Differential staining of merozoites and DVs. (A) Giemsa stain of late-stage pRBCs undergoing schizont rupture. (B) Vital stain of the same culture with BCECF-AM showing intense staining of DVs (large arrow) alongside staining of merozoites (small arrow). (C) Same culture stained with Hoechst 33342, which detects merozoites (small arrow) but spares the DVs (large arrow). (D) Giemsa stain of isolated DVs. Insert shows the recording of analysis in a hemocytometer with DVs presenting as particles with the size of platelets (PLT). (E) Staining of isolated DVs with BCECF. (F) No staining with Hoechst 33342. (G) Phase-contrast image of DVs to which RBCs were added. (H) Same smear fluorescently stained for RBC band 3 protein. (I) Giemsa stain of purified merozoite preparation used in Western blots. (J) Western blots of RBC membranes, merozoites, and DVs probed with Abs against RBC band 3 protein and MSP1 (K). DV preparations contained very scant contaminations with MSP1. Arrows depict the positions of the respective molecular weight markers. Scale bars indicate 10 μm.