Easy alignment strategy for multi-level micropatterning

In recent years, there has been a strong demand in the biological community for multiplexed protein patterns for the control and study of cellular growth at subcellular and cellular length scales. Here, an easy alignment strategy for multi-level micropatterning is proposed enabling the broadening of patterning possibilities and the increase of architecturing complexity. In practice, our method consists of a set of custom-made machined holders for standardized fabrication of polymeric stamps and for controlled alignment of successive printing steps. With this approach, three interdigitated protein patterns of 5 mu m width were successfully obtained with an angular alignment precision below 0.55 degrees and a lateral alignment accuracy of less than 35 mu m over areas of 1 cm(2). After growth of primary neurons onto these multiplexed substrates, guided neurite extension was observed along the patterns. (C) 2018 Elsevier B.V. All rights reserved.