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Visualizing acto-myosin dynamics and vortices at a membrane surface using interferometric scattering microscopy

View ORCID ProfileDarius Köster, View ORCID ProfileNikolas Hundt, View ORCID ProfileGavin Young, Adam Fineberg, View ORCID ProfilePhilipp Kukura, View ORCID ProfileSatyajit Mayor
doi: https://doi.org/10.1101/199778
Darius Köster
1National Centre for Biological Sciences, Tata Institute for Fundamental Research, GKVK, Bellary Road, Bangalore 560065, India
2Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry CV4 7AL, UK
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  • For correspondence: d.koester@warwick.ac.uk
Nikolas Hundt
3Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3QZ, UK
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Gavin Young
3Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3QZ, UK
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Adam Fineberg
3Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3QZ, UK
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Philipp Kukura
3Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3QZ, UK
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Satyajit Mayor
1National Centre for Biological Sciences, Tata Institute for Fundamental Research, GKVK, Bellary Road, Bangalore 560065, India
4Institute for Stem Cell Biology and Regenerative Medicine, Bangalore 560065, India
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Abstract

The plasma membrane and the underlying cytoskeletal cortex constitute active platforms for many cellular processes. Recent work has shown that acto-myosin dynamics modify the local membrane organization, but the molecular details are not well understood due to difficulties with experimentally accessing the relevant time and length scales. Here, we use interferometric scattering (iSCAT) microscopy to investigate a minimal acto-myosin network linked to a supported lipid bilayer membrane. Using the magnitude of the interferometric contrast, which is proportional to molecular mass, we detect, image and distinguish actin and myosin filaments. As a result, we can follow single, membrane attached actin filaments diffusing within the acto-myosin network, revealing differing types of motion depending on filament length. We go on to quantify myosin II filament dwell times and processivity as a function of ATP concentration, providing evidence for the predicted ensemble behavior of myosin head domains. Simultaneous observation of long-term network flow and organization enables us to link changes in myosin II filament dynamics with decreasing ATP concentrations to a switch in the acto-myosin network from a remodeling, fluid state to contractile behavior, and to observe the formation of vortices so far only predicted by theory.

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Posted February 06, 2018.
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Visualizing acto-myosin dynamics and vortices at a membrane surface using interferometric scattering microscopy
Darius Köster, Nikolas Hundt, Gavin Young, Adam Fineberg, Philipp Kukura, Satyajit Mayor
bioRxiv 199778; doi: https://doi.org/10.1101/199778
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Visualizing acto-myosin dynamics and vortices at a membrane surface using interferometric scattering microscopy
Darius Köster, Nikolas Hundt, Gavin Young, Adam Fineberg, Philipp Kukura, Satyajit Mayor
bioRxiv 199778; doi: https://doi.org/10.1101/199778

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