Prusicki Maria Ada, Keizer Emma Mathilde, Van Rosmalen Rik Peter, Fleck Christian, Schnittger Arp
Department of Developmental Biology, University of Hamburg, Hamburg, Germany.
Department of Mathematical and Statistical Methods, Wageningen University and Research, Wageningen, The Netherlands.
Bio Protoc. 2020 May 5;10(9):e3611. doi: 10.21769/BioProtoc.3611.
Live cell imaging has tremendously promoted our understanding of cellular and subcellular processes such as cell division. Here, we present a step-by-step protocol for a robust and easy-to-use live cell imaging approach to study male meiosis in the plant as recently established. Our method relies on the concomitant analysis of two reporter genes that highlight chromosome configurations and microtubule dynamics. In combination, these reporter genes allowed the discrimination of five cellular parameters: cell shape, microtubule array, nucleus position, nucleolus position, and chromatin condensation. These parameters can adopt different states, , the nucleus position can be central or lateral. Analyzing how tightly these states are associated gives rise to landmark stages that in turn allow a quantitative and qualitative dissection of meiotic progression. We envision that such an approach can also provide valuable criteria for the analysis of cell differentiation processes outside of meiosis.
活细胞成像极大地促进了我们对细胞和亚细胞过程（如细胞分裂）的理解。在此，我们展示了一种稳健且易于使用的活细胞成像方法的逐步方案，用于研究植物中的雄性减数分裂，该方法是最近建立的。我们的方法依赖于对两个报告基因的同步分析，这两个报告基因突出了染色体构型和微管动力学。结合起来，这些报告基因能够区分五个细胞参数：细胞形状、微管阵列、细胞核位置、核仁位置和染色质凝聚。这些参数可以呈现不同的状态，例如，细胞核位置可以是中央的或侧面的。分析这些状态之间的紧密关联程度会产生标志性阶段，进而允许对减数分裂进程进行定量和定性的剖析。我们设想，这样的方法也可以为减数分裂之外的细胞分化过程分析提供有价值的标准。
