Luteolin-7-O-β-D-glucoside is one of the most common flavonoids with diverse biological activity. Its application is limited due to its low-water-solubility. Three types of microemulsions were prepared, and the solubility of luteolin-7-O-β-D-glucoside in these molecular aggregates was determined by HPLC. UV-Vis absorption and NMR spectroscopy were used to study the properties of luteolin-7-O-β-D-glucoside in cetyltrimethylammonium bromide (CTAB) and Tween80 micelles in order to gain the information about the binding of luteolin-7-O-β-D-glucoside with micelles. The results showed that luteolin-7-O-β-D-glucoside could form conjugation in micelles, and the binding constants in CTAB and Tween80 micelles were 2326, 362 L/mol, respectively. Luteolin-7-O-β-D-glucoside was located at the interface of hydrophillic layers of CTAB micelles and embedded in the palisade of Tween80 micelles.