FLUORESCENCE CORRELATIONS, SINGLE-MOLECULE DETECTION AND LARGE NUMBER SCREENING - APPLICATIONS IN BIOTECHNOLOGY
FLUORESCENCE CORRELATIONS, SINGLE-MOLECULE DETECTION AND LARGE NUMBER SCREENING - APPLICATIONS IN BIOTECHNOLOGY
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DOI：
10.1016/0168-1656(95)00054-t
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发表时间：
1995-07-31
影响因子：
4.1
通讯作者：
RIGLER, R
中科院分区：
文献类型：
--
作者：
RIGLER, R
荧光相关光谱（FCS），当在非常小的体积元素中获得的低背景条件下进行时，是检查分子相互作用以及它们的时间依赖性的有力工具。可以分析的具有生物学重要性的相互作用是核酸引物与DNA或RNA靶标之间、肽配体与分离的以及细胞结合的受体之间、抗原与抗体之间的杂交。由于可以在小体积中快速分析相互作用，而不需要将未结合的配体与结合的配体分离，因此可以设想FCS在大规模药物筛选中的重要应用。灵敏度已经提高到可以在亚毫秒范围内检测单个染料分子的程度。这开辟了检测罕见事件的可能性，例如在感染的早期阶段出现病原体或在筛选组合文库时表现出不寻常特性的突变体。
Fluorescence correlation spectroscopy (FCS), when carried out under conditions with low background as obtained in very small volume elements, is a powerful tool for examining molecular interactions as well as their time dependence. Interactions of biological importance which can be analyzed are hybridization between nucleic acid primers and DNA or RNA targets, between peptide ligands and isolated as well as cell-bound receptors, between antigen and antibodies. Since the interaction can be analyzed rapidly in small volumes without the need for separating unbound from bound ligand, an important application of FCS is envisaged in large-scale drug screening. The sensitivity has been advanced to the point that detection of single dye molecules is possible in the submillisecond range. This opens up the possibility for detecting rare events such as the appearance of pathogens in the early phase of infection or mutants exhibiting unusual properties when screening combinatorial libraries.