流式细胞术多色方案的方法学探讨

摘要

Objective To establish a method for the identification of multi cell sub populations and to ex-plore the optimization of flow cytometry.Methods The different fluorescent antibody markers from dif-ferent cell sub populations were used to mark mouse peripheral blood cells and establish the multi-color scheme of flow cytometry by flow cytometry,voltage regulation,fluorescence compensation regulation. Then,these instrument parameters were recorded to set the instrument and the different multi-color marking results were compared and analysied.Results Optimizing the detection voltage,the correct ad-justment of compensation,the correct choice of fluorescein,the same cell marker dye overlap minimiza-tion,can effectively reduce the deviation of the experiment.The distribution of cell subsets was clear,and the ratio of fluorescence intensity was expressed correctly,which was consistent with the results obtained by single staining method.Conclusion Multi color matching can effectively reduce the experimental cost, the amount of samples used,experiment duration,and make full use of flow cytometry.%目的：建立流式细胞仪多细胞亚群鉴定和多色标记技术平台，探讨流式细胞术多色配色的优化方案。方法利用不同荧光抗体进行小鼠外周血标本细胞亚群的标记，通过流式细胞仪，建立流式不同配色方案，调节电压，调节荧光补偿，同时以获得的参数重新设置仪器，比较多色标记得到的不同实验结果并分析。结果优化检测电压、补偿正确调节、荧光素的正确选择、同种细胞的标记染料重叠最小化能有效降低实验的偏差。细胞亚群分布清晰，荧光强度比例表达正确，与通常采用的实验样品单染色方法得到的结果一致。结论多色配色能有效降低实验费用，减少标本使用量，减少实验用时，充分利用流式细胞仪性能。