A novel method of protein patterning based on anodic oxidation of Si surface by atomic force microscope (AFM) is presented. An oxide pattern is drawn on the Si surface by applying d.c. voltage between the Au-coated AFM tip and the Si substrate. The oxide patterns can be used as a template for patterning of biomolecules. Protocols for both positive and negative patterning of protein were established. (C) 2003 Elsevier Ltd. All rights reserved.