The new method investigation for more effective utilization of flow cytometry

本课题的目的是寻找更有效地利用流式细胞术（FCM）的方法。流式细胞术最常用于细胞表面抗原分析。目前，该分析方法仅用于计算阳性细胞的相对百分比，而对抗原的定量分析还没有建立起来。本课题设计了一种新的细胞表面抗原定量分析指标，将荧光染料染色的细胞前向散射（FSC）图分为5块，计算每块的FSC和荧光强度（FI）的平均值。将FSC的平均值绘制在X轴上，FI的平均值绘制在Y轴上，并将回归线的倾斜度称为FF系数作为新的指标。当FF系数为常数时，表明细胞的荧光强度随细胞大小的变化而不断变化，细胞表面单位面积上的抗原值为常数。 ...更多信息 t.我们用这种方法进行了检验，得到了如下结果. FSC与FI在CD 4阳性T淋巴细胞的CD 4抗原上呈线性正相关。其FF系数为0.40（μ SY ±）。健康成年男性为0.04，正常成年男性为0.36 ± 0.05。[）0.04，两者之间有显著性差异. T淋巴细胞上IL-2 α抗原的FF-系数增加（1.60（μ S·± 1·））。[）0.40，对照：0.06（]SY. ±. ConA刺激淋巴细胞后，CD_4的FF系数无明显变化（0.19 ± 0.16）。[）0.04，对照：0.20（]SY. ±. [）0.05）.3.记忆细胞（CD 45 RO强阳性和CD 45 RA阴性细胞）上的CD 4抗原的FF系数大于幼稚细胞（CD 45 RA强阳性和CD 45 RO阴性细胞）上的FF系数。计算出的CD_8抗原的FF系数值基本一致，我们认为我们提出的新指标FF系数可用于用细胞表面抗原定量分析的方法对疾病进行详细的检查。少

The purpose of this project is to find the methods to utilize flow cytometry (FCM) more effectively. FCM has been used most frequently for cell-surface antigen analysis. Hitherto the analysis is usually utilized only to calculate relative percentages of positive cells and the quantitative analysis of antigens has not been vetermined. In this project we have devised the new index for cell-surface antigens-quantitative analysis to study more detail.The forward scatter (FSC) pattern of cells stained with flurescent dye is divided into 5 pieces, and mean values of FSC and fluorescence intensity (FI) are calculated on each piece. The mean values of FSC are plotted on the X-axis, those of FI are on the Y-axis and an inclination of the recurrent line is named FF-coefficient as the new index. When FF-coefficient is constant, it indicates that cells' fluorscence intensity must change constantly according to change of the cell size and antigen values per unit area on the cell surface are constan … More t. We have examined and got the following results with this method.1. Lineal positive relationship between FSC and FI is recognized on CD4 antigens of CD4 positive T-lymphocytes. Its FF-coefficient is 0.40(]SY.+-。[)0.04 in healthy adult male, 0.36(]SY.+-。[)0.04 in healthy adult female and significant difference is recognized between them.2. The FF-coefficient of IL-2alpha antigens on T-lymphocytes increases (1.60(]SY.+-。[)0.40, control : 0.06(]SY.+-。[)0.16) after stimulating lymphocytes by ConA.To the contrary the FF-coefficient of CD4 dose not change (0.19(]SY.+-。[)0.04, control : 0.20(]SY.+-。[)0.05).3. The FF-coefficient of CD4 antigens on memory cells (CD45RO strong positive and CD45RA negative cells) is larger than it on naive cells (CD45RA strong positive and CD45RO negative cells). Almost same values of the FF-coefficient of CD8 antigens are counted.We conclude that our proposed new index, FF-coefficient, must be useful for detail examination of diseases with the method of quntitative analysis of cell surface antigens. Less