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Live Cell Painting: image-based profiling in live cells using Acridine Orange
Biorxiv : the Preprint Server for Biology
|September 11, 2024
Summary
Live Cell Painting (LCP) offers a new way to study cell health in real-time without fixation. This high-content imaging method uses Acridine orange to track cellular changes, aiding drug and nanoparticle assessments.
Area of Science:
- Cell biology
- High-content imaging
- Drug discovery
Background:
- Image-based profiling, including Cell Painting, analyzes cellular responses but requires cell fixation, limiting live-cell studies.
- Existing methods like MTT assays may miss subtle cellular changes induced by certain treatments.
Purpose of the Study:
- Introduce Live Cell Painting (LCP), a high-content, image-based assay for real-time cell health assessment.
- Demonstrate LCP's utility in tracking cellular responses to inhibitors, nanoparticles, and drug treatments.
Main Methods:
- Developed Live Cell Painting (LCP), a method utilizing Acridine orange, a metachromatic dye, for live-cell imaging.
- Applied LCP to monitor acidic vesicle redistribution, cellular responses to silver nanoparticles, and drug-induced liver injury.
Main Results:
- LCP successfully tracked acidic vesicle redistribution in response to inhibitors.
- LCP detected subtle cellular changes induced by silver nanoparticles, outperforming MTT assays.
- LCP facilitated dose-response assessments and clustering of drugs causing liver injury.
Conclusions:
- Live Cell Painting (LCP) provides an affordable, easy-to-use method for assessing overall cell health in live cells.
- LCP shows promise for screening drugs and nanoparticles, and for investigating new biological questions.
