Label-free Imaging of Microtubules with Sub-nm Precision Using Interferometric Scattering Microscopy.

ABSTRACT: Current in vitro optical studies of microtubule dynamics tend to rely on fluorescent labeling of tubulin, with tracking accuracy thereby limited by the quantum yield of fluorophores and by photobleaching. Here, we demonstrate label-free tracking of microtubules with nanometer precision at kilohertz frame rates using interferometric scattering microscopy (iSCAT). With microtubules tethered to a glass substrate using low-density kinesin, we readily detect sequential 8 nm steps in the microtubule center of mass, characteristic of a single kinesin molecule moving a microtubule. iSCAT also permits dynamic changes in filament length to be measured with <5 nm precision. Using the arbitrarily long observation time enabled by label-free iSCAT imaging, we demonstrate continuous monitoring of microtu

SUBMITTER: Andrecka J 

PROVIDER: S-EPMC4806212 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature