Department of Biotechnology and Biophysics, Biozentrum, Julius Maximilian University Würzburg, Am Hubland, 97074 Würzburg, Germany.
Department of Electron Microscopy, Biozentrum, Julius Maximilian University Würzburg, Am Hubland, 97074 Würzburg, Germany.
J Cell Sci. 2014 Oct 15;127(Pt 20):4351-5. doi: 10.1242/jcs.156620. Epub 2014 Aug 21.
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.
在这里，我们将超分辨率荧光定位显微镜与扫描电子显微镜相结合，以在分子分辨率下分别对分离的非洲爪蟾卵母细胞核包膜中的核孔复合体蛋白进行成像，并对这两种成像模式下的蛋白位置进行绘图。我们利用核孔复合体的周期性分子结构，将直接随机光学重建显微镜图像与电子显微镜以小于 20nm 的精度进行叠加。相关图像证明了通过一级和二级抗体进行的标准免疫细胞化学对核孔复合体蛋白进行了定量分子标记和定位，并揭示了核孔复合体由八个 gp210（也称为 NUP210）蛋白同源二聚体组成。此外，我们还发现了具有九倍对称性的核孔复合体亚群，这些亚群偶尔会出现在更典型的八倍对称结构中。
