Fig. 2
From: Detection of early-stage lung cancer in sputum using automated flow cytometry and machine learning
Automated gating of FCM data. A Bead size exclusion gate (BSE) parameters are set on the entire sputum sample with the bottom left threshold derived from automated NIST bead flowClust analysis (i inset). The top right corner of the BSE gate is set at 2.5 × 105 based on the observation that events above that threshold on both forward (FSC) and side (SSC) scatter area are dead cells (ii inset; FVS510 viability histogram). B In some samples, events appear in the lower righthand corner of FSC-H vs SSC-H plots. These events are gated out to avoid including them as live, viability marker negative, TCPP dull cells as shown in (iii). C A viability threshold is set on the Non-debris events based on FVS510 fluorescence (details in Fig. 3 and Additional file 1). D A singlets gate is then set on viable events (details in Fig. 4 and Additional file 1). Viable singlets are used for downstream numerical analysis. FI: Fluorescence Intensity