title: Ultra-High Resolution 3D Imaging of Whole Cells.
abstract: Fluorescence nanoscopy, or super-resolution microscopy, has become an important tool in cell biological research. However, because of its usually inferior resolution in the depth direction (50-80 nm) and rapidly deteriorating resolution in thick samples, its practical biological application has been effectively limited to two dimensions and thin samples. Here, we present the development of whole-cell 4Pi single-molecule switching nanoscopy (W-4PiSMSN), an optical nanoscope that allows imaging of three-dimensional (3D) structures at 10- to 20-nm resolution throughout entire mammalian cells. We demonstrate the wide applicability of W-4PiSMSN across diverse research fields by imaging complex molecular architectures ranging from bacteriophages to nuclear pores, cilia, and synaptonemal complexes in large 3D cellular volumes.
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Huang
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; The Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK.
authors.forename: George
authors.initials: G
authors.lastname: Sirinakis
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; The Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, UK.
authors.forename: Edward S
authors.initials: ES
authors.lastname: Allgeyer
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA.
authors.forename: Lena K
authors.initials: LK
authors.lastname: Schroeder
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Department of Chemistry, Harvey Mudd College, Claremont, CA 91711, USA.
authors.forename: Whitney C
authors.initials: WC
authors.lastname: Duim
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Department of Biomedical Engineering, Yale University, CT 06520, USA.
authors.forename: Emil B
authors.initials: EB
authors.lastname: Kromann
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA.
authors.forename: Thomy
authors.initials: T
authors.lastname: Phan
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA.
authors.forename: Felix E
authors.initials: FE
authors.lastname: Rivera-Molina
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA.
authors.forename: Jordan R
authors.initials: JR
authors.lastname: Myers
authors.affiliation: Microbial Sciences Institute, Yale University, West Haven, CT 06516, USA; Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520, USA.
authors.forename: Irnov
authors.initials: I
authors.lastname: Irnov
authors.affiliation: The Jackson Laboratory, Bar Harbor, ME 04609, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Lessard
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA.
authors.forename: Yongdeng
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: The Jackson Laboratory, Bar Harbor, ME 04609, USA.
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Handel
authors.affiliation: Microbial Sciences Institute, Yale University, West Haven, CT 06516, USA; Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520, USA; Howard Hughes Medical Institute, Yale University, New Haven, CT 06520, USA; Department of Microbial Pathogenesis, Yale School of Medicine, New Haven, CT 06520, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Jacobs-Wagner
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA.
authors.forename: C Patrick
authors.initials: CP
authors.lastname: Lusk
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Nanobiology Institute, Yale University, West Haven, CT 06516, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Rothman
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Nanobiology Institute, Yale University, West Haven, CT 06516, USA.
authors.forename: Derek
authors.initials: D
authors.lastname: Toomre
authors.affiliation: Department of Engineering Science, University of Oxford, Oxford OX1 3PJ, UK; Centre for Neural Circuits and Behaviour, University of Oxford, Oxford OX1 3SR, UK.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Booth
authors.affiliation: Department of Cell Biology, School of Medicine, Yale University, New Haven, CT 06520, USA; Department of Biomedical Engineering, Yale University, CT 06520, USA; Nanobiology Institute, Yale University, West Haven, CT 06516, USA. Electronic address: joerg.bewersdorf@yale.edu.
authors.forename: Joerg
authors.initials: J
authors.lastname: Bewersdorf
mesh_terms: D000818:Animals; D001435:Bacteriophages / Q000648:ultrastructure; D022181:COP-Coated Vesicles / Q000648:ultrastructure; D003584:Cytological Techniques / Q000295:instrumentation / Q000379:methods*; D006056:Golgi Apparatus / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D000072760:Single Molecule Imaging / Q000295:instrumentation / Q000379:methods*; D013090:Spermatocytes / Q000648:ultrastructure; D013573:Synaptonemal Complex / Q000648:ultrastructure