High throughput quantitative tracking of single parasite in Plasmodium falciparum

New systematic profiling of drug effects is in urgent demand due to limitations in existing drug assessment approaches to evaluate comprehensive drug effects and distinguish heterogeneity in mixed populations. One challenge is the underdeveloped methods for investigation of causal association between artemisinin induced dormancy and recrudescence in Plasmodium falciparum . We developed Quantitative Tracking After Chemotherapy Exposure (qTRACE) and its artificial intelligence (AI) mode to evaluate cytotoxic and cytostatic effects simultaneously from single parasite resolution in a high throughput manner. Applying qTRACE, we revealed true parasite-drug killing dynamics after artemisinin exposure and found that up to 50% of viable parasites originate from recovery of artemisinin-induced dormancy. We further developed next generation qTRACE integrated with deep learning-based segmentation and analysis, thereby directly linking time-lapse phenotypes of label-free live cell. qTRACE is a versatile platform for in-depth single cell level drug effects evaluation, recovery after drug exposure studies and complex long-term growth experiments.