Allen Terence D, Rutherford Sandra A, Murray Stephen, Drummond Sheona P, Goldberg Martin W, Kiseleva Elena
Department of Structural Cell Biology, Paterson Institute for Cancer Research, University of Manchester, Manchester M20 4BX, United Kingdom.
Methods Cell Biol. 2008;88:389-409. doi: 10.1016/S0091-679X(08)00420-2.
Accessing internal structure and retaining relative three dimensional (3D) organization within the nucleus has always proved difficult in the electron microscope. This is due to the overall size and largely fibrous nature of the contents, making large scale 3D reconstructions difficult from thin sections using transmission electron microscopy. This chapter brings together a number of methods developed for visualization of nuclear structure by scanning electron microscopy (SEM). These methods utilize the easily accessed high resolution available in field emission instruments. Surface imaging has proved particularly useful to date in studies of the nuclear envelope and pore complexes, and has also shown promise for internal nuclear organization, including the dynamic and radical reorganization of structure during cell division. Consequently, surface imaging in the SEM has the potential to make a significant contribution to our understanding of nuclear structure.
在电子显微镜下,要观察细胞核内部结构并保持其相对三维(3D)组织一直颇具难度。这是因为细胞核内容物的整体尺寸较大且主要为纤维状,使得利用透射电子显微镜从薄切片进行大规模3D重建变得困难。本章汇集了多种通过扫描电子显微镜(SEM)观察细胞核结构所开发的方法。这些方法利用了场发射仪器中易于获取的高分辨率。迄今为止,表面成像在核膜和核孔复合体的研究中已证明特别有用,并且在细胞核内部组织研究方面也展现出前景,包括细胞分裂过程中结构的动态和根本性重组。因此,扫描电子显微镜中的表面成像有潜力对我们理解细胞核结构做出重大贡献。