Abstract

Cellular morphology has been recognized as a rich source of information. Using phenotypic profiling methods, changes in cellular morphology can be quantified by the generation of a morphological profile or fingerprint. 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. The comparison of phenotypic profiles between agents with related mode of action has shown that similar perturbations lead to similar changes in cellular morphology. Thus, by using a set of reference compounds, phenotypic profiling can also deliver information about possible mechanisms of action of small molecules.
At the Compound Management and Screening Center of the Max Planck Society (COMAS), the Cell Painting Assay was established and 3600 reference compounds as well as nearly 10.000 proprietary compounds were measured. Custom analysis tools have been developed which include similarity searching between phenotypic profiles, searching for commonly found references for a set of compounds or visualization of profiles as heat maps or line plots. These tools allow every chemist to analyze the phenotypic profiles of his compounds to identify biological activity in a new series, and to guide synthetic decision making by comparing the profiles within a given compound class. Furthermore, the generation of target hypotheses regarding the mode of action of the compounds is supported.