Multiparametric phenotypic screening based on cellular morphology interrogates many biological pathways simultaneously and is therefore a valuable screening tool for the discovery of new biological activities. The cell painting assay stains various cellular features using six different dyes in one well. By automated image analysis, hundreds of parameters are calculated from the images which deliver a phenotypic profile of the cell. It has been shown that compounds with similar modes of action deliver similar phenotypic profiles. Using a reference set of compounds with known modes of action, it is possible to assign probable modes of action to new compounds and to discover compounds with potentially new modes of action.Here we describe the cell painting assay as a screening tool using a hit identification workflow which has been implemented using open-source software.
Keywords: Cell painting assay; Hit identification; Morphological profiling; Phenotypic profile; Screening.