Assay Development and Screening
Advanced Imaging & High-Content Assays
Having historically looked at characterising our compound collection for cytotoxicity and cytostaticity or performed the routine annotation of the AstraZeneca compound collection with frequent hitters, we are looking into alternative compound annotation solutions (e.g. imaging or transcriptomics output) and the added benefits they may provide. The Cell Painting imaging assay multiplexes six fluorescent stains to enable the visualisation of up to eight cellular compartments in U2OS cells. We will present how we have established the Cell Painting imaging assay in 384-well plates and made it compatible with our CoLab automation platform. The use of a centrifugal plate washer afforded the reduction of staining solution needed and combined with a microvalve-based microplate dispenser allowed the miniaturisation of the assay to 1536-well plates. Driven by sustainability, we have also explored the use of a microplate cleaning system to wash and reuse plates for this assay. Specific collections we have applied the Cell Painting imaging assay include a set of compounds also tested with a metabolomics output, compounds that are part of a phenotypic set or compounds from a nuisance compound set. On the analysis side, we will share our experiences using a traditional analysis pipeline (segmentation, working with ca. 1500 features) and the work we are planning to do using deep learning models to best exploit images and data generated. The latter part will touch on the work we have been doing to segment compartments from images.