Olziersky Anna-Maria, Smith Chris A, Burroughs Nigel, McAinsh Andrew D, Meraldi Patrick
Department of Cell Physiology and Metabolism, University of Geneva, Geneva, Switzerland.
Centre for Mechanochemical Cell Biology & Division of Biomedical Science, Warwick Medical School, University of Warwick, Coventry, United Kingdom.
Methods Cell Biol. 2018;145:1-27. doi: 10.1016/bs.mcb.2018.03.009.
Mitosis is a highly dynamic and choreographed process in which chromosomes are captured by the mitotic spindle and physically segregated into the two daughter cells to ensure faithful transmission of the genetic material. Live-cell fluorescence microscopy enables these dynamics to be analyzed over diverse temporal scales. Here we present the methodologies to study chromosome segregation at three timescales: we first show how automated tracking of kinetochores enables investigation of mitotic spindle and chromosome dynamics in the seconds-to-minutes timescale; next we highlight how new DNA live dyes allow the study of chromosome segregation over a period of several hours in any cell line; finally, we demonstrate how image sequences acquired over several days can reveal the fate of whole cell populations over several consecutive cell divisions.
有丝分裂是一个高度动态且精心编排的过程,在此过程中,染色体被有丝分裂纺锤体捕获并物理分离到两个子细胞中,以确保遗传物质的忠实传递。活细胞荧光显微镜技术能够在不同的时间尺度上分析这些动态过程。在这里,我们展示了在三个时间尺度上研究染色体分离的方法:我们首先展示了着丝粒的自动跟踪如何能够在秒到分钟的时间尺度上研究有丝分裂纺锤体和染色体的动态;接下来,我们强调了新型DNA活染料如何能够在任何细胞系中研究长达数小时的染色体分离过程;最后,我们展示了在几天内获取的图像序列如何能够揭示整个细胞群体在连续几个细胞分裂过程中的命运。