Figure 1.

Visualization of Plasmodium falciparum (Pf)-derived extracellular vesicles (EVs) by imaging flow cytometry (IFC). (A) Visualization of single-stained Pf-derived EVs by IFC. Pf-derived EVs stained with lipid (DiI, DiD, and DiO), RNA [thiazole orange (TO)], or protein (GO) dyes. Insert shows percentage of TO-positive EVs (43%), gated according to unlabeled EVs. Representative results from at least three experiments are shown. Abbreviations: BF, bright field; SSC, side scatter. (B) Internalization (uptake) of Pf-derived EVs into monocytes as visualized by IFC. EVs were stained with lipid (DiI, DiD, and DiO), RNA (TO), or protein (GO) dyes and then 7.5*10⁷ dyed EVs were introduced into 1.5*10⁶ THP-1 cells for 5 min. The cells were fixated as described in Section “Method” and vesicle uptake was imaged using IFC. EVs are detected as spots inside recipient cells. Insert shows percentage of EV-positive cells (72.5%), gated according to unlabeled EVs. Representative results from at least three experiments are shown. Abbreviations: BF, bright field; SSC, side scatters.