Sporozoites were seeded with 3% BSA/ RPMI into an 8‐well Labtek chambered cover glass. After fixation with 4% PFA for 20 min, cells were permeabilized with 0.5% TritonX for 15 min. Primary antibodies were incubated for 1 h and washed twice with PBS. Secondary antibodies together with Hoechst were incubated for 1 h. Cells were washed twice with PBS and observed under the microscope. Images were either taken on a Zeiss CellObserver widefield (63×) or Nikon/PerkinElmer spinning disc (100×) microscope. Image processing was performed with ImageJ. Antibodies: rabbit anti‐GFP for IFA 1/40 (abfinity 0.4 µg/µl), mouse anti CSP (mAb 3D11, 1/100, Yoshida et al, 1980 (link)), goat anti‐mouse Alexa 594 1/1,000 (Invitrogen 2 mg/ml), goat anti‐rabbit Alexa 594 1/1,000 (Invitrogen 2 mg/ml). For STED imaging, anti‐mouse Atto 594 1/300 (Sigma) and anti‐rabbit Atto 647N 1/300 (Sigma) was used. The staining of sporozoites with antibodies or SiR‐tubulin was performed as described previously (Spreng et al, 2019 (link)).