Calvert Meredith E K, Lannigan Joanne
Temasek Life Sciences Laboratory, National University of Singapore, Singapore.
Curr Protoc Cytom. 2010 Apr;Chapter 9:Unit 9.32.1-16. doi: 10.1002/0471142956.cy0932s52.
In studies of eukaryotic cell cycle regulation, the budding yeast Saccharoymyces cerevisiae offers many advantages as a model system. Due to its simple growth requirements and genetic tractability, this organism is a powerful tool for investigating the molecular regulation of cell cycle control. One earlier disadvantage to performing cell cycle analyses in yeast was that existing methods were restricted to either visual analysis or flow cytometry, both of which present limitations in the scope and accuracy of the data obtained. This unit demonstrates the combined use of DNA content measurements and bright-field image analysis using multispectral imaging flow cytometry (MIFC) to provide a more precise quantitation of yeast cell cycle distribution. The advantage of this method is the ability to analyze large numbers of cells using multiple cell cycle indicators in a relatively short amount of time.
在真核细胞周期调控的研究中，芽殖酵母酿酒酵母作为一种模式系统具有诸多优势。由于其简单的生长需求和遗传易操作性，这种生物体是研究细胞周期控制分子调控的有力工具。早期在酵母中进行细胞周期分析的一个缺点是，现有方法仅限于视觉分析或流式细胞术，这两种方法在获取数据的范围和准确性方面都存在局限性。本单元展示了结合使用DNA含量测量和利用多光谱成像流式细胞术（MIFC）的明场图像分析，以更精确地定量酵母细胞周期分布。该方法的优点是能够在相对较短的时间内使用多个细胞周期指标分析大量细胞。
