Ciliary contact interactions dominate surface scattering of swimming eukaryotes.
ABSTRACT: 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.
Project description:Active self-propelled colloidal populations induce time-dependent three-dimensional fluid flows, which alter the rheological (viscoelastic) properties of their fluidic media. Researchers have also studied passive colloids mixed with bacterial suspensions to understand the hydrodynamic coupling between active and passive colloids. With recent developments in biological cell-driven biohybrid microswimmers, different type biological microswimmer (e.g., bacteria and algae) populations need to interact fluidically with each other in the same fluidic media, while such interactions have not been studied experimentally yet. Therefore, we report the swimming behavior of two opposite types of biological microswimmer (active colloid) populations: Chlamydomonas reinhardtii (C. reinhardtii) algae (puller-type microswimmers) population in coculture with Escherichia coli (E. coli) bacteria (pusher-type microswimmers) population. We observed noticeable fluidic coupling deviations from the existing understanding of passive colloids mixed with bacterial suspensions previously studied in the literature. The fluidic coupling among puller- and pusher-type microswimmers led to nonequilibrium fluctuations in the fluid flow due to their opposite swimming patterns. Such coupling could be the main reason behind the shift in motility behaviors of these two opposite-type swimmer populations suspended in the same fluidic media.
Project description:Caulobacter crescentus, a monotrichous bacterium, swims by rotating a single right-handed helical filament. CW motor rotation thrusts the cell forward 1, a mode of motility known as the pusher mode; CCW motor rotation pulls the cell backward, a mode of motility referred to as the puller mode 2. The situation is opposite in E. coli, a peritrichous bacterium, where CCW rotation of multiple left-handed filaments drives the cell forward. The flagellar motor in E. coli generates more torque in the CCW direction than the CW direction in swimming cells 3,4. However, monotrichous bacteria including C. crescentus swim forward and backward at similar speeds, prompting the assumption that motor torques in the two modes are the same 5,6. Here, we present evidence that motors in C. crescentus develop higher torques in the puller mode than in the pusher mode, and suggest that the anisotropy in torque-generation is similar in two species, despite the differences in filament handedness and motor bias (probability of CW rotation).
Project description: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.
Project description:Fission and fusion processes of particle clusters occur in many areas of physics and chemistry from subnuclear to astronomic length scales. Here we study fission and fusion of magnetic microswimmer clusters as governed by their hydrodynamic and dipolar interactions. Rich scenarios are found that depend crucially on whether the swimmer is a pusher or a puller. In particular a linear magnetic chain of pullers is stable while a pusher chain shows a cascade of fission (or disassembly) processes as the self-propulsion velocity is increased. Contrarily, magnetic ring clusters show fission for any type of swimmer. Moreover, we find a plethora of possible fusion (or assembly) scenarios if a single swimmer collides with a ringlike cluster and two rings spontaneously collide. Our predictions are obtained by computer simulations and verifiable in experiments on active colloidal Janus particles and magnetotactic bacteria.
Project description:Motivated by recent experiments demonstrating that motile algae get trapped in draining foams, we study the trajectories of microorganisms confined in model foam channels (section of a Plateau border). We track single Chlamydomonas reinhardtii cells confined in a thin three-circle microfluidic chamber and show that their spatial distribution exhibits strong corner accumulation. Using empirical scattering laws observed in previous experiments (scattering with a constant scattering angle), we next develop a two-dimension geometrical model and compute the phase space of trapped and periodic trajectories of swimmers inside a three-circles billiard. We find that the majority of cell trajectories end up in a corner, providing a geometrical mechanism for corner accumulation. Incorporating the distribution of scattering angles observed in our experiments and including hydrodynamic interactions between the cells and the surfaces into the geometrical model enables us to reproduce the experimental probability density function of micro-swimmers in microfluidic chambers. Both our experiments and models demonstrate therefore that motility leads generically to trapping in complex geometries.
Project description:The journey of spermatozoa through the female genital tract is facilitated by rheotaxis, or the cell's preference to swim against a flow, as well as thigmotaxis, the wall tracking behaviour, which guides them to the site of fertilisation. The aim of this study was to characterise the rheotactic and thigmotactic response of stallion sperm within a microfluidic channel. Stallion sperm rheotaxis was assessed within the microfluidic channel with regard to: (i) A range of flow velocities, (ii) Varying media viscosity and (iii) Sperm hyperactivation. Sperm distribution across the microfluidic channel was also studied and compared to human and ram sperm. Stallion sperm progressed furthest at a velocity range of 10-30?µm/s, with an optimum velocity of 20?µm/s. A flow viscosity of 2.5cP or greater reduced sperm rheotaxis (P?<?0.05). Stallion sperm that were hyperactivated were unable to exhibit rheotaxis within the microfluidic channel, whereas, both hyperactivated human and ram sperm did exhibit positive rheotaxis under the same conditions. The number of sperm swimming near the microfluidic channel walls was higher than in the microfluidic channel centre (P?<?0.05). This is the first study to illustrate that stallion sperm are rheotactically responsive and increasing viscosity reduces this response. We also demonstrated that sperm are predominantly inclined to swim along a surface and uniquely, hyperactivated stallion sperm are non-progressive and do not exhibit a rheotactic response unlike other species.
Project description:Cilia-driven motility and fluid transport are ubiquitous in nature and essential for many biological processes, including swimming of eukaryotic unicellular organisms, mucus transport in airway apparatus or fluid flow in the brain. The-biflagellated micro-swimmer Chlamydomonas reinhardtii is a model organism to study the dynamics of flagellar synchronization. Hydrodynamic interactions, intracellular mechanical coupling or cell body rocking is believed to play a crucial role in the synchronization of flagellar beating in green algae. Here, we use freely swimming intact flagellar apparatus isolated from a wall-less strain of Chlamydomonas to investigate wave dynamics. Our analysis on phase coordinates shows that when the frequency difference between the flagella is high (10-41% of the mean), neither mechanical coupling via basal body nor hydrodynamics interactions are strong enough to synchronize two flagella, indicating that the beating frequency is perhaps controlled internally by the cell. We also examined the validity of resistive force theory for a flagellar apparatus swimming freely in the vicinity of a substrate and found quantitative agreement between the experimental data and simulations with a drag anisotropy of ratio 2. Finally, using a simplified wave form, we investigated the influence of phase and frequency differences, intrinsic curvature and wave amplitude on the swimming trajectory of flagellar apparatus. Our analysis shows that by controlling the phase or frequency differences between two flagella, steering can occur.
Project description:Field-driven direct assembly of nanoscale matter has impact in disparate fields of science. In microscale systems, such concept has been recently exploited to optimize propulsion in viscous fluids. Despite the great potential offered by miniaturization, using self-assembly to achieve transport at the nanoscale remains an elusive task. Here we show that a hybrid propeller, composed by a ferromagnetic nanorod and a paramagnetic microsphere, can be steered in a fluid in a variety of modes, from pusher to puller, when the pair is dynamically actuated by a simple oscillating magnetic field. We exploit this unique design to build more complex structures capable of carrying several colloidal cargos as microscopic trains that quickly disassemble at will under magnetic command. In addition, our prototype can be extended to smaller nanorods below the diffraction limit, but still dynamically reconfigurable by the applied magnetic field.
Project description:Sperm competition, in which the ejaculates of multiple males compete to fertilize a female's ova, results in strong selection on sperm traits. Although sperm size and swimming velocity are known to independently affect fertilization success in certain species, exploring the relationship between sperm length, swimming velocity and fertilization success still remains a challenge. Here, we use the zebra finch (Taeniopygia guttata), where sperm size influences sperm swimming velocity, to determine the effect of sperm total length on fertilization success. Sperm competition experiments, in which pairs of males whose sperm differed only in length and swimming speed, revealed that males producing long sperm were more successful in terms of (i) the number of sperm reaching the ova and (ii) fertilizing those ova. Our results reveal that although sperm length is the main factor determining the outcome of sperm competition, complex interactions between male and female reproductive traits may also be important. The mechanisms underlying these interactions are poorly understood, but we suggest that differences in sperm storage and utilization by females may contribute to the outcome of sperm competition.
Project description:Motile cilia, also called flagella, are found across a broad range of species; some cilia propel prokaryotes and eukaryotic cells like sperm, while cilia on epithelial surfaces create complex fluid patterns e.g., in the brain or lung. For sperm, the picture has emerged that the flagellum is not only a motor but also a sensor that detects stimuli from the environment, computing the beat pattern according to the sensory input. Thereby, the flagellum navigates sperm through the complex environment in the female genital tract. However, we know very little about how environmental signals change the flagellar beat and, thereby, the swimming behavior of sperm. It has been proposed that distinct signaling domains in the flagellum control the flagellar beat. However, a detailed analysis has been mainly hampered by the fact that current comprehensive analysis approaches rely on complex microscopy and analysis systems. Thus, knowledge on sperm signaling regulating the flagellar beat is based on custom quantification approaches that are limited to only a few aspects of the beat pattern, do not resolve the kinetics of the entire flagellum, rely on manual, qualitative descriptions, and are only a little comparable among each other. Here, we present SpermQ, a ready-to-use and comprehensive analysis software to quantify sperm motility. SpermQ provides a detailed quantification of the flagellar beat based on common time-lapse images acquired by dark-field or epi-fluorescence microscopy, making SpermQ widely applicable. We envision SpermQ becoming a standard tool in flagellar and motile cilia research that allows to readily link studies on individual signaling components in sperm and distinct flagellar beat patterns.