The protocol can be used to create and extrude phospholipid liposomes. This extrusion protocol roughly follows some guidelines published by the manufacturer. However, we found that the manufacturer's directions were insufficient in several areas. In particular, the manufacturer guidance on initial preparation of lipids was incomplete; 'good' liposomes cannot be prepared directly from dry lipid mixtures. In addition, our protocol provides supplementary guidance for extrusion and suspension of different classes of polar lipids; efficacy of the various matrixes we recommend was largely determined through trial and error. The manufacturer directions (downloaded March 4, 2017) are attached to this protocol as a .pdf document.This protocol was original created by