The monomolecular organization of the main archaeal tetraether lipds from the thermoplasma acidophilum and sulfolobus metallicus were studied by means of a Langmuir film balance. With pure water as the subphase, the experiments were performed at a temperature of 19?C ± 1?C . For the balance experiments the archaeal tetraether lipids were dissolved in 30µl 1 mMol concentrated chloroform. Monolayer films were transferred on silicon wafers substrate. After transferred for several kinds of silicon wafers substrate (Hydrophobic, Hydrophilic, and Aminosilanization) the monolayer films were further (Hydrophobic, Hydrophilic and Aminosilanization) ) investigated using ellipsometry and atomic force microscopy to find out whether bolaamphiphilic might be differently oriented at the water-air interface including “horseshoe” and “upright standing” configuration. Using Ellipsometry, the thickness of monolayer films was represented as a mean value combination of domains. Meanwhile, Atomic Force Microscopy (AFM) was employed to observe the surface morphology. The heat capacity measured with DSC indicates a lower value was obtained in the asymmetric tetraether llipids.