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Signal and Sensitivity Enhancement Through Optical Interference Coating for DNA and Protein Microarray Applications

机译:通过光学干涉涂层增强信号和灵敏度适用于DNA和蛋白质微阵列应用

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摘要

Optical inteference (OI) coated slides with unique optical properties were utilized in microarray analyses, demonstrating their enhanced detection sensitivity over traditional microarray substrates. The OI coating is comprised of a proprietary multilayered, dielectric, thin-film interference coating located beneath the functional coating (aminosilane or epoxysilane). It is designed to enhance the fluorescence in the Cy3 and Cy5 channel by increasing the light absorption of the dyes by about 6-fold and by redirecting emitted fluorescence into the detector during scanning, resulting in a theoretical limit of about 12-fold signal amplification. Two-color DNA microarray experiments conducted on the OI slides showed over 8-fold signal amplification, conservation of gene expression ratios, and increased signal-to-noise ratio when compared to control slides, indicating enhanced detection sensitivity. Protein microarray assays also exhibited over 8-fold signal amplification at three different target concentrations, demonstrating the versatility of the OI slides for different microarray applications. Further, the DNA and protein assays performed on the OI slides exhibited excellent detection sensitivity even at the low target amounts essential for diagnostic applications. The OI slides are compatible with commonly used protocols, printers, scanners and other microarray equipment. Therefore, the OI slides offer an attractive alternative to traditional microarray substrates, where enhanced detection sensitivity is desired.
机译:具有独特光学特性的光学干扰(OI)涂层载玻片用于微阵列分析,证明了其相对于传统微阵列底物的增强的检测灵敏度。 OI涂层由位于功能涂层(氨基硅烷或环氧硅烷)下方的专有多层电介质薄膜干涉涂层组成。通过将染料的光吸收提高约6倍,并在扫描过程中将发射的荧光重定向到检测器中,可以增强Cy3和Cy5通道中的荧光,从而导致信号放大的理论极限达到约12倍。与对照玻片相比,在OI玻片上进​​行的双色DNA微阵列实验显示信号扩增超过8倍,基因表达率保持不变,信噪比提高,表明检测灵敏度得到了提高。蛋白质微阵列分析在三种不同的目标浓度下也显示出超过8倍的信号放大,证明了OI载玻片对于不同微阵列应用的多功能性。此外,即使在诊断应用必不可少的低目标量下,在OI载玻片上进行的DNA和蛋白质测定也表现出出色的检测灵敏度。 OI幻灯片与常用协议,打印机,扫描仪和其他微阵列设备兼容。因此,OI载玻片为传统的微阵列基板提供了有吸引力的替代方法,在传统的微阵列基板上,需要提高检测灵敏度。

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