title: Choosing dyes for cw-STED nanoscopy using self-assembled nanorulers.
abstract: Superresolution microscopy is currently revolutionizing optical imaging. A key factor for getting images of highest quality is - besides a well-performing imaging system - the labeling of the sample. We compared the fluorescent dyes Abberior Star 488, Alexa 488, Chromeo 488 and Oregon Green 488 for use in continuous wave (cw-)STED microscopy in aqueous buffer and in a durable polymer matrix. To optimize comparability, we designed DNA origami standards labeled with the fluorescent dyes including a bead-like DNA origami with dyes focused on one spot and a DNA origami with two marks at a designed distance of ∼100 nm. Our data show that all dyes are well suited for cw-STED microscopy but that the optimal dye depends on the embedding medium. The precise comparison enabled by DNA origami nanorulers indicates that these structures have matured from the proof-of-concept to easily applicable tools in fluorescence microscopy.
authors.affiliation: NanoBioSciences Group, Institute for Physical and Theoretical Chemistry, TU Braunschweig, Hans-Sommer-Str. 10, 38106 Braunschweig, Germany. b.lalkens@tu-bs.de p.tinnefeld@tu-bs.de.
authors.forename: Susanne
authors.initials: S
authors.lastname: Beater
authors.forename: Phil
authors.initials: P
authors.lastname: Holzmeister
authors.forename: Enrico
authors.initials: E
authors.lastname: Pibiri
authors.forename: Birka
authors.initials: B
authors.lastname: Lalkens
authors.forename: Philip
authors.initials: P
authors.lastname: Tinnefeld
mesh_terms: D004247:DNA / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D049329:Nanostructures / Q000737:chemistry*; D013499:Surface Properties
chemical_list: D005456:Fluorescent Dyes; D004247:DNA