Localization of TRAP and CSP was visualized by immunofluorescence staining of hemolymph sporozoites. Hemolymph was isolated from P. berghei‐infected mosquitoes as described previously (Klug et al, 2020 (link)) and collected in plastic reaction tubes. Obtained hemolymph sporozoites were transferred into an 8‐well glass‐bottom imaging chamber (Nunc Lab‐Tek), activated with 3% BSA in RPMI medium, and forced to attach to the bottom by centrifugation at 800 rpm for 8 min. Subsequently sporozoites were allowed to glide for 15 min at RT. Afterwards, samples were fixed using 4% paraformaldehyde (PFA) in PBS for 1 h at room temperature (RT). Fixed samples were washed three times with PBS and treated with primary antibody solutions for 1 h at RT. Subsequently samples were washed three times with PBS and treated with secondary antibody solutions for 1 h at RT in the dark. Finally, samples were washed three times in PBS and the supernatant was discarded. Samples were examined directly using a spinning disc confocal microscope (Nikon Ti series) with 100‐fold magnification (Plan Apo VC 100×/1.4 N.A. oil immersion) and a Hamamatsu sCMOS ORCA Flash 4.0 camera.