Ahn Sunmin, Freedman David S, Zhang Xirui, Unlü M Selim
Department of Biomedical Engineering, Boston University, Boston, MA, USA.
Methods Mol Biol. 2013;1039:181-200. doi: 10.1007/978-1-62703-535-4_16.
Optical label-free biosensors have demonstrated advantages over fluorescent-based detection methods by allowing accurate quantification while also being capable of measuring dynamic bimolecular interactions. A simple, high-throughput, solid-phase, and label-free technique, interferometric reflectance imaging sensor (IRIS), can quantify the mass density of DNA with pg/mm(2) sensitivity by measuring the optical path difference. We present the design of the IRIS instrument and complementary microarrays that can be used to perform a quantitative analysis of DNA microarrays. Finally, we present methods to accurately calculate the hybridization efficiency and identify SNPs from dynamic measurements, as well as supporting software algorithms needed for robust data processing.
无光学标记生物传感器已显示出优于基于荧光的检测方法的优势，它能够进行精确量化，同时还能测量动态双分子相互作用。干涉反射成像传感器（IRIS）是一种简单、高通量、固相且无标记的技术，通过测量光程差，它能够以pg/mm(2)的灵敏度对DNA的质量密度进行量化。我们展示了IRIS仪器和互补微阵列的设计，这些可用于对DNA微阵列进行定量分析。最后，我们介绍了从动态测量中准确计算杂交效率和识别单核苷酸多态性（SNP）的方法，以及稳健的数据处理所需的支持软件算法。
