We report on methods to fabricate robust micro- and nanopatterned platforms, comprising high functional group densities and quasi three-dimensional structures, for possible applications in biochip, array technologies. For this purpose, amine-terminated poly(amidoamine) (PAMAM) dendrimers were immobilized via amide linkage formation on 11,11'-dithiobis(N-hydroxysuccinimidylundecanoate) (NHS-C-10) self-assembled monolayers (SAMs) on gold surfaces. The coupling reaction and the resulting assemblies were characterized by grazing incidence reflection Fourier transform infrared spectroscopy, contact angle measurements, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy; the obtained surface coverage values were successfully fitted with a Langmuir isotherm. The fraction of unreacted peripheral primary amine groups of the surface-immobilized PAMAM dendrimers was 28% as determined by XPS analysis of trifluoroacetic anhydride-labeled assemblies. Patterning of the PAMAM dendrimers on NHSC10 SAMs on the micrometer and sub-100-nm scale was achieved by microcontact printing and dip pen nanolithography. The resulting patterns are characterized by their high degree of order and stability of the transferred molecules due to covalent attachment.