A simple self-calibrating method to measure the height of fluorescent molecules and beads at nanoscale resolution.

Kerssemakers, Jacob W J; Blosser, Timothy R; Dekker, Cees · Nano Lett · 2014

basic_science · Level V

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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.