High-Content Screening of Schistosomula Phenotypes

The larval High-Content Screening (HCS) was run as described previously [31 (link)]. Briefly cultured schistosomula were cultured for 72 hours in absence or presence of compound. After a gentle re-suspension using a Biomek FxP to generated an even distribution of parasites across the well, images were collected at 10x for phenotype analysis and at 4x over a time-series for motility analysis using an Image Xpress micro (Molecular devices, USA). Image analysis was then conducted using Pipeline Pilot 9.0 (Biovia, USA). Phenotype analysis was conducted using a previously validated two class Laplacian-modified Bayesian categorization model built using standard schistosomacides and motility analysis by individual parasite area displacement over the time series [31 (link)]. To generate larval IC50s the percent inhibition at specific compound concentrations were calculated by comparison to the mean DMSO scores (32 wells) on each plate. These were then imported into Prism 6 (Graphpad) to generate IC50 values.