Nanoliposome, or submicron bilayer lipid vesicle, is a new technology for the encapsulation and delivery of bioactive agents. The list of bioactive material that can be incorporated to nanoliposomes is immense, ranging from pharmaceuticals to cosmetics and nutraceuticals. Because of their biocompatibility and biodegradability, along with their nanosize, nanoliposomes have potential applications in a vast range of fields, including nanotherapy (e.g. diagnosis, cancer therapy, gene delivery), cosmetics, food technology and agriculture. Nanoliposomes are able to enhance the performance of bioactive agents by improving their solubility and bioavailability, in vitro and in vivo stability, as well as preventing their unwanted interactions with other molecules. Another advantage of nanoliposomes is cell-specific targeting, which is a prerequisite to attain drug concentrations required for optimum therapeutic efficacy in the target site while minimising adverse effects on healthy cells and tissues. This chapter covers nanoliposomes, particularly with respect to their properties, preparation methods and analysis.