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Recent years have seen a dramatic increase in the application of organoids to developmental biology, biomedical and translational studies. Organoids are large structures with high phenotypic complexity and are imaged on a wide range of platforms, from simple benchtop stereoscopes to high-content confocal-based imaging systems. The large volumes of images, resulting from hundreds of organoids cultured at once, are becoming increasingly difficult to inspect and interpret. Hence, there is a pressing demand for a coding-free, intuitive and scalable solution that analyses such image data in an automated yet rapid manner. Here, we present MOrgAna, a Python-based software that implements machine learning to segment images, quantify and visualize morphological and fluorescence information of organoids across hundreds of images, each with one object, within minutes. Although the MOrgAna interface is developed for users with little to no programming experience, its modular structure makes it a customizable package for advanced users. We showcase the versatility of MOrgAna on several in vitro systems, each imaged with a different microscope, thus demonstrating the wide applicability of the software to diverse organoid types and biomedical studies.

MOrgAna: accessible quantitative analysis of organoids with machine learning