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              <p>Abstract Three-dimensional multicellular aggregates like organoids and spheroids have become essential tools to study the biological mechanisms involved in the progression of diseases. In cancer research, they are now widely used as in vitro models for drug testing. However, their analysis still relies on tedious manual procedures, which hinders their routine use in large-scale biological assays. Here, we introduce a novel drop millifluidic approach to screen and sort large populations containing over one thousand multicellular aggregates. Our system utilizes real-time image processing to detect pheno-morphological traits in cellular aggregates. They are then encapsulated in millimetric drops, actuated on-demand using the acoustic radiation force. We demonstrate the performance of our system by sorting spheroids with uniform sizes from a heterogeneous population, and by isolating organoids from spheroids with different phenotypes. We anticipate that this work offers the potential to standardize drug testing on multicellular aggregates, which promises accelerated progress in biomedical research.</p>
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