Archaeosomes are a novel generation of liposomes that are made from polar ether lipids extracted from the Archaea. Archaeosomes are more stable in the presence of acidic or alkaline pH, bile salts, high temperatures and phospholipase in comparison with liposomes and have found application in drug, vaccine and gene delivery. The aim of this study was to formulate archaeosomes using lipid extracted from Acidianus brierleyi and evaluation of physic-chemical properties. The lipids were extracted from A.brierleyi and analyzed by High Performance Thin-Layer Chromatography (HPTLC). Archaeosomes were prepared using film method and methylene blue was used as a drug model. Then they characterized by Differential Scanning Calorimetry (DSC), and their particle sizes were also determined. The release and permeation of methylene blue was carried out using dialysis membrane and rat skin, subsequently. HPTLC analysis of the extracted lipids revealed that the glycerol ether was the major lipids with more than 70 percent probability. Results of particle size assay showed a mean size of 109.83 nm. The results of DSC showed the shift in melting point of methylene blue on loading that is a good indication of methylene blue loaded archaeosome. The addition of cholesterol improved the ability of archaeosome to encapsulate methylene blue. Encapsulation efficiency was 81.66 ± 2.88% and there was about 100% release after 24 h. Also the drug release pattern followed Peppas model. The results demonstrated that archaeosome may be a suitable new type of nano-particle that can be used as drug delivery system.