We took advantage of the microflow hydrodynamics in the evaporation of sessile droplets to increase the height uniformity of thin lipid films for the subsequent electroformation of defect-free giant unilamellar vesicles (GUV). By serially casting progressively larger liposome suspension droplets on the same spot of an indium-tin-oxide (ITO) electrode, we managed to leverage the coffee ring effect (CRE) in the evaporation of each droplet to generate a smeared multilayer film of uniform thickness. This multidroplet technique of lipid film formation outperformed the traditional single-droplet deposition, improving the final quality of electroformed GUV samples. The proposed film formation technique constitutes a solvent-free method that results in a dramatic reduction (∼20×) in the appearance of undesirable structures like nonspherical (NSV), multilamellar (MLV), and multivesicular (MVV) vesicles.
An environmentally friendly and straightforward dehydration/rehydration method for glycerophospholipid mixing that avoids the use of organic solvents, cosolvents, or additives was developed. We prepared binary mixtures of zwitterionic and anionic glycerophospholipids using only deionized water in the entire mixing process. The resulting lipid films were subsequently reconstituted in vesicular form and compared to controls using differential scanning calorimetry. The calorimetric scans revealed no significant differences between mixing methods for any of the studied cases. These findings suggest that the developed dehydration/rehydration procedure creates a sample with equivalent compositional uniformity than the conventional solvent evaporation technique.
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