Extended Data Figure 10: Flow cytometry gating strategy.
From: Quantitative microbiome profiling links gut community variation to microbial load
A fixed gating and staining approach was applied28. Both blank and sample solutions were stained with SYBR Green I. a, The FL1-A/FL3-A acquisition plot of a blank sample (0.85% w/v physiological solution) with gate boundaries indicated. A threshold value of 2,000 was applied on the FL1 channel. b, Secondary gating was performed on the FSC-A/SSC-A channels to further discriminate between debris or background and microbial events. c, d, FL1-A/FL3-A count acquisition of a faecal sample (c) with secondary gating on FSC-A/SSC-A channels on the basis of blank analyses (d). Total counts were defined as events registered in the FL1-A/FL3-A gating area, excluding debris or background events observed in the FSC-A/SSC-A R1 gate. The flow rate was set at 14 μl per minute and the acquisition rate did not exceed 10,000 events per second. Each panel reflects events registered over the course of a 30-s acquisition period.
