Ferrandiz Nuria, Royle Stephen J
Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Gibbet Hill Road, Coventry, CV4 7AL, UK.
Bio Protoc. 2023 Jul 5;13(13):e4708. doi: 10.21769/BioProtoc.4708.
Errors in chromosome segregation during mitosis lead to chromosome instability, resulting in an unbalanced number of chromosomes in the daughter cells. Light microscopy has been used extensively to study chromosome missegregation by visualizing errors of the mitotic spindle. However, less attention has been paid to understanding spindle function in the broader context of intracellular structures and organelles during mitosis. Here, we outline a protocol to visualize chromosomes and endomembranes in mitosis, combining light microscopy and 3D volume electron microscopy, serial block-face scanning electron microscopy (SBF-SEM). SBF-SEM provides high-resolution imaging of large volumes and subcellular structures, followed by image analysis and 3D reconstruction. This protocol allows scientists to visualize the whole subcellular context of the spindle during mitosis.
有丝分裂期间染色体分离错误会导致染色体不稳定，从而使子细胞中的染色体数量失衡。光学显微镜已被广泛用于通过观察有丝分裂纺锤体的错误来研究染色体错分离。然而，在有丝分裂期间细胞内结构和细胞器的更广泛背景下，对纺锤体功能的理解却较少受到关注。在这里，我们概述了一种在有丝分裂中可视化染色体和内膜的方案，该方案结合了光学显微镜和三维体积电子显微镜——连续块面扫描电子显微镜（SBF-SEM）。SBF-SEM可对大体积和亚细胞结构进行高分辨率成像，随后进行图像分析和三维重建。该方案使科学家能够在有丝分裂期间可视化纺锤体的整个亚细胞背景。
