Resolving Fast, Confined Diffusion in Bacteria with Image Correlation Spectroscopy.

ABSTRACT: By following single fluorescent molecules in a microscope, single-particle tracking (SPT) can measure diffusion and binding on the nanometer and millisecond scales. Still, although SPT can at its limits characterize the fastest biomolecules as they interact with subcellular environments, this measurement may require advanced illumination techniques such as stroboscopic illumination. Here, we address the challenge of measuring fast subcellular motion by instead analyzing single-molecule data with spatiotemporal image correlation spectroscopy (STICS) with a focus on measurements of confined motion. Our SPT and STICS analysis of simulations of the fast diffusion of confined molecules shows that image blur affects both STICS and SPT, and we find biased diffusion rate measurements for STICS ana

SUBMITTER: Rowland DJ 

PROVIDER: S-EPMC4880800 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature