Curved ratchets improve bacteria rectification in microfluidic devices.

We study how bacteria rectification in microfluidics devices can be optimized by performing experiments with eight ratchets of different shape and size. Results show that curved ratchets perform best and that their radius of curvature influences how well they perform, as it affects the time bacteria spend on the ratchet surface. We find that the optimal bacterial ratchet is a 60µm radius semicircle witch 15µm concavities. We also show that the angle at which bacteria leave the ratchets can play an important role in their efficiency. Lastly, we reproduce our experimental conditions in a simple numerical simulation to confirm our findings.

Green algae scatter off sharp viscosity gradients.

We study the behaviour of the green alga Chlamydomonas reinhardtii (CR) in the presence of neighbouring regions of different viscosity. We show that the velocity and angular diffusion of the algae decreases when the viscosity of the surrounding medium is increased. We report on a phenomenon occurring when the algae try to cross from a region of low viscosity to a highly viscous one, which causes CR to re-orient and scatter away from the interface if it is approached at a sufficiently small angle. We highlight that the effect does not occur for CR crossing from high to low viscosity regions. Lastly we show that algae do not concentrate in the region of high viscosity despite them swimming slower there. On the contrary, they concentrate in the region of low viscosity or maintain a uniform concentration profile, depending on the viscosity ratio between the two regions.

Pattern Engineering of Living Bacterial Colonies Using Meniscus-Driven Fluidic Channels.

Creating adaptive, sustainable, and dynamic biomaterials is a forthcoming mission of synthetic biology. Engineering spatially organized bacterial communities has a potential to develop such bio-metamaterials. However, generating living patterns with precision, robustness, and a low technical barrier remains as a challenge. Here we present an easily implementable technique for patterning live bacterial populations using a controlled meniscus-driven fluidics system, named as MeniFluidics. We demonstrate multiscale patterning of biofilm colonies and swarms with submillimeter resolution. Utilizing the faster bacterial spreading in liquid channels, MeniFluidics allows controlled bacterial colonies both in space and time to organize fluorescently labeled Bacillus subtilis strains into a converged pattern and to form dynamic vortex patterns in confined bacterial swarms. The robustness, accuracy, and low technical barrier of MeniFluidics offer a tool for advancing and inventing new living materials that can be combined with genetically engineered systems, and adding to fundamental research into ecological, evolutional, and physical interactions between microbes.

The effect of flow on swimming bacteria controls the initial colonization of curved surfaces.

The colonization of surfaces by bacteria is a widespread phenomenon with consequences on environmental processes and human health. While much is known about the molecular mechanisms of surface colonization, the influence of the physical environment remains poorly understood. Here we show that the colonization of non-planar surfaces by motile bacteria is largely controlled by flow. Using microfluidic experiments with Pseudomonas aeruginosa and Escherichia coli, we demonstrate that the velocity gradients created by a curved surface drive preferential attachment to specific regions of the collecting surface, namely the leeward side of cylinders and immediately downstream of apexes on corrugated surfaces, in stark contrast to where nonmotile cells attach. Attachment location and rate depend on the local hydrodynamics and, as revealed by a mathematical model benchmarked on the observations, on cell morphology and swimming traits. These results highlight the importance of flow on the magnitude and location of bacterial colonization of surfaces.

Geometric control of bacterial surface accumulation.

Controlling and suppressing bacterial accumulation at solid surfaces is essential for preventing biofilm formation and biofouling. Whereas various chemical surface treatments are known to reduce cell accumulation and attachment, the role of complex surface geometries remains less well understood. Here, we report experiments and simulations that explore the effects of locally varying boundary curvature on the scattering and accumulation dynamics of swimming Escherichia coli bacteria in quasi-two-dimensional microfluidic channels. Our experimental and numerical results show that a concave periodic boundary geometry can decrease the average cell concentration at the boundary by more than 50% relative to a flat surface.

Scattering of biflagellate microswimmers from surfaces.

We use a three-bead-spring model to investigate the dynamics of biflagellate microswimmers near a surface. While the primary dynamics and scattering are governed by geometric-dependent direct contact, the fluid flows generated by the swimmer locomotion are important in orienting it toward or away from the surface. Flagellar noise and in particular cell spinning about the main axis help a surface-trapped swimmer escape, whereas the time a swimmer spends at the surface depends on the incident angle. The dynamics results from a nuanced interplay of direct collisions, hydrodynamics, noise, and the swimmer geometry. We show that to correctly capture the dynamics of a biflagellate swimmer, minimal models need to resolve the shape asymmetry.

Entrainment dominates the interaction of microalgae with micron-sized objects.

The incessant activity of swimming microorganisms has a direct physical effect on surrounding microscopic objects, leading to enhanced diffusion far beyond the level of Brownian motion with possible influences on the spatial distribution of non-motile planktonic species and particulate drifters. Here we study in detail the effect of eukaryotic flagellates, represented by the green microalga Chlamydomonas reinhardtii, on microparticles. Macro- and microscopic experiments reveal that microorganism-colloid interactions are dominated by rare close encounters leading to large displacements through direct entrainment. Simulations and theoretical modelling show that the ensuing particle dynamics can be understood in terms of a simple jump-diffusion process, combining standard diffusion with Poisson-distributed jumps. This heterogeneous dynamics is likely to depend on generic features of the near-field of swimming microorganisms with front-mounted flagella.

Bimodal rheotactic behavior reflects flagellar beat asymmetry in human sperm cells.

Rheotaxis, the directed response to fluid velocity gradients, has been shown to facilitate stable upstream swimming of mammalian sperm cells along solid surfaces, suggesting a robust physical mechanism for long-distance navigation during fertilization. However, the dynamics by which a human sperm orients itself relative to an ambient flow is poorly understood. Here, we combine microfluidic experiments with mathematical modeling and 3D flagellar beat reconstruction to quantify the response of individual sperm cells in time-varying flow fields. Single-cell tracking reveals two kinematically distinct swimming states that entail opposite turning behaviors under flow reversal. We constrain an effective 2D model for the turning dynamics through systematic large-scale parameter scans, and find good quantitative agreement with experiments at different shear rates and viscosities. Using a 3D reconstruction algorithm to identify the flagellar beat patterns causing left or right turning, we present comprehensive 3D data demonstrating the rolling dynamics of freely swimming sperm cells around their longitudinal axis. Contrary to current beliefs, this 3D analysis uncovers ambidextrous flagellar waveforms and shows that the cell's turning direction is not defined by the rolling direction. Instead, the different rheotactic turning behaviors are linked to a broken mirror symmetry in the midpiece section, likely arising from a buckling instability. These results challenge current theoretical models of sperm locomotion.

Microalgae Scatter off Solid Surfaces by Hydrodynamic and Contact Forces.

Interactions between microorganisms and solid boundaries play an important role in biological processes, such as egg fertilization, biofilm formation, and soil colonization, where microswimmers move within a structured environment. Despite recent efforts to understand their origin, it is not clear whether these interactions can be understood as being fundamentally of hydrodynamic origin or hinging on the swimmer's direct contact with the obstacle. Using a combination of experiments and simulations, here we study in detail the interaction of the biflagellate green alga Chlamydomonas reinhardtii, widely used as a model puller microorganism, with convex obstacles, a geometry ideally suited to highlight the different roles of steric and hydrodynamic effects. Our results reveal that both kinds of forces are crucial for the correct description of the interaction of this class of flagellated microorganisms with boundaries.

Rheotaxis facilitates upstream navigation of mammalian sperm cells.

A major puzzle in biology is how mammalian sperm maintain the correct swimming direction during various phases of the sexual reproduction process. Whilst chemotaxis may dominate near the ovum, it is unclear which cues guide spermatozoa on their long journey towards the egg. Hypothesized mechanisms range from peristaltic pumping to temperature sensing and response to fluid flow variations (rheotaxis), but little is known quantitatively about them. We report the first quantitative study of mammalian sperm rheotaxis, using microfluidic devices to investigate systematically swimming of human and bull sperm over a range of physiologically relevant shear rates and viscosities. Our measurements show that the interplay of fluid shear, steric surface-interactions, and chirality of the flagellar beat leads to stable upstream spiralling motion of sperm cells, thus providing a generic and robust rectification mechanism to support mammalian fertilisation. A minimal mathematical model is presented that accounts quantitatively for the experimental observations.DOI: http://dx.doi.org/10.7554/eLife.02403.001.

Membrane viscosity determined from shear-driven flow in giant vesicles.

The viscosity of lipid bilayer membranes plays an important role in determining the diffusion constant of embedded proteins and the dynamics of membrane deformations, yet it has historically proven very difficult to measure. Here we introduce a new method based on quantification of the large-scale circulation patterns induced inside vesicles adhered to a solid surface and subjected to simple shear flow in a microfluidic device. Particle image velocimetry based on spinning disk confocal imaging of tracer particles inside and outside of the vesicle and tracking of phase-separated membrane domains are used to reconstruct the full three-dimensional flow pattern induced by the shear. These measurements show excellent agreement with the predictions of a recent theoretical analysis, and allow direct determination of the membrane viscosity.

Ciliary contact interactions dominate surface scattering of swimming eukaryotes.

Interactions between swimming cells and surfaces are essential to many microbiological processes, from bacterial biofilm formation to human fertilization. However, despite their fundamental importance, relatively little is known about the physical mechanisms that govern the scattering of flagellated or ciliated cells from solid surfaces. A more detailed understanding of these interactions promises not only new biological insights into structure and dynamics of flagella and cilia but may also lead to new microfluidic techniques for controlling cell motility and microbial locomotion, with potential applications ranging from diagnostic tools to therapeutic protein synthesis and photosynthetic biofuel production. Due to fundamental differences in physiology and swimming strategies, it is an open question of whether microfluidic transport and rectification schemes that have recently been demonstrated for pusher-type microswimmers such as bacteria and sperm cells, can be transferred to puller-type algae and other motile eukaryotes, because it is not known whether long-range hydrodynamic or short-range mechanical forces dominate the surface interactions of these microorganisms. Here, using high-speed microscopic imaging, we present direct experimental evidence that the surface scattering of both mammalian sperm cells and unicellular green algae is primarily governed by direct ciliary contact interactions. Building on this insight, we predict and experimentally verify the existence of optimal microfluidic ratchets that maximize rectification of initially uniform Chlamydomonas reinhardtii suspensions. Because mechano-elastic properties of cilia are conserved across eukaryotic species, we expect that our results apply to a wide range of swimming microorganisms.

Human spermatozoa migration in microchannels reveals boundary-following navigation.

The migratory abilities of motile human spermatozoa in vivo are essential for natural fertility, but it remains a mystery what properties distinguish the tens of cells which find an egg from the millions of cells ejaculated. To reach the site of fertilization, sperm must traverse narrow and convoluted channels, filled with viscous fluids. To elucidate individual and group behaviors that may occur in the complex three-dimensional female tract environment, we examine the behavior of migrating sperm in assorted microchannel geometries. Cells rarely swim in the central part of the channel cross-section, instead traveling along the intersection of the channel walls ("channel corners"). When the channel turns sharply, cells leave the corner, continuing ahead until hitting the opposite wall of the channel, with a distribution of departure angles, the latter being modulated by fluid viscosity. If the channel bend is smooth, cells depart from the inner wall when the curvature radius is less than a threshold value close to 150 µm. Specific wall shapes are able to preferentially direct motile cells. As a consequence of swimming along the corners, the domain occupied by cells becomes essentially one-dimensional, leading to frequent collisions, and needs to be accounted for when modeling the behavior of populations of migratory cells and considering how sperm populate and navigate the female tract. The combined effect of viscosity and three-dimensional architecture should be accounted for in future in vitro studies of sperm chemoattraction.

Fluctuations, dynamics, and the stretch-coil transition of single actin filaments in extensional flows.

Semiflexible polymers subject to hydrodynamic forcing play an important role in cytoskeletal motions in the cell, particularly when filaments guide molecular motors whose motions create flows. Near hyperbolic stagnation points, filaments experience a competition between bending elasticity and tension and are predicted to display suppressed thermal fluctuations in the extensional regime and a buckling instability under compression. Using a microfluidic cross-flow geometry, we verify these predictions in detail, including a fluctuation-rounded stretch-coil transition of actin filaments.

Fluid velocity fluctuations in a suspension of swimming protists.

In dilute suspensions of swimming microorganisms the local fluid velocity is a random superposition of the flow fields set up by the individual organisms, which in turn have multipole contributions decaying as inverse powers of distance from the organism. Here we show that the conditions under which the central limit theorem guarantees a Gaussian probability distribution function of velocities are satisfied when the leading force singularity is a Stokeslet, but are not when it is any higher multipole. These results are confirmed by numerical studies and by experiments on suspensions of the alga Volvox carteri, which show that deviations from Gaussianity arise from near-field effects.