Modification for substrate surface morphology is a kind of effective technique for controlling thin film growth orientation. In this work, continuous V-grooves are fabricated on monocrystalline Si(100) substrate using TiO2 resist mask by anisotropic wet etching. Influence of thickness of TiO2 resist mask on etching patterns is investigated. The integrity and cross section of V-grooves are observed by laser scanning confocal microscope (LSCM) and scanning electronic microscopy (SEM), respectively. The floccules precipitating in V-grooves are verified by an energy dispersive spectrometer (EDS). Results indicate that the Si(100) substrate surface is modified to the array of V-grooves along the Si direction with Si(110) sidewalls.