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In designing new biomaterials, specific chemical and topographical cues will be important in guiding cell response. Filopodia are actin-driven structures produced by cells and speculated to be involved in cell sensing of the three-dimensional environment. This report quantifies filopodia response to cylindrical nano-columns (100nm diameter, 160nm high) produced by colloidal lithography. Also observed were actin cytoskeleton morphology by fluorescence microscopy and filopodia morphology by electron microscopy (scanning and transmission). The results showed that the fibroblasts used produced more filopodia per μm of cell perimeter and that filopodia could often be seen to interact with the cells’ nano-environment. By understanding as to which features evoke spatial reactions in cells, it may be possible to design better biomaterials.