Institute of Nanotechnology (INT) and Karlsruhe, Nano Micro Facility (KNMF), Karlsruhe Institute of Technology (KIT), Germany; Physical Institute and Center for Nanotechnology (CeNTech), University of Münster, Germany.
Small. 2013 Oct 11;9(19):3266-75. doi: 10.1002/smll.201203183. Epub 2013 Apr 2.
Multiplexing, i.e., the application and integration of more than one ink in an interdigitated microscale pattern, is still a challenge for microcontact printing (μCP) and similar techniques. On the other hand there is a strong demand for interdigitated patterns of more than one protein on subcellular to cellular length scales in the lower micrometer range in biological experiments. Here, a new integrative approach is presented for the fabrication of bioactive microarrays and complex multi-ink patterns by polymer pen lithography (PPL). By taking advantage of the strength of microcontact printing (μCP) combined with the spatial control and capability of precise repetition of PPL in an innovative way, a new inking and writing strategy is introduced for PPL that enables true multiplexing within each repetitive subpattern. Furthermore, a specific ink/substrate platform is demonstrated that can be used to immobilize functional proteins and other bioactive compounds over a biotin-streptavidin approach. This patterning strategy aims specifically at application by cell biologists and biochemists addressing a wide range of relevant pattern sizes, easy pattern generation and adjustment, the use of only biofriendly, nontoxic chemicals, and mild processing conditions during the patterning steps. The retained bioactivity of the fabricated cm(2) area filling multiprotein patterns is demonstrated by showing the interaction of fibroblasts and neurons with multiplexed structures of fibronectin and laminin or laminin and ephrin, respectively.
多重处理，即多种墨水在叉指微图案中的应用和集成，对于微接触印刷（μCP）和类似技术来说仍然是一个挑战。另一方面，在生物实验中，对于亚细胞到细胞长度尺度的、数量超过一种的蛋白质的叉指图案有着强烈的需求，其尺寸在几微米到几十微米之间。在这里，我们提出了一种新的综合方法，用于通过聚合物笔式光刻（PPL）来制作生物活性微阵列和复杂的多墨水图案。通过利用微接触印刷（μCP）的优势，结合 PPL 的空间控制和精确重复能力，我们以一种创新的方式引入了一种新的 PPL 着墨和书写策略，从而能够在每个重复的子图案中实现真正的多重处理。此外，还展示了一种特定的墨水/基底平台，该平台可用于通过生物素-链霉亲和素方法固定功能蛋白和其他生物活性化合物。这种图案化策略特别针对细胞生物学家和生物化学家应用，其涵盖了广泛的相关图案尺寸、易于生成和调整图案、仅使用生物友好、无毒的化学物质以及图案化步骤中的温和处理条件等特点。通过展示成纤维细胞和神经元与纤连蛋白和层粘连蛋白或层粘连蛋白和 ephrin 的多重结构的相互作用，证明了所制造的大面积填充多蛋白图案的保留生物活性。
