How to Track Protists in Three Dimensions

We present an apparatus optimized for tracking swimming microorganisms in the size range 10-1000 microns, in three dimensions (3D), far from surfaces, and with negligible background convective fluid motion. CCD cameras attached to two long working distance microscopes synchronously image the sample from two perpendicular directions, with narrowband dark-field or bright-field illumination chosen to avoid triggering a phototactic response. The images from the two cameras can be combined to yield 3D tracks of the organism. Using additional, highly directional broad-spectrum illumination with millisecond timing control the phototactic trajectories in 3D of organisms ranging from Chlamydomonas to Volvox can be studied in detail. Surface-mediated hydrodynamic interactions can also be investigated without convective interference. Minimal modifications to the apparatus allow for studies of chemotaxis and other taxes.Comment: 8 pages, 7 figure

Mixing by Swimming Algae

In this fluid dynamics video, we demonstrate the microscale mixing enhancement of passive tracer particles in suspensions of swimming microalgae, Chlamydomonas reinhardtii. These biflagellated, single-celled eukaryotes (10 micron diameter) swim with a "breaststroke" pulling motion of their flagella at speeds of about 100 microns/s and exhibit heterogeneous trajectory shapes. Fluorescent tracer particles (2 micron diameter) allowed us to quantify the enhanced mixing caused by the swimmers, which is relevant to suspension feeding and biogenic mixing. Without swimmers present, tracer particles diffuse slowly due solely to Brownian motion. As the swimmer concentration is increased, the probability density functions (PDFs) of tracer displacements develop strong exponential tails, and the Gaussian core broadens. High-speed imaging (500 Hz) of tracer-swimmer interactions demonstrates the importance of flagellar beating in creating oscillatory flows that exceed Brownian motion out to about 5 cell radii from the swimmers. Finally, we also show evidence of possible cooperative motion and synchronization between swimming algal cells.Comment: 1 page, APS-DFD 2009 Gallery of Fluid Motio

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.)

Antiphase Synchronization in a Flagellar-Dominance Mutant of Chlamydomonas

Groups of beating flagella or cilia often synchronize so that neighboring filaments have identical frequencies and phases. A prime example is provided by the unicellular biflagellate Chlamydomonas reinhardtii, which typically displays synchronous in-phase beating in a low-Reynolds number version of breaststroke swimming. We report here the discovery that ptx1, a flagellar dominance mutant of C. reinhardtii, can exhibit synchronization in precise antiphase, as in the freestyle swimming stroke. Long-duration high-speed imaging shows that ptx1 flagella switch stochastically between in-phase and antiphase states, and that the latter has a distinct waveform and significantly higher frequency, both of which are strikingly similar to those found during phase slips that stochastically interrupt in-phase beating of the wildtype. Possible mechanisms underlying these observations are discussed.Comment: 5 pages, 4 figure

Aerotaxis in the closest relatives of animals

As the closest unicellular relatives of animals, choanoflagellates serve as useful model organisms for understanding the evolution of animal multicellularity. An important factor in animal evolution was the increasing ocean oxygen levels in the Precambrian, which are thought to have influenced the emergence of complex multicellular life. As a first step in addressing these conditions, we study here the response of the colony-forming choanoflagellate $\textit{Salpingoeca rosetta}$ to oxygen gradients. Using a microfluidic device that allows spatio-temporal variations in oxygen concentrations, we report the discovery that $\textit{S. rosetta}$ displays positive aerotaxis. Analysis of the spatial population distributions provides evidence for logarithmic sensing of oxygen, which enhances sensing in low oxygen neighborhoods. Analysis of search strategy models on the experimental colony trajectories finds that choanoflagellate aerotaxis is consistent with stochastic navigation, the statistics of which are captured using an effective continuous version based on classical run-and-tumble chemotaxis.Engineering and Physical Sciences Research Council, St John’s College, European Research Council (Advanced Investigator Grant ID: 247333), Wellcome Trust (Senior Investigator Award

Antiphase Synchronization in a Flagellar-Dominance Mutant of Chlamydomonas

This is the final version. Available on open access from American Physical Society via the DOI in this record. Groups of beating flagella or cilia often synchronize so that neighboring filaments have identical frequencies and phases. A prime example is provided by the unicellular biflagellate Chlamydomonas reinhardtii, which typically displays synchronous in-phase beating in a low-Reynolds number version of breaststroke swimming. We report the discovery that ptx1, a flagellar-dominance mutant of C. reinhardtii, can exhibit synchronization in precise antiphase, as in the freestyle swimming stroke. High-speed imaging shows that ptx1 flagella switch stochastically between in-phase and antiphase states, and that the latter has a distinct waveform and significantly higher frequency, both of which are strikingly similar to those found during phase slips that stochastically interrupt in-phase beating of the wild-type. Possible mechanisms underlying these observations are discussed.Spanish Ministerio de Ciencia y InnovaciónEngineering and Physical Sciences Research Council (EPSRC)Biotechnology and Biological Sciences Research Council (BBSRC)European Research CouncilWellcome Trus

Antiphase synchronization in a flagellar-dominance mutant of Chlamydomonas

Groups of beating flagella or cilia often synchronize so that neighboring filaments have identical frequencies and phases. A prime example is provided by the unicellular biflagellate Chlamydomonas reinhardtii, which typically displays synchronous in-phase beating in a low-Reynolds number version of breaststroke swimming. We report the discovery that ptx1, a flagellar-dominance mutant of C. reinhardtii, can exhibit synchronization in precise antiphase, as in the freestyle swimming stroke. High-speed imaging shows that ptx1 flagella switch stochastically between in-phase and antiphase states, and that the latter has a distinct waveform and significantly higher frequency, both of which are strikingly similar to those found during phase slips that stochastically interrupt in-phase beating of the wild-type. Possible mechanisms underlying these observations are discussed. Published by the American Physical Society under the terms of the Creative Commons Attribution 3.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article's title, journal citation, and DOI.Support is acknowledged from the Spanish Ministerio de Ciencia y Innovación Grant No. FIS2010-22322-C01 and a Ramón y Cajal Fellowship (I. T.), an EPSRC postdoctoral Fellowship (M. P.), the BBSRC, the EPSRC, ERC Advanced Investigator Grant No. 247333, and a Senior Investigator Award from the Wellcome Trust (R. E. G.)Peer Reviewe