In single-dose screening all data points for a ligand have to show a consistent significant deviation from the references for the ligand to be considered a Hit. The minimum required response is defined on the “Data” page of DI.Screening Analysis, either as a manual minimum or as a multiple of the global standard deviation over all references of the assay. The mean response for one replicate well of the ligand must not deviate from the ligand average by more than 20 %. Otherwise, the Inhomogeneity Quality is set to negative.