A regression model approach to enable cell morphology correction in high-throughput flow cytometry.
A regression model approach to enable cell morphology correction in high-throughput flow cytometry.
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DOI：
10.1038/msb.2011.64
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发表时间：
2011-09-27
影响因子：
9.9
通讯作者：
Shmulevich, Ilya
中科院分区：
文献类型：
--
作者：
Knijnenburg, Theo A.;Roda, Oriol;Wan, Yakun;Nolan, Garry P.;Aitchison, John D.;Shmulevich, Ilya
暴露于刺激物的细胞表现出广泛的反应，确保整个群体的表型变异性。这种单细胞行为通常通过流式细胞术进行检查;然而，通常用于选择具有相似形态学特征的一小部分细胞亚群的门控程序使得在多种实验条件下定量比较细胞变得困难，甚至是不可能的，因为这些条件可能导致细胞的变化。深刻的形态学变化。为了克服这些限制，我们开发了一种回归方法来校正由于细胞大小和粒度的差异而导致的荧光强度的变化，而不丢弃任何细胞，这是门控本身所做的。这种方法能够在涉及数千个样品的高通量实验中定量研究细胞异质性和转录噪声。我们使用这种方法来分析酵母敲除菌株的文库，并揭示了该群体建立对油酸诱导的双峰反应所需的基因。我们确定了一组表观遗传调节和核孔蛋白，通过保持一个“反应迟钝的人口”，可能会提供人口的优势，多样化的赌注对冲。
Cells exposed to stimuli exhibit a wide range of responses ensuring phenotypic variability across the population. Such single cell behavior is often examined by flow cytometry; however, gating procedures typically employed to select a small subpopulation of cells with similar morphological characteristics make it difficult, even impossible, to quantitatively compare cells across a large variety of experimental conditions because these conditions can lead to profound morphological variations. To overcome these limitations, we developed a regression approach to correct for variability in fluorescence intensity due to differences in cell size and granularity without discarding any of the cells, which gating ipso facto does. This approach enables quantitative studies of cellular heterogeneity and transcriptional noise in high-throughput experiments involving thousands of samples. We used this approach to analyze a library of yeast knockout strains and reveal genes required for the population to establish a bimodal response to oleic acid induction. We identify a group of epigenetic regulators and nucleoporins that, by maintaining an ‘unresponsive population,’ may provide the population with the advantage of diversified bet hedging.
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