A novel supramolecular thin film in which a non-linear optical (NLO) chromophore is uniaxially included in the helical cavity of amylose, revealed a spontaneous self-assembly with a long-range polar order of the chromophores perpendicular to the film surface. This was manifested by a significantly large second harmonic generation (SHG) observed from solution-cast solid supramolecular thin films on a glass substrate without any external field. Such self-assembly/self-poling and the long-term polar stability of the supramolecular chromophores are thought to be due to the helical rigid-rod structure, their dipolar and/or chiral recognition in the ordering process following the polar initiation through the interface interaction of the chromophore with the glass substrate, and H-bonding among the ordered supramolecules.