Schäferling Michael, Nagl Stefan
Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, 93040 Regensburg, Germany.
Anal Bioanal Chem. 2006 Jun;385(3):500-17. doi: 10.1007/s00216-006-0317-5. Epub 2006 Apr 12.
Microarray formats have become an important tool for parallel (or multiplexed) monitoring of biomolecular interactions. Surface-immobilized probes like oligonucleotides, cDNA, proteins, or antibodies can be used for the screening of their complementary targets, covering different applications like gene or protein expression profiling, analysis of point mutations, or immunodiagnostics. Numerous reviews have appeared on this topic in recent years, documenting the intriguing progress of these miniaturized assay formats. Most of them highlight all aspects of microarray preparation, surface chemistry, and patterning, and try to give a systematic survey of the different kinds of applications of this new technique. This review places the emphasis on optical technologies for microarray analysis. As the fluorescent read out of microarrays is dominating the field, this topic will be the focus of the review. Basic principles of labeling and signal amplification techniques will be introduced. Recent developments in total internal reflection fluorescence, resonance energy transfer assays, and time-resolved imaging are addressed, as well as non-fluorescent imaging methods. Finally, some label-free detection modes are discussed, such as surface plasmon microscopy or ellipsometry, since these are particularly interesting for microarray development and quality control purposes.
微阵列形式已成为用于并行(或多重)监测生物分子相互作用的重要工具。诸如寡核苷酸、cDNA、蛋白质或抗体等表面固定化探针可用于筛选其互补靶标,涵盖不同应用,如基因或蛋白质表达谱分析、点突变分析或免疫诊断。近年来,关于这一主题出现了大量综述,记录了这些小型化检测形式的有趣进展。其中大多数都强调了微阵列制备、表面化学和图案化的各个方面,并试图对这项新技术的不同应用进行系统综述。本综述重点关注用于微阵列分析的光学技术。由于微阵列的荧光读出在该领域占主导地位,因此该主题将成为综述的重点。将介绍标记和信号放大技术的基本原理。讨论了全内反射荧光、共振能量转移分析和时间分辨成像的最新进展,以及非荧光成像方法。最后,讨论了一些无标记检测模式,如表面等离子体显微镜或椭偏仪,因为这些对于微阵列开发和质量控制目的特别有意义。