title: Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes.
abstract: We introduce MINFLUX, a concept for localizing photon emitters in space. By probing the emitter with a local intensity minimum of excitation light, MINFLUX minimizes the fluorescence photons needed for high localization precision. In our experiments, 22 times fewer fluorescence photons are required as compared to popular centroid localization. In superresolution microscopy, MINFLUX attained ~1-nm precision, resolving molecules only 6 nanometers apart. MINFLUX tracking of single fluorescent proteins increased the temporal resolution and the number of localizations per trace by a factor of 100, as demonstrated with diffusing 30Sribosomal subunits in livingEscherichia coliAs conceptual limits have not been reached, we expect this localization modality to break new ground for observing the dynamics, distribution, and structure of macromolecules in living cells and beyond.
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany. stefan.hell@mpibpc.mpg.de francisco.balzarotti@mpibpc.mpg.de.
authors.forename: Francisco
authors.identifier: 0000-0003-1471-8120
authors.initials: F
authors.lastname: Balzarotti
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Yvan
authors.identifier: 0000-0001-6209-4864
authors.initials: Y
authors.lastname: Eilers
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Klaus C
authors.identifier: 0000-0002-8897-2335
authors.initials: KC
authors.lastname: Gwosch
authors.affiliation: Department of Cell and Molecular Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
authors.forename: Arvid H
authors.identifier: 0000-0001-8087-7715
authors.initials: AH
authors.lastname: Gynnå
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Westphal
authors.affiliation: Centro de Investigaciones en Bionanociencias (CIBION), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.|Departamento de Física, Facultad de Ciencias Exactas y Naturales, University of Buenos Aires, Buenos Aires, Argentina.
authors.forename: Fernando D
authors.identifier: 0000-0002-3277-7215
authors.initials: FD
authors.lastname: Stefani
authors.affiliation: Department of Cell and Molecular Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.
authors.forename: Johan
authors.identifier: 0000-0001-5522-1810
authors.initials: J
authors.lastname: Elf
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany. stefan.hell@mpibpc.mpg.de francisco.balzarotti@mpibpc.mpg.de.|Department of Optical Nanoscopy, Max Planck Institute for Medical Research, Heidelberg, Germany.|Optical Nanoscopy Division, German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Stefan W
authors.identifier: 0000-0002-9638-5077
authors.initials: SW
authors.lastname: Hell
mesh_terms: D004247:DNA / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry; D008164:Luminescent Proteins / Q000032:analysis*; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*; D017785:Photons; D054680:Ribosome Subunits, Small, Bacterial / Q000737:chemistry; D000072760:Single Molecule Imaging / Q000379:methods*
chemical_list: D008164:Luminescent Proteins; D004247:DNA