Migrating Epithelial Monolayer Flows Like a Maxwell Viscoelastic Liquid.

We perform a bidimensional Stokes experiment in an active cellular material: an autonomously migrating monolayer of Madin-Darby canine kidney epithelial cells flows around a circular obstacle within a long and narrow channel, involving an interplay between cell shape changes and neighbor rearrangements. Based on image analysis of tissue flow and coarse-grained cell anisotropy, we determine the tissue strain rate, cell deformation, and rearrangement rate fields, which are spatially heterogeneous. We find that the cell deformation and rearrangement rate fields correlate strongly, which is compatible with a Maxwell viscoelastic liquid behavior (and not with a Kelvin-Voigt viscoelastic solid behavior). The value of the associated relaxation time is measured as τ=70±15 min, is observed to be independent of obstacle size and division rate, and is increased by inhibiting myosin activity. In this experiment, the monolayer behaves as a flowing material with a Weissenberg number close to one which shows that both elastic and viscous effects can have comparable contributions in the process of collective cell migration.

Plastic and viscous dissipations in foams: cross-over from low to high shear rates.

Soft glassy materials made of deformable cells, such as liquid foams, simultaneously display elastic, plastic and viscous behaviours. Bubble deformation is elastic until the material plastically yields and bubbles swap neighbours, then bubbles relax dissipatively towards a new energy minimum. This relaxation occurs in a finite time, and shearing a foam at a fast strain rate compared to that time leads to a viscous flow. To describe such an elastic, plastic and viscous behaviour we introduce a simplified scalar model of foam deformation and flow with a periodic pinning potential. The continuum mechanics behaviour of the foam emerges as an ensemble average over disordered units without requiring that they are coupled. Our model captures surprisingly well various features of the viscous dissipation during plastic deformation. At low shear rates, the time averaged stress is smaller than the static yield stress. A critical shear rate exists: any flow at fixed stress has a shear rate above this critical value. Moreover, the model only involves measurable parameters, which enables us to compare it with existing experiments and simulations.

Comparative study of non-invasive force and stress inference methods in tissue.

In the course of animal development, the shape of tissue emerges in part from mechanical and biochemical interactions between cells. Measuring stress in tissue is essential for studying morphogenesis and its physical constraints. For that purpose, a possible new approach is force inference (up to a single prefactor) from cell shapes and connectivity. It is non-invasive and can provide space-time maps of stress in a whole tissue, unlike existing methods. To validate this approach, three force-inference methods, which differ in their approach of treating indefiniteness in an inverse problem between cell shapes and forces, were compared. Tests using two artificial and two experimental data sets consistently indicate that our Bayesian force inference, by which cell-junction tensions and cell pressures are simultaneously estimated, performs best in terms of accuracy and robustness. Moreover, by measuring the stress anisotropy and relaxation, we cross-validated the force inference and the global annular ablation of tissue, each of which relies on different prefactors. A practical choice of force-inference methods in different systems of interest is discussed.

Understanding and predicting viscous, elastic, plastic flows.

Foams, gels, emulsions, polymer solutions, pastes and even cell assemblies display both liquid and solid mechanical properties. On a local scale, such "soft glassy" systems are disordered assemblies of deformable rearranging units, the complexity of which gives rise to their striking flow behaviour. On a global scale, experiments show that their mechanical behaviour depends on the orientation of their elastic deformation with respect to the flow direction, thus requiring a description by tensorial equations for continuous materials. However, due to their strong non-linearities, the numerous candidate models have not yet been solved in a general multi-dimensional geometry to provide stringent tests of their validity. We compute the first solutions of a continuous model for a discriminant benchmark, namely the flow around an obstacle. We compare it with experiments of a foam flow and find an excellent agreement with the spatial distribution of all important features: we accurately predict the experimental fields of velocity, elastic deformation, and plastic deformation rate in terms of magnitude, direction, and anisotropy. We analyse the role of each parameter, and demonstrate that the yield strain is the main dimensionless parameter required to characterize the materials. We evidence the dominant effect of elasticity, which explains why the stress does not depend simply on the shear rate. Our results demonstrate that the behaviour of soft glassy materials cannot be reduced to an intermediate between that of a solid and that of a liquid: the viscous, the elastic and the plastic contributions to the flow, as well as their couplings, must be treated simultaneously. Our approach opens the way to the realistic multi-dimensional prediction of complex flows encountered in geophysical, industrial and biological applications, and to the understanding of the link between structure and rheology of soft glassy systems.

Fast determination of coarse-grained cell anisotropy and size in epithelial tissue images using Fourier transform.

Mechanical strain and stress play a major role in biological processes such as wound healing or morphogenesis. To assess this role quantitatively, fixed or live images of tissues are acquired at a cellular precision in large fields of views. To exploit these data, large numbers of cells have to be analyzed to extract cell shape anisotropy and cell size. Most frequently, this is performed through detailed individual cell contour determination, using so-called segmentation computer programs, complemented if necessary by manual detection and error corrections. However, a coarse-grained and faster technique can be recommended in at least three situations: first, when detailed information on individual cell contours is not required; for instance, in studies which require only coarse-grained average information on cell anisotropy. Second, as an exploratory step to determine whether full segmentation can be potentially useful. Third, when segmentation is too difficult, for instance due to poor image quality or too large a cell number. We developed a user-friendly, Fourier-transform-based image analysis pipeline. It is fast (typically 10^{4} cells per minute with a current laptop computer) and suitable for time, space, or ensemble averages. We validate it on one set of artificial images and on two sets of fully segmented images, one from a Drosophila pupa and the other from a chicken embryo; the pipeline results are robust. Perspectives include in vitro tissues, nonbiological cellular patterns such as foams and xyz stacks.

Dripping of a crystal.

Dripping is usually associated with fluid motion, but here we describe the analogous phenomenon of a 3He crystal growing and melting under the influence of surface tension and gravity. The pinch-off of the crystal is described by a purely geometric equation of motion, viscous dissipation or inertia being negligible. In analogy to fluid pinch-off, the minimum neck radius R{n} goes to zero like a power law, but with a new scaling exponent of 12 . However, for a significant part of the neck's macroscopic evolution the scaling exponent is found to be much closer to 13 . This observation may be consistent with simulations and theoretical results showing a very slow approach to the asymptotic pinch solution, making the "critical region" very small, both in time and space. After pinch-off, we observe a similar 13 -scaling for the recoil of a crystal tip, both in simulation and experiment. For very early times our experiments are consistent with an approximate theory predicting an asymptotic regime with exponent 12 . Future experiments must show whether the transient 13 scaling is a universal feature of crystal melting, or perhaps an artifact of our experimental setup.

Quasicrystalline three-dimensional foams.

We present a numerical study of quasiperiodic foams, in which the bubbles are generated as duals of quasiperiodic Frank-Kasper phases. These foams are investigated as potential candidates to the celebrated Kelvin problem for the partition of three-dimensional space with equal volume bubbles and minimal surface area. Interestingly, one of the computed structures falls close to (but still slightly above) the best known Weaire-Phelan periodic candidate. In addition we find a correlation between the normalized bubble surface area and the root mean squared deviation of the number of faces, giving an additional clue to understanding the main geometrical ingredients driving the Kelvin problem.

Coarsening of three-dimensional grains in crystals, or bubbles in dry foams, tends towards a universal, statistically scale-invariant regime.

We perform extensive Potts model simulations of three-dimensional dry foam coarsening. Starting with 2.25 million bubbles, we have enough statistics to fulfill the three constraints required for the study of statistical scale invariance: first, enough time for the transient to end and reach the scaling state; then, enough time in the scaling state itself to characterize its properties; and finally, enough bubbles at the end to avoid spurious finite size effects. In the scaling state, we find that the average surface area of the bubbles increases linearly with time. The geometry (bubble shape and size) and topology (number of faces and edges), as well as their correlations, become constant in time. Their distributions agree with the data of the literature. We present an analytical model (universal, up to parameters extracted from the simulations) for a disordered foam minimizing its free energy, which agrees with the simulations. We discuss the limitations of the simulations and of the model.

Interaction of the third helix of Antennapedia homeodomain and a phospholipid monolayer, studied by ellipsometry and PM-IRRAS at the air-water interface.

The penetratin peptide, a 16 amino acid sequence extracted from Antennapedia homeodomain, is able to translocate across a neural cell membrane through an unknown mechanism, most likely a non-specific interaction with membrane lipids. Beyond its potential application as vector targeting small hydrophilic molecules and enabling them to reach a cell nucleus, this observation raises intriguing questions concerning the physico-chemistry of peptide-lipid interactions. Here we present a study of the role of lipid surface pressure and head charge on the mechanism of interaction. This was performed using optical techniques: surface infrared spectroscopy and ellipsometry, applied to a monolayer of phospholipids deposited at the air-water interface. Determination of the structure and orientation of peptides and lipids (separately or together) evidenced that electrostatic rather than amphiphilic interactions determine the peptide adsorption and its action on lipids.

Three-dimensional bubble clusters: shape, packing, and growth rate.

We consider three-dimensional clusters of equal-volume bubbles packed around a central bubble and calculate their energy and optimal shape. We obtain the surface area and bubble pressures to improve on existing growth laws for three-dimensional bubble clusters. We discuss the possible number of bubbles that can be packed around a central one: the "kissing problem," here adapted to deformable objects.

Equilibrium states and ground state of two-dimensional fluid foams.

We study the equilibrium energies of two-dimensional (2D) noncoarsening fluid foams, which consist of bubbles with fixed areas. The equilibrium states correspond to local minima of the total perimeter. We present a theoretical derivation of energy minima; experiments with ferrofluid foams, which can be either highly distorted, locally relaxed, or globally annealed; and Monte Carlo simulations using the extended large-Q Potts model. For a dry foam with small size variance we develop physical insight and an electrostatic analogy, which enables us to (i) find an approximate value of the global minimum perimeter, accounting for (small) area disorder, the topological distribution, and physical boundary conditions; (ii) conjecture the corresponding pattern and topology: small bubbles sort inward and large bubbles sort outward, topological charges of the same signs "repel" while charges of the opposite signs "attract;" (iii) define local and global markers to determine directly from an image how far a foam is from its ground state; (iv) conjecture that, in a local perimeter minimum at prescribed topology, the pressure distribution and thus the edge curvature are unique. Some results also apply to 3D foams.

Discrete rearranging disordered patterns: prediction of elastic and plastic behavior, and application to two-dimensional foams.

We study the elasto-plastic behavior of materials made of individual (discrete) objects such as a liquid foam made of bubbles. The evolution of positions and mutual arrangements of individual objects is taken into account through statistical quantities such as the elastic strain of the structure, the yield strain, and the yield function. The past history of the sample plays no explicit role except through its effect on these statistical quantities. They suffice to relate the discrete scale with the collective global scale. At this global scale, the material behaves as a continuous medium; it is described with tensors such as elastic strain, stress, and velocity gradient. We write the differential equations which predict their elastic and plastic behavior in both the general case and the case of simple shear. An overshoot in the shear strain or shear stress is interpreted as a rotation of the deformed structure, which is a purely tensorial effect that exists only if the yield strain is at least of order 0.3. We suggest practical applications including the following: when to choose a scalar formalism rather than a tensorial one; how to relax trapped stresses; and how to model materials with a low, or a high, yield strain.

Discrete rearranging disordered patterns, part II: 2D plasticity, elasticity and flow of a foam.

The plastic flow of a foam results from bubble rearrangements. We study their occurrence in experiments where a foam is forced to flow in 2D: around an obstacle; through a narrow hole; or sheared between rotating disks. We describe their orientation and frequency using a topological matrix defined in the companion paper (F. Graner, B. Dollet, C. Raufaste, and P. Marmottant, this issue, 25 (2008) DOI 10.1140/epje/i2007-10298-8), which links them with continuous plasticity at large scale. We then suggest a phenomenological equation to predict the plastic strain rate: its orientation is determined from the foam's local elastic strain; and its rate is determined from the foam's local elongation rate. We obtain a good agreement with statistical measurements. This enables us to describe the foam as a continuous medium with fluid, elastic and plastic properties. We derive its constitutive equation, then test several of its terms and predictions.

Discrete rearranging disordered patterns, part I: robust statistical tools in two or three dimensions.

Discrete rearranging patterns include cellular patterns, for instance liquid foams, biological tissues, grains in polycrystals; assemblies of particles such as beads, granular materials, colloids, molecules, atoms; and interconnected networks. Such a pattern can be described as a list of links between neighbouring sites. Performing statistics on the links between neighbouring sites yields average quantities (hereafter "tools") as the result of direct measurements on images. These descriptive tools are flexible and suitable for various problems where quantitative measurements are required, whether in two or in three dimensions. Here, we present a coherent set of robust tools, in three steps. First, we revisit the definitions of three existing tools based on the texture matrix. Second, thanks to their more general definition, we embed these three tools in a self-consistent formalism, which includes three additional ones. Third, we show that the six tools together provide a direct correspondence between a small scale, where they quantify the discrete pattern's local distortion and rearrangements, and a large scale, where they help describe a material as a continuous medium. This enables to formulate elastic, plastic, fluid behaviours in a common, self-consistent modelling using continuous mechanics. Experiments, simulations and models can be expressed in the same language and directly compared. As an example, a companion paper (P. Marmottant, C. Raufaste, and F. Graner, this issue, 25 (2008) DOI 10.1140/epje/i2007-10300-7) provides an application to foam plasticity.

Numerical modelling of foam Couette flows.

A numerical computation based on a tensorial visco-elasto-plastic model based on continuous mechanics is compared to experimental measurements on liquid foams for a bidimensional Couette flow between two glass plates, both in stationary and transient cases. The main features of the model are elasticity up to a plastic yield stress, and viscoelasticity above it. The effect of the friction of the plates is taken into account. The numerical modelling is based on a small set of standard material parameters that are fully characterised. Shear localisation as well as acute transient observations are reproduced and agree with experimental measurements. The plasticity appears to be the fundamental mechanism of the localisation of the flow. Finally, the present approach could be extended from liquid foams to similar materials such as emulsions, colloids or wet granular materials, that exhibit localisation.

Tensile forces govern germ-layer organization in zebrafish.

Understanding the factors that direct tissue organization during development is one of the most fundamental goals in developmental biology. Various hypotheses explain cell sorting and tissue organization on the basis of the adhesive and mechanical properties of the constituent cells. However, validating these hypotheses has been difficult due to the lack of appropriate tools to measure these parameters. Here we use atomic force microscopy (AFM) to quantify the adhesive and mechanical properties of individual ectoderm, mesoderm and endoderm progenitor cells from gastrulating zebrafish embryos. Combining these data with tissue self-assembly in vitro and the sorting behaviour of progenitors in vivo, we have shown that differential actomyosin-dependent cell-cortex tension, regulated by Nodal/TGFbeta-signalling (transforming growth factor beta), constitutes a key factor that directs progenitor-cell sorting. These results demonstrate a previously unrecognized role for Nodal-controlled cell-cortex tension in germ-layer organization during gastrulation.

An elastic, plastic, viscous model for slow shear of a liquid foam.

We suggest a scalar model for deformation and flow of an amorphous material such as a foam or an emulsion. To describe elastic, plastic and viscous behaviours, we use three scalar variables: elastic deformation, plastic deformation rate and total deformation rate; and three material-specific parameters: shear modulus, yield deformation and viscosity. We obtain equations valid for different types of deformations and flows slower than the relaxation rate towards mechanical equilibrium. In particular, they are valid both in transient or steady flow regimes, even at large elastic deformation. We discuss why viscosity can be relevant even in this slow shear (often called "quasi-static") limit. Predictions of the storage and loss moduli agree with the experimental literature, and explain with simple arguments the non-linear large amplitude trends.

Yield drag in a two-dimensional foam flow around a circular obstacle: effect of liquid fraction.

We study the two-dimensional flow of foams around a circular obstacle within a long channel. In experiments, we confine the foam between liquid and glass surfaces. In simulations, we use a deterministic software, the Surface Evolver, for bubble details and a stochastic one, the extended Potts model, for statistics. We adopt a coherent definition of liquid fraction for all studied systems. We vary it in both experiments and simulations, and determine the yield drag of the foam, that is, the force exerted on the obstacle by the foam flowing at very low velocity. We find that the yield drag is linear over a large range of the ratio of obstacle to bubble size, and is independent of the channel width over a large range. Decreasing the liquid fraction, however, strongly increases the yield drag; we discuss and interpret this dependence.

Structure and fluctuations of a single floating lipid bilayer.

A single lipid molecular bilayer of 17 or 18 carbon chain phosphocholines, floating in water near a flat wall, is prepared in the bilayer gel phase and then heated to the fluid phase. Its structure (electron density profile) and height fluctuations are determined by using x-ray reflectivity and non-specular scattering. By fitting the off-specular signal to that calculated for a two-dimensional membrane using a Helfrich Hamiltonian, we determine the three main physical quantities that govern the bilayer height fluctuations: The wall attraction potential is unexpectedly low; the surface tension, roughly independent on chain length and temperature, is moderate (approximately 5 x 10(-4) J.m(-2)) but large enough to dominate the intermediate range of the fluctuation spectrum; and the bending modulus abruptly decreases by an order-of-magnitude from 10(-18) J to 10(-19) J at the bilayer gel-to-fluid transition.