Nanoscale separation of molecular species based on their rotational mobility

Abstract

We combine far-field fluorescence nanoscopy through serializedrecording of switchable emitters with polarization-sensitive fluorescencedetection. In addition to imaging with nanoscale spatial resolution, thistechnique allows determination of the fluorescence anisotropy of eachdetected dipole emitter and thus an estimate of its rotational mobility. Subpopulationsof fluorescent markers can thus be separated based on theirinteraction with the sample. We applied this new functional nanoscopy toimaging of living mammalian cells.