Two-color, laser excitation improves temporal resolution for detecting the dynamic, plasmonic coupling between...

The ability of two, scattering gold nanoparticles (GNPs) to plasmonically couple in a manner that is dependent on the interparticle separation has been exploited to measure nanometer-level displacements. However, despite broad applicability to monitoring biophysical dynamics, the long time scales (<5 Hz) with which plasmonic coupling are typically measured are not suitable for many dynamic molecular processes, generally occurring over several milliseconds. Here, we introduce a new technique intended to overcome this technical limitation: ratiometric analysis using monochromatic, evanescent darkfield illumination (RAMEDI). As a proof-of-principle, we monitored dynamic, plasmonic coupling arising from the binding of single biotin- and neutravidin-GNPs with a temporal resolution of 38 ms. We also show that the observable bandwidth is extendable to faster time scales by demonstrating that RAMEDI is capable of achieving a signal-to-noise ratio greater than 20 from individual GNPs observed with 200 Hz bandwidth.