%0 Journal Article %A Pedro M. Pereira %A Nils Gustafsson %A Mark Marsh %A Musa M. Mhlanga %A Ricardo Henriques %T Super-Beacons: open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy %D 2020 %R 10.1101/2020.01.20.912311 %J bioRxiv %P 2020.01.20.912311 %X 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. %U https://www.biorxiv.org/content/biorxiv/early/2020/02/04/2020.01.20.912311.full.pdf