Third Institute of Physics, University of Göttingen , 37077 Göttingen, Germany.
Universitätsmedizin Göttingen, University of Göttingen, Department of Molecular Biology, GZMB , 37073 Göttingen, Germany.
ACS Nano. 2017 Dec 26;11(12):11839-11846. doi: 10.1021/acsnano.7b04671. Epub 2017 Sep 20.
The nuclear envelope, comprising the inner and the outer nuclear membrane, separates the nucleus from the cytoplasm and plays a key role in cellular functions. Nuclear pore complexes (NPCs), which are embedded in the nuclear envelope, control transport of macromolecules between the two compartments. Here, using dual-color metal-induced energy transfer (MIET), we determine the axial distance between Lap2β and Nup358 as markers for the inner nuclear membrane and the cytoplasmic side of the NPC, respectively. Using MIET imaging, we reconstruct the 3D profile of the nuclear envelope over the whole basal area, with an axial resolution of a few nanometers. This result demonstrates that optical microscopy can achieve nanometer axial resolution in biological samples and without recourse to complex interferometric approaches.
核膜由内、外核膜组成,将细胞核与细胞质隔开,在细胞功能中发挥着关键作用。核孔复合物(NPC)嵌入核膜中,控制着这两个隔室之间的大分子物质的运输。在这里,我们使用双色金属诱导能量转移(MIET)来确定 Lap2β 和 Nup358 之间的轴向距离,它们分别作为内核膜和 NPC 细胞质侧的标志物。使用 MIET 成像,我们重建了整个基底区域的核膜的 3D 轮廓,轴向分辨率为几个纳米。这一结果表明,光学显微镜可以在生物样本中实现纳米级的轴向分辨率,而无需采用复杂的干涉方法。