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.
Footnotes
Competing interests
The authors declare no competing interests.
Author contributions
A.L., G.K., S.v.d.L., C.F. and M.S. conceived of and designed the experiments; A.L. and G.K. performed the experiments; C.F. and S.v.d.L. analyzed the data; M.S wrote the paper.
Funding
This work was supported by the Biophotonics Initiative of the Bundesministerium für Bildung und Forschung [grant numbers 13N11019 and 13N12781].
Supplementary material available online at http://jcs.biologists.org/lookup/suppl/doi:10.1242/jcs.156620/-/DC1
- Received May 15, 2014.
- Accepted August 6, 2014.
- © 2014. Published by The Company of Biologists Ltd
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