Commercial microelectrode arrays (MEAs) for in vitro neuroelectrophysiology studies rely on conventional two dimensional (2D) neuronal cultures that are seeded on the planar surface of such MEAs and thus form a random neuronal network. The cells attaching on these types of surfaces grow in 2D and lose their native morphology, which may also influence their neuroelectrical behavior. Besides, a random neuronal network formed on this planar surface in vitro also lacks comparison to the in vivo state of brain tissue. In order to improve the present MEA platform with the above mentioned concerns, in this paper, the authors introduce a three dimensional platform for neuronal cell culturing, where a linear nanoscaffold is patterned on a microsieve array by displacement Talbot lithography (DTL) and reactive ion etching. Good pattern uniformity is achieved by the DTL method on the topographically prepatterned nonflat surface of the microsieve array. Primary cortical cells cultured on the nanopatterned microsieve array show an organized network due to the contact guidance provided by the nanoscaffold, presenting 47% of the total outgrowths aligning with the nanogrooves in the observed view of field. Hence, the authors state that this nanopatterned microsieve array can be further integrated into microsieve-based microelectrode arrays to realize an advanced Brain-on-Chip model that allows us to investigate the neurophysiology of cultured neuronal networks with specifically organized architectures.