title: Binding-activated localization microscopy of DNA structures.
abstract: Many nucleic acid stains show a strong fluorescence enhancement upon binding to double-stranded DNA. Here we exploit this property to perform superresolution microscopy based on the localization of individual binding events. The dynamic labeling scheme and the optimization of fluorophore brightness yielded a resolution of ∼14 nm (fwhm) and a spatial sampling of 1/nm. We illustrate our approach with two different DNA-binding dyes and apply it to visualize the organization of the bacterial chromosome in fixed Escherichia coli cells. In general, the principle of binding-activated localization microscopy (BALM) can be extended to other dyes and targets such as protein structures.
authors.affiliation: Laboratory for Biologically Oriented Materials, ETH Zurich, Zurich, Switzerland. ingmar.schoen@mat.ethz.ch
authors.forename: Ingmar
authors.initials: I
authors.lastname: Schoen
authors.forename: Jonas
authors.initials: J
authors.lastname: Ries
authors.forename: Enrico
authors.initials: E
authors.lastname: Klotzsch
authors.forename: Helge
authors.initials: H
authors.lastname: Ewers
authors.forename: Viola
authors.initials: V
authors.lastname: Vogel
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D004247:DNA / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D005456:Fluorescent Dyes / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007364:Intercalating Agents / Q000494:pharmacology; D008853:Microscopy / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology / Q000379:methods; D009930:Organic Chemicals / Q000494:pharmacology; D011485:Protein Binding
chemical_list: D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D009930:Organic Chemicals; C099476:PicoGreen; D049452:Green Fluorescent Proteins; D004247:DNA