Single-molecule detection technologies in miniaturized high-throughput screening:: Fluorescence intensity distribution analysis

被引：31

作者：

Haupts, U ^{[1
]}

Rüdiger, M ^{[1
]}

Ashman, S ^{[1
]}

Turconi, S ^{[1
]}

Bingham, R ^{[1
]}

Wharton, C ^{[1
]}

Hutchinson, J ^{[1
]}

Carey, C ^{[1
]}

Moore, KJ ^{[1
]}

Pope, AJ ^{[1
]}

机构：

[1] GlaxoSmithKline, Harlow CM19 5AW, Essex, England

来源：

JOURNAL OF BIOMOLECULAR SCREENING | 2003年 / 8卷 / 01期

关键词：

single molecule; confocal; fluorescence; miniaturization; uHTS;

D O I：

10.1177/1087057102239669

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Single-molecule detection technologies are becoming a powerful readout format to support ultra-high-throughput screening. These methods are based on the analysis of fluorescence intensity fluctuations detected from a small confocal volume element. The fluctuating signal contains information about the mass and brightness of the different species in a mixture. The authors demonstrate a number of applications of fluorescence intensity distribution analysis (FIDA), which discriminates molecules by their specific brightness. Examples for assays based on brightness changes induced by quenching/dequenching of fluorescence, fluorescence energy transfer, and multiple-binding stoichiometry are given for important drug targets such as kinases and proteases, FIDA also provides a powerful method to extract correct biological data in the presence of compound fluorescence.

引用

页码：19 / 33

页数：15

共 40 条

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