Wang Kai, Leong Kam W, Yang Yong
Department of Chemical and Biomedical Engineering, West Virginia University.
Department of Biomedical Engineering, Columbia University.
J Vis Exp. 2016 Dec 8(118):54840. doi: 10.3791/54840.
Substrate nanotopography has been shown to be a potent modulator of cell phenotype and function. To dissect nanotopography modulation of cell behavior, a large area of nanopatterned substrate is desirable so that enough cells can be cultured on the nanotopography for subsequent biochemical and molecular biology analyses. However, current nanofabrication techniques have limitations to generate highly defined nanopatterns over a large area. Herein, we present a method to expand nanopatterned substrates from a small, highly defined nanopattern to a large area using stitch technique. The method combines multiple techniques, involving soft lithography to replicate poly(dimethylsiloxane) (PDMS) molds from a well-defined mold, stitch technique to assemble multiple PDMS molds to a single large mold, and nanoimprinting to generate a master mold on polystyrene (PS) substrates. With the PS master mold, we produce PDMS working substrates and demonstrate nanotopographical modulation of cell spreading. This method provides a simple, affordable yet versatile avenue to generate well-defined nanopatterns over large areas, and is potentially extended to create micro-/nanoscale devices with hybrid components.
底物纳米拓扑结构已被证明是细胞表型和功能的有效调节因子。为了剖析纳米拓扑结构对细胞行为的调节作用，需要大面积的纳米图案化底物，以便在纳米拓扑结构上培养足够数量的细胞，用于后续的生化和分子生物学分析。然而，目前的纳米制造技术在大面积上生成高度精确的纳米图案方面存在局限性。在此，我们提出一种使用拼接技术将小面积、高度精确的纳米图案扩展到大面积纳米图案化底物的方法。该方法结合了多种技术，包括利用软光刻从一个定义明确的模具复制聚二甲基硅氧烷（PDMS）模具，采用拼接技术将多个PDMS模具组装成一个大模具，以及通过纳米压印在聚苯乙烯（PS）底物上生成母模。利用PS母模，我们制备了PDMS工作底物，并证明了纳米拓扑结构对细胞铺展的调节作用。该方法提供了一种简单、经济且通用的途径，可在大面积上生成定义明确的纳米图案，并有可能扩展用于制造具有混合组件的微/纳米级器件。
