应用CompBeads建立流式多色分析补偿的方法  

Establishment of a compensation method for polychromatic flow cytometry based on compbeads

作　　者：傅晓岚[1] 周伟[1] 王晴[1] 杨空明 谢谆怡[1] 吴玉章[1] 

机构地区：[1]第三军医大学全军免疫学研究所,重庆400038

出　　处：《免疫学杂志》2008年第3期340-345,共6页Immunological Journal

摘　　要：目的探讨一种快速简便的流式细胞仪多色分析时的关键参数(荧光补偿)设置方法。方法以BDComp-Beads、实验样品(本实验为慢性乙型肝炎患者外周全血)分别与单克隆荧光抗体染色,结合流式细胞仪Diva软件调节细胞仪光电倍增管(PMT)电压,计算建立荧光补偿矩阵。同时以获得的参数设置仪器,运行慢性乙型肝炎患者外周全血6色荧光标记实验样品。结果用CompBeads建立的补偿参数稳定可靠,能有效消除荧光光谱之间的交叉重叠。6色荧光标记实验样品检测的结果,可见细胞亚群分布清晰,荧光强度比例表达适当,与通常采用的实验样品单染色方法补偿结果一致(P>0.05)。结论该方法能快速方便进行流式细胞仪检测时仪器参数最佳化设置调整,在多色免疫标记流式检测中有较大的应用价值。Objective To establish a method for rapid and easy setting of the fluorescence compensation parameters, which is critical for polychromatic flow cytometry analysis. Methods BD compheads and samples [ peripheral whole blood of chronic hepatitis B patients （CHB） ] were stained with fluorochrome conjugated monoclonal antibody separately. Then, the voltage of photomultiplier tube （PMT） was regulated using Diva software and fluorescence compensation matrix was established. The peripheral lymphocyte subsets of patients with chronic hepatitis B （CHB） were detected by 6-color staining with the established parameters. Results The established parameters were stable and reliable, which can eliminate spectral overlaps hetwecn different fluorochromes. The test showed that the distribution of subsets was clearly and the intensity of fluorescence was moderate, which was not significantly different from the results of usual compensation method （ P 〉0.05）. Conclusion This method based on compheads can adjust the parameter perfectly, and can he widely used in the polychromatic flow cytometry.

关 键 词：流式细胞仪 多色 补偿 

分 类 号：R446.6[医药卫生—诊断学]