An effective analysis of individual cells using flow cytometry and its clinical applicatjion.

本研究旨在探索流式细胞术（FCM）在临床医学中的有效应用，利用单个细胞的信息。到目前为止，利用流式细胞术进行细胞免疫分型主要集中在定量具有一定分子组合的细胞比例，荧光强度（FI）的变化不能直接反映抗原密度。为了确定细胞表面分子密度，FCM在单个细胞上获得的前向光散射（FSC）和FI被绘制成散点图，其中x轴和y轴分别表示FSC和FI。当FI和FSC之间可以看出成比例关系时，可以用回归曲线的斜率（或其绝对值）作为分子密度的指标，称为FF指数。通过这种方式，FI与FSC散点图可以评估细胞大小与抗原表达量之间的关系。此外，我们还探索了利用FCM的列表模式自动进行这些分析的计算机软件。我们分析了正常外周血淋巴细胞、血液学恶性细胞和血液病患者的淋巴细胞，发现使用该软件可以更精确地进行临床分析。未来还需要更多的临床研究、开发的软件的评价和标准化以及细胞表面抗原的实际定量。

The purpose of this study is to explore an effective application of flow cytometry (FCM) to clinical medicine by using the information of individual cells. Until now, cytometric immunophenotyping using FCM has focused mainly on the quantification of the proportion of cells bearing certain combinations of molecules and the change of fluorescence intensity (FI) dose not reflect the antigen density directly. For the qualification of cell surface molecule density, the forward light scatter (FSC) and FI, obtained by FCM on an individual cell basis were plotted in a scattergram where x-and y-axis show FSC and FI respectively. Where a proportional relationship between FI and FSC can been seen, the slope of the line of regression (or its absolute value) may be used as an indicator of the molecule density, which is referred to as the FF index. In this way, the FI vs FSC scattergram enables the evaluation of the relationship between the size of the cell and the amount of antigen expressed. Additionally we explored the computer software which enables to practice automatically these analysis by using the listmode of FCM. We analyzed normal peripheral lymphocytes, hematological malignant cells and lymphocytes of hemodyalisis patients and found the potent usefulness of more precise clinical analysis using this software. In future, more clinical studies, the evaluation and standardization of the developed software and actual quantification of cell surface antigens are needed.