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Phenotyping single-cell motility in microfluidic confinement

Samuel A. Bentley, Vasileios Anagnostidis, Hannah Laeverenz Schlogelhofer, Fabrice Gielen, View ORCID ProfileKirsty Y. Wan
doi: https://doi.org/10.1101/2021.12.24.474109
Samuel A. Bentley
Living Systems Institute, Stocker Road, University of Exeter, United Kingdom, EX4 4QD
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Vasileios Anagnostidis
Living Systems Institute, Stocker Road, University of Exeter, United Kingdom, EX4 4QD
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Hannah Laeverenz Schlogelhofer
Living Systems Institute, Stocker Road, University of Exeter, United Kingdom, EX4 4QD
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Fabrice Gielen
Living Systems Institute, Stocker Road, University of Exeter, United Kingdom, EX4 4QD
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  • For correspondence: K.Y.Wan2@exeter.ac.uk F.Gielen@exeter.ac.uk
Kirsty Y. Wan
Living Systems Institute, Stocker Road, University of Exeter, United Kingdom, EX4 4QD
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  • ORCID record for Kirsty Y. Wan
  • For correspondence: K.Y.Wan2@exeter.ac.uk F.Gielen@exeter.ac.uk
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Abstract

At all scales, the movement patterns of organisms serve as dynamic read-outs of their behaviour and physiology. We devised a novel droplet microfluidics assay to encapsulate single algal microswimmers inside closed arenas, and comprehensively studied their roaming behaviour subject to a large number of environmental stimuli. We compared two model species, Chlamydomonas reinhardtii (freshwater alga, 2 cilia), and Pyramimonas octopus (marine alga, 8 cilia), and detailed their highly-stereotyped behaviours and the emergence of a trio of macroscopic swimming states (smooth-forward, quiescent, tumbling or excitable backward). Harnessing ultralong timeseries statistics, we reconstructed the species-dependent reaction network that underlies the choice of locomotor behaviour in these aneural organisms, and discovered the presence of macroscopic non-equilibrium probability fluxes in these active systems. We also revealed for the first time how microswimmer motility changes instantaneously when a chemical is added to their microhabitat, by inducing deterministic fusion between paired droplets - one containing a trapped cell, and the other, a pharmacological agent that perturbs cellular excitability. By coupling single-cell entrapment with unprecedented tracking resolution, speed and duration, our approach offers unique and potent opportunities for diagnostics, drug-screening, and for querying the genetic basis of micro-organismal behaviour.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted December 24, 2021.
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Phenotyping single-cell motility in microfluidic confinement
Samuel A. Bentley, Vasileios Anagnostidis, Hannah Laeverenz Schlogelhofer, Fabrice Gielen, Kirsty Y. Wan
bioRxiv 2021.12.24.474109; doi: https://doi.org/10.1101/2021.12.24.474109
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Phenotyping single-cell motility in microfluidic confinement
Samuel A. Bentley, Vasileios Anagnostidis, Hannah Laeverenz Schlogelhofer, Fabrice Gielen, Kirsty Y. Wan
bioRxiv 2021.12.24.474109; doi: https://doi.org/10.1101/2021.12.24.474109

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