A combination of spontaneous reactive chemical domains bounded by non-fouling zones provides a means to covalently immobilize biomolecules in structured, spatially defined arrays. These arrays have application in a wide range of biotechnologies including tissue engineering, proteomics, and diagnostics. In this paper, we describe the fabrication of multi-chemistry micropatterns from plasma polymers. X-ray photoelectron spectroscopy (XPS), together with Time-of-Flight Static Secondary Ion Mass Spectrometry (ToF-SSIMS) and confocal imaging has been utilized to confirm the reactivity and integrity of micropatterns fabricated from amine-reactive maleic anhydride (ppMA) on non-fouling tetraglyme (ppTg). The covalent immobilization of antibodies via the formation of amide linkages with the anhydride groups occurs only in the ppMA domains, while antibody activity is confirmed via their ability to attract specific fluorescent antigens. These micropatterns therefore provide a convenient and effective platform for covalently immobilizing biomolecules in spatially defined areas without the need for multiple step wet chemical immobilization strategies. Crown Copyright (C) 2011 Published by Elsevier Ltd. All rights reserved.