SpeedyTrack: Direct microsecond wide-field single-molecule tracking and super-resolution mapping via CCD vertical shift.

ABSTRACT: We introduce spatially-encoded dynamics tracking (SpeedyTrack), a strategy to enable direct microsecond wide-field single-molecule tracking/imaging on common microscopy setups. Capitalizing on the native sub-microsecond vertical charge shifting capability of popular electron-multiplying charge-coupled devices (EM-CCDs), SpeedyTrack staggers wide-field single-molecule images along the CCD chip at ~10-row spacings between consecutive timepoints, effectively projecting the time domain to the spatial domain. Wide-field tracking is thus achieved for freely diffusing molecules at down to 50 μs temporal resolutions for >30 timepoints, permitting trajectory analysis to quantify diffusion coefficients up to 1,000 μm²/s. Concurrent acquisition of single-molecule diffusion trajectories and

SUBMITTER: Steves MA 

PROVIDER: S-EPMC12026894 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature