Natural swelling: in this method soluble lipids in chloroform are pipetted on a Teflon ring. The chloroform is allowed to evaporate and the ring is then placed under the vacuum for several hours. Next the aqueous buffer is added gently over the Teflon ring and lipids are allowed to naturally swell to form GUVs overnight. the disadvantage of this method is that a large amount of multilamellar vesicles and lipid debris are formed.
Electroformation: In this method lipids are placed on a conductive cover glass (indium tin oxide or ITO coated glass) or on Pt wires instead of a Teflon ring and after vacuuming, buffer is placed on the dried lipids and it is sandwiched using a second conductive cover glass. Next an electrical field with certain frequency and voltage is applied which promotes formation of GUVs. For polyunsaturated lipids, this technique can induce a significant oxidation effect on the vesicles. Nevertheless, it is a very common and reliable technique to generate GUVs. Modified approaches exist that employ gel-assisted swelling (agarose-assisted swelling or PVA-assisted swelling) for the formation of GUVs under more biologically relevant conditions.
A variety of methods exist to encapsulate biological reactants within GUVs by using water-oil interfaces as a scaffold to assemble lipid layers. This allows the use GUVs as cell-like membrane containers for the in vitro recreation (and investigation) of biological functions. These encapsulation methods include microfluidic methods, which allow for a high-yield production of vesicles with consistent sizes.