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

DOI :

10.3791/63124-v

8:26 min

November 23rd, 2021

November 23rd, 2021

2,106 Views

1Institute of Applied Physics, TU Wien, 2Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna

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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Automated Analysis

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