Extended Data Fig. 4: Flow cytometry Gating strategy.
From: Evaluating totipotency using criteria of increasing stringency
a, Displayed is an example of the gating strategy implemented to generate flow cytometry data. The data displayed uses the iCdx2 Elf5.2A.mCherry ESCs, which are eGFP+, contain the Elf5.2A.mCherry reporter, and are stained for both CD40 and Plet1.The plots are pseudo-colour plots and the x-axis labels state markers and fluorochromes used. Laser (V=violet; B=blue; G=green; R=red) and filter bandpass information is also included on x-axis in brackets. Ancestry plots are included to show how cells were gated: majority of cells were included in FSC/SSC gate due to size discrepancy between cells of interest and underlying feeder cells. Single cells were determined by FSC-W x FSC-H and SSC-W x SSC-H plots. Live cells were gated using dead cell stain Sytox blue. Cells of interest were discriminated from underlying feeder cells (EGFP negative) by expression of eGFP. Fluorescence minus one (FMO) controls were used to establish gates in order to separate positive and negative populations.
