BD Influx流式细胞分选仪参数调试和最佳条件的设定

Research on the setting of optimal conditions and adjustment of sorting parameters for BD Influx flow cytometer

目的:探索BD Influx流式细胞仪分选细胞亚群的调试参数和最佳设定条件。方法:使用两种质控试剂Rainbow Beads和Accudrop Beads作为实验材料,对BD Influx流式细胞仪的针孔、液流、荧光通道、前向角散射光信号、液滴频率、振幅和液滴延迟等参数进行优化。结果:使用型号为100μm的喷嘴,在液流聚焦清楚、液柱打入废液收集管的正中央、荧光光斑进入针孔中央且最亮、前向角散射(FSC)光光斑进入针孔中央且最亮、断点最高、液流分叉呈稳定的亮点、振幅为5.20±1.00以及液滴延迟为35.20±2.00等最佳设定条件下,其细胞样本分选纯度可达99%,工作效率可达95%。结论:流式细胞仪分选参数条件的设定,可为获得高纯度、高活性的细胞提供快速调试方法。

Objective：To explore the optimal conditions and adjustment of sorting parameters for BD Influx flow cytometer. Methods： Using Rainbow Beads and Accudrop Beads as experimental materials, optimization of pinhole, stream, fluorescence channel, forward scatter signal, drop frequency, piezo amplitude and drop delay was conducted. Results： The optimal condition was as follows, using the 100 μm nozzle, the stream was centered on the pinholes and sharply focused, the stream was aimed at the drain, the brightest signal was in the center of the pinhole after optimizing the fluorescence channel, the brightest signal was in the center of the FSC pinhole after optimizing the forward scatter signal, the breakoff point was highest after adjusting the drop frequency, and the droplet shape remained constant. When the piezo amplitude was around 5.20＆#177;1.00 and the drop delay value was around 35.20＆#177;2.00, the adjustment efficiency achieved its optimal condition, with its purity at 99%and sorting efficiency at 95%. Conclusion：Rapid setting of optimal conditions and adjustment of sorting parameters for BD Influx flow cytometer guarantees the success of gathering cells of high purity and activity after cell sorting.