喷嘴型号的选择对流式分选效果的影响

Influence of nozzle type on sorting efficiency of flow cytometry

目的用70、100两种型号的喷嘴对不同颗粒进行流式分选,观察喷嘴型号对流式分选的影响。方法BD Calibiite Bead,人单核细胞白血病细胞株Thp-1、白血病细胞株H6和人外周血分离单核细胞染色后进行流式分选,观察70、100两种不同型号的喷嘴对分选时液流的稳定性、分选纯度、细胞活性的影响。结果BD Calibiite Bead、人外周血单核细胞,喷嘴的选择对其液流稳定性、分选纯度没有影响;人外周血单核细胞两种喷嘴分选后PI染色显示其细胞活性都在90%以上,使用100喷嘴分选活性较使用70喷嘴高;Thp-1、H6凋亡染色后进行分选,100喷嘴的分选纯度较70喷嘴高。结论不同大小的喷嘴对小微球颗粒及小细胞分选影响不大。对脆弱、易激惹细胞及荧光抗体与细胞结合不稳定的实验选用较大的喷嘴,分选纯度高,分选后细胞的活性更好。

Objective To sort different sizes of particles with flow cytometry with two types of nozzles(type 70 and type100),and to detect the effect of different type of nozzles on cell sorting.Methods The stability of liquid flow,sorting purity and cell viability were observed of human monocyte leukemia cell line Thp-1,leukemia cell line H6 and the separation of human peripheral blood mononuclear cells in sorting by BD Calibiite Bead with using two types of nozzles.Results BD caliber Bead,human peripheral blood mononuclear cells,and nozzle selection had no effect on the stability of liquid flow and sorting purity.PI staining showed that the viability of human peripheral blood mononuclear cells was above 90%after sorting with two nozzles,and the viability of sorting with 100 nozzles was higher than that with 70 nozzles.After Thp-1 and H6 apoptosis staining,the separation purity of 100 nozzles was better than that of 70 nozzles.Conclusion Different type of nozzles have little effect on the purity of small particles and cell sorting,while smaller nozzles have a certain effect on cell viability.