Dip-pen nanolithography of reactive alkoxysilanes on glass.

The use of organofunctional silane chemistry is a flexible and general method for immobilizing biomolecules on silicon oxide surfaces, including fabricating DNA, small-molecule, and protein microarrays. The biggest hurdle in employing dip-pen nanolithography (DPN) for extending this general approach to the nanoscopic domain is the tendency of trialkoxy- and trichlorosilanes to rapidly polymerize due to hydrolysis reactions. The control of the local water concentration between the substrate surface and the scanning AFM tip is critical, both to the physical and chemical processes involved in DPN writing and to the ability to form well-defined thin layers of reactive silanes without extensive polymerization induced disorder. We found that we could control the degree of polymerization through careful choice of the alkoxysilane used as the "ink" for DPN and through control of the relative humidity during inking and writing with the coated AFM tip. As a proof-of-principle, we demonstrate that areas patterned with an alkoxysilane on glass with DPN are functional for subsequent immobilization of fluorescently labeled streptavidin via covalent attachment of biotin. This preliminary result sets the stage for the ability to capture proteins in their fully hydrated state from buffered solution, by molecular recognition onto previously written reactive nanoscopic regions on oxidized silicon and glass.