Micromirror Total Internal Reflection Microscopy for High-Performance Single Particle Tracking at Interfaces

Single particle tracking has found broad applications in the life and physical sciences, enabling the observation and characterization of nano- and microscopic motion. Fluorescence-based approaches are ideally suited for high-background environments, such as tracking lipids or proteins in or on cells, due to superior background rejection. Scattering-based detection is preferable when localization precision and imaging speed are paramount due to the in principle infinite photon budget. Here, we show that micromirror-based total internal reflection dark field microscopy enables background suppression previously only reported for interferometric scattering microscopy, resulting in nanometer localization precision at 6 mu s exposure time for 20 nm gold nanoparticles with a 25 x 25 mu m(2) field of view. We demonstrate the capabilities of our implementation by characterizing sub-nanometer deterministic flows of 20 nm gold nanoparticles at liquid-liquid interfaces. Our results approach the optimal combination of background suppression, localization precision, and temporal resolution achievable with pure scattering-based imaging and tracking of nanoparticles at interfaces.

Automated summary

Single particle tracking has broad applications in life and physical sciences, allowing observation and characterization of nano- and microscopic motion. This study demonstrates that micromirror-based total internal reflection dark field microscopy achieves efficient background suppression, with nanometer localization precision and high temporal resolution.