使用细胞分选仪进行流式细胞术中的不确定性量化
Uncertainty quantification in flow cytometry using a cell sorter
AI总结：
提出一种利用细胞分选仪进行简单实验的方法，通过分选并重测低于阈值的微珠来量化流式细胞术中的仪器噪声，并利用局部仿射变换将估计值转移到无分选仪的细胞仪上。
摘要：
在流式细胞术中，很难将群体方差和仪器噪声对总测量变异的贡献区分开来。从根本上说，这是因为人们无法对同一个粒子进行多次测量。我们提出了一种使用细胞分选仪来区分仪器特定变异的简单实验。对于强度分布在单峰周围的微珠群体，使用分选仪收集测量强度低于某个阈值的微珠。然后对这个粒子子集进行重新测量。如果测量值中的变异仅来自样本，那么第二组测量值也应该完全低于我们的阈值。因此，任何“溢出”都是由于仪器特定效应引起的——我们证明了分选后测量值的分布如何足以提取由仪器引起的累积变异性的估计值。我们工作的一个显著特点是我们没有对上述噪声的来源做任何假设。然后，我们展示了“局部仿射变换”如何让我们将这些估计值转移到未配备分选仪的流式细胞仪上。我们使用我们的分析估计了一组三种仪器和两种微珠类型在一系列样本流速下的噪声。最后，我们讨论了仪器噪声对最优分类的影响以及其他应用。
英文摘要：
In cytometry, it is difficult to disentangle the contributions of population variance and instrument noise towards total measured variation. Fundamentally, this is due to the fact that one cannot measure the same particle multiple times. We propose a simple experiment that uses a cell sorter to distinguish instrument-specific variation. For a population of beads whose intensities are distributed around a single peak, the sorter is used to collect beads whose measured intensities lie below some threshold. This subset of particles is then remeasured. If the variation in the measured values is only due to the sample, the second set of measurements should also lie entirely below our threshold. Any 'spillover' is therefore due to instrument specific effects - we demonstrate how the distribution of the post-sort measurements is sufficient to extract an estimate of the cumulative variability induced by the instrument. A distinguishing feature of our work is that we do not make any assumptions about the sources of said noise. We then show how 'local affine transformations' let us transfer these estimates to cytometers not equipped with a sorter. We use our analysis to estimate noise for a set of three instruments and two bead types, across a range of sample flow rates. Lastly, we discuss the implications of instrument noise on optimal classification, as well as other applications.
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