CD4单阳参考品辅助检测九色标记人血样品的流式分析技术

Flow Pattern Analysis Technique for CD4 Single-positive Reference Substance-assisted Detection of Nine-color Labeled Human Blood Samples

目的探索流式多色分析中确定检测器最优参数的客观方法。方法以BD FACSymphony流式细胞仪检测CD3、CD4、CD8、CD25、CD27、CD45RA、CD56、CD127、CD197九色标记人外周血样品,选用中等表达强度各荧光素标记CD4单阳对照品,以递增检测参数重复分析获得各通道检测器性能曲线,计算获得流式仪器检测优化参数,再以优化参数完成样品检测和补偿调节。结果补偿前检测群体模糊不清,无法确定检测数据质量;补偿后可见细胞群体的位移,各通道荧光溢漏扩散均较理想,阴性、阳性群体区分明显,各细胞群体指标显示清晰。结论在多参数流式分析中可以采用适当强度光谱匹配的单阳样品客观、快速的确定检测参数,进而完成补偿调节,可避免操作人员主观判断失误,提高检测质量。

Objective To explore the method to determine the optimal parameters of detector in polychromatic flow cytometry.Methods CD3,CD4,CD8,CD25,CD27,CD45RA,CD56,CD127,CD197 in human peripheral blood were detected by BD FACSymphony flow cytometry.CD4 single positive control was labeled with moderate expression of each fluorescein.The performance curve of each channel detector was obtained by repeated analysis with increasing detection parameters.The optimized parameters of flow cytometry instrument were calculated,and then the sample detection and compensation adjustment were completed with the optimized parameters.Results The detection group before compensation was ambiguous and did not determine the quality of detection data.After compensation,the displacement of cell population was visible,and the fluorescence leakage diffusion of each channel was ideal.The negative and positive groups were clearly distinguished,and the indicators of each cell population were clear.Conclusion In the multi-parameter flow pattern analysis,the single-positive sample with appropriate intensity spectrum matching can be used to determine the detection parameters objectively and quickly,and then complete the compensation adjustment,which can avoid the subjective judgment error of operators and improve the detection quality.