%0 Generic %A Pereira, P %A Gustafsson, N %A Marsh, M %A Mhlanga, M %A Henriques, R %C Cold Spring Harbor, NY, USA %D 2020 %F discovery:10090448 %I BioRxiv %T Super-Beacons: open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy %U https://discovery.ucl.ac.uk/id/eprint/10090448/ %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. %Z 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 (http://creativecommons.org/licenses/by/4.0/).