The method and optimized condition for enriching empty plasmids transfected specific macrophages by flow cytometry sorting
WU Hao1, WANG Min1, LUO Xianqiang1, GUO Jianchun1, ZHENG Binjiao1,2
1.Zhejiang Key Laboratory of Medical Genetics, Wenzhou Medical University, Wenzhou 325035, China; 2.Attardi Institute of Mitochondrial Biomedicine, Wenzhou Medical University, Wenzhou 325035, China
WU Hao,WANG Min,LUO Xianqiang, et al. The method and optimized condition for enriching empty plasmids transfected specific macrophages by flow cytometry sorting[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2019, 49(6): 408-411,417.
Abstract：Objective: To establish and optimize the method for enriching PX458 empty plasmid transfected J774A.1 cells by BD FACSAria II flow cytometer in order to figure out the optimum condition for screening J774A.1 cells. Methods: PX458 empty plasmid transfected J774A.1 cell line was constructed and stained with 7-AAD. GFP-positive transfectants of J774A.1 cells were sorted by 100 μm nozzle and initial, 4-way purity, purity, yield, fine tune, single cell sort modes respectively. The sorting efficiency was compared in different instrument parameters. Results: Under the stable fluid flow condition, the purity of the cells obtained by each sort mode of 100 μm nozzle was over 80%, and the sort mode with highest efficiency is purity mode, reaching 95.7%±1.0%. Conclusion: When the PX458 empty plasmid transfected J774A.1 cell line was shorted by BD FACSAria II flow cytometer with 100 μm nozzle, the purity sort mode is preferred.
