Optimisation of a silicon/silicon dioxide substrate for a fluorescence DNA microarray
Optimisation of a silicon/silicon dioxide substrate for a fluorescence DNA microarray
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DOI：
10.1016/j.bios.2004.03.018
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发表时间：
2004-11-01
影响因子：
12.6
通讯作者：
Garrigues, M
中科院分区：
文献类型：
--
作者：
Bras, M;Dugas, V;Garrigues, M
本文提出了一个全面的理论和实验表征的调制的荧光强度的光干涉用于DNA微阵列的氧化硅基板上的建设。该模型预测的荧光信号的90倍的变化取决于氧化物的厚度。对于Cy 3染料，信号在90 nm氧化物厚度下最大，对应于标准玻璃基底的7.5倍增强。对于实验验证的模型，我们已经准备了Si/SiO2基板上的不同的平行步骤，减少氧化物厚度在同一样品上使用缓冲氧化物蚀刻（BOE）蚀刻工艺热氧化后。SiO2表面已被官能化的硅烷单层之前，在原位合成的L 185寡核苷酸探针。在与互补靶杂交后，荧光强度随氧化物厚度的变化与理论模型非常一致。对玻璃基板的实验比较显示了10倍的检测灵敏度增强。我们的研究结果表明，Si/SiO2基板是一个有吸引力的替代标准的玻璃载玻片荧光DNA微阵列的实现时，检测灵敏度是一个重要的问题。(C)2004 Elsevier B. V.保留所有权利。
This paper presents a comprehensive theory and experimental characterisation of the modulation of the fluorescence intensity by the construction of optical interferences on oxidised silicon substrates used for DNA microarrays. The model predicts a 90-fold variation of the fluorescence signal depending on the oxide thickness. For a Cy3 dye, the signal is maximal for a 90 nm oxide thickness corresponding to a 7.5-fold enhancement with respect to a standard glass substrate. For experimental validation of the model, we have prepared Si/SiO2 substrates with different parallel steps of decreasing oxide thicknesses on the same sample using a buffered oxide etch (BOE) etching process after thermal oxidation. The SiO2 surface has been functionalised by a silane monolayer before in situ synthesis of L 185 oligonucleotide probes. After hybridisation with complementary targets, the variations of the fluorescence intensity versus oxide thickness are in very good accordance with the theoretical model. The experimental comparison against a glass substrate shows a 10-fold enhancement of the detection sensitivity. Our results demonstrate that a Si/SiO2 substrate is an attractive alternative to standard glass slides for the realisation of fluorescence DNA microarrays whenever detection sensitivity is an important issue. (C) 2004 Elsevier B.V. All rights reserved.