Abstract:

Objective To sort green fluorescent protein (GFP) positive population of breast cancer cell line MCF-7 by flow cytometry sorter FACSAriaⅡ, and investigate the effect of different setup on the purity, yield and viability of collected cells. Methods GFP positive population of MCF-7 cells was sorted by 70 μm, 85 μm and 100 μm nozzles and 4-way purity, purity and yield sort modes respectively. The yield was calculated with cell counting, the purity was analyzed by flow cytometry, and the viability was determined by trypan blue staining. Results FACSAriaⅡ gained purity of more than 99% with all three nozzles. The yields of 70 μm, 85 μm and 100 μm nozzles were (37.67±2.05)%, (56.33±2.62)% and (43.33±0.47)% respectively, while the percentages of live cells were (88.23±0.97)%, (91.87±1.05)% and (95.33±0.90)% respectively. Sorting cells by 4-way purity, purity and yield sort modes with 85 μm nozzle, the yields were (51.00±2.45)%, (67.33±2.05)% and (74.00±0.82)% respectively, and the purities were 99.6%, 99.3% and 96.2% respectively. Conclusion In sorting GFP positive population of breast cancer cell line MCF-7 by FACSAriaⅡ, 85 μm nozzle and purity sort mode may be the favorable setup.

Key words: cytometry sorting, nozzle, sort mode, purity, yield