Dissection of molecular assembly dynamics by tracking orientation and position of single molecules in live cells.

ABSTRACT: Regulation of order, such as orientation and conformation, drives the function of most molecular assemblies in living cells but remains difficult to measure accurately through space and time. We built an instantaneous fluorescence polarization microscope, which simultaneously images position and orientation of fluorophores in living cells with single-molecule sensitivity and a time resolution of 100 ms. We developed image acquisition and analysis methods to track single particles that interact with higher-order assemblies of molecules. We tracked the fluctuations in position and orientation of molecules from the level of an ensemble of fluorophores down to single fluorophores. We tested our system in vitro using fluorescently labeled DNA and F-actin, in which the ensemble orientation of po

SUBMITTER: Mehta SB 

PROVIDER: S-EPMC5081662 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature