Evaluating the application of Cell Painting to phenotypic screening

Session

Poster Session Three

Authors

Dr Rebecca Kelly (1), Dr Adam Rolt (1), Dr Nina Vyas (1), Dr Hervé Barjat (1), Dr Eve Corrie (1), Dr Emma Jones (1), Dr Lorna FitzPatrick (1), Dr Emily Offer (1)

Affiliations

1. Medicines Discovery Catapult

Keywords

Cell painting, high content imaging, spinning disk confocal

Abstract text

Introduction 

Cell painting is a high-content image-based technique which is used to generate complex, information-rich, and interdependent measurements of cellular phenotype in vitro. Because of the unbiased, hypothesis free, nature of cell painting, it can be utilised in many areas of preclinical drug discovery, such as analysing the impact of genetic variants, profiling drug mechanism of action, and elucidating the phenotypic response of cells to of novel therapeutic candidates.

Using a set of fluorescence dyes, cell painting allows visualisation of cell components such as nucleus, nucleolus, ER/golgi, mitochondria, actin cytoskeleton and plasma membrane. Morphological changes within the cells mediated by disease or drug treatment can be quantified using analysis pipelines and used to study drug response based on the cell phenotypic response. 

Methods/Materials

Here cell painting protocols were established on the Opera Phenix Plus high content imaging system. Sample data was generated using a commonly used cell line, treated with well-annotated compound set. Analysis pipelines were developed to QC data, understand robustness of the assays and to automate image analysis.

Results and Discussion

A cell painting high content workflow allows approximately 1,500 measurements to be extracted from each cell, based on changes in size, shape, texture and fluorescence intensity. Combining all this data creates morphological profiles which can be evaluated in response to drug treatments, allowing characterisation of drug mechanisms on phenotypic cell responses. Here in this study, we have established a cell painting workflow at MDC, from high content imaging on the Opera Phenix Plus to analysis with Harmony and Cell Profiler software.

Conclusion

Cell painting is a powerful tool to analyse morphological profiles, and phenotypic characteristics of cells.  Application to drug discovery can provide invaluable mechanistic insights, and phenotypic profile information on new drugs, or drug classes.

References