Organoid brightfield identification-based therapy screening

We developed a high-throughput live-cell imaging-based organoid analysis platform, Organoid Brightfield Identification-based Therapy Screening (Orbits), that allows for kinetic monitoring of organoid growth and drug responses in routine extracellular matrix domes, high-throughput 384-well microplates and advanced microcavity plates, solely based on brightfield imaging. The label-free Orbits deep learning analysis approach was validated against current standard assays for kinetic imaging (fluorescent viability dyes) and high-throughput analysis of organoid viability (CellTiter-Glo 3D assay). By incorporating a fluorescent cell death marker, intra-well normalization for organoid death could be achieved providing further insight into the mechanistic action of the drugs (cytotoxic vs. cytostatic), a feature not achievable with the CellTiter-Glo 3D assay. Our findings validate that Orbits, as a scalable, high-throughput technology, would facilitate the use of patient-derived organoids for drug development and ex vivo therapy screening. The developed platform also has broad application potential, from providing a launching point for further brightfield-based assay development to be used for fundamental research, to guiding clinical decisions for personalized medicine.