Parallelized bright-field and fluorescence imaging platform for organoids.

Organoid viability, maturation, and growth are commonly assayed through bright-field and fluorescence microscopy using a single-objective lens. However, standard imaging systems pose significant limitations for high-throughput applications, particularly in large-scale experiments requiring simultaneous imaging of organoids, necessitating tools that can rapidly and consistently capture organoid features while minimizing disturbances to culture conditions. Here, we present a multi-camera array scanner (MCAS) that parallelizes imaging through the simultaneous use of 48 objective lenses and sensors, resulting in a 98% reduction in acquisition times compared to commercial high-content imagers. We demonstrate and validate this system in multiple-well plate formats, in both 2D and 3D neural cell cultures, and in bright field and fluorescence. The MCAS improves the efficiency for measuring organoid growth rates, assessing responses to morphogens and drugs, and measuring viral transduction rates. Together, these findings establish the MCAS as a scalable and versatile imaging platform for rapid phenotyping in organoid research.