title: A simple self-calibrating method to measure the height of fluorescent molecules and beads at nanoscale resolution.
abstract: We describe a simple self-calibrating technique, incident-beam interference sweeping, for measuring the height of fluorescent labels. Using a tilted back-reflecting mirror and a scanning laser beam, a modulated fluorescence emission allows height determination of a label from a surface with a resolution of ∼ 3 nm. In addition, we show that the absolute distance of a label from the top-mounted mirror can be determined with a resolution of a few tens of nanometers over a micrometer range.
authors.affiliation: Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology , Lorentzweg 1, 2628 CJ Delft, The Netherlands.
authors.forename: Jacob W J
authors.initials: JW
authors.lastname: Kerssemakers
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Blosser
authors.forename: Cees
authors.initials: C
authors.lastname: Dekker