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Tracking of Individual Cell Cohorts in AsynchronousSaccharomyces cerevisiaePopulations

机译:异步酿酒酵母种群中单个细胞队列的跟踪

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AbstractA novel flow cytometric procedure has been developed with the aim to obtain the growth properties of individualSaccharomyces cerevisiaecells in asynchronous culture. The method is based on labeling of the cell surface with FITC‐conjugated concanavalin A and detection of the single‐cell fluorescence with flow cytometry after cell exposure to growth conditions. Because the formation of new cell wall material in budded cells is restricted to the bud tip, exposure of the stained cells to growth conditions results in three cell types: (i) stained cells, (ii) partially stained cells, and (iii) unstained cells Analysis of the staining pattern over time permits the determination of the specifice growth rate of the cell population, the length of the budded cell cycle phase, and the growth pattern during the cell cycle of newly formed, partially stained daughter cells. The procedure has been tested with yeast cell populations growing at different rates. The data suggest an exponential increase in the size of individual cells during the cell cycle, as reflected by the forward angle light scattering (FALS) signals. It has been found that the apparent single‐cell specific cell size growth rates, determined by FALS intensity, are significantly lower than the specific growth rates of the overall population. This could indicate that the tracking of a cohort of cells is significantly perturbed by a distribution of staining levels of daughter cells at cell division and that FALS may not be a good indicator of the cell
机译:摘要 开发了一种新的流式细胞术方法,旨在获得异步培养中酿酒酵母个体细胞的生长特性。该方法基于用 FITC 偶联的刀豆球蛋白 A 标记细胞表面,并在细胞暴露于生长条件下后用流式细胞术检测单细胞荧光。由于出芽细胞中新细胞壁物质的形成仅限于芽尖,因此将染色细胞暴露于生长条件下会导致三种细胞类型:(i)染色细胞,(ii)部分染色细胞和(iii)未染色细胞 分析染色模式随时间的变化可以确定细胞群的特定生长速率, 出芽细胞周期阶段的长度,以及新形成的、部分染色的子细胞在细胞周期中的生长模式。该程序已经过酵母细胞群以不同速率生长的测试。数据表明,在细胞周期中,单个细胞的大小呈指数级增长,正如前向角光散射(FALS)信号所反映的那样。已经发现,由 FALS 强度决定的表观单细胞比细胞大小增长率显着低于总体群体的比生长率。这可能表明细胞群的跟踪受到细胞分裂时子细胞染色水平分布的显着干扰,并且 FALS 可能不是细胞的良好指标

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