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Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions

Author

Listed:
  • Christian Niederauer

    (AMOLF)

  • Chikim Nguyen

    (AMOLF)

  • Miles Wang-Henderson

    (AMOLF)

  • Johannes Stein

    (Max Planck Institute of Biochemistry)

  • Sebastian Strauss

    (Max Planck Institute of Biochemistry)

  • Alexander Cumberworth

    (AMOLF)

  • Florian Stehr

    (Max Planck Institute of Biochemistry)

  • Ralf Jungmann

    (Max Planck Institute of Biochemistry
    Ludwig Maximilian University)

  • Petra Schwille

    (Max Planck Institute of Biochemistry)

  • Kristina A. Ganzinger

    (AMOLF)

Abstract

DNA-PAINT based single-particle tracking (DNA-PAINT-SPT) has recently significantly enhanced observation times in in vitro SPT experiments by overcoming the constraints of fluorophore photobleaching. However, with the reported implementation, only a single target can be imaged and the technique cannot be applied straight to live cell imaging. Here we report on leveraging this technique from a proof-of-principle implementation to a useful tool for the SPT community by introducing simultaneous live cell dual-color DNA-PAINT-SPT for quantifying protein dimerization and tracking proteins in living cell membranes, demonstrating its improved performance over single-dye SPT.

Suggested Citation

  • Christian Niederauer & Chikim Nguyen & Miles Wang-Henderson & Johannes Stein & Sebastian Strauss & Alexander Cumberworth & Florian Stehr & Ralf Jungmann & Petra Schwille & Kristina A. Ganzinger, 2023. "Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions," Nature Communications, Nature, vol. 14(1), pages 1-8, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-40065-8
    DOI: 10.1038/s41467-023-40065-8
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    References listed on IDEAS

    as
    1. Titiwat Sungkaworn & Marie-Lise Jobin & Krzysztof Burnecki & Aleksander Weron & Martin J. Lohse & Davide Calebiro, 2017. "Single-molecule imaging reveals receptor–G protein interactions at cell surface hot spots," Nature, Nature, vol. 550(7677), pages 543-547, October.
    2. Florian Stehr & Johannes Stein & Julian Bauer & Christian Niederauer & Ralf Jungmann & Kristina Ganzinger & Petra Schwille, 2021. "Tracking single particles for hours via continuous DNA-mediated fluorophore exchange," Nature Communications, Nature, vol. 12(1), pages 1-8, December.
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