Fig. 3

Variation among replicate measurements for each dataset. a Gating strategy for the determination of CFSE MFI for each time point. Cells were gated apart from debris in SSC-A × FCS-A plot. Considering only P1 gated events, single cells were gated apart from the doublets and clumps in FCS-H × FSC-A plot, and the CFSE MFI was measured considering only single cells. The analysis considered 4000 events into the single cells gate. b The presence of debris, doublet and clumps after cell harvesting and fixation interfere the final results of counting based methods, as well as those cells that remain in the plate after harvesting. On the other hand, these influences does not interfere on fluorescence-based method, which is based on the CFSE signal intensity of a small sample of single cells from each time point. c The coefficient of variation of three replicates for each time point. Statistical analysis was carried out by paired Wilcoxon test