title: Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy.
abstract: We demonstrate how a conventional confocal spinning-disk (CSD) microscope can be converted into a doubly resolving image scanning microscopy (ISM) system without changing any part of its optical or mechanical elements. Making use of the intrinsic properties of a CSD microscope, we illuminate stroboscopically, generating an array of excitation foci that are moved across the sample by varying the phase between stroboscopic excitation and rotation of the spinning disk. ISM then generates an image with nearly doubled resolution. Using conventional fluorophores, we have imaged single nuclear pore complexes in the nuclear membrane and aggregates of GFP-conjugated Tau protein in three dimensions. Multicolor ISM was shown on cytoskeletal-associated structural proteins and on 3D four-color images including MitoTracker and Hoechst staining. The simple adaptation of conventional CSD equipment allows superresolution investigations of a broad variety of cell biological questions.
authors: Third Institute of Physics-Biophysics and Institute for X-Ray Physics, Georg-August University, 37077 Göttingen, Germany.
authors: Olaf
authors: O
authors: Schulz
authors: Christoph
authors: C
authors: Pieper
authors: Michaela
authors: M
authors: Clever
authors: Janine
authors: J
authors: Pfaff
authors: Aike
authors: A
authors: Ruhlandt
authors: Ralph H
authors: RH
authors: Kehlenbach
authors: Fred S
authors: FS
authors: Wouters
authors: Jörg
authors: J
authors: Großhans
authors: Gertrude
authors: G
authors: Bunt
authors: Jörg
authors: J
authors: Enderlein
mesh_terms: D007089:Image Enhancement / Q000295:instrumentation* / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods* / Q000592:standards*