DNA micropatterning on polycrystalline diamond via one-step direct amination.
DNA micropatterning on polycrystalline diamond via one-step direct amination.
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DOI：
10.1021/la050883d
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发表时间：
2006-03
期刊：
影响因子：
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通讯作者：
Guojun Zhang;K. Song;Yusuke Nakamura;T. Ueno;T. Funatsu;I. Ohdomari;H. Kawarada
中科院分区：
文献类型：
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作者：
Guojun Zhang;K. Song;Yusuke Nakamura;T. Ueno;T. Funatsu;I. Ohdomari;H. Kawarada
我们报告了一种在多晶金刚石上一步直接胺化的新方法，通过光刻产生用于 DNA 微图案化的功能化表面。多晶金刚石在氨气中受到紫外线照射，直接生成胺基。图案化后，光学显微镜证实覆盖有金掩模的微图案的尺寸和形状是规则的。微图案外部的区域通过 C3F8 等离子体进行氟封端钝化，并通过空间分辨 X 射线光电子能谱 (XPS) 来表征两个不同表面上的化学变化（通过一步直接胺化形成图案区域内的胺基团和图案区域外部的氟封端）。然后，用活性胺基团终止的图案化区域通过双功能分子用探针 DNA 固定。通过将荧光标记的目标 DNA 与附着在微图案内的互补和非互补探针 DNA 杂交来进行序列特异性。通过落射荧光显微镜观察到的荧光微图案与通过光学显微镜成像的荧光微图案相对应。对 DNA 修饰钻石进行的 DNA 杂交和变性实验表明，钻石表面具有卓越的稳定性。比较不同胺化时间对荧光强度的影响。研究了作为钝化层的不同终端，结果发现氟终端具有最大的信噪比。
We report a novel method of one-step direct amination on polycrystalline diamond to produce functionalized surfaces for DNA micropatterning by photolithography. Polycrystalline diamond was exposed to UV irradiation in ammonia gas to generate amine groups directly. After patterning, optical microscopy confirmed that micropatterns covered with an Au mask were regular in size and shape. The regions outside the micropatterns were passivated with fluorine termination by C3F8 plasma, and the chemical changes on the two different surfaces--the amine groups inside the patterned regions by one-step direct amination and fluorine termination outside the patterned regions--were characterized by spatially resolved X-ray photoelectron spectroscopy (XPS). The patterned areas terminated with active amine groups were then immobilized with probe DNA via a bifunctional molecule. The sequence specificity was conducted by hybridizing fluorescently labeled target DNA to both complementary and noncomplementary probe DNA attached inside the micropatterns. The fluorescence micropatterns observed by epifluorescence microscopy corresponded to those imaged by optical microscopy. DNA hybridization and denaturation experiments on a DNA-modified diamond show that the diamond surfaces reveal superior stability. The influence of a different amination time on fluorescence intensity was compared. Different terminations as passivated layers were investigated, and as a result, fluorine termination points to the greatest signal-to-noise ratio.