Two-dimensional (2D)-supramolecular assemblies of pi-conjugated chromophores are relatively less common compared to a large number of recent examples on their low dimensional (0D or 1D) assemblies or 3D architectures. This article reports a rational design for the 2D supramolecular assembly of an amphiphilic core-substituted naphthalene-diimide derivative (cNDI-1). The building block contains a naphthalene-diimide (NDI) chromophore, symmetrically substituted with two dodecyl chains from the aromatic core while the imide positions are functionalized with two hydrophilic wedges containing oligo-oxyethylene chains. In water, it exhibits entropically favorable self-assembly with a critical aggregation concentration of 1.5 x 10(-5) M and a lower critical solution temperature of 55 degrees C. The UV/vis absorption spectrum in water shows bathochromically shifted absorption bands compared to that of the monomeric dye in THF, indicating offset it-stacking among the NDI chromophores. C-H symmetric and asymmetric stretching frequencies in the FT-IR spectrum support the presence of organized hydrocarbon chains in trans conformation in the self-assembled state, similar to that in the crystalline n-alkanes, which is further supported by studying the general polarization (GP) values of a noncovalently entrapped Laurdan dye. The atomic force microscopy (AFM) image shows the formation of ultrathin (height < 2.0 nm) ribbons for the spontaneously assembled sample which eventually produces a large-area 2D nanosheet by the lateral organization. The powder X-ray diffraction pattern of the drop-tasted film, prepared from the preformed aggregates, reveals sharp peaks that indicate a crystalline lamellar packing along the direction of the 2D growth. Differential scanning calorimetry trace shows the melting of the crystalline alkyl chain domain at T > 75 degrees C, which destroys the 2D assembly. Local-scale photoconductivity of the ordered 2D assembly, studied by the flash-photolysis time-resolved microwave conductivity (FP-TRMC) technique, reveals an anisotropic conductivity with similar to 3 times larger conductivity along the parallel direction compared to that along the perpendicular one.