Real-time 3D single molecule tracking.

ABSTRACT: To date, single molecule studies have been reliant on tethering or confinement to achieve long duration and high temporal resolution measurements. Here, we present a 3D single-molecule active real-time tracking method (3D-SMART) which is capable of locking on to single fluorophores in solution for minutes at a time with photon limited temporal resolution. As a demonstration, 3D-SMART is applied to actively track single Atto 647 N fluorophores in 90% glycerol solution with an average duration of ~16 s at count rates of ~10 kHz. Active feedback tracking is further applied to single proteins and nucleic acids, directly measuring the diffusion of various lengths (99 to 1385 bp) of single DNA molecules at rates up to 10 µm²/s. In addition, 3D-SMART is able to quantify the occupancy o

SUBMITTER: Hou S 

PROVIDER: S-EPMC7368020 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature