Department of Chemistry, Pennsylvania State University , University Park, Pennsylvania 16802, United States.
Langmuir. 2013 Sep 10;29(36):11535-45. doi: 10.1021/la402362u. Epub 2013 Aug 29.
Combining biological molecules with integrated circuit technology is of considerable interest for next generation sensors and biomedical devices. Current lithographic microfabrication methods, however, were developed for compatibility with silicon technology rather than bioorganic molecules, and consequently it cannot be assumed that biomolecules will remain attached and intact during on-chip processing. Here, we evaluate the effects of three common photoresists (Microposit S1800 series, PMGI SF6, and Megaposit SPR 3012) and two photoresist removers (acetone and 1165 remover) on the ability of surface-immobilized DNA oligonucleotides to selectively recognize their reverse-complementary sequence. Two common DNA immobilization methods were compared: adsorption of 5'-thiolated sequences directly to gold nanowires and covalent attachment of 5'-thiolated sequences to surface amines on silica coated nanowires. We found that acetone had deleterious effects on selective hybridization as compared to 1165 remover, presumably due to incomplete resist removal. Use of the PMGI photoresist, which involves a high temperature bake step, was detrimental to the later performance of nanowire-bound DNA in hybridization assays, especially for DNA attached via thiol adsorption. The other three photoresists did not substantially degrade DNA binding capacity or selectivity for complementary DNA sequences. To determine whether the lithographic steps caused more subtle damage, we also tested oligonucleotides containing a single base mismatch. Finally, a two-step photolithographic process was developed and used in combination with dielectrophoretic nanowire assembly to produce an array of doubly contacted, electrically isolated individual nanowire components on a chip. Postfabrication fluorescence imaging indicated that nanowire-bound DNA was present and able to selectively bind complementary strands.
将生物分子与集成电路技术相结合对于下一代传感器和生物医学设备具有重要意义。然而,当前的光刻微加工方法是为了与硅技术兼容而开发的,而不是与生物有机分子兼容,因此不能假设生物分子在芯片处理过程中会保持附着和完整。在这里,我们评估了三种常见的光刻胶(Microposit S1800 系列、PMGI SF6 和 Megaposit SPR 3012)和两种光刻胶去除剂(丙酮和 1165 去除剂)对表面固定的 DNA 寡核苷酸选择性识别其反向互补序列的能力的影响。比较了两种常见的 DNA 固定方法:直接吸附到金纳米线上的 5'-巯基化序列和通过表面胺共价连接到涂有二氧化硅的纳米线上的 5'-巯基化序列。我们发现与 1165 去除剂相比,丙酮对选择性杂交有有害影响,可能是由于不完全去除光刻胶。使用 PMGI 光刻胶,其中涉及高温烘烤步骤,对杂交测定中纳米线结合 DNA 的后续性能不利,特别是对于通过硫醇吸附附着的 DNA。其他三种光刻胶不会显著降低 DNA 结合能力或对互补 DNA 序列的选择性。为了确定光刻步骤是否造成更微妙的损害,我们还测试了含有单个碱基错配的寡核苷酸。最后,开发了两步光刻工艺,并与介电泳纳米线组装结合使用,在芯片上生产出一系列双接触、电隔离的单个纳米线组件。制备后荧光成像表明存在纳米线结合的 DNA,并且能够选择性地结合互补链。