Image segmentation of cytosolic and nucleoplasmic markers allows for volume calculations.

Exemplary single image slices from time points pre-schizogony (t−2), schizogony start (t0) and schizogony end (tend) showing cytoplasmic marker pARL-GFP (green), nuclear marker NLS-mCherry (cyan), and thresholding masks generated for measuring respective areas and volumes. GFP mask was determined by automatic thresholding and mCherry by manual and visual adjustment. Shown is only z slice 11 of a stack of 20 slices to demonstrate segmentation, while analysis was carried out on the entire stack, which then contains all the nuclei. Contrast was adjusted for better visualization. Scale bar is 1 μm. (TIF)