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nature research | reporting summary
April 2018Instrument 4 lasers (405nm, 488nm, 561nm, 638nm). A 70-micron nozzle running at 70 psi and 90kHz was used as the setup for each sort
session.
Software FACSDiva (version 8.0.1) and FlowJo (version 10) were used to collect and analyze the flow cytometry data.
Cell population abundance Post-sort samples were analyzed for purity using the same FACS sorter. Using the same gating strategy , the purity of the
samples was determined and calculated to be 97–99%
Gating strategy Before gating on fluorescence, live, single cells were gated using FSC-A and SSC-A (for intact cells) and SSC-W/SSC-H and FSC-W/
FSC-H (to ensure that only singlets were sorted). FACS gates were drawn to include only live single cells based on Calcein Blue
AM+ and 7-AAD (Thermo Fisher Scientific). Further gates were drawn to arrive at CD3+CD45+EpCAM- (for CD3+ selected
samples) or CD45+EpCAM- cells (for CD45+ selected samples). Boundaries between positive and negative cell fractions were
drawn based on single-color stains.
Tick this box to confirm that a figure exemplifying the gating strategy is provided in the Supplementary Information.