O'Toole Eileen, Müller-Reichert Thomas
The Boulder Laboratory for 3-D Electron Microscopy of Cells, University of Colorado, Boulder, CO, USA.
Methods Mol Biol. 2009;545:135-44. doi: 10.1007/978-1-60327-993-2_8.
In this chapter we describe the preparation of early mitotic C. elegans embryos for the tomographic reconstruction of end-morphologies of spindle microtubules. Early embryos are prepared by high-pressure freezing and freeze-substitution for thin-layer embedding in Epon/Araldite. We further describe data acquisition, tomographic reconstruction, and 3-D modeling of microtubules in serially sectioned mitotic spindles. The presented techniques are applicable to other model systems.
在本章中，我们描述了制备早期有丝分裂期秀丽隐杆线虫胚胎的方法，用于纺锤体微管末端形态的断层重建。早期胚胎通过高压冷冻和冷冻置换法制备，用于在环氧树脂/阿拉底树脂中进行薄层包埋。我们还进一步描述了在连续切片的有丝分裂纺锤体中微管的数据采集、断层重建和三维建模。所介绍的技术适用于其他模型系统。
