The new method investigation for more effective utilization of flow cytometry

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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