Improved Flow Cytometric Analysis of the Budding Yeast Cell Cycle
Improved Flow Cytometric Analysis of the Budding Yeast Cell Cycle
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DOI：
10.4161/cc.1.2.114
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发表时间：
2002-03-01
期刊：
影响因子：
4.3
通讯作者：
Reed, Steven I.
中科院分区：
文献类型：
--
作者：
Haase, Steven B.;Reed, Steven I.
发芽酵母酿酒酵母是研究真核细胞周期调控的一个非常有用的模型系统。流式细胞术分析发芽酵母中的DNA含量已成为分析细胞周期进程的标准工具。然而，使用DNA结合染料-碘化丙啶的流行方案存在一些缺点，这些缺点混淆了流式细胞术的准确分析。在这里，我们展示了DNA结合染料SYTOX绿在酵母细胞周期分析中的应用。SYTOX Green分析的样品表现出更好的变异系数，改善了DNA含量和荧光之间的线性关系，并减少了与染料浓度、生长条件或细胞大小变化相关的峰漂移。
The budding yeast, Saccharomyces cerevisiae has been a remarkably useful model system for the study of eukaryotic cell cycle regulation. Flow cytometric analysis of DNA content in budding yeast has become a standard tool for the analysis of cell cycle progression. However, popular protocols utilizing the DNA binding dye, propidium iodide, suffer from a number of drawbacks that confound accurate analysis by flow cytometry. Here we show the utility of the DNA binding dye, SYTOX Green, in the cell cycle analysis of yeast. Samples analyzed using SYTOX Green exhibited better coefficients of variation, improved linearity between DNA content and fluorescence, and decreased peak drift associated with changes in dye concentration, growth conditions or cell size.