[1]沈链链,刘艳青,管强东.FACSAriaⅢ细胞分选参数选择及条件优化[J].常州大学学报(自然科学版),2022,34(04):75-81.[doi:10.3969/j.issn.2095-0411.2022.04.010]
　SHEN Lianlian,LIU Yanqing,GUAN Qiangdong.Optimization of Sorting Parameters and Conditions with FACS Aria Ⅲ[J].Journal of Changzhou University(Natural Science Edition),2022,34(04):75-81.[doi:10.3969/j.issn.2095-0411.2022.04.010]
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FACSAriaⅢ细胞分选参数选择及条件优化()
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常州大学学报(自然科学版)[ISSN:2095-0411/CN:32-1822/N]

卷:
第34卷
期数:
2022年04期
页码:
75-81
栏目:
生物医药工程
出版日期:
2022-07-28

文章信息/Info

Title:
Optimization of Sorting Parameters and Conditions with FACS Aria Ⅲ
文章编号:
2095-0411(2022)04-0075-07
作者:
沈链链; 刘艳青; 管强东
(南京医科大学公共卫生学院,江苏南京211166)
Author(s):
SHEN Lianlian;  LIU Yanqing;  GUAN Qiangdong
(School of Public Health, Nanjing Medical University, Nanjing 211166, China)
关键词:
细胞分选;  FACS Aria Ⅲ;  参数;  优化
Keywords:
sorting;  FACS Aria Ⅲ;  parameters;  optimization
分类号:
R 331
DOI:
10.3969/j.issn.2095-0411.2022.04.010
文献标志码:
A
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100 μm喷嘴和5×106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100 μm喷嘴,5×106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。
Abstract:
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 100 μm nozzle and cell concentration at 5×106cells/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 100 μm nozzle, cell concentrations at 5×106cells/mL and 4-way purity mode is recommended for tube sorting.

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(责任编辑:李艳,周安迪)

备注/Memo

备注/Memo:
收稿日期: 2021-12-17。
基金项目: 南京医科大学科技发展基金资助项目(NMUB2019010)。
作者简介: 沈链链(1984—), 女, 江苏吴江人, 硕士, 实验师。通信联系人: 管强东(1992—), E-mail: gqd@njmu.edu.cn
更新日期/Last Update: 1900-01-01