Growth and fission of the mitochondrion (a) Parasites with an smHA-tagged copy of the ATP Synthase F0 Subunit D (ATPd, Pf3D7_0311800) as a mitochondrial marker were prepared by U-ExM, stained with NHS ester (greyscale), BODIPY TRc (white), SYTOX (cyan) and anti-HA (mitochondrion; magenta) antibodies and imaged using Airyscan microscopy across the asexual blood stage. Maximum intensity projections of both a subsection of the cell (partial mito) and the full cell (full mito) are shown. (b) ATPd staining was compared against Mitotracker orange CMTMRos (yellow), which showed discontinuous staining in looped regions. (c) Area of the mitochondrion was quantified for parasites of varying nucleus number. 73 cells were counted across 4 biological replicates. **** = p<0.001, ns = p>0.05 by one-way ANOVA, error bars = SD. (d) Schizont with mitochondria that have undergone fission (yellow zoom), mitochondria that are shared between two nascent merozoites (black zoom), and mitochondria left outside merozoites in the forming residual body (grey). (e) During fission, mitochondria associate with the cytoplasmic extension of the MTOC. Images are maximum-intensity projections, number on image = Z-axis thickness of projection in µm. White scale bars = 2 µm, yellow scale bars = 500 nm.

Anaguano D, Frölich S, Muralidharan V, Wilson DW, Dvorin J, Absalon S. Atlas of Plasmodium falciparum intraerythrocytic development using expansion microscopy. bioRxiv [Preprint]. 2023 24:2023.03.22.533773.