title: Breaking the diffraction limit of light-sheet fluorescence microscopy by RESOLFT.
abstract: We present a plane-scanning RESOLFT [reversible saturable/switchable optical (fluorescence) transitions] light-sheet (LS) nanoscope, which fundamentally overcomes the diffraction barrier in the axial direction via confinement of the fluorescent molecular state to a sheet of subdiffraction thickness around the focal plane. To this end, reversibly switchable fluorophores located right above and below the focal plane are transferred to a nonfluorescent state at each scanning step. LS-RESOLFT nanoscopy offers wide-field 3D imaging of living biological specimens with low light dose and axial resolution far beyond the diffraction barrier. We demonstrate optical sections that are thinner by 5-12-fold compared with their conventional diffraction-limited LS analogs.
authors.affiliation: Optical Nanoscopy Division, German Cancer Research Center, 69120 Heidelberg, Germany; Bioquant, 69120 Heidelberg, Germany; Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany;
authors.forename: Patrick
authors.initials: P
authors.lastname: Hoyer
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany;
authors.forename: Gustavo
authors.initials: G
authors.lastname: de Medeiros
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany; Faculty of Information Technology and Bionics, Pázmány Péter Catholic University, 1083 Budapest, Hungary;
authors.forename: Bálint
authors.initials: B
authors.lastname: Balázs
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany;
authors.forename: Nils
authors.initials: N
authors.lastname: Norlin
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany;
authors.forename: Christina
authors.initials: C
authors.lastname: Besir
authors.affiliation: Optical Nanoscopy Division, German Cancer Research Center, 69120 Heidelberg, Germany; Bioquant, 69120 Heidelberg, Germany; Department of Infectious Diseases, Virology, University of Heidelberg, 69120 Heidelberg, Germany;
authors.forename: Janina
authors.initials: J
authors.lastname: Hanne
authors.affiliation: Department of Infectious Diseases, Virology, University of Heidelberg, 69120 Heidelberg, Germany;
authors.forename: Hans-Georg
authors.initials: HG
authors.lastname: Kräusslich
authors.affiliation: Optical Nanoscopy Division, German Cancer Research Center, 69120 Heidelberg, Germany; Bioquant, 69120 Heidelberg, Germany;
authors.forename: Johann
authors.initials: J
authors.lastname: Engelhardt
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
authors.forename: Steffen J
authors.initials: SJ
authors.lastname: Sahl
authors.affiliation: Optical Nanoscopy Division, German Cancer Research Center, 69120 Heidelberg, Germany; Bioquant, 69120 Heidelberg, Germany; Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany shell@gwdg.de hufnagel@embl.de.
authors.forename: Stefan W
authors.initials: SW
authors.lastname: Hell
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany; shell@gwdg.de hufnagel@embl.de.
authors.forename: Lars
authors.initials: L
authors.lastname: Hufnagel
keywords: 3D; RESOLFT; light-sheet microscopy; live-cell imaging; optical nanoscopy