Fig. 7.

Three‐step strategy for sorting single cells by FACS. First step: set the drop delay by manually modifying the model to obtain a high yield. Second step: PBS could be used as loading buffer, which has no significant effect on cell viability in 4 h. Third step: 10% FBS in culture medium, 2% BSA in PBS, or 5% FBS in PBS could be selected as the collecting buffer depending on the different requirements of the experiment. Specifically, 5% FBS in PBS is optimal for single‐cell RNA sequencing because it affects gene expression less than PBS alone.