Strategy for Generating Giant Unilamellar Vesicles with Tunable Size Using the Modified cDICE Method.

Our study investigates an optimization strategy that uses a size cutoff in giant unilamellar vesicles (GUVs) generated from water-in-oil (W/O) emulsion droplets using the modified continuous droplet interface crossing encapsulation method. While this method is rapid and cost-effective and yields high encapsulation efficiency, it suffers from a broad, poorly controlled size distribution of vesicles, a significant drawback for the construction of artificial cells. We address this by systematically varying key parameters, such as chamber rotation time, angular frequency, and inner solution density, to refine the GUV size distribution. Our study highlights the importance of these parameters as practical experimental knobs for the refinement of GUVs size. Our results are supported by a physical model, which helps explain the observed size selection phenomena. We also examine how the salinity of the inner solution affects the encapsulation efficiency, finding that a high efficiency is maintained even at physiologically relevant salt concentrations. Our approach offers a practical method for selecting vesicle sizes, thereby facilitating the creation of cell-sized compartments with biologically relevant properties for synthetic biology applications.