This poster presents Live Cell Painting with a new non-toxic dye (ChromaLIVE) to probe cell physiology in high-content screening. It addresses the limitations of traditional HCS performed on fixed cells, which miss dynamic physiological changes.

This work highlights a novel fluorescent non-toxic dye (ChromaLIVE), enabling real-time phenotypic profiling in live cells and the quantification of disease-relevant phenotypes like apoptosis, autophagy, and ER stress. ChromaLIVE's unique properties, such as its multi-chromatic nature and non-interference with cell physiology, allow for extended monitoring. Its mix-and-read format also simplifies screening procedures by obviating washing steps.

The study validates ChromaLIVE's biocompatibility at the genome, proteome, and cell levels, confirming its high compatibility with live-cell imaging. This allows for the precise quantification of time-sensitive cellular events, as demonstrated through analyses of apoptosis kinetics (Staurosporine), autophagy kinetics (Rapamycin), and ER stress (Thapsigargin).

Leveraging progress in data science and high-content screening tools, ChromaLIVE emerges as a versatile tool for exploring cell phenotypes in drug discovery, capable of nuanced quantification of subtle, disease-relevant phenotypes and predicting MoA.