Fig. 1.

Workflow and visualization of critical protocol steps. a Optimal density of seeded HeLa cells 24 h post seeding. b Representative images of dissection of mosquito salivary glands (red arrow) attached to the head (left) or removed (right). c HeLa cells infected with P. berghei sporozoites at a density representing sporozoite numbers used in the here described protocol. d HeLa cells infected with PbmCherry_Hsp70 65 hpi, prior to detached cell (red) collection. e Collection of detached cells at 65 hpi. f Representative image of collected PbmCherry_Hsp70 detached cells (red) at 65 hpi and uninfected HeLa cell debris (unlabelled). g Amaxa^® Nucleofector^® II device with transfection program U33. h Intravenous injection of transfected parasites into a naïve mouse. i Drug selection of transgenic parasites. j Checking the parasitaemia by Wright stain or fluorescence. k Monitor transgenic parasites by fluorescence. l Preparation of blood stabilates. Scale bars in all images are 50 µm