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JoVE Journal
Bioengineering

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Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
 

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Article DOI: 10.3791/63124-v
November 23rd, 2021

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Summary November 23rd, 2021

This article presents a method for spatiotemporal analysis of mobile, single-molecule Förster resonance energy transfer (smFRET)-based probes using widefield fluorescence microscopy. The newly developed software toolkit allows the determination of smFRET time traces of moving probes, including the correct FRET efficiency and the molecular positions, as functions of time.

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