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。

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.