We theoretically investigate the membrane fluctuations of red blood cells with focus laid on the role of the cytoskeleton, viewing the system as a membrane coupled to sparse spring network. This model is exactly solvable and enables us to examine the coupling strength dependence of the membrane undulation. We find that the coupling modifies the fluctuation spectrum at wavelengths longer than the mesh size of the network, while leaving the fluid-like behavior of the membrane intact at shorter wavelengths. The fluctuation spectra can be markedly different, depending on not only the relative amplitude of the bilayer bending energy with respect to the cytoskeleton deformation energy but also the bilayer-cytoskelton coupling strength.
We theoretically investigate the dynamics of a floating lipid bilayer membrane coupled with a two-dimensional cytoskeleton network, taking into explicitly account the intermonolayer friction, the discrete lattice structure of the cytoskeleton, and its prestress. The lattice structure breaks lateral continuous translational symmetry and couples Fourier modes with different wavevectors. It is shown that within a short time interval a long-wavelength deformation excites a collection of modes with wavelengths shorter than the lattice spacing. These modes relax slowly with a common renormalized rat…
We analyze both the static and dynamic fluctuation spectrum of the red-blood cell in a unified manner, using a simple model of the composite membrane. In this model, the two-dimensional spectrin network that forms the cytoskeleton is treated as a rigid shell which is located at some constant average separation from the lipid bilayer. The cytoskeleton thereby confines both the static and dynamic fluctuations of the lipid bilayer. The predictions of the model account for the wavevector and frequency dependence of the experimental data. The observed amplitude of the thermal fluctuations is relate…
The plasma membrane of human red blood cells consists of a lipid bilayer attached to a regular network of underlying cytoskeletal polymers. We model this system at a dynamic coarse-grained level, treating the bilayer as an elastic sheet and the cytoskeletal network as a series of phantom entropic springs. In contrast to prior simulation efforts, we explicitly account for dynamics of the cytoskeletal network, both via motion of the protein anchors that attach the cytoskeleton to the bilayer and through breaking and reconnection of individual cytoskeletal filaments. Simulation results are explai…
We address the relationship between membrane microheterogeneity and anomalous subdiffusion in cell membranes by carrying out Monte Carlo simulations of two-component lipid membranes. We find that near-critical fluctuations in the membrane lead to transient subdiffusion, while membrane-cytoskeleton interaction strongly affects phase separation, enhances subdiffusion, and eventually leads to hop diffusion of lipids. Thus, we present a minimum realistic model for membrane rafts showing the features of both microscopic phase separation and subdiffusion.
Blebs are balloon-shaped membrane protrusions that form during many physiological processes. Using computer simulation of a particle-based model for self-assembled lipid bilayers coupled to an elastic meshwork, we investigated the phase behavior and kinetics of blebbing. We found that blebs form for large values of the ratio between the areas of the bilayer and the cytoskeleton. We also found that blebbing can be induced when the cytoskeleton is subject to a localized ablation or a uniform compression. The results obtained are qualitatively in agreement with the experimental evidence and the m…
Internalization of particles by cells plays a crucial role for adsorbing nutrients and fighting infection. Endocytosis is one of the most important mechanisms of the particles uptake which encompass multiple pathways. Although endocytosis is a complex mechanism involving biochemical signaling and active force generation, the energetic cost associated to the large deformations of the cell membrane wrapping around the foreign particle is an important factor controlling this process, which can be studied using quantitative physical models. Of particular interest is the competition between membran…
When a ligand that is bound to an integral membrane receptor is pulled, the membrane and the underlying cytoskeleton can deform before either the membrane delaminates from the cytoskeleton or the ligand detaches from the receptor. If the membrane delaminates from the cytoskeleton, it may be further extruded and form a membrane tether. We develop a phenomenological model for this processes by assuming that deformations obey Hooke's law up to a critical force at which the cell membrane locally detaches from the cytoskeleton and a membrane tether forms. We compute the probability of tether format…
We identify a class of composite membranes: fluid bilayers coupled to an elastic meshwork, that are such that the meshwork's energy is a function $F_\mathrm{el}[A_ξ]$ \textit{not} of the real microscopic membrane area $A$, but of a \textit{smoothed} membrane's area $A_ξ$, which corresponds to the area of the membrane coarse-grained at the mesh size $ξ$. We show that the meshwork modifies the membrane tension $σ$ both below and above the scale $ξ$, inducing a tension-jump $Δσ=dF_\mathrm{el}/dA_ξ$. The predictions of our model account for the fluctuation spectrum of red blood cells membranes cou…
The functions and elasticities of the cell are largely related to the structures of the cytoskeletons underlying the lipid bi-layer. Among various cell types, the Red Blood Cell (RBC) possesses a relatively simple cytoskeletal structure. Underneath the membrane, the RBC cytoskeleton takes the form of a two dimensional triangular network, consisting of nodes of actins (and other proteins) and edges of spectrins. Recent experiments focusing on the malaria infected RBCs (iRBCs) showed that there is a correlation between the elongation of spectrins in the cytoskeletal network and the stiffening of…
Eukaryotic cells adhere to extracellular matrix during the normal development of the organism, forming static adhesion as well as during cell motility. We study this process by considering a simplified coarse-grained model of a vesicle that has uniform adhesion energy with a flat substrate, mobile curved membrane proteins and active forces. We find that a high concentration of curved proteins alone increases the spreading of the vesicle, by the self-organization of the curved proteins at the high curvature vesicle-substrate contact line, thereby reducing the bending energy penalty at the vesic…
The ability of cells to undergo reversible shape changes is often crucial to their survival. For Red Blood Cells (RBCs), irreversible alteration of the cell shape and flexibility often causes anemia. Here we show theoretically that RBCs may react irreversibly to mechanical perturbations because of tensile stress in their cytoskeleton. The transient polymerization of protein fibers inside the cell seen in sickle cell anemia or a transient external force can trigger the formation of a cytoskeleton-free membrane protrusion of micrometer dimensions. The complex relaxation kinetics of the cell shap…
In this work, we review previously developed coarse-grained (CG) particle models for biological membrane and red blood cells (RBCs) and discuss the advantages of the CG particle method over the continuum and atomic simulations on modeling biological phenomena. CG particle models can largely increase the length scale and time scale of atomic simulations by eliminating fast degrees of freedom while preserving the mesoscopic structures and properties of the simulated system. One the other hand, CG particle models can be used to capture microstructural alternations in diseased RBCs and simulate to…
We study a Gaussian model of the membrane of red blood cells: a "phantom" triangular network of springs attached at its vertices to a fluid bilayer with curvature elasticity and tension. We calculate its fluctuation spectrum and we discuss the different regimes and non-monotonic features, including the precise crossover at the mesh size between the already known limits with two different tensions and the renormalisation of the bending rigidity at low wavevectors. We also show that the non-diagonal correlations reveal, in "dark field", the cytoskeletal defects. As a first step toward a non-inva…
Cellular uptake through the lipid membranes plays an important role in adsorbing nutrients and fighting infection and can be used for drug delivery and nanomedicine developments. Endocytosis is one of the known pathways of the cellular uptake which associate with elastic deformation of the membrane wrapping around the foreign particle. The deformability of the membrane itself is strongly regulated by the presence of a cortical cytoskeleton placed underneath the membrane. It has been shown that size, shape, and orientation of the particles influence on their entry into the cell. Here, we study …
Reduced blood flow, as occurring in ischemia or resulting from exposure to microgravity such as encountered in space flights, induces a decrease in the level of shear stress sensed by the endothelial cells forming the inner part of blood vessels. In the present study, we use a microvasculature-on-a-chip device in order to investigate in vitro the effect of such a reduction in shear stress on shear-adapted endothelial cells. We find that, within one hour of exposition to reduced wall shear stress, human umbilical vein endothelial cells undergo a reorganization of their actin skeleton, with a de…
Phagocytosis is the process of engulfment and internalization of comparatively large particles by the cell, that plays a central role in the functioning of our immune system. We study the process of phagocytosis by considering a simplified coarse grained model of a three-dimensional vesicle, having uniform adhesion interaction with a rigid particle, in the presence of curved membrane proteins and active cytoskeletal forces. Complete engulfment is achieved when the bending energy cost of the vesicle is balanced by the gain in the adhesion energy. The presence of curved (convex) proteins reduces…
Adhesion between a T cell and an antigen presenting cell is achieved by TCR-pMHC and LFA1-ICAM1 protein complexes. These segregate to form a special pattern, known as the immunological synapse (IS), consisting of a central quasi-circular domain of TCR-pMHC bonds surrounded by a peripheral domain of LFA1-ICAM1 complexes. Insights gained from imaging studies had led to the conclusion that the formation of the central adhesion domain in the IS is driven by active (ATP-driven) mechanisms. Recent studies, however, suggested that passive (thermodynamic) mechanisms may also play an important role in …
Cellular wrapping of elastic particles by a supported lipid membrane
Université Paris Cité, CNRS, Institut Jacques Monod（巴黎西岱大学，法国国家科学研究中心，雅克·莫诺研究所）；Institut Curie, Université PSL, Sorbonne Université, CNRS UMR168（居里研究所，巴黎文理研究大学，索邦大学，法国国家科学研究中心联合研究实验室）；Department of Physics, Institute for Advanced Studies in Basic Sciences (IASBS)（基础科学研究高级学院物理系）；Department of Physics and Astronomy, University of Pennsylvania（宾夕法尼亚大学物理与天文学系）
We highlight changes to cell signalling under virus invasion (SARS-CoV-2), involving disturbance of membranes and of nanodomains, modulated by the cytoskeleton. Virus alters the mechanical properties of the membranes, impairing mesophase structures mediated by the fractal architecture initiated by actomyosin.
We consider the motion of red blood cells and other non-spherical microcapsules dilutely suspended in a simple shear flow. Our analysis indicates that depending on the viscosity, membrane elasticity, geometry and shear rate, the particle exhibits either tumbling, tank-treading of the membrane about the viscous interior with periodic oscillations of the orientation angle, or intermittent behavior in which the two modes occur alternately. For red blood cells, we compute the complete phase diagram and identify a novel tank-treading-to-tumbling transition at low shear rates. Observations of such m…
In biological settings membranes typically interact locally with other membranes or the extracellular matrix in the exterior, as well as with internal cellular structures such as the cytoskeleton. Characterization of the dynamic properties of such interactions presents a difficult task. Significant progress has been achieved through simulations and experiments, yet analytical progress in modelling pinned membranes has been impeded by the complexity of governing equations. Here we circumvent these difficulties by calculating analytically the time-dependent Green's function of the operator gover…
The unique ability of a red blood cell to flow through extremely small microcapillaries depends on the viscoelastic properties of its membrane. Here, we study in vitro the response time upon flow startup exhibited by red blood cells confined into microchannels. We show that the characteristic transient time depends on the imposed flow strength, and that such a dependence gives access to both the effective viscosity and the elastic modulus controlling the temporal response of red cells. A simple theoretical analysis of our experimental data, validated by numerical simulations, further allows us…
The dynamics of red blood cells (RBCs) in simple shear flow was studied using a theoretical approach based on three variables: a shape parameter, the inclination angle $θ$, and phase angle $ϕ$ of the membrane rotation. At high shear rate and low viscosity contrast of internal fluid, RBCs exhibit tank-treading motion, where $ϕ$ rotates with swinging oscillation of shape and $θ$. At low shear rate, tumbling motion occurs and $θ$ rotates. In the middle region between these two phases, it is found that synchronized rotation of $ϕ$ and $θ$ with integer ratios of the frequencies occurs in addition t…
Eukaryote cells have a flexible shape, which dynamically changes according to the function performed by the cell. One mechanism for deforming the cell membrane into the desired shape is through the expression of curved membrane proteins. Furthermore, these curved membrane proteins are often associated with the recruitment of the cytoskeleton, which then applies active forces that deform the membrane. This coupling between curvature and activity was previously explored theoretically in the linear limit of small deformations, and low dimensionality. Here we explore the unrestricted shapes of ves…
Biologically driven non-equilibrium fluctuations are often characterized by their non-Gaussianity or by an "effective temperature", which is frequency dependent and higher than the ambient temperature. We address these two measures theoretically by examining a randomly kicked "particle", with a variable number of kicking "motors", and show how these two indicators of non-equilibrium behavior can contradict. Our results are compared with new experiments on shape fluctuations of red-blood cell membranes, and demonstrate how the physical nature of the motors in this system can be revealed using t…
Cell membranes are anchored to the cytoskeleton via immobile inclusions. We investigate the effect of such anchors on the in-plane dynamics of a fluid membrane and mobile inclusions (proteins) embedded in it. The immobile particles lead to a decreased diffusion coefficient of mobile ones and suppress the correlated diffusion of particle pairs. Due to the long-range, quasi-two-dimensional nature of membrane flows, these effects become significant at a low area fraction (below one percent) of immobile inclusions.
Central to understanding membrane bound cell signaling is to quantify how the membrane ultra-structure consisting of transient spatial domains modulates signaling and how the signaling influences this ultra-structure. Yet, measuring the association of membrane proteins with domains in living, intact cells poses considerable challenges. Here, we describe a non-destructive method to quantify protein-lipid domain and protein cytoskeleton interactions in single, intact cells enabling continuous monitoring of the protein domains interaction over time during signaling.
We present rheology experiments on dilute solutions of vesicles and red blood cells (RBC). Varying the viscosity ratio $λ$ between internal and external fluids, the microscopic dynamics of suspended objects can be qualitatively changed from tank-treading ($tt$) to tumbling ($tb$). We find that in the tt regime the viscosity $η$, decreases when $λ$ increases, in contrast with droplet emulsions and elastic capsule theories which are sometimes invoked to model RBC dynamics. At a critical $λ$ (close to the tt-tb transition) $η$ exhibits a minimum before it increases in the tb regime. This is consi…
Eukaryotic cells intrinsically change their shape, by changing the composition of their membrane and by restructuring their underlying cytoskeleton. We present here further studies and extensions of a minimal physical model, describing a closed vesicle with mobile curved membrane protein complexes. The cytoskeletal forces describe the protrusive force due to actin polymerization which is recruited to the membrane by the curved protein complexes. We characterize the phase diagrams of this model, as function of the magnitude of the active forces, nearest-neighbor protein interactions and the pro…
We perform an analytical investigation of the cell interface dynamics in the framework of a minimal phase field model of cell motility suggested in [1], which consists of two coupled evolution equations for the order parameter and a two-dimensional vector field describing the actin network polarization (orientation). We derive a closed evolutionary integro-differential equation governing the cell interface dynamics. The equation includes the normal velocity of the membrane, curvature, volume relaxation, and a parameter that is determined by the non-equilibrium effects in the cytoskeleton. This…
Blood exhibits a heterogeneous nature of hematocrit, velocity, and effective viscosity in microcapillaries. Microvascular bifurcations have a significant influence on the distribution of the blood cells and blood flow behavior. This paper presents a simulation study performed on the two-dimensionalmotions and deformation of multiple red blood cells in microvessels with diverging and converging bifurcations. Fluid dynamics and membrane mechanics were incorporated. Effects of cell shape, hematocrit, and deformability of the cell membrane on rheological behavior of the red blood cells and the hem…
We solve the dynamic equations of motion for a growing semi-flexible polymer, or fiber, approaching a fluctuating membrane at an angle. At late times we find three different regimes: fiber {\em stalling}, when fiber growth stops due to membrane resistence, {\em run-away}, in which the polymer bends away from the membrane, and another regime in which the membrane response is nonlinear and tubular membrane {\em spicules} are formed. We discuss which regions of the resulting `phase diagram' are explored by (i) single and bundled actin fibers in living cells, (ii) sickle hemoglobin fibers in red b…
We use mesoscale numerical simulations to investigate the unsteady dynamics of a single red blood cell (RBC) subjected to an external mechanical load. We carry out a detailed comparison between the {\it loading} (L) dynamics, following the imposition of the mechanical load on the RBC at rest, and the {\it relaxation} (R) dynamics, allowing the RBC to relax to its original shape after the sudden arrest of the mechanical load. Such a comparison is carried out by analyzing the characteristic times of the two corresponding dynamics, i.e., $t_L$ and $t_R$. When the intensity of the mechanical load …
Nanoparticles delivering drugs, disseminating cancer cells, and red blood cells (RBCs) during splenic filtration must deform and pass through the sub-micrometer and high aspect ratio interstices between the endothelial cells lining blood vessels. The dynamics of passage of particles/cells through these slit-like interstices remain poorly understood because the in vitro reproduction of slits with physiological dimensions in devices compatible with optical microscopy observations requires expensive technologies. Here, novel microfluidic PDMS devices containing high aspect ratio slits with sub-mi…
Biconcave-discoid (empirical shape of a red blood cell) capsule finds numerous applications in the field of bio-fluid dynamics and rheology, apart from understanding the behavior of red blood cells (RBC) in blood flow. A detailed analysis is, therefore, carried out to understand the effects of the uniaxial extensional flow and electric field on the deformation of a RBC and biconcave-discoid polymeric capsule in the axisymmetric regime. The transient deformation is computed numerically using axisymmetric boundary integral method for Stokes flow considering the Skalak membrane constitutive law a…
We set up the statistical mechanics for a nearly flat, thermally equilibrated fluid membrane, attached to an elastic network through one of its sides. We predict that the resulting structural (inversion) asymmetry of the membrane, notably due to the elastic network attached to one of its sides, can generate a local spontaneous curvature $C_0$, that may in turn destabilize the otherwise flat membrane. As $C_0$ rises above a threshold at a fixed temperature, a flat tethered membrane in the thermodynamic limit becomes structurally unstable, signaling {\em crumpling} of the flat membrane. In-vitro…
The use of drugs derived from benzothiadiazine, which is a bicyclic heterocyclic benzene derivative, has become a widespread treatment for diseases such as hypertension (treated with diuretics such as bendroflumethiazide or chlorothiazide), low blood sugar (treated with non-diuretic diazoxide) or the human immunodeficiency virus, among others. In this work we have investigated the interactions of benzothiadiazine with the basic components of cell membranes and solvents such as phospholipids, cholesterol, ions and water. The analysis of the mutual microscopic interactions is of central importan…
Via computer simulations, we provide evidence that the shear rate induced red blood cell tumbling-to-tank-treading transition also occurs at quite high volume fractions, where collective effects are important. The transition takes place as the ratio of suspension stress to the characteristic cell membrane stress exceeds a certain value, independent of volume fraction and cell deformability. This value coincides with that for a transition from an orientationally less ordered to a highly ordered phase. The average cell deformation does not show any signature of the transition, but rather follows…
We present a rigorous procedure to derive coarse-grained red blood cell (RBC) models, which lead to accurate mechanical properties of realistic RBCs. Based on a semi-analytic theory linear and non-linear elastic properties of the RBC membrane can be matched with those obtained in optical tweezers stretching experiments. In addition, we develop a nearly stress-free model which avoids a number of pitfalls of existing RBC models, such as non-biconcave equilibrium shape and dependence of RBC mechanical properties on the triangulation quality. The proposed RBC model is suitable for use in many exis…
We discuss thermal and active fluctuations of a compressible bilayer vesicle by using the results of hydrodynamic theory for vesicles. Coupled Langevin equations for the membrane deformation and the density fields are employed to calculate the power spectral density matrix of membrane fluctuations. Thermal contribution is obtained by means of the fluctuation dissipation theorem, whereas active contribution is calculated from exponentially decaying time correlation functions of active random forces. We obtain the total power spectral density as a sum of thermal and active contributions. An appa…
This paper considers the effect of electrostatics on the stability of a charged membrane. We show that at low ionic strength and high surface charge density, repulsion between charges on the membrane renders it unstable to the formation of holes. A free straight edge is unstable to modulations with wavelength longer than the Debye screening length. Hence at low ionic strength, membranes will disintegrate into vesicles. We use these results to interpret stable holes in red blood cell ghosts (Steck et. al., Science 168:255(1970)).
The relation between thermal fluctuations and the mechanical response of a free membrane has been explored in great detail, both theoretically and experimentally. However, understanding this relationship for membranes, locally pinned by proteins, is significantly more challenging. Given that the coupling of the membrane to the cell cytoskeleton, the extracellular matrix and to other internal structures is crucial for the regulation of a number of cellular processes, understanding the role of the pinning is of great interest. In this manuscript we consider a single protein (elastic spring of a …
Directly examining subcellular mechanics whilst avoiding excessive strain of a live cell requires the precise control of light stress on very small areas, which is fundamentally difficult. Here we use a glass nanopipet out of contact with the plasma membrane to both exert the stress on the cell and also accurately monitor cellular compression. This allows the mapping of cell stiffness at a lateral resolution finer than 100 nm. We calculate the stress a nanopipet exerts on a cell as the sum of the intrinsic pressure between the tip face and the plasma membrane plus its direct pressure on any gl…
We extend previous work on homogeneous bilayers to calculate the barriers to fusion of planar bilayers which contain two different amphiphiles, a lamellae-former and a hexagonal former, with different compositions of the twoin each leaf. Self-consistent field theory is employed, and both standard and alternative pathways are explored. We first calculate these barriers as the amount of hexagonal former is increased equally in both leaves to levels appropriate to the plasma membrane of human red blood cells. We follow these barriers as the composition of hexagonal-formers is then increased in th…
The dynamics of single red blood cells (RBCs) determine microvascular blood flow by adapting their shape to the flow conditions in the narrow vessels. In this study, we explore the dynamics and shape transitions of RBCs on the cellular scale under confined and unsteady flow conditions using a combination of microfluidic experiments and numerical simulations. Tracking RBCs in a comoving frame in time-dependent flows reveals that the mean transition time from the symmetric croissant to the off-centered, non-symmetric slipper shape is significantly faster than the opposite shape transition, which…
We study deformability-based sorting of same-size RBCs via DLD using an in-house integral equation solver. Our goal is to quantitatively characterize the physical mechanisms that enable the cell separation. To this end, we systematically investigate the effects of the interior fluid viscosity and membrane elasticity of a cell on its behavior. In particular, we consider deep devices in which a cell can show rich dynamics such as taking a particular angular orientation depending on its mechanical property. We have found out that cells moving with a sufficiently high positive inclination angle wi…
The dynamics of red blood cells (RBCs) in oscillatory shear flow was studied using differential equations of three variables: a shape parameter, the inclination angle $θ$, and phase angle $ϕ$ of the membrane rotation. In steady shear flow, three types of dynamics occur depending on the shear rate and viscosity ratio. i) tank-treading (TT): $ϕ$ rotates while the shape and $θ$ oscillate. ii) tumbling (TB): $θ$ rotates while the shape and $ϕ$ oscillate. iii) intermediate motion: both $ϕ$ and $θ$ rotate synchronously or intermittently. In oscillatory shear flow, RBCs show various dynamics based on…
We preset a computational study of bending models for the curvature elasticity of lipid bilayer membranes that are relevant for simulations of vesicles and red blood cells. We compute bending energy and forces on triangulated meshes and evaluate and extend four well established schemes for their approximation: Kantor and Nelson 1987, Phys. Rev. A 36, 4020, Jülicher 1996, J. Phys. II France 6, 1797, Gompper and Kroll 1996, J. Phys. I France 6, 1305, and Meyer et. al. 2003 in Visualization and Mathematics III, Springer, p35, termed A, B, C, D. We present a comparative study of these four schemes…
