This paper describes the shrinkage and re-expansion of individual femtoliter-volume aqueous droplets that were suspended in an organic medium and held in an optical vortex trap. To elucidate the mechanism behind this phenomenon, we constructed a heat- and mass-transfer model and carried out experimental verifications of our model. From these studies, we conclude that an evaporation mechanism sufficiently describes the shrinkage of aqueous droplets held in a vortex trap, whereas a mechanism based on the supersaturation of the organic phase by water that surrounds the droplet adequately explains the re-expansion of the shrunk droplet. The proposed mechanisms correlated well with experimental observations using different organic media, when H2O was replaced with D2O and when an optical tweezer was used to induce droplet shrinkage rather than an optical vortex trap. For H2O droplets, the temperature rise within the droplet during shrinkage was on the order of 1 K or less, owing to the rapid thermal conduction of heat away from the droplet at the microscale and the sharp increase in solubility for water by the organic phase with slight elevations in temperature. Because most chemical species confined to droplets can be made impenetrable to the aqueous/organic interface, a change in the volume of aqueous droplets translates into a change in concentration of the dissolved species within the droplets. Therefore, this phenomenon should find use in the study of fundamental chemical processes that are sensitive to concentration, such as macromolecular crowding and protein nucleation and crystallization.