Scientific Research
Conditions for high efficient sorting experiments by flow cytometry
Huang Yingying, Xu Qin, Jiang Yu
Published 2021-01-01
Cite as IMHGN, 2021, 27(1): 24-29. DOI: 10.3760/cma.j.issn.1007-1245.2021.01.006
Abstract
ObjectiveTo improve the yield and purity of sorting experiments by flow cytometry, to establish efficient sorting conditions, and to provide references for the application of fluorescence-activated cell sorting.
MethodsThe beads were used as the test samples, and the BD FACS Sorp Aria II was used as the test instrument from October, 2019 to June, 2020. Different sorting velocities, sorting modes, and nozzles were investigated, and the influencing factors were analyzed. The naïve T cells were sorted under the optimized conditions.
ResultsThe BD FACS Sorp Aria II gained purity of more than 98% with four different sorting velocities, flow rates of 1.0, 3.0, 5.0, and 7.0, by using a 70 μm nozzle and the purity mode. However, the higher the flow rate, the lower the sorting yield. Sorting samples by different sorting modes, the efficiency and yield of the yield sorting mode were (99.67±0.33)% and (79.78±7.14)%, respectively, which were higher than those of the other two modes, but the purity was the lowest one by the yield sorting mode (both P＜0.05). The instrument gained a purity of more than 98% with all the three nozzles, and the yields of the three nozzles were not statistically different (P＞0.05).
ConclusionWhen naïve T cells are sorted by BD FACS Sorp Aria II flow cytometer with a 70 μm nozzle, a flow rate of 1.0, and the purity sorting mode are preferred.
Key words:
Sorting velocities; Sorting modes; Nozzles; Sorting yield; Purity
Contributor Information
Huang Yingying
Core Facilities, School of Medicine, Zhejiang University, Hangzhou 310058, China
Xu Qin
Department of Rheumatology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China
Jiang Yu
School of Basic Medical Sciences, Zhejiang University, Hangzhou 310058, China
