Journal
JOURNAL OF CELL SCIENCE
Volume 127, Issue 20, Pages 4351-4355Publisher
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.156620
Keywords
Correlative electron and super-resolution fluorescence microscopy; Nuclear pore complex; dSTORM; Localization microscopy; Quantification
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Funding
- Biophotonics Initiative of the Bundesministerium fur Bildung und Forschung [13N11019, 13N12781]
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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.
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