ML-guided robotic microinjection of single neurons in human brain organoids

Abstract

Human brain organoids have emerged as powerful models for studying the physiology, pathology, and evolution of the human brain. When combined with single-cell approaches, they offer the potential to directly probe the dynamics and mechanisms underlying cell fate decisions. A major technical bottleneck, however, remains the ability to reliably visualize and manipulate individual cells within their dense and heterogeneous tissue environment. Microinjection has proven effective for this purpose, allowing direct delivery of membrane-impermeable probes into single cells within intact tissue. Despite its versatility, widespread adoption of microinjection has been limited by its technically demanding and low-throughput nature. Automated microinjection systems developed for murine tissue have demonstrated that robotics can overcome these limitations, enabling systematic single-cell lineage tracing at scale. Here, we present a vision-guided robotic system capable of imaging organoid slices, identifying tissue boundaries, and targeting specific single cells for microinjection. This approach is generalizable across murine and human tissues and enables high-throughput single-cell manipulation, opening new avenues for studying human brain development at scale.