Live-imaging rate-of-kill compound profiling for Chagas disease drug discovery with a new automated high-content assay
Fig 1
Rate-of-kill assay development: a) Rate-of-kill assay outline; b) Live cell imaging of Tc-X10/7-E2Crimson-infected H9C2-eGFP cells (left panel: brightfield, middle panel: fluorescence image) and image algorithm segmentation segmentation/quantification of E2Crimson-labeled parasites (right panel, segmented parasites are circled white); c) time-course data. Left panel: mean number of Tc-X10/7-E2Crimson-infected H9C2-eGFP cells versus time. Middle panel: total number of intracellular amastigotes detected by the image analysis algorithm per well versus time. Right panel: percent infected host cells versus time. The small number of infected cells seen at early timepoints in the uninfected control is due to image analysis artefacts while the system equilibrates. A representative experiment of 3 biological repeats is shown. Error bars represent standard deviation of 12 technical replicates.