Characterization of liposome suspensions by flow cytometry

Novel approaches to drug delivery and induction of immune responses using liposomes have received much attention in recent years. Liposomes, however, are not a singular entity, but can be produced with a diverse group of phospholipids that form microspheres of different sizes, physical structure, electrochemical characteristics, and most importantly, physiologic properties. The purpose of this study was to establish the usefulness of flow cytometry as a convenient, rapid method for assessing the relative size and uniformity of liposomal preparations. Liposomes were made from phospholipid suspensions by sonication alone, or sonication followed by microemulsification. Forward laser light scatter (FSC) analysis of liposomal preparations by flow cytometry indicated that microemulsification produced homogeneous, small vesicles which were less than 1 micron in diameter, compared to the more heterogeneous sized liposomes generated by sonication alone. Transmission electron micrographs of the liposomal preparations were used to confirm the FSC results and showed that liposomes prepared by microemulsification were homogeneous, unilamellar vesicles which exhibited a mean diameter of 99.8 nm, whereas the sonicated-only preparation was more heterogeneous in size, exhibiting a mean diameter of 154.1 nm. Analysis of various liposome preparations by FSC during a 9 week storage period showed that small vesicles were relatively stable. We conclude that flow cytometry using FSC analysis provides a rapid, reproducible and convenient method to evaluate the relative size, uniformity and stability of liposomes.