Institute of Science and Technology Austria (IST Austria), Klosterneuburg, Austria.
Department of Experimental Plant Biology, Faculty of Science, Charles University, Prague, Czechia.
Methods Mol Biol. 2022;2382:105-114. doi: 10.1007/978-1-0716-1744-1_6.
The analysis of dynamic cellular processes such as plant cytokinesis stands and falls with live-cell time-lapse confocal imaging. Conventional approaches to time-lapse imaging of cell division in Arabidopsis root tips are tedious and have low throughput. Here, we describe a protocol for long-term time-lapse simultaneous imaging of multiple root tips on a vertical-stage confocal microscope with automated root tracking. We also provide modifications of the basic protocol to implement this imaging method in the analysis of genetic, pharmacological or laser ablation wounding-mediated experimental manipulations. Our method dramatically improves the efficiency of cell division time-lapse imaging by increasing the throughput, while reducing the person-hour requirements of such experiments.
对植物胞质分裂等动态细胞过程的分析取决于活细胞延时共聚焦成像。拟南芥根尖细胞分裂的延时成像传统方法繁琐,通量低。本文描述了一种在垂直载物台共聚焦显微镜上对多个根尖进行长期延时共聚焦成像并自动跟踪根尖的方法。我们还对基本方案进行了修改,以将这种成像方法应用于遗传、药理学或激光消融损伤介导的实验操作的分析。我们的方法通过增加通量、减少此类实验的人时需求,显著提高了细胞分裂延时成像的效率。