FACS Aria Ⅲ细胞分选参数选择及条件优化  

Optimization of Sorting Parameters and Conditions with FACS Aria Ⅲ

作　　者：沈链链[1] 刘艳青[1] 管强东 SHEN Lianlian;LIU Yanqing;GUAN Qiangdong(School of Public Health,Nanjing Medical University,Nanjing 211166,China)

机构地区：[1]南京医科大学公共卫生学院,江苏南京211166

出　　处：《常州大学学报（自然科学版）》2022年第4期75-81,共7页Journal of Changzhou University：Natural Science Edition

基　　金：南京医科大学科技发展基金资助项目(NMUB2019010)。

摘　　要：以BGC-823细胞分选为例,探讨FACS Aria Ⅲ细胞分选仪分选参数选择及条件优化的方法。依次采用2种不同规格的喷嘴、3种不同浓度的细胞以及5种不同的分选模式进行分选,比较各种条件下96孔板分选的单细胞入孔率和克隆形成率以及试管式分选的细胞纯度和细胞状态,来确定最优方案。结果显示对于BCG-823类似的细胞,96孔板分选采用100μm喷嘴和5×10^(6)个/mL细胞浓度能获得更高的单细胞入孔率和克隆形成率,对于试管式分选采用100μm喷嘴,5×10^(6)个/mL细胞浓度和4-way purity模式将获得更高的分选后细胞纯度。The production of Green Fluorescent Protein in recombinant BGC823 cells was used as a model to investigate how to optimize sorting parameters and conditions with FACS Aria Ⅲ.Cells were sorted with nozzles with different orifice diameters,cell samples at different cell concentrations were sorted,and five sorting modes were employed,to compare the proportions of single cells and the clone formation rates in different wells for 96-well plate sorting;and compare the purities of sorted cells and cell growth rates for tube sorting,respectively.The use of the 100μm nozzle and cell concentration at 5×10^(6)cells/mL is recommended for 96-well plate sorting of the cell types similar to BGC-823,to increase the proportion of wells which contain single cells and the clone formation rate.Taking into account both the purity of the sorted cells and time consumption for sorting,the use of the 100μm nozzle,cell concentrations at 5×10^(6)cells/mL and 4-way purity mode is recommended for tube sorting.

关 键 词：细胞分选 FACS AriaⅢ 参数 优化 

分 类 号：R331[医药卫生—人体生理学]