Rhoptries undergo biogenesis near the centriolar plaque and are segregated during nuclear division.

3D7 parasites were prepared by ultrastructural expansion microscopy (U-ExM), stained with N-hydroxysuccinimide (NHS) ester (grayscale), BODIPY TRc (white), SYTOX (cyan), and an anti-rhoptry antibodies and imaged using Airyscan microscopy. (a) Images of whole parasites throughout schizogony stained using an anti-RAP1 (rhoptry bulb; magenta) antibody. (b) Zoom into rhoptry pairs of 3D7 parasites that were prepared for U-ExM and stained with NHS ester (grayscale) along with antibodies against RAMA (rhoptry bulb; magenta) and RON4 (rhoptry neck; yellow) to assess rhoptry neck biogenesis. We observed that the rhoptry neck begins as a single focus inside each rhoptry. Rhoptries then get duplicated and segregated alongside the centriolar plaque. During the final mitosis, the rhoptry neck begins to elongate and the rhoptries separate from centriolar plaque. Images are maximum-intensity projections, number on image = Z-axis thickness of projection in µm. Scale bars = 2 µm. . Liffner B, Cepeda Diaz AK, Blauwkamp J, Anaguano D, Frolich S, Muralidharan V, Wilson DW, Dvorin JD, Absalon S. Atlas of Plasmodium falciparum intraerythrocytic development using expansion microscopy. Elife. 2023 12:RP88088.