We present a procedure for high volume fabrication of nanopore membrane chips, combining low cost hot embossing for nanopore replication and conventional photolithography for manufacture of the membranes, alignment marks and break lines. The embossing masters are fabricated by electron beam lithography allowing customisation of the nanopore design. Nanopore membrane chips containing four membranes were fabricated and three of the membranes were structured with arrays of pores with three different diameters. Several different nanopore periods were tested. For pillar sizes in the embossing master of 460, 250 and 95 nm, optimization of the pattern transfer process resulted in nanopore sizes of 550, 330 and 140 nm. For the minimum periods employed for these three pore sizes of 1000, 500 and 300 nm, respectively, the membranes were found to be stable. This fabrication technology opens the way for high volume batch processing of nanostructured membranes, facilitating new avenues for research and technology. (C) 2003 Elsevier Science B.V. All rights reserved.