Fig 1. Initial gating strategy to analyze Blastocystis cells.

Cells were gated for focused cells using brightfield channel, then selection of single cells using aspect ratio and area units, and finally to classify viable and non-viable cells using PI-staining characteristics. The above graphs shows the analysis for Blastocystis ST1-NUH9 isolate (A). Subsequent analyses made use of features arising from Hoechst and CFSE staining characteristics as well as features from brightfield and side-scatter channels (B).