Label-Free Live-Cell Imaging improves Mode of Action Classification
Published: 2025-04-22
Formatted citation
Forsgren E, Rietdijk J, Holmberg D, Johansson M, Carreras-Puigvert J, Trygg J, Lovell G, Spjuth O, and Jonsson P..
Label-Free Live-Cell Imaging improves Mode of Action Classification.
bioRxiv.
2025.04.22.649936 (2025).
DOI: 10.1101/2025.04.22.649936
Abstract
Morphological profiling is a powerful method for identifying the modes of action (MOAs) of chemical compounds. However, most approaches rely on fixed-cell assays that capture only a single time point, missing the dynamic nature of cellular responses. While live-cell imaging captures these temporal effects, its potential for MOA classification using high-dimensional features remains unexplored. We strategically convert large-scale time-series image data (>82,000 images) into interpretable phenotypic trajectories and show that label-free live-cell imaging captures meaningful temporal phenotypic signatures that improve MOA classification compared to single-time-point analyses. The results are consistent across two cell lines and six MOA groups. This workflow enables mechanistic insight from live-cell images and supports the growing interest in label-free methods, which offer advantages in preparation time and cost, and preserve the native state of cells. Our findings demonstrate that live-cell imaging, combined with data-driven analysis, offers a powerful tool for high-throughput drug screening.