title: Live-cell multiplane three-dimensional super-resolution optical fluctuation imaging.
abstract: Super-resolution optical fluctuation imaging (SOFI) provides an elegant way of overcoming the diffraction limit in all three spatial dimensions by computing higher-order cumulants of image sequences of blinking fluorophores acquired with a classical widefield microscope. Previously, three-dimensional (3D) SOFI has been demonstrated by sequential imaging of multiple depth positions. Here we introduce a multiplexed imaging scheme for the simultaneous acquisition of multiple focal planes. Using 3D cross-cumulants, we show that the depth sampling can be increased. The simultaneous acquisition of multiple focal planes significantly reduces the acquisition time and thus the photobleaching. We demonstrate multiplane 3D SOFI by imaging fluorescently labelled cells over an imaged volume of up to 65 × 65 × 3.5 μm(3) without depth scanning. In particular, we image the 3D network of mitochondria in fixed C2C12 cells immunostained with Alexa 647 fluorophores and the 3D vimentin structure in living Hela cells expressing the fluorescent protein Dreiklang.
authors.affiliation: cole Polytechnique Fédérale de Lausanne, Laboratoire d'Optique Biomédicale, 1015 Lausanne, Switzerland.
authors.forename: Stefan
authors.initials: S
authors.lastname: Geissbuehler
authors.affiliation: cole Polytechnique Fédérale de Lausanne, Laboratoire d'Optique Biomédicale, 1015 Lausanne, Switzerland.
authors.forename: Azat
authors.initials: A
authors.lastname: Sharipov
authors.affiliation: cole Polytechnique Fédérale de Lausanne, Laboratory of Bioorganic Chemistry and Molecular Imaging, Institute of Chemical Sciences and Engineering (ISIC), 1015 Lausanne, Switzerland.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Godinat
authors.affiliation: cole Polytechnique Fédérale de Lausanne, Laboratoire d'Optique Biomédicale, 1015 Lausanne, Switzerland.
authors.forename: Noelia L
authors.initials: NL
authors.lastname: Bocchio
authors.affiliation: cole Polytechnique Fédérale de Lausanne, Global Health Institute, 1015 Lausanne, Switzerland.
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Sandoz
authors.affiliation: Georg August University, III. Institute of Physics, 37077 Göttingen, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Huss
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, 37077 Göttingen, Germany.
authors.forename: Nickels A
authors.initials: NA
authors.lastname: Jensen
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, 37077 Göttingen, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Jakobs
authors.affiliation: Georg August University, III. Institute of Physics, 37077 Göttingen, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Enderlein
authors.affiliation: cole Polytechnique Fédérale de Lausanne, Global Health Institute, 1015 Lausanne, Switzerland.
authors.forename: F
authors.initials: F
authors.lastname: Gisou van der Goot
authors.affiliation: cole Polytechnique Fédérale de Lausanne, Laboratory of Bioorganic Chemistry and Molecular Imaging, Institute of Chemical Sciences and Engineering (ISIC), 1015 Lausanne, Switzerland.
authors.forename: Elena A
authors.initials: EA
authors.lastname: Dubikovskaya
authors.affiliation: cole Polytechnique Fédérale de Lausanne, Laboratoire d'Optique Biomédicale, 1015 Lausanne, Switzerland.
authors.forename: Theo
authors.initials: T
authors.lastname: Lasser
authors.affiliation: cole Polytechnique Fédérale de Lausanne, Laboratoire d'Optique Biomédicale, 1015 Lausanne, Switzerland.
authors.forename: Marcel
authors.initials: M
authors.lastname: Leutenegger
mesh_terms: D000818:Animals; D002232:Carbocyanines; D002460:Cell Line; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000295:instrumentation* / Q000379:methods; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D008928:Mitochondria / Q000502:physiology / Q000648:ultrastructure*; D057054:Molecular Imaging / Q000295:instrumentation* / Q000379:methods; D032446:Myoblasts / Q000502:physiology / Q000648:ultrastructure*; D014746:Vimentin / Q000737:chemistry / Q000378:metabolism
chemical_list: C569686:Alexa Fluor 647; D002232:Carbocyanines; D005456:Fluorescent Dyes; D014746:Vimentin; D049452:Green Fluorescent Proteins