A central characteristic of DV biochemistry and physiology is the high pH gradient (acid inside) that the organelle has. Regulation of DV pH is not fully understood, but it includes a V -type H + ATPase that hydrolyzes cytosolic ATP to pump H + into the DV [80] . Interestingly, although conflicting data have been published [81] it is now generally accepted that changes in DV pH and volume are linked to CQ resistance caused by PfCRT [21, 22, 74, 76] . These are further predicted to affect drug, lipid, metabolite and osmolyte traffic in and out of the DV. However, since the DV is a subcellular organelle for an intracellular parasite, precise quantification of these perturbations is quite challenging. It requires the development of novel, overlapping approaches, as described later.