Cortical Spheroid Development Tracked with a Novel Imaging Platform (DNA-PRISM) Reveals Culture Complexity and Establishes Reproducible Phenotype Baseline for Compound Screens and Disease Modeling

Moving personalized medicine and drug candidates from the lab to the clinic will require addressing the current hurdles that impede high-throughput generation of experimental and clinical data. One such challenge with current imaging technology is its limited ability to provide large cross-sections of protein expression within the same sample for in-depth analysis of cellular processes. Traditional immunofluorescence enables acquisition in up to five channels before having to rely on specialized and expensive equipment that is not readily usable without extensive training or financial commitment. Recent advances in multiplexed imaging techniques now allow tracking large numbers of markers at once, which creates exciting possibilities for teasing out pathways and generating more refined cell phenotypes. One such technique, optimized in our lab, uses DNA-conjugated antibodies (DNA-PRISM) to acquire highly multiplexed images without harsh antibody stripping, or other destructive treatments. DNA-PRISM provides comparable results to traditional immunofluorescence, while eliminating some of its more frustrating limitations, such as long secondary antibody incubation and the need to match animal hosts to avoid cross-reactivity. It is readily adaptable using standard imaging microscopes and without the need for specialized equipment or extensive training. Here we use PRISM to stain for multiple markers in spinner flask 3D cortical neural spheroids, derived from human iPSCs, looking at cell subtypes within the complex culture. In conjunction with automation, DNA-PRISM can establish an effective high-throughput platform for drug and compound screening, as well as neurodevelopmental disease phenotyping of complex model systems by eliminating variability due to culture conditions or staining efficacy.