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‘Simultaneous tracking of cell motility in liquid and at the solid-liquid interface’

Andrew L. Hook, James L. Flewellen, Irwin M. Zaid, Richard M. Berry, Jean-Frédéric Dubern, Alessandro Carabelli, Ricky Wildman, Noah Russell, Paul Williams, Morgan R. Alexander
doi: https://doi.org/10.1101/541615
Andrew L. Hook
1Advanced Materials and Healthcare Technologies Division, School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, United Kingdom
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James L. Flewellen
2Immune Receptor Activation Laboratory, The Francis Crick Institute, London NW1 1AT, United Kingdom
3Division of Immunology and Inflammation, Department of Medicine, Imperial College London, London SW7 2A2, United Kingdom
4Department of Physics, Clarendon Laboratory, University of Oxford, Parks Road, Oxford OX1 3PU, United Kingdom
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Irwin M. Zaid
4Department of Physics, Clarendon Laboratory, University of Oxford, Parks Road, Oxford OX1 3PU, United Kingdom
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Richard M. Berry
4Department of Physics, Clarendon Laboratory, University of Oxford, Parks Road, Oxford OX1 3PU, United Kingdom
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Jean-Frédéric Dubern
5Centre for Biomolecular Sciences, School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, United Kingdom
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Alessandro Carabelli
1Advanced Materials and Healthcare Technologies Division, School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, United Kingdom
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Ricky Wildman
6Department of Chemical and Environmental Engineering, School of Engineering, University of Nottingham, Nottingham NG7 2RD, United Kingdom
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Noah Russell
7Department of Electrical and Electronic Engineering, School of Engineering, University of Nottingham, Nottingham NG7 2RD, United Kingdom
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Paul Williams
5Centre for Biomolecular Sciences, School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, United Kingdom
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  • For correspondence: williams@nottingham.ac.uk morgan.alexander@nottingham.ac.uk
Morgan R. Alexander
1Advanced Materials and Healthcare Technologies Division, School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, United Kingdom
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  • For correspondence: williams@nottingham.ac.uk morgan.alexander@nottingham.ac.uk
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1. Abstract

To better understand key behaviors of living cells, such as bacterial biofilm formation, they must be observed above surfaces and at the interface between the surface and liquid medium. We have established a methodology for label-free imaging and tracking of individual cells simultaneously at both the solid-liquid interface and within the bulk, utilizing imaging modes of digital holographic microscopy (DHM) in 3D, differential interference contrast (DIC) and total internal reflectance microscopy (TIRM) in 2D as well as analysis protocols using a bespoke software package. We illustrate the power of this method by making detailed single cell measurements of Pseudomonas aeruginosa in the first minutes of their interaction with a glass surface, focusing on the role of the flagella stators, motAB and motCD. Using this new method we have determined their relative contributions to bulk and near surface motion for populations of cells at the single cell level.

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Posted February 05, 2019.
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‘Simultaneous tracking of cell motility in liquid and at the solid-liquid interface’
Andrew L. Hook, James L. Flewellen, Irwin M. Zaid, Richard M. Berry, Jean-Frédéric Dubern, Alessandro Carabelli, Ricky Wildman, Noah Russell, Paul Williams, Morgan R. Alexander
bioRxiv 541615; doi: https://doi.org/10.1101/541615
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‘Simultaneous tracking of cell motility in liquid and at the solid-liquid interface’
Andrew L. Hook, James L. Flewellen, Irwin M. Zaid, Richard M. Berry, Jean-Frédéric Dubern, Alessandro Carabelli, Ricky Wildman, Noah Russell, Paul Williams, Morgan R. Alexander
bioRxiv 541615; doi: https://doi.org/10.1101/541615

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