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用于微阵列扫描和分析的创新型仪器:寡核苷酸双链体行为表征的应用

Innovative instrumentation for microarray scanning and analysis: application for characterization of oligonucleotide duplexes behavior.

作者信息

Khomyakova E B, Dreval E V, Tran-Dang M, Potier M C, Soussaline F P

机构信息

IMSTAR, 60, rue Notre Dame des Champs, 75006 Paris, France.

出版信息

Cell Mol Biol (Noisy-le-grand). 2004 May;50(3):217-24.

Abstract

Accuracy in microarray technology requires new approaches to microarray reader development. A microarray reader system (optical scanning array or OSA reader) based on automated microscopy with large field of view, high speed 3 axis scanning at multiple narrow-band spectra of excitation light has been developed. It allows fast capture of high-resolution, multi-fluorescence images and is characterized by a linear dynamic range and sensitivity comparable to commonly used photo-multiplier tube (PMT)-based laser scanner. Controlled by high performance software, the instrument can be used for scanning and quantitative analysis of any type of dry microarray. Studies implying temperature-controlled hybridization chamber containing a microarray can also be performed. This enables the registration of kinetics and melting curves. This feature is required in a wide range of on-chip chemical and enzymatic reactions including on-chip PCR amplification. We used the OSA reader for the characterization of hybridization and melting behaviour of oligonucleotide:oligonucleotide duplexes on three-dimensional Code Link slides.

摘要

微阵列技术的准确性需要微阵列读取器开发的新方法。已经开发出一种基于自动显微镜的微阵列读取器系统(光学扫描阵列或OSA读取器),具有大视野、在激发光的多个窄带光谱下进行高速3轴扫描。它能够快速捕获高分辨率的多荧光图像,其线性动态范围和灵敏度与常用的基于光电倍增管(PMT)的激光扫描仪相当。该仪器由高性能软件控制,可用于扫描和定量分析任何类型的干式微阵列。也可以进行包含微阵列的温控杂交室的研究。这使得能够记录动力学和熔解曲线。在包括芯片上PCR扩增在内的广泛的芯片上化学和酶促反应中都需要这一特性。我们使用OSA读取器来表征三维Code Link载玻片上寡核苷酸:寡核苷酸双链体的杂交和熔解行为。

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