been investigating the fundamental physics of this system through a variety of model system approaches, both experimental and computational. In the last year we have started measurements on living human cells, observing cilia shape during beating, and measuring speed and coherence of the collective dynamics. We report on significant differences in the collective motion in ciliated cell carpets from a variety of diseases, and we attempt to reconcile the collective dynamical phenotypes to the properties of individual filaments and the mechanics of the environment.   

Active matter clusters at interfaces.

Collective and directed motility or swarming is an emergent phenomenon displayed by many self-organized assemblies of active biological matter such as clusters of embryonic cells during tissue development and flocks of birds. Such clusters typically encounter very heterogeneous environments. What happens when a cluster encounters an interface between two different environments has implications for its function and fate. Here we study this problem by using a mathematical model of a cluster that treats it as a single cohesive unit whose movement depends on the nature of the local environment. We find that low speed clusters which exert forces but no active torques, encountering an interface with a moderate difference in properties can lead to refraction or even total internal reflection of the cluster. For large speeds and clusters with active torques, they show more complex behaviors crossing the interface multiple times, becoming trapped at the interface and deviating from the predictable refraction and reflection of the low velocity clusters. Our results show a wide range of behaviors that occur when collectively moving active biological matter moves across interfaces and these insights can be used to control motion by patterning environments.