RT Journal Article
SR Electronic
T1 Interferometric scattering microscopy reveals microsecond nanoscopic protein motion on a live cell membrane
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 401133
DO 10.1101/401133
A1 Richard W. Taylor
A1 Reza Gholami Mahmoodabadi
A1 Verena Rauschenberger
A1 Andreas Giessl
A1 Alexandra Schambony
A1 Vahid Sandoghdar
YR 2018
UL http://biorxiv.org/content/early/2018/08/27/401133.abstract
AB Much of the biological functions of a cell are dictated by the intricate motion of proteins within its membrane over a spatial range of nanometers to tens of micrometers and time intervals of microseconds to minutes. While this rich parameter space is not accessible to fluorescence microscopy, it can be within reach of interferometric scattering (iSCAT) particle tracking. Being sensitive even to single unlabeled proteins, however, iSCAT is easily accompanied by a large speckle-like background, which poses a substantial challenge for its application to cellular imaging. Here, we show that these difficulties can be overcome and demonstrate tracking of transmembrane epidermal growth factor receptors (EGFR) with nanometer precision in all three dimensions at up to microsecond speeds and tens of minutes duration. We provide unprecedented examples of nanoscale motion and confinement in ubiquitous processes such as diffusion in the plasma membrane, transport on filopodia, and endocytosis.