SearcharxivSearch

arXiv subjects

Thierry Darnige

Publications and source records attributed to Thierry Darnige.

13 recordsLinked to original sources

Deep learning-enhanced Lagrangian 3D Tracking of motile microorganisms

How microorganisms respond to and interact with their environment can vary significantly from individual to individual, which can have important microbiological and ecological implications. However, most microscopy techniques can only observe motile microorganisms for short times because of their limited fields of view. Using Lagrangian tracking, a single microorganism can be followed in 3D, potentially indefinitely, allowing to decipher individual phenotypical traits. Current Lagrangian tracking methods use the fluorescence signal emitted by the microorganism as feedback to keep it in focus. However, over long times, epifluorescent imaging can induce photobleaching and photodamage, and importantly, not all microorganisms can easily be made fluorescent. Additionally, traditional algorithms used in feedback loops to determine microorganism position are prone to errors, especially in optically complex media. Here, we present a faster, more reliable, and versatile Lagrangian tracking method that uses deep learning to determine the 3D position of the microorganism. This new method demonstrates enhanced accuracy and speed in tracking fluorescent bacteria with fluorescence microscopy also in optically complex media. Furthermore, we track bacteria with other microscopy modalities, such as brightfield microscopy -- for example, this enables us to track magnetotactic bacteria, which cannot be made fluorescent without degrading their magnetotactic properties. These novel capabilities allow to extract previously inaccessible quantitative information, significantly advancing the study of microorganism behavior -- and thus opening new avenues for research in complex biological and ecological systems.

physics.bio-ph

Confinement controls bacterial spreading at all scales

Navigation of microorganisms is controlled by internal processes ultimately sensitive to mechanical or chemical signaling encountered along the path. In many natural environments, such as porous soils or physiological ducts, motile species alternate between bulk and surface motion displaying in each case, distinct kinematics. This inherent complexity is key to many practical biological and ecological issues involving spreading and contamination, essential for understanding the spatiotemporal structuring of populations in their environment. However grasping the interplay between geometrical confinement and kinematics driven by internal biological responses remains poorly understood from a physical and biological standpoint. Here, we address this question through experimental and theoretical analysis in the heuristic situation of two parallel confining surfaces. We track wild-type E. coli - a model peritrichous flagellated bacterium - in 3D over extended periods of time. We obtain the first experimental measurements of the emerging diffusivity and bulk/surface residence times as a function of confinement height and the specific chiral kinematics at surfaces. All experimental results are quantitatively reproduced, without parametric adjustment, by a non-Markovian stochastic (BV) model that incorporates the internal biochemical memory carried by a phosphorylated protein switching the motor rotation. By matching the results with a Markovian (memoryless) companion model, we derive an analytical expression for the diffusivity and demonstrate how confining walls influence microbial long-range dispersion. This approach also provides a general conceptual basis for understanding how microorganisms navigate complex environments, in which their movement alternates between bulk and surfaces.

physics.bio-ph

The 2024 Motile Active Matter Roadmap

Activity and autonomous motion are fundamental aspects of many living and engineering systems. Here, the scale of biological agents covers a wide range, from nanomotors, cytoskeleton, and cells, to insects, fish, birds, and people. Inspired by biological active systems, various types of autonomous synthetic nano- and micromachines have been designed, which provide the basis for multifunctional, highly responsive, intelligent active materials. A major challenge for understanding and designing active matter is their inherent non-equilibrium nature due to persistent energy consumption, which invalidates equilibrium concepts such as free energy, detailed balance, and time-reversal symmetry. Furthermore, interactions in ensembles of active agents are often non-additive and non-reciprocal. An important aspect of biological agents is their ability to sense the environment, process this information, and adjust their motion accordingly. It is an important goal for the engineering of micro-robotic systems to achieve similar functionality. With many fundamental properties of motile active matter now reasonably well understood and under control, the ground is prepared for the study of physical aspects and mechanisms of motion in complex environments, of the behavior of systems with new physical features like chirality, of the development of novel micromachines and microbots, of the emergent collective behavior and swarming of intelligent self-propelled particles, and of particular features of microbial systems. The vast complexity of phenomena and mechanisms involved in the self-organization and dynamics of motile active matter poses major challenges, which can only be addressed by a truly interdisciplinary effort involving scientists from biology, chemistry, ecology, engineering, mathematics, and physics.

cond-mat.soft

Sedimentation of a single soluble particle at low Reynolds and high P\'eclet numbers

We investigate experimentally the dissolution of an almost spherical butyramide particle during its sedimentation, in the low Reynolds high P\'eclet regime. The particle sediments in a quiescent aqueous solution, and its shape and position are measured simultaneously by a camera attached to a translation stage. The particle is tracked in real time, and the translation stage moves accordingly to keep the particle in the field of the camera. The measurements from the particle image show that the radius shrinking rate is constant with time, and independent of the initial radius of the particle. We explain this with a simple model, based on the sedimentation law in the Stokes' regime and the mass transfer rate at low Reynolds and high P\'eclet numbers. The theoretical and experimental results are consistent within $20\%$. We introduce two correction factors to take into account the non-sphericity of the particle and the inclusions of air bubbles inside the particle, and reach quantitative agreement. With these corrections, the indirect measurement of the radius shrinking rate deduced from the position measurement is also in agreement with the model. We discuss other correction factors, and explain why there are negligible in the present experiment. We also compute the effective Sherwood number as a function of an effective P\'eclet number.

physics.flu-dyn

Frustrated run and tumble of swimming E-coli bacteria in nematic liquid crystals

In many situations bacteria move in complex environments, as for example in soils, oceans or the human gut-track microbiome. In these natural environments, carrier fluids such as mucus or reproductive fluids show complex structure associated with non-Newtonian rheology. Many fundamental questions concerning the the ability to navigate in such environments remain unsolved due to the inherent complexity of the natural surroundings. Recently, the interaction of swimming bacteria with nematic liquid crystals has attracted lot of attention. In these structured fluids, the kinetics of bacterial motion is constrained by the orientational molecular order of the liquid crystal (or director field) and novel spatio-temporal patterns arise from this orientational constraint, as well as from the interactions with topological defects. A question unaddressed so far is how bacteria are able to change swimming direction in such an environment. In this work, we study the swimming mechanism of a single bacterium, E. coli, constrained to move along the director field of a lyotropic chromonic liquid crystal (LCLC) that is confined to a planar cell. In such an environment, the spontaneous run and tumble motion of the bacterium gets frustrated: the elasticity of the liquid crystal prevents flagella from unbundling. Interestingly, in order to change direction, bacteria execute a reversal motion along the director field, driven by the relocation of a single flagellum to the other side of the bacterial body, coined as a frustrated tumble. We present a detailed experimental characterization of this phenomenon, exploiting exceptional spatial and temporal resolution of bacteria and flagella dynamics during swimming, obtained using a two color Lagrangian tracking technique. We suggest a possible mechanism behind the frustrated run and tumble motion, accounting for these observations.

cond-mat.soft

Run-to-Tumble Variability Controls the Surface Residence Times of ${\it E.~coli}$ Bacteria

Motile bacteria are known to accumulate at surfaces, eventually leading to changes in bacterial motility and bio-film formation. We use a novel two-colour, three-dimensional Lagrangian tracking technique, to follow simultaneously the body and the flagella of a wild-type ${\it Escherichia~coli}$. We observe long surface residence times and surface escape corresponding mostly to immediately antecedent tumbling. A motility model accounting for a large behavioural variability in run-time duration, reproduces all experimental findings and gives new insights into surface trapping efficiency.

cond-mat.soft

Optimised hyperbolic microchannels for the mechanical characterisation of bio-particles

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 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 the dynamics of objects with a wide range of sizes, characteristics and behaviours of relevance in the biological world.

cond-mat.soft

3D spatial exploration by E. coli echoes motor temporal variability

Unraveling bacterial strategies for spatial exploration is crucial for understanding the complexity in the organization of life. Bacterial motility determines the spatio-temporal structure of microbial communities, controls infection spreading and the microbiota organization in guts or in soils. Most theoretical approaches for modeling bacterial transport rely on their run-and-tumble motion. For Escherichia coli, the run time distribution was reported to follow a Poisson process with a single characteristic time related to the rotational switching of the flagellar motors. However, direct measurements on flagellar motors show heavy-tailed distributions of rotation times stemming from the intrinsic noise in the chemotactic mechanism. Currently, there is no direct experimental evidence that the stochasticity in the chemotactic machinery affect the macroscopic motility of bacteria. In stark contrast with the accepted vision of run-and-tumble, here we report a large behavioral variability of wild-type \emph{E. coli}, revealed in their three-dimensional trajectories. At short observation times, a large distribution of run times is measured on a population and attributed to the slow fluctuations of a signaling protein triggering the flagellar motor reversal. Over long times, individual bacteria undergo significant changes in motility. We demonstrate that such a large distribution of run times introduces measurement biases in most practical situations. Our results reconcile the notorious conundrum between run time observations and motor switching statistics. We finally propose that statistical modeling of transport properties currently undertaken in the emerging framework of active matter studies, should be reconsidered under the scope of this large variability of motility features.

physics.bio-ph

Swimming bacteria in Poiseuille flow: the quest for active Bretherton-Jeffery trajectories

Using a 3D Lagrangian tracking technique, we determine experimentally the trajectories of non-tumbling E. coli mutants swimming in a Poiseuille flow. We identify a typology of trajectories in agreement with a kinematic "active Bretherton-Jeffery" model, featuring an axi-symmetric self-propelled ellipsoid. In particular, we recover the "swinging" and "shear tumbling" kinematics predicted theoretically by Zöttl et al. Moreover using this model, we derive analytically new features such as quasi-planar piece-wise trajectories, associated with the high aspect ratio of the bacteria, as well as the existence of a drift angle around which bacteria perform closed cyclic trajectories. However, the agreement between the model predictions and the experimental results remains local in time, due to the presence of Brownian rotational noise.

physics.flu-dyn

Mechanical fluctuations suppress the threshold of soft-glassy solids : the secular drift scenario

We propose a dynamical mechanism leading to the fluidization of soft-glassy amorphous mate-rial driven below the yield-stress by external mechanical fluctuations. The model is based on the combination of memory effect and non-linearity, leading to an accumulation of tiny effects over a long-term. We test this scenario on a granular packing driven mechanically below the Coulomb threshold. We bring evidences for an effective viscous response directly related to small stress modulations in agreement with the theoretical prediction of a generic secular drift.

cond-mat.soft

Creep and fluidity of a real granular packing near jamming

We study the internal dynamical processes taking place in a granular packing below yield stress. At all packing fractions and down to vanishingly low applied shear, a logarithmic creep is evidenced. The experiments are analyzed under the scope of a visco-elastic model introducing an internal "fluidity" variable. For all experiments, the creep dynamics can be rescaled onto a unique curve which displays jamming at the random-close-packing limit. At each packing fraction, a stress value is evidenced, corresponding to the onset of internal granular reorganisation leading to a slowing down the creep dynamics before the final yield.

cond-mat.soft

Intruder mobility in a vibrated granular packing

We study experimentally the dynamics of a dense intruder sinking under gravity inside a vibrated 2D granular packing. The surrounding flow patterns are characterized and the falling trajectories are interpreted in terms of an effectivive friction coefficient related to the intruder mean descent velocity (flow rules). At higher confining pressures i.e. close to jamming, a transition to intermittent dynamics is evidenced and displays anomalous "on-off" blockade statistics. A systematic analysis of the flow rules, obtained for different intruder sizes, either in the flowing regime or averaged over the flowing and blockade regimes, strongly suggest the existence of non-local properties for the vibrated packing rheology.}

cond-mat.soft

Enhanced diffusion due to active swimmers at a solid surface

We consider two systems of active swimmers moving close to a solid surface, one being a living population of wild-type \textit{E. coli} and the other being an assembly of self-propelled Au-Pt rods. In both situations, we have identified two different types of motion at the surface and evaluated the fraction of the population that displayed ballistic trajectories (active swimmers) with respect to those showing random-like behavior. We studied the effect of this complex swimming activity on the diffusivity of passive tracers also present at the surface. We found that the tracer diffusivity is enhanced with respect to standard Brownian motion and increases linearly with the activity of the fluid, defined as the product of the fraction of active swimmers and their mean velocity. This result can be understood in terms of series of elementary encounters between the active swimmers and the tracers.

physics.bio-ph