Fig. 1: The experimental workflow for continuous single-cell imaging of dynamic processes over the entire intraerythrocytic cycle from invasion to egress.

A Acquisition of 3D-stacks of single cell images using differential interference contrast (DIC) and the fluorescence Airyscan mode. B Training a neural network based on Cellpose on DIC images of uninfected and infected erythrocytes, yielding models for erythrocytes and parasite cell recognition and segmentation. In the case of infected erythrocyte, three parasite models were trained, on ring stages, trophozoites/schizonts and all stages. C Automatic cell segmentation via the trained models, delineating the erythrocyte plasma membrane, the erythrocyte cytosol and the parasite. D Analyzing the spatial and temporal dynamics of the process under investigation throughout the replicative cycle and 3D rendering of the captured images for visualization.