Simultaneous Multicolor Single-Molecule Tracking with Single-Laser Excitation via Spectral Imaging.

ABSTRACT: Single-molecule tracking (SMT) offers rich information on the dynamics of underlying biological processes, but multicolor SMT has been challenging due to spectral cross talk and a need for multiple laser excitations. Here, we describe a single-molecule spectral imaging approach for live-cell tracking of multiple fluorescent species at once using a single-laser excitation. Fluorescence signals from all the molecules in the field of view are collected using a single objective and split between positional and spectral channels. Images of the same molecule in the two channels are then combined to determine both the location and the identity of the molecule. The single-objective configuration of our approach allows for flexible sample geometry and the use of a live-cell incubation chamber requi

SUBMITTER: Huang T 

PROVIDER: S-EPMC5984991 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature