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

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

English

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2,278 Views

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08:26 min

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November 23, 2021

DOI:

10.3791/63124-v

DOI:
10.3791/63124-v

•

08:26 min
November 23, 2021

•

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

Summary

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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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Cite this Article

Schrangl, L., Göhring, J., Kellner, F., Huppa, J. B., Schütz, G. J. Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes. J. Vis. Exp. (177), e63124, doi:10.3791/63124 (2021).

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