Batchelor Prize Lecture Fluid dynamics at the scale of the cell

The world of cellular biology provides us with many fascinating fluid dynamical phenomena that lie at the heart of physiology, development, evolution and ecology. Advances in imaging, micromanipulation and microfluidics over the past decade have made possible high-precision measurements of such flows, providing tests of microhydrodynamic theories and revealing a wealth of new phenomena calling out for explanation. Here I summarize progress in four areas within the field of ‘active matter’: cytoplasmic streaming in plant cells, synchronization of eukaryotic flagella, interactions between swimming cells and surfaces and collective behaviour in suspensions of microswimmers. Throughout, I emphasize open problems in which fluid dynamical methods are key ingredients in an interdisciplinary approach to the mysteries of life.The work described here has also been generously supported by the Engineering and Physical Sciences Research Council, the Biology and Biotechnology Research Council, the Leverhulme Trust, the European Research Council, and the Wellcome Trust.This is the final version of the article. It first appeared from Cambridge University Press via http://dx.doi.org/10.1017/jfm.2016.58

Coordinated beating of algal flagella is mediated by basal coupling

This is the final version of the article. Available from National Academy of Sciences via the DOI in this recordCilia and flagella often exhibit synchronized behavior; this includes phase locking, as seen in Chlamydomonas, and metachronal wave formation in the respiratory cilia of higher organisms. Since the observations by Gray and Rothschild of phase synchrony of nearby swimming spermatozoa, it has been a working hypothesis that synchrony arises from hydrodynamic interactions between beating filaments. Recent work on the dynamics of physically separated pairs of flagella isolated from the multicellular alga Volvox has shown that hydrodynamic coupling alone is sufficient to produce synchrony. However, the situation is more complex in unicellular organisms bearing few flagella. We show that flagella of Chlamydomonas mutants deficient in filamentary connections between basal bodies display markedly different synchronization from the wild type. We perform micromanipulation on configurations of flagella and conclude that a mechanism, internal to the cell, must provide an additional flagellar coupling. In naturally occurring species with 4, 8, or even 16 flagella, we find diverse symmetries of basal body positioning and of the flagellar apparatus that are coincident with specific gaits of flagellar actuation, suggesting that it is a competition between intracellular coupling and hydrodynamic interactions that ultimately determines the precise form of flagellar coordination in unicellular algae.This work is supported by a Junior Research Fellowship from Magdalene College Cambridge (to K.Y.W.) and a Wellcome Trust Senior Investigator Award (to R.E.G.)

Time Irreversibility and Criticality in the Motility of a Flagellate Microorganism

This is the final version of the article. Available from American Physical Society via the DOI in this record.Active living organisms exhibit behavioral variability, partitioning between fast and slow dynamics. Such variability may be key to generating rapid responses in a heterogeneous, unpredictable environment wherein cellular activity effects continual exchanges of energy fluxes. We demonstrate a novel, noninvasive strategy for revealing nonequilibrium control of swimming—specifically, in an octoflagellate microalga. These organisms exhibit surprising features of flagellar excitability and mechanosensitivity, which characterize a novel, time-irreversible “run-stop-shock” motility comprising forward runs, knee-jerk shocks with dramatic beat reversal, and long stops during which cells are quiescent yet continue to exhibit submicron flagellar vibrations. Entropy production, associated with flux cycles arising in a reaction graph representation of the gait-switching dynamics, provides a direct measure of detailed balance violation in this primitive alga.Financial support is acknowledged from Magdalene College, Cambridge, through a Junior Research Fellowship (K. Y. W.), and Senior Investigator Grants No. 097855MA and No. 207510/Z/17/Z from the Wellcome Trust (R. E. G.). We thank Robert G. Endres and Eric Lauga for the discussions

Rhythmicity, Recurrence, and Recovery of Flagellar Beating

This is the final version of the article. Available from American Physical Society via the DOI in this recordThe eukaryotic flagellum beats with apparently unfailing periodicity, yet responds rapidly to stimuli. Like the human heartbeat, flagellar oscillations are now known to be noisy. Using the alga C. reinhardtii, we explore three aspects of nonuniform flagellar beating. We report the existence of rhythmicity, waveform noise peaking at transitions between power and recovery strokes, and fluctuations of interbeat intervals that are correlated and even recurrent, with memory extending to hundreds of beats. These features are altered qualitatively by physiological perturbations. Further, we quantify the recovery of periodic breaststroke beating from transient hydrodynamic forcing. These results will help constrain microscopic theories on the origins and regulation of flagellar beating.Financial support is acknowledged from the Engineering and Physical Sciences Research Council, European Research Council Advanced Investigator Grant No. 247333, and a Senior Investigator Award from the Wellcome Trust

Directed collective motion of bacteria under channel confinement

Dense suspensions of swimming bacteria are known to exhibit collective behaviour arising from the interplay of steric and hydrodynamic interactions. Unconfined suspensions exhibit transient, recurring vortices and jets, whereas those confined in circular domains may exhibit order in the form of a spiral vortex. Here we show that confinement into a long and narrow macroscopic ‘racetrack’ geometry stabilises bacterial motion to form a steady unidirectional circulation. This motion is reproduced in simulations of discrete swimmers that reveal the crucial role that bacteria-driven fluid flows play in the dynamics. In particular, cells close to the channel wall produce strong flows which advect cells in the bulk against their swimming direction.Weexamine in detail the transition from a disordered state to persistent directed motion as a function of the channel width, and show that the width at the crossover point is comparable to the typical correlation length of swirls seen in the unbounded system. Our results shed light on the mechanisms driving the collective behaviour of bacteria and other active matter systems, and stress the importance of the ubiquitous boundaries found in natural habitats.This is the final published version. It first appeared at http://dx.doi.org/10.1088/1367-2630/18/7/075002

Lag, lock, sync, slip: the many 'phases' of coupled flagella

This is the final version of the article. Available from the Royal Society via the DOI in this recordIn a multitude of life's processes, cilia and flagella are found indispensable. Recently, the biflagellated chlorophyte alga Chlamydomonas has become a model organism for the study of ciliary motility and synchronization. Here, we use high-speed, high-resolution imaging of single pipette-held cells to quantify the rich dynamics exhibited by their flagella. Underlying this variability in behaviour are biological dissimilarities between the two flagella—termed cis and trans, with respect to a unique eyespot. With emphasis on the wild-type, we derive limit cycles and phase parametrizations for self-sustained flagellar oscillations from digitally tracked flagellar waveforms. Characterizing interflagellar phase synchrony via a simple model of coupled oscillators with noise, we find that during the canonical swimming breaststroke the cis flagellum is consistently phase-lagged relative to, while remaining robustly phase-locked with, the trans flagellum. Transient loss of synchrony, or phase slippage, may be triggered stochastically, in which the trans flagellum transitions to a second mode of beating with attenuated beat envelope and increased frequency. Further, exploiting this alga's ability for flagellar regeneration, we mechanically induced removal of one or the other flagellum of the same cell to reveal a striking disparity between the beatings of the cis and trans flagella, in isolation. These results are evaluated in the context of the dynamic coordination of Chlamydomonas flagella.Financial support is acknowledged from the EPSRC, ERC Advanced Investigator Grant 247333, and a Senior Investigator Award from the Wellcome Trust (R.E.G.)

Nonlinear concentration patterns and bands in autochemotactic suspensions

In suspensions of microorganisms, pattern formation can arise from the interplay of chemotaxis and the fluid flows collectively generated by the organisms themselves. Here we investigate the resulting pattern formation in square and elongated domains in the context of two distinct models of locomotion in which the chemoattractant dynamics is fully coupled to the fluid flows and swimmer motion. Analyses for both models reveal an aggregative instability due to chemotaxis, independent of swimmer shape and type, and a hydrodynamic instability for "pusher" swimmers. We discuss the similarities and differences between the models. Simulations reveal a critical length scale of the swimmer aggregates and this feature can be utilized to stabilize swimmer concentration patterns into quasi-one-dimensional bands by varying the domain size. These concentration bands transition to traveling pulses under an external chemoattractant gradient, as observed in experiments with chemotactic bacteria.E.L. acknowledges a New Jersey Institute of Technology faculty seed grant award. R.E.G. was supported in part by Established Career Fellowship EP/M017982/1 from the Engineering and Physical Sciences Research Council and the Schlumberger Chair Fund. M.J.S. acknowledges support from NSF Grants No. DMS-1463962 and No. DMS-1620331, as well as the NSF Grant No. DMR-1420073 awarded to the MRSEC at NYU

A Compact Eulerian Representation of Axisymmetric Inviscid Vortex Sheet Dynamics

A classical problem in fluid mechanics is the motion of an axisymmetric vortex sheet evolving under the action of surface tension, surrounded by an inviscid fluid. Lagrangian descriptions of these dynamics are well-known, involving complex nonlocal expressions for the radial and longitudinal velocities in terms of elliptic integrals. Here we use these prior results to arrive at a remarkably compact and exact Eulerian evolution equation for the sheet radius $r(z,t)$ in an explicit flux form associated with the conservation of enclosed volume. The flux appears as an integral involving the pairwise mutual induction formula for vortex loop pairs first derived by Helmholtz and Maxwell. We show how the well-known linear stability results for cylindrical vortex sheets in the presence of surface tension and streaming flows [A.M. Sterling and C.A. Sleicher, $J.~Fluid~Mech.$ ${\bf 68}$, 477 (1975)] can be obtained directly from this formulation. Furthermore, the inviscid limit of the empirical model of Eggers and Dupont [$J.~Fluid~Mech.$ $\textbf{262}$ 205 (1994); $SIAM~J.~Appl.~Math.$ ${\bf 60}$, 1997 (2000)], which has served as the basis for understanding singularity formation in droplet pinchoff, is derived within the present formalism as the leading order term in an asymptotic analysis for long slender axisymmetric vortex sheets, and should provide the starting point for a rigorous analysis of singularity formation.This work was supported in part by Established Career Fellowship EP/M017982/1 from the EPSRC (REG & AIP). REG and AIP are grateful to the I.H.E.S., and especially Patrick Gourdon, for hospitality during an extended visit supported by the Schlumberger Visiting Professorship (REG)

Elastohydrodynamic synchronization of adjacent beating flagella

It is now well established that nearby beating pairs of eukaryotic flagella or cilia typically synchronize in phase. A substantial body of evidence supports the hypothesis that hydrodynamic coupling between the active filaments, combined with waveform compliance, provides a robust mechanism for synchrony. This elastohydrodynamic mechanism has been incorporated into bead-spring models in which the beating flagella are represented by microspheres tethered by radial springs as they are driven about orbits by internal forces. While these low-dimensional models reproduce the phenomenon of synchrony, their parameters are not readily relatable to those of the filaments they represent. More realistic models, which reflect the underlying elasticity of the axonemes and the active force generation, take the form of fourth-order nonlinear partial differential equations (PDEs). While computational studies have shown the occurrence of synchrony, the effects of hydrodynamic coupling between nearby filaments governed by such continuum models have been examined theoretically only in the regime of interflagellar distances d large compared to flagellar length $\textit{L}$. Yet in many biological situations $\textit{d/L}$≪1. Here we present an asymptotic analysis of the hydrodynamic coupling between two extended filaments in the regime $\textit{d/L}$≪1 and find that the form of the coupling is independent of the microscopic details of the internal forces that govern the motion of the individual filaments. The analysis is analogous to that yielding the localized induction approximation for vortex filament motion, extended to the case of mutual induction. In order to understand how the elastohydrodynamic coupling mechanism leads to synchrony of extended objects, we introduce a heuristic model of flagellar beating. The model takes the form of a single fourth-order nonlinear PDE whose form is derived from symmetry considerations, the physics of elasticity, and the overdamped nature of the dynamics. Analytical and numerical studies of this model illustrate how synchrony between a pair of filaments is achieved through the asymptotic coupling.This work was supported by Wellcome Trust Senior Investigator Award 097855MA (R.E.G. and A.I.P.) and by a Marie Curie Career Integration Grant (E.L.)

Flagellar synchronization through direct hydrodynamic interactions

This is the final version of the article. Available from eLife Sciences Publications via the DOI in this recordFlows generated by ensembles of flagella are crucial to development, motility and sensing, but the mechanisms behind this striking coordination remain unclear. We present novel experiments in which two micropipette-held somatic cells of Volvox carteri, with distinct intrinsic beating frequencies, are studied by high-speed imaging as a function of their separation and orientation. Analysis of time series shows that the interflagellar coupling, constrained by lack of connections between cells to be hydrodynamical, exhibits a spatial dependence consistent with theory. At close spacings it produces robust synchrony for thousands of beats, while at increasing separations synchrony is degraded by stochastic processes. Manipulation of the relative flagellar orientation reveals in-phase and antiphase states, consistent with dynamical theories. Flagellar tracking with exquisite precision reveals waveform changes that result from hydrodynamic coupling. This study proves unequivocally that flagella coupled solely through a fluid can achieve robust synchrony despite differences in their intrinsic properties.Funding. European Research Council (Advanced Investigator Grant 247333): Douglas R Brumley, Kirsty Y Wan, Marco Polin, Raymond E Goldstein. Wellcome Trust (Senior Investigator Award): Douglas R Brumley, Kirsty Y Wan, Raymond E Goldstein. Engineering and Physical Sciences Research Council: Kirsty Y Wan, Marco Polin, Raymond E Goldstein. Human Frontier Science Program: Douglas R Brumle