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bioRxiv · 10.1101/199778

Revealing intricate acto-myosin dynamics at a membrane surface using interferometric scattering microscopy

Koester, D.·Hundt, N.·Young, G. C.·Fineberg, A.·Kukura, P.·Mayor, S.
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Abstract

The plasma membrane and the underlying cytoskeletal cortex constitute active platforms for a variety of cellular processes. Recent work has shown that the remodeling acto-myosin network modifies local membrane organization, but the molecular details are only partly 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, and fast acquisition rates, we detect, and image individual membrane attached actin filaments diffusing within the acto-myosin network and follow individual myosin II filament dynamics. We 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. Our results show how decreasing ATP concentrations lead to both increasing dwell times of individual myosin II filaments and a global change from a remodeling to a contractile state of the acto-myosin network. Statement of SignificanceHere, we show that interferometric scattering microscopy in combination with single particle tracking enables label-free, high contrast imaging of filament dynamics on surfaces, while distinguishing different species based on their mass. These results significantly broaden the available toolkit, and associated capabilities of researchers studying dynamics of biological machines at interfaces.

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Koester, D., Hundt, N., Young, G. C., Fineberg, A., Kukura, P., Mayor, S.. 2017-10-07. Revealing intricate acto-myosin dynamics at a membrane surface using interferometric scattering microscopy. https://doi.org/10.1101/199778

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