(9) Set up the Avanti Polar Lipids, Inc extrusion kit with a 200 nm polycarbonate membrane between two support membranes. Ensure all membranes have been wetted with type 1 Milli-Q water before assembling, that the hex nut apex points downwards, and that all components are assembled tightly. This ensures full insertion to reduce undue pressure on the syringes and mitigates the chance of leakages (Figure ). (10) Fill one syringe with 1 mL of the lipid solution and align the tip with one side of the extrusion kit (Figure ). Place the other empty syringe into the opposite side of the kit, ensuring both syringes have tightened tips to minimise leakage. (14) Elute the lipids with the external solution, ensuring the column does not run dry.