In this work, a simple, reproducible, and inexpensive fabrication route for moldless micropatterning of a colloidal assembly on a conducting substrate surface is actualized by means of electrophoretic deposition (EPD) technique. A synergetic combination of photolithography and EPD is employed for fabricating micropatterns of barium titanate (BaTiO 3 ) nanoparticles on an indium-tin-oxide (ITO) coated glass slide or a Pt/ITO substrate. At first, high quality resist molds with various micropatterns are fabricated on ITO glass slide by photolithography technique, which is used for providing a controlled local electric field during the EPD process. Then, BaTiO 3 nanoparticles suspension is prepared in KCl aqueous solution and is demonstrated the BaTiO 3 nanoparticles are negative charged in the suspension. At last, EPD of various BaTiO 3 micropatterns is accomplished successfully on the anodic ITO glass slide or Pt/ITO substrate using the micropatterned ITO glass slide as the cathode, indicating that it is a simple and potential route for micropatterning of colloidal assembly on a non-modified conducting substrate by virtue of EPD process.