Maco Bohumil, Fahrenkrog Birthe, Huang Ning-Ping, Aebi Ueli
M. E. Müller Institute for Structural Biology, Biozentrum, University of Basel, Switzerland.
Methods Mol Biol. 2006;322:273-88. doi: 10.1007/978-1-59745-000-3_19.
To study the ultrastructure of nuclear pore complexes (NPCs), a wide spectrum of different electron microscopy (EM) or atomic force microscopy (AFM) techniques can be employed. The combination of these methods can reveal new insights into the structural and functional organization of this important supramolecular machine through which nucleocytoplasmic transport occurs. Negative staining, quick freezing/freeze-drying/rotary metal shadowing, embedding and thin sectioning, cryoelectron microscopy and tomography, scanning electron microscopy, or combination with immunolabeling techniques are tools for collecting data and information about the three-dimensional structure and architecture of the NPCs. AFM enables investigation of the functional dynamics of native NPCs under physiological conditions.
为了研究核孔复合体（NPCs）的超微结构，可以采用多种不同的电子显微镜（EM）或原子力显微镜（AFM）技术。这些方法的结合能够揭示这个重要超分子机器的结构和功能组织的新见解，核质运输正是通过该机器发生的。负染色、快速冷冻/冷冻干燥/旋转金属阴影法、包埋和超薄切片、冷冻电子显微镜和断层扫描、扫描电子显微镜，或与免疫标记技术相结合，都是用于收集有关NPCs三维结构和构造的数据和信息的工具。原子力显微镜能够在生理条件下研究天然NPCs的功能动态。
