Davidson Reshma, Liu Yajun, Gerien Kenneth S, Wu Jian-Qiu
Department of Molecular Genetics, The Ohio State University, 615 Biological Sciences Building, 484 W. 12th Ave, Columbus, OH, 43210, USA.
Department of Molecular and Cellular Biochemistry, The Ohio State University, 333 Hamilton Hall, 1645 Neil avenue, Columbus, OH, 43210, USA.
Methods Mol Biol. 2016;1369:9-23. doi: 10.1007/978-1-4939-3145-3_2.
Single-cell microscopy provides a powerful tool to visualize cellular and subcellular processes in wild-type and mutant cells by observing fluorescently tagged proteins. Here, we describe three simple methods to visualize fission yeast cells: gelatin slides, coverslip-bottom dishes, and tetrad fluorescence microscopy. These imaging methods and data analysis using free software make it possible to quantify protein localization, dynamics, and concentration with high spatial and temporal resolution. In fission yeast, the actomyosin contractile ring is essential for cytokinesis. We use the visualization and quantification of contractile ring proteins as an example to demonstrate how to use these methods.
单细胞显微镜技术提供了一种强大的工具，可通过观察荧光标记的蛋白质来可视化野生型和突变细胞中的细胞及亚细胞过程。在此，我们描述三种可视化裂殖酵母细胞的简单方法：明胶载玻片、底部有盖玻片的培养皿和四分体荧光显微镜。这些成像方法以及使用免费软件进行的数据分析，使得以高空间和时间分辨率对蛋白质定位、动态变化及浓度进行量化成为可能。在裂殖酵母中，肌动球蛋白收缩环对于胞质分裂至关重要。我们以收缩环蛋白的可视化和量化为例，来说明如何使用这些方法。
