Two-dimensional (2D) colloidal crystals have intriguing properties, which make them quite useful in a wide range of applications. For example, 2D colloidal crystals have been widely used as physical masks in, for example, nanosphere lithography, [1] for preparation of metal nanoparticle arrays with a variety of particle morphologies, [2] as templates for growth of binary colloidal crystals, [3] and recently even as templating substrates for preparation of novel plasmonic crystals. [4] Up to now, several methods have been developed to grow 2D colloidal crystals, including self-assembly by capillary force, [5] spincoating, [2a,6] electrophoretic deposition, [7] and LangmuirBlodgett deposition.[8] Among them, convective self-assembly induced by solvent evaporation has been shown to be an effective approach for self-organizing colloidal beads into 2D crystals with hexagonal close-packing geometry. [5] In a typical process, [5a] immersion capillary force at the suspension drying front tends to bring particles into contact when the liquid suspension thickness becomes comparable with the colloidal sphere diameter. Evaporation-induced flow brings more particles towards the suspension front and the crystal grows larger.A patterned substrate provides an effective way to control the packing symmetry and orientation of colloidal crystallizations, [9,10] which is quite important for further extending colloidal crystal applications. Several research groups have reported using a 2D-patterned substrate to drive the growth of colloidal crystals. [9,10a] Self-assembly in the presence of one-dimensional (1D) patterned substrates has also been studied in combination with thermal equilibrium assembly, [10a] convective assembly, [10b] electrostatic assembly, [10c] and electrophoretic deposition process.[10d] Although 2D centered-rectangular lattice colloidal crystals have been fabricated on a 1D-patterned substrate, high defect densities and short-range ordering were often observed in those previously reported results. [10a,10b,10d] To the best of our knowledge, an efficient method to prepare high-quality monolayer square-lattice colloidal crystals has not been reported yet.In this Communication, we report our study of self-assembly of colloidal polystyrene (PS) spheres on a 1D-patterned substrate. Previous studies showed that colloidal structures formed on a 1D-patterned substrate are highly sensitive to the ratio between sphere diameter (d) and groove period (p).[10a]Here, we explore other factors that could have a key influence on the formation of 2D square symmetric colloidal crystals using a 1D-patterned substrate. By introducing an experimental cell that allows simultaneous actions and strong competition of forces on colloids exerted by the suspension drying front and the patterned substrate, we have identified distinct effects of convective assembly on the growth of colloidal crystal by controlling the angle orientation (a) between the colloidal suspension drying front and 1D template grooves. We find that those experi...