Cold field emission gun-scanning electron microscopy: A new tool for morphological and ultrastructural analysis of liposomes

Liposomes are lipid vesicles largely investigated in the past 30 years as pharmaceutical carriers. In the development of new liposome-based formulations, the study of liposome surface properties remains a crucial step. For this purpose, microscopy techniques can provide useful information, although each such technique suffers from some limitations. Here, we have used cold field emission gun-scanning electron microscopy (cFEG-SEM) to acquire detailed images of liposome surface. In particular, we observed PEGylated and non-PEGylated liposomes in different size ranges. In the case of nanosized liposomes (mean diameter about 200 nm), a morphological evaluation of the whole preparation was obtained. On the other hand, in the case of giant liposomes (mean diameter about 2 μm), it was possible to observe the different surface ultrastructures of the two formulations. In particular, a regular and only slightly wrinkled surface was observed in the case of non-PEGylated liposomes, while a very irregular surface ultrastructure was visible in the case of PEGylated liposomes. This study shows, for the first time, the potential of cFEG-SEM as a new and powerful tool to obtain information on liposome morphology and, at least in the case of giant liposomes, on ultrastructure of the liposome surface.