Experience with Single Cell Sorting on a BD FACSAria Flow Cytometer and Subsequent Single Cell Culture

The key points of single cell sorting on a BD FACSAria flow cytometer were elucidated, including a good state of cells, an appropriate cell concentration, the selection of nozzle size, drop delay determination and adjustment of the home location of ACDU (automatic cell deposition unit). The mean fluorescence intensity and purity of cells obtained by single cell sorting and subsequent cell culture were better than cells that were sorted into a tube and cultured. In order to obtain cells with both high mean fluorescence intensity and purity, more wells containing only one single cell should be selected and cells in these wells should be cultured individually. When the number of cells was sufficient, cells with high mean fluorescence intensity and purity should be selected by flow cytometry. The whole process was introduced in detail, offering valuable references for researchers in this field.