Physical and chemical properties of solid materials are modified by introducing defects, which disarrange the atomic periodic structure. Typical example is oxygen vacancies on titanium dioxide (TiO2) surfaces. Oxygen vacancies on TiO2 surfaces provide new physical and chemical surface properties, such as conductivity, catalytic activity, hydrophilicity, etc. To date, annealing, electron-/photo-stimulated desorption, and chemical reaction have been reported to create oxygen vacancies on TiO2 surfaces. However, these techniques do not allow position control of the defects at the atomic scale. We report the creation of single oxygen vacancy using a scanning tunneling microscope (STM). This technique creates oxygen vacancy at desired site. In addition, based on the experimental findings, we discuss the mechanism of manipulating atomic defects using the STM.