title: Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution.
abstract: Here, we combine super-resolution fluorescence localization microscopy with scanning electron microscopy to map the position of proteins of nuclear pore complexes in isolated Xenopus laevis oocyte nuclear envelopes with molecular resolution in both imaging modes. We use the periodic molecular structure of the nuclear pore complex to superimpose direct stochastic optical reconstruction microscopy images with a precision of <20 nm on electron micrographs. The correlative images demonstrate quantitative molecular labeling and localization of nuclear pore complex proteins by standard immunocytochemistry with primary and secondary antibodies and reveal that the nuclear pore complex is composed of eight gp210 (also known as NUP210) protein homodimers. In addition, we find subpopulations of nuclear pore complexes with ninefold symmetry, which are found occasionally among the more typical eightfold symmetrical structures.
authors.affiliation: Department of Biotechnology and Biophysics, Biozentrum, Julius Maximilian University Würzburg, Am Hubland, 97074 Würzburg, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Löschberger
authors.affiliation: Department of Biotechnology and Biophysics, Biozentrum, Julius Maximilian University Würzburg, Am Hubland, 97074 Würzburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Franke
authors.affiliation: Department of Electron Microscopy, Biozentrum, Julius Maximilian University Würzburg, Am Hubland, 97074 Würzburg, Germany.
authors.forename: Georg
authors.initials: G
authors.lastname: Krohne
authors.affiliation: Department of Biotechnology and Biophysics, Biozentrum, Julius Maximilian University Würzburg, Am Hubland, 97074 Würzburg, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: van de Linde
authors.affiliation: Department of Biotechnology and Biophysics, Biozentrum, Julius Maximilian University Würzburg, Am Hubland, 97074 Würzburg, Germany m.sauer@uni-wuerzburg.de.
authors.forename: Markus
authors.initials: M
authors.lastname: Sauer
mesh_terms: D000818:Animals; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008855:Microscopy, Electron, Scanning / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D015394:Molecular Structure; D009685:Nuclear Envelope / Q000648:ultrastructure*; D022022:Nuclear Pore / Q000737:chemistry / Q000648:ultrastructure*; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry / Q000648:ultrastructure*; D009865:Oocytes / Q000648:ultrastructure; D055503:Protein Multimerization; D029867:Xenopus Proteins / Q000737:chemistry / Q000648:ultrastructure*; D014982:Xenopus laevis
chemical_list: C512903:NUP210 protein, Xenopus; D028861:Nuclear Pore Complex Proteins; D029867:Xenopus Proteins
keywords: Correlative electron and super-resolution fluorescence microscopy; Localization microscopy; Nuclear pore complex; Quantification; dSTORM