To demonstrate the utility of this high-throughput workflow screening, human rectal organoids were generated and cultured in Gri3D plates. For proof-of-concept,
organoids were exposed to staurosporine and its effects on organoid morphology were monitored over time on the ImageXpress® Micro Confocal High-Content Imager.
For image analysis, a deep learning-based model was developed using IN Carta image analysis software to segment label-free organoids grown in Gri3D plates. This
approach allowed for robust automated image analysis of organoids for monitoring purposes. In addition, this approach can also be modified for end-point fluorescencebased viability analysis. Using Gri3D plates combined with the ImageXpress Micro Confocal system and IN Carta Image Analysis Software, we followed the development and self-organization of human rectal organoids over time with transmitted light (TL) and fluorescence imaging and quantified the cytotoxic effects of staurosporine.