Oillic C, Mur P, Blanquet E, Delapierre G, Vinet F, Billon T
CEA-DRT-LETI/CEA Grenoble, 17 rue des Martyrs, 38054 Grenoble Cedex 9, France.
Biosens Bioelectron. 2007 Apr 15;22(9-10):2086-92. doi: 10.1016/j.bios.2006.09.022. Epub 2006 Nov 21.
To improve the sensitivity of fluorescence detection in DNA microarrays, the use of silicon nanostructures based on chemical vapor deposition (CVD) processes adopted for the growth of rough polycrystalline silicon was investigated. These substrates present advantages of two main properties which could lead to an enhancement of the fluorescence detection, i.e. (i) the increase of the available surface area in order to achieve a high loading capacity of biomolecules and (ii) the optimization of the stack of silicon nanostructures support. Indeed, the structures were elaborated on an initial thermal oxide layer and then covered with a silicon oxide layer, obtained by oxidation and allowing the functionalization for the subsequent grafting of DNA probes. Moreover, these oxide layers play a part in the fluorescence detection. The influence of the silicon oxide layer thickness above and below the silicon grains in close relation with the density of nanostructures on the emitted fluorescence was emphasized. This paper presents an experimental characterization of the fluorescence intensity and the optimization of the different layers that composed the substrate used for DNA microarrays. The performances of the microarrays were investigated by means of hybridization experiments using complementary fluorescent labeled-oligonucleotides targets. Our results indicate that an optimized substrate can be designed and that the use of oxidized silicon nanostructures for support of biochip could be a strategy for improving the sensitivity of fluorescence detection.
为提高DNA微阵列中荧光检测的灵敏度,研究了基于化学气相沉积(CVD)工艺生长粗糙多晶硅的硅纳米结构的应用。这些基板具有两个主要特性的优势,这可能会增强荧光检测,即:(i)增加可用表面积以实现生物分子的高负载能力,以及(ii)优化硅纳米结构载体的堆叠。实际上,这些结构是在初始热氧化层上精心制作的,然后覆盖一层氧化硅层,该氧化硅层通过氧化获得,并允许对后续DNA探针的接枝进行功能化。此外,这些氧化层在荧光检测中起作用。强调了与纳米结构密度密切相关的硅晶粒上方和下方的氧化硅层厚度对发射荧光的影响。本文介绍了荧光强度的实验表征以及构成DNA微阵列所用基板的不同层的优化。通过使用互补荧光标记的寡核苷酸靶标的杂交实验研究了微阵列的性能。我们的结果表明,可以设计出优化的基板,并且使用氧化硅纳米结构来支撑生物芯片可能是提高荧光检测灵敏度的一种策略。