上海交通大学学报(医学版) ›› 2018, Vol. 38 ›› Issue (7): 845-.doi: 10.3969/j.issn.1674-8115.2018.07.025

• 技术与方法 • 上一篇    

流式细胞仪MoFlo AstriosEQ 96孔板单细胞分选方法的条件优化

张小翠,符蓉,赵犇鹏   

  1. 上海交通大学基础医学院基础医学公共技术平台,上海 200025
  • 出版日期:2018-07-28 发布日期:2018-07-30
  • 通讯作者: 张小翠,电子信箱:zhangcui1986921@163.com。
  • 作者简介:张小翠(1985—),女,实验师,博士;电子信箱: zhangcui1986921@163.com。

Condition optimization of the flow cytometry MoFlo Astrios EQ on single-cell sorting in 96-well plate

ZHANG Xiao-cui, FU Rong, ZHAO Ben-peng   

  1. Core Facility of Basic Medical Sciences, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China
  • Online:2018-07-28 Published:2018-07-30

摘要: 目的·优化流式细胞仪MoFlo Astrios EQ 96孔板单细胞分选方法,为单细胞技术的应用奠定基础。方法·通过对仪器精确调节及各项参数优化,采用不同孔径喷嘴、不同分选方式对32D、U937、iBMDM和293T共4种细胞进行单细胞分选,观察分选后单细胞孔数以及单细胞所形成克隆孔数。结果·在单细胞分选模式下,70、100 μm喷嘴分选获得单细胞的孔数分别为83~91、87~93孔。分选后的单细胞经7~10 d培养后,70 μm喷嘴分选获得的单细胞所形成克隆的孔数为36~58孔;100 μm喷嘴条件下电荷式分选、“无电”直接分选获得的单细胞所形成克隆的孔数分别为53~78、69~81孔。结论·进行单细胞分选时,选择100 μm喷嘴和“无电”直接分选方式可得到较好的细胞活性与较高的细胞克隆率。

关键词: 流式细胞仪, 96孔板, 单细胞分选, 喷嘴, &, ldquo, 无电&, rdquo, 直接分选, 单细胞培养

Abstract:

Objective · To optimize the method of the flow cytometry MoFlo Astrios EQ on single-cell sorting in 96-well plate. Methods · Using different aperture nozzles and sorting ways, the 32D, U937, iBMDM and 293T cells were used for single-cell sorting after the precise adjustment of the instrument and various parameters. The hole numbers with single cell and single-cell clones were detected after sorting. Results · In the single-cell sorting mode, the hole numbers with single cell were 83-9170 μm nozzle and 87-93100 μm nozzle. After 7-10 days of culture, the hole numbers with single-cell clones were 36-5870 μm nozzle. In 100μmnozzle, the hole numbers with single-cell clones were53-78electrostatic charge sorting and 69-81straight-down sorting, respectively. Conclusion · In single-cell sorting, a better cell viability and higher cloning rate are observed in 100 μm nozzle and straight-down sorting.

Key words: flow cytometry, 96-well plate, single-cell sorting, nozzle, straight-down sorting, single-cell culture

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