This application note explores how combining 3D single spheroid models with live-cell analysis can enhance compound screening and drug discovery workflows. Using a library of 880 FDA-approved compounds, researchers evaluated changes in spheroid growth, viability, and morphology over time to gain a more comprehensive understanding of compound activity than traditional endpoint assays can provide. The study demonstrates how live-cell imaging enables continuous, non-invasive monitoring of 3D cell models, delivering rich kinetic data while maintaining cells in a physiologically relevant environment.

The application note highlights the value of phenotypic screening in complex 3D models, allowing researchers to identify cytotoxic, cytostatic, and disruptive compound responses through simultaneous measurement of spheroid size and viability. It also compares findings from 2D monolayer cultures with 3D spheroid assays, revealing important differences in compound sensitivity and response that may improve the translation of in vitro results to in vivo outcomes. By integrating advanced imaging, automated analysis, and 3D cell culture, this workflow provides a powerful approach for identifying promising drug candidates and generating more predictive biological insights early in the drug development process.