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PubMed · 12854995

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

Ulrich Haupts·Martin Rüdiger·Stephen Ashman·Sandra Turconi·Ryan Bingham·Charlotte Wharton·Jonathan Hutchinson·Charlotte Carey·Keith J Moore·Andrew J Pope
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Abstract

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.

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Alcohol DehydrogenaseAlkaline PhosphataseAntigens, Human PlateletEndopeptidasesFluorescenceLigandsMicroscopy, ConfocalRNAJournal Article

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BibTeXRIS

Ulrich Haupts, Martin Rüdiger, Stephen Ashman, Sandra Turconi, Ryan Bingham, Charlotte Wharton, Jonathan Hutchinson, Charlotte Carey, Keith J Moore, Andrew J Pope. 2003. Single-molecule detection technologies in miniaturized high-throughput screening: fluorescence intensity distribution analysis.. https://doi.org/10.1177/1087057102239669

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