Ramdas Latha, Zhang Wei
Cancer Genomics Core Laboratory, University of Texas, M.D. Anderson Cancer Center, Houston, USA.
Methods Mol Biol. 2006;319:261-73. doi: 10.1007/978-1-59259-993-6_13.
Of the technologies available for measuring gene expression, microarrays using cDNA targets is one of the most common and well-developed high-throughput techniques. With this technique, the expression levels of thousands of genes are measured simultaneously. DNA probes are immobilized on solid surfaces, either membrane-based or chemically coated glass surfaces. On glass arrays, the probes are hybridized with fluorescent-labeled target samples. Fluorescence intensities, which reflect gene expression levels, are detected by imaging the array using a laser or white-light source and capturing the image using photomultiplier tube detection or a charge-coupled device camera. Different laser-based scanners are used in laboratories to scan microarray images. This chapter discusses the imaging process and the protocols being developed.
在可用于测量基因表达的技术中，使用cDNA靶标的微阵列是最常见且发展完善的高通量技术之一。通过该技术，可以同时测量数千个基因的表达水平。DNA探针固定在固体表面，这些表面可以是基于膜的，也可以是化学涂层的玻璃表面。在玻璃微阵列上，探针与荧光标记的靶标样本杂交。反映基因表达水平的荧光强度通过使用激光或白光源对阵列进行成像并使用光电倍增管检测或电荷耦合器件相机捕获图像来检测。实验室中使用不同的基于激光的扫描仪来扫描微阵列图像。本章将讨论成像过程以及正在开发的方案。
