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Super-Beacons: open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy

Pedro M. Pereira, Nils Gustafsson, View ORCID ProfileMark Marsh, View ORCID ProfileMusa M. Mhlanga, View ORCID ProfileRicardo Henriques
doi: https://doi.org/10.1101/2020.01.20.912311
Pedro M. Pereira
1MRC-Laboratory for Molecular Cell Biology. University College London, London, UK
2The Francis Crick Institute, London, UK
4Bacterial Cell Biology, MOSTMICRO, ITQB-NOVA, Oeiras, Portugal
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Nils Gustafsson
1MRC-Laboratory for Molecular Cell Biology. University College London, London, UK
5Department für Physik and CeNS, Ludwig-Maximilians-Universität, Munich, Germany
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Mark Marsh
1MRC-Laboratory for Molecular Cell Biology. University College London, London, UK
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Musa M. Mhlanga
4Bacterial Cell Biology, MOSTMICRO, ITQB-NOVA, Oeiras, Portugal
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Ricardo Henriques
1MRC-Laboratory for Molecular Cell Biology. University College London, London, UK
2The Francis Crick Institute, London, UK
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  • For correspondence: r.henriques@ucl.ac.uk
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Abstract

Localization based super-resolution microscopy relies on the detection of individual molecules cycling between fluorescent and non-fluorescent states. These transitions are commonly regulated by high-intensity illumination, imposing constrains to imaging hardware and producing sample photodamage. Here, we propose single-molecule self-quenching as a mechanism to generate spontaneous photoswitching independent of illumination. To demonstrate this principle, we developed a new class of DNA-based open-source Super-Resolution probes named Super-Beacons, with photoswitching kinetics that can be tuned structurally, thermally and chemically. The potential of these probes for live-cell friendly Super-Resolution Microscopy without high-illumination or toxic imaging buffers is revealed by imaging Interferon Inducible Transmembrane proteins (IFITMs) at sub-100nm resolutions.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted February 04, 2020.
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Super-Beacons: open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy
Pedro M. Pereira, Nils Gustafsson, Mark Marsh, Musa M. Mhlanga, Ricardo Henriques
bioRxiv 2020.01.20.912311; doi: https://doi.org/10.1101/2020.01.20.912311
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Super-Beacons: open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy
Pedro M. Pereira, Nils Gustafsson, Mark Marsh, Musa M. Mhlanga, Ricardo Henriques
bioRxiv 2020.01.20.912311; doi: https://doi.org/10.1101/2020.01.20.912311

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