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Concurrent diffusion of nicotinic acetylcholine receptors and fluorescent cholesterol disclosed by two-colour sub-millisecond MINFLUX-based single-molecule tracking

Author

Listed:
  • Francesco Reina

    (Friedrich-Schiller-Universität Jena
    Leibniz Institute of Photonic Technologies
    University of Vienna)

  • Lucas A. Saavedra

    (UCA-CONICET)

  • Christian Eggeling

    (Friedrich-Schiller-Universität Jena
    Leibniz Institute of Photonic Technologies
    Jena Centre for Soft Matter (JCSM)
    Leibniz Centre for Photonics in Infection Research (LPI))

  • Francisco J. Barrantes

    (UCA-CONICET)

Abstract

The diffusion and interaction dynamics of membrane proteins and lipids are key for cell function, but their disclosure is hampered by limited temporal and spatial resolution of conventional observation technologies. Here we exploit the capabilities of minimal fluorescence emission photon fluxes (MINFLUX) microscopy in single-molecule co-tracking experiments of an important membrane protein and cholesterol with enhanced spatiotemporal resolution. Specifically, we interrogate the 2D translational mobility of a ubiquitous cell-surface protein, the nicotinic acetylcholine receptor, in tandem with a fluorescent cholesterol analogue for minute-long periods, reaching nanometric precision and sub-millisecond time resolution. To this end, we implement a multiplexing procedure that enables the simultaneous excitation of the two fluorescent-labelled molecules using a single wavelength, followed by discrimination of their emissions via differential ratiometric recording. We disclose a cholesterol-dependent heterogeneous spectrum of diffusive behaviours with regions of joint translational motion.

Suggested Citation

  • Francesco Reina & Lucas A. Saavedra & Christian Eggeling & Francisco J. Barrantes, 2025. "Concurrent diffusion of nicotinic acetylcholine receptors and fluorescent cholesterol disclosed by two-colour sub-millisecond MINFLUX-based single-molecule tracking," Nature Communications, Nature, vol. 16(1), pages 1-16, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-61489-4
    DOI: 10.1038/s41467-025-61489-4
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    References listed on IDEAS

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    1. 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.
    2. Yonatan Golan & Eilon Sherman, 2017. "Resolving mixed mechanisms of protein subdiffusion at the T cell plasma membrane," Nature Communications, Nature, vol. 8(1), pages 1-15, August.
    3. Abhishek Sau & Sebastian Schnorrenberg & Ziqiang Huang & Debolina Bandyopadhyay & Ankith Sharma & Clara-Marie Gürth & Sandeep Dave & Siegfried M. Musser, 2025. "Overlapping nuclear import and export paths unveiled by two-colour MINFLUX," Nature, Nature, vol. 640(8059), pages 821-827, April.
    4. Eva Sevcsik & Mario Brameshuber & Martin Fölser & Julian Weghuber & Alf Honigmann & Gerhard J. Schütz, 2015. "GPI-anchored proteins do not reside in ordered domains in the live cell plasma membrane," Nature Communications, Nature, vol. 6(1), pages 1-10, November.
    5. Gonzalo Almarza & Francisco Sánchez & Francisco J Barrantes, 2014. "Transient Cholesterol Effects on Nicotinic Acetylcholine Receptor Cell-Surface Mobility," PLOS ONE, Public Library of Science, vol. 9(6), pages 1-18, June.
    6. Roman Schmidt & Tobias Weihs & Christian A. Wurm & Isabelle Jansen & Jasmin Rehman & Steffen J. Sahl & Stefan W. Hell, 2021. "MINFLUX nanometer-scale 3D imaging and microsecond-range tracking on a common fluorescence microscope," Nature Communications, Nature, vol. 12(1), pages 1-12, December.
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