Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution

Anna Löschberger, Christian Franke, Georg Krohne, Sebastian van de Linde, Markus Sauer

Research output: Contribution to journalArticle

68 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)4351-4355
Number of pages5
JournalJournal of Cell Science
Volume127
Issue number20
Early online date21 Aug 2014
DOIs
Publication statusPublished - 14 Oct 2014
Externally publishedYes

Keywords

  • correlative electron and super-resolution fluorescence microscopy
  • localization microscopy
  • nuclear pore complex
  • dSTORM
  • immunocytochemistry

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