A new method is demonstrated, where three-dimensional protein structures are made by employing multi-photon polymerization and photobiotin photolysis. The technique enables the construction of arbitrary two- and three-dimensional shapes with submicron resolution. The integrity of the immobilized biotin is confirmed by derivatization with fluorescently labeled streptavidin. Fluorescence microscopy is used in order to visualize the distribution of fluorescent streptavidin on the 3D structure. (c) 2007 Elsevier B.V. All rights reserved.