Archives de l'auteur: oduroure

Non-linear elastic properties of actin patches to partially rescue yeast endocytosis efficiency in the absence of the cross-linker Sac6.

Belbahri Reda, Michelot Alphée, Heuvingh Julien, du Roure Olivia

Special issue on « Force transmission by nonlinear biomaterials » in Soft Matter. Soft Matter, 2022,18, 1479-1488

Clathrin mediated endocytosis is an essential and complex cellular process involving more than 60 proteins. In yeast, successful endocytosis requires counteracting a large turgor pressure. To this end, yeasts assemble actin patches, which accumulate elastic energy during their assembly. We investigated the material properties of reconstituted actin patches from a wild-type (WT) strain and a mutant strain lacking the cross-linker Sac6 (sac6D), which has reduced endocytosis efficiency in live cells. We hypothesized that a change in the viscous properties of the actin patches, which would dissipate more mechanical energy, could explain this reduced efficiency. There was however no significant difference in the viscosity of both types of patches. However, we discovered a significantly different non-linear elastic response. While WT patches had a constant elastic modulus at different stresses, sac6D patches had a lower elastic modulus at low stresses, before stiffening at higher ones, up to values similar to WT patches. To understand the consequences of this discovery, we performed, in-vivo, a precise analysis of actin patch dynamics. Our analysis reveals that a small fraction of actin patches successfully complete endocytosis in sac6D cells, provided that those assemble an excess of actin at the membrane compared to WT. This observation indicates that non-linear elastic properties of actin networks in sac6D cells contribute to rescue endocytosis, requiring nevertheless more actin material to build-up the necessary stored elastic energy.

Signatures of elastoviscous buckling in the dilute rheology of stiff polymers.

Chakrabarti B., Liu Y., du Roure O., Lindner A. and Saintillan D.

Journal of Fluid Mechanics. 2021. 919 A12.

As a stiff polymer tumbles in shear flow, it experiences compressive viscous forces that can cause it to buckle and undergo a sequence of morphological transitions with increasing flow strength. We use numerical simulations to uncover the effects of these transitions on the steady shear rheology of a dilute suspension of stiff polymers. Our results agree with classic scalings for Brownian rods in relatively weak flows but depart from them above the buckling threshold. Signatures of elastoviscous buckling include enhanced shear thinning and an increase in the magnitude of normal stress differences. We discuss our findings in the light of past work on rigid Brownian rods and non-Brownian elastic fibres and highlight the subtle role of thermal fluctuations in triggering instabilities.

Pinching the cortex of live cells reveals thickness instabilities caused by myosin II motors

Valentin Laplaud, Nicolas Levernier, Judith Pineau, Mabel San Roman, Lucie Barbier, Pablo J. Sáez, Ana-Maria Lennon-Duménil, Pablo Vargas, Karsten Kruse, Olivia du Roure, Matthieu Piel and Julien Heuvingh

Science Advances, Vol 7(27)

The cell cortex is a contractile actin meshwork, which determines cell shape and is essential for cell mechanics, migration, and division. Because its thickness is below optical resolution, there is a tendency to consider the cortex as a thin uniform two-dimensional layer. Using two mutually attracted magnetic beads, one inside the cell and the other in the extracellular medium, we pinch the cortex of dendritic cells and provide an accurate and time-resolved measure of its thickness. Our observations draw a new picture of the cell cortex as a highly dynamic layer, harboring large fluctuations in its third dimension because of actomyosin contractility. We propose that the cortex dynamics might be responsible for the fast shape-changing capacity of highly contractile cells that use amoeboid-like migration.

Programmable Design and Performance of Modular Magnetic Microswimmers

Christoph Pauer, Olivia du Roure, Julien Heuvingh, Tim Liedl, Joe Tavacoli

Advanced Materials (2021) 2006237. https://doi.org/10.1002/adma.202006237

Synthetic biomimetic microswimmers are promising agents for in vivo healthcare and important frameworks to advance the understanding of locomotion strategies and collective motion at the microscopic scale. Nevertheless, constructing these devices with design flexibility and in large numbers remains a challenge. Here, a step toward meeting this challenge is taken by assembling such swimmers via the programmed shape and arrangement of superparamagnetic micromodules. The method’s capacity for design flexibility is demonstrated through the assembly of a variety of swimmer architectures. On their actuation, strokes characterized by a balance of viscous and magnetic forces are found in all cases, but swimmers formed from a series of size‐graded triangular modules swim quicker than more traditional designs comprising a circular “head” and a slender tail. Linking performance to design, rules are extracted informing the construction of a second‐generation swimmer with a short tail and an elongated head optimized for speed. Its fast locomotion is attributed to a stroke that better breaks beating symmetry and an ability to beat fully with flex at high frequencies. Finally, production at scale is demonstrated through the assembly and swimming of a flock of the triangle‐based architectures to reveal four types of swimmer couplings.

Optimised hyperbolic microchannels for the mechanical characterisation of bio-particles

Yanan Liu, Konstantinos Zografos, Joana Fidalgo, Charles Duchêne, Clément Quintard, Thierry Darnige, Vasco Filipe, Sylvain Huille, Olivia du Roure, Mónica SN Oliveira, Anke Lindner

Soft Matter, 2020,16, 9844-9856

Highlighted by the inside font cover

The transport of bio-particles in viscous flows exhibits a rich variety of dynamical behaviour, such as morphological transitions, complex orientation dynamics or deformations. Characterising such complex behaviour under well controlled flows is key to understanding the microscopic mechanical properties of biological particles as well as the rheological properties of their suspensions. While generating regions of simple shear flow in microfluidic devices is relatively straightforward, generating straining flows in which the strain rate is maintained constant for a sufficiently long time to observe the objects’ morphologic evolution is far from trivial. In this work, we propose an innovative approach based on optimised design of microfluidic converging–diverging channels coupled with a microscope-based tracking method to characterise the dynamic behaviour of individual bio-particles under homogeneous straining flow. The tracking algorithm, combining a motorised stage and a microscopy imaging system controlled by external signals, allows us to follow individual bio-particles transported over long-distances with high-quality images. We demonstrate experimentally the ability of the numerically optimised microchannels to provide linear velocity streamwise gradients along the centreline of the device, allowing for extended consecutive regions of homogeneous elongation and compression. We selected three test cases (DNA, actin filaments and protein aggregates) to highlight the ability of our approach for investigating dynamics of objects with a wide range of sizes, characteristics and behaviours of relevance in the biological world.

Programmed Wrapping and Assembly of Droplets with Mesoscale Polymers

Dylan M. Barber, Zhefei Yang, Lucas Prévost, Olivia du Roure, Anke Lindner, Todd Emrick, Alfred J. Crosby

Advanced Functional Materials 2002704

Nature is remarkably adept at using interfaces to build structures, encapsulate reagents, and regulate biological processes. Inspired by nature, flexible polymer‐based ribbons, termed “mesoscale polymers” (MSPs), are described to modulate interfacial interactions with liquid droplets. This produces unprecedented hybrid assemblies in the forms of flagellum‐like structures and MSP‐wrapped droplets. Successful preparation of these hybrid structures hinges on interfacial interactions and tailored MSP compositions, such as MSPs with domains possessing distinctly different affinity for fluid–fluid interfaces as well as mechanical properties. In situ measurements of MSP–droplet interactions confirm that MSPs possess a negligible bending stiffness, allowing interfacial energy to drive mesoscale assembly. By exploiting these interfacial driving forces, mesoscale polymers are demonstrated as a powerful platform that underpins the preparation of sophisticated hybrid structures in fluids.

Microfluidic In-Situ Measurement of Poisson’s Ratio of Hydrogels

Jean Cappello, Vincent d’Herbemont, Anke Lindner, Olivia Du Roure

Micromachines (2020) 11(3) 318

Being able to precisely characterize the mechanical properties of soft microparticles is essential for numerous situations, from the understanding of the flow of biological fluids to the development of soft micro-robots. Here, we present a simple measurement technique for determining Poisson’s ratio of soft micron-sized hydrogels in the presence of a surrounding liquid. This method relies on the measurement of the deformation, in two orthogonal directions, of a rectangular hydrogel slab compressed uni-axially inside a microfluidic channel. Due to the in situ character of the method, the sample does not need to be dried, allowing for the measurement of the mechanical properties of swollen hydrogels. Using this method, we determined Poisson’s ratio of hydrogel particles composed of polyethylene glycol (PEG) and varying solvents fabricated using a lithography technique. The results demonstrate, with high precision, the dependence of the hydrogel compressibility on the solvent fraction and character. The method is easy to implement and can be adapted for the measurement of a variety of soft and biological materials.

Flexible filaments buckle into helicoidal shapes in strong compressional flows

Brato Chakrabarti, Yanan Liu, John Lagrone, Ricardo Cortez, Lisa Fauci, Olivia du Roure, David Saintillan, and Anke Lindner

Nature Physics (2020) 16 (6), 689-694

The occurrence of coiled or helical morphologies is common in nature, from plant roots to DNA packaging into viral capsids, as well as in applications such as oil drilling processes. In many examples, chiral structures result from the buckling of a straight fiber either with intrinsic twist or to which end moments have been applied in addition to compression forces. Here, we elucidate a generic way to form regular helicoidal shapes from achiral straight filaments transported in viscous flows with free ends. Through a combination of experiments using fluorescently labeled actin filaments in microfluidic divergent flows and of two distinct sets of numerical simulations, we demonstrate the robustness of helix formation. A nonlinear stability analysis is performed and explains the emergence of such chiral structures from the nonlinear interaction of perpendicular planar buckling modes, an effect that solely requires a strong compressional flow, independent of the exact nature of the fiber or type of flow field. The fundamental mechanism for the uncovered morphological transition and characterization of the emerging conformations advance our understanding of several biological and industrial processes and can also be exploited for the controlled microfabrication of chiral objects.

Mechanical stiffness of reconstituted actin patches correlates tightly with endocytosis efficiency

Jessica Planade*, Reda Belbahri*, Micaela Boiero Sanders, Audrey Guillotin, Olivia du Roure#, Alphée Michelot#, Julien Heuvingh#

PLoS Biology 2019 17(10): e3000500. https://doi.org/10.1371/journal.pbio.3000500

Clathrin-mediated endocytosis involves the sequential assembly of more than 60 proteins at the plasma membrane. An important fraction of these proteins regulates the assembly of an actin-related protein 2/3 (Arp2/3)-branched actin network, which is essential to generate the force during membrane invagination. We performed, on wild-type (WT) yeast and mutant strains lacking putative actin crosslinkers, a side-by-side comparison of in vivo endocytic phenotypes and in vitro rigidity measurements of reconstituted actin patches. We found a clear correlation between softer actin networks and a decreased efficiency of endocytosis. Our observations support a chain-of-consequences model in which loss of actin crosslinking softens Arp2/3-branched actin networks, directly limiting the transmission of the force. Additionally, the lifetime of failed endocytic patches increases, leading to a larger number of patches and a reduced pool of polymerizable actin, which slows down actin assembly and further impairs endocytosis.

Transport of flexible fibers in confined microchannels

Jean Cappello, Mathias Bechert, Camille Duprat, Olivia du Roure, Fran ̧cois Gallaire, and Anke Lindner.

Physical Review Fluids 4, 034202. (Selected as Editor’s Suggestion)

When transported in confined geometries rigid fibers show interesting transport dynamics induced by friction with the top and bottom walls. Fiber flexibility causes an additional coupling between fiber deformation and transport and is expected to lead to more complex dynamics. A first crucial step for their understanding is the characterization of the deformed fiber shape. Here we characterize this shape for a fiber transported in a confined plug flow perpendicular to the flow direction using a combination of microfluidic experiments and numerical simulations. In the experiments, size, initial orientation, and mechanical properties of the fibers are controlled using microfabrication techniques and in situ characterization methods. The numerical simulations use modified Brinkman equations as well as full three-dimensional simulations. We show that the bending of a perpendicular fiber results from the force distribution acting on the elongated object and is proportional to the elasto-viscous number, which compares viscous to elastic forces. We quantitatively characterize the influence of the confinement on the fiber deformation. The precise understanding of the deformation of a flexible fiber in a confined geometry can also be used in future to understand the deformation and transport of more complex deformable particles in confined flows, such as vesicles or red blood cell