title: Fluorescence Imaging with One-Nanometer Accuracy (FIONA) of Cy3-DNA under Deoxygenation Conditions.
abstract: INTRODUCTIONFIONA, short for fluorescence imaging with one-nanometer accuracy, is a simple method for achieving localization of single (or single groups of) fluorophores with nanometer accuracy in the xy plane. This protocol provides the steps necessary to run a control experiment, using DNA labeled with Cy3, to assess the efficacy of the FIONA setup and the level of attainable resolution. The Cy3-DNA is immobilized on a coverslip and imaged under deoxygenation conditions. It is important that the fluorophores remain photostable throughout the experiment. This requires an oxygen-scavenging system (e.g., glucose oxidase and catalase) in the medium.
authors.affiliation: Biophysics Center and Physics Department, University of Illinois, Urbana, IL 61801, USA.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Selvin
authors.forename: Tyler
authors.initials: T
authors.lastname: Lougheed
authors.forename: Melinda
authors.initials: M
authors.lastname: Tonks Hoffman
authors.forename: Hyokeun
authors.initials: H
authors.lastname: Park
authors.forename: Hamza
authors.initials: H
authors.lastname: Balci
authors.forename: Benjamin H
authors.initials: BH
authors.lastname: Blehm
authors.forename: Erdal
authors.initials: E
authors.lastname: Toprak