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  Tracking single particles for hours via continuous DNA-mediated fluorophore exchange

Stehr, F., Stein, J., Bauer, J., Niederauer, C., Jungmann, R., Ganzinger, K., et al. (2021). Tracking single particles for hours via continuous DNA-mediated fluorophore exchange. Nature Communications, 12(1): 4432. doi:10.1038/s41467-021-24223-4.

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 Creators:
Stehr, Florian1, Author              
Stein, Johannes1, Author              
Bauer, Julian1, Author
Niederauer, Christian2, Author
Jungmann, Ralf3, Author              
Ganzinger, Kristina2, Author
Schwille, Petra1, Author              
Affiliations:
1Schwille, Petra / Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565169              
2external, ou_persistent22              
3Jungmann, Ralf / Molecular Imaging and Bionanotechnology, Max Planck Institute of Biochemistry, Max Planck Society, ou_2149679              

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Free keywords: FLUORESCENCE; TRAJECTORIES; MICROSCOPY; MOBILITY; BEHAVIOR; CELLSScience & Technology - Other Topics;
 Abstract: Monitoring biomolecules in single-particle tracking experiments is typically achieved by employing fixed organic dyes or fluorescent fusion proteins linked to a target of interest. However, photobleaching typically limits observation times to merely a few seconds, restricting downstream statistical analysis and observation of rare biological events. Here, we overcome this inherent limitation via continuous fluorophore exchange using DNA-PAINT, where fluorescently-labeled oligonucleotides reversibly bind to a single-stranded DNA handle attached to the target molecule. Such versatile and facile labeling allows uninterrupted monitoring of single molecules for extended durations. We demonstrate the power of our approach by observing DNA origami on membranes for tens of minutes, providing perspectives for investigating cellular processes on physiologically relevant timescales. The length of single-particle tracking experiments are limited due to photobleaching. Here the authors achieve long-term single-particle tracking with continuous fluorophore exchange in DNA-PAINT and use this to observe DNA origami on lipid bilayers for tens of minutes.

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Language(s): eng - English
 Dates: 2021
 Publication Status: Published online
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Degree: -

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Title: Nature Communications
  Abbreviation : Nat. Commun.
Source Genre: Journal
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Publ. Info: London : Nature Publishing Group
Pages: - Volume / Issue: 12 (1) Sequence Number: 4432 Start / End Page: - Identifier: ISSN: 2041-1723
CoNE: https://pure.mpg.de/cone/journals/resource/2041-1723