Automated Analysis of Single-Molecule Photobleaching Data by Statistical Modeling of Spot Populations.

ABSTRACT: The number of fluorophores within a molecule complex can be revealed by single-molecule photobleaching imaging. A widely applied strategy to analyze intensity traces over time is the quantification of photobleaching step counts. However, several factors can limit and bias the detection of photobleaching steps, including noise, high numbers of fluorophores, and the possibility that several photobleaching events occur almost simultaneously. In this study, we propose a new approach, to our knowledge, to determine the fluorophore number that correlates the intensity decay of a population of molecule complexes with the decay of the number of visible complexes. We validated our approach using single and fourfold Atto-labeled DNA strands. As an example we estimated the subunit stoichiometry of so

SUBMITTER: Liesche C 

PROVIDER: S-EPMC4675885 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature