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Microbial stir bars: light-activated rotation of tethered bacterial cells to enhance mixing in stagnant fluids

Jyoti P Gurung, Moein N Kashani, Charitha M de Silva, View ORCID ProfileMatthew AB Baker
doi: https://doi.org/10.1101/2023.01.26.525760
Jyoti P Gurung
1School of Biotechnology and Biomolecular Science, UNSW Sydney, Sydney, NSW, 2052, Australia
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Moein N Kashani
2Future Industries Institute, University of South Australia, Mawson Lakes, SA, 5095, Australia
3Australian National Fabrication Facility – South Australia Node, Mawson Lakes, SA, 5095, Australia
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Charitha M de Silva
4School of Mechanical and Manufacturing Engineering, UNSW Sydney, Sydney, NSW, 2052, Australia
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Matthew AB Baker
1School of Biotechnology and Biomolecular Science, UNSW Sydney, Sydney, NSW, 2052, Australia
5ARC Centre of Excellence in Synthetic Biology, UNSW Sydney, Sydney, NSW, 2052, Australia
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  • ORCID record for Matthew AB Baker
  • For correspondence: matthew.baker@unsw.edu.au
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Abstract

Microfluidics devices are gaining significant interest in biomedical applications. However, in a micron-scale device, reaction speed is often limited by the slow rate of diffusion of the reagents. Several active and passive micro-mixers have been fabricated to enhance mixing in microfluidic devices. Here, we demonstrate external control of mixing by rotating a rodshaped bacterial cell. This rotation is driven by ion transit across the bacterial flagellar stator complex. We first measured the flow fields generated by rotating a single bacterial cell rotationally locked to rotate either clockwise (CW) or counterclockwise (CCW). Micro-Particle Image Velocimetry (μPIV) and Particle Tracking Velocimetry results showed that a bacterial cell of ~ 2.75 μm long, rotating at 5.75 ± 0.39 Hz in a counterclockwise direction could generate distinct micro-vortices with circular flow fields with a mean velocity of 4.72 ± 1.67 μm/s and maximum velocity of 7.90 μm/s in aqueous solution. We verified our experimental data with a numerical simulation at matched flow conditions which revealed vortices of similar dimensions and speed. We observed that the flow-field diminished with increasing z-height above the plane of the rotating cell. Lastly, we showed we could activate and tune rotational mixing remotely using strains engineered with Proteorhodopsin (PR), where rotation could be activated by controlled external illumination using green laser light (561 nm).

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • We have added additional Supplementary Figures for simulation data and more discussion on limitations.

Copyright 
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 4.0 International license.
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Posted March 11, 2023.
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Microbial stir bars: light-activated rotation of tethered bacterial cells to enhance mixing in stagnant fluids
Jyoti P Gurung, Moein N Kashani, Charitha M de Silva, Matthew AB Baker
bioRxiv 2023.01.26.525760; doi: https://doi.org/10.1101/2023.01.26.525760
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Microbial stir bars: light-activated rotation of tethered bacterial cells to enhance mixing in stagnant fluids
Jyoti P Gurung, Moein N Kashani, Charitha M de Silva, Matthew AB Baker
bioRxiv 2023.01.26.525760; doi: https://doi.org/10.1101/2023.01.26.525760

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