High-throughput histopathology for complex in vitro models

Histology has been a cornerstone for complex in vitro model (CIVM) characterization for decades. However, it remains a low-throughput method with time-consuming workflows. Here, we introduce a holistic "histo-workflow," utilizing 3D-printed histomolds that facilitate co-planar embedding of CIVMs at high throughput, resulting in up to 80 samples in one section. We developed a variety of model-specific histomold designs that enable spatially controlled histological sectioning and downstream analyses. We describe these workflows, including mold generation, high-plex staining, and image analysis, and exemplify their application to histological analyses of various CIVMs. Altogether, the histomolds introduced here afford opportunities for CIVM processing and analysis, while significantly reducing labor and reagent resources, thereby democratizing high-throughput handling of CIVMs in histopathology.