PT  - JOURNAL ARTICLE
AU  - Pereira, Pedro M.
AU  - Gustafsson, Nils
AU  - Marsh, Mark
AU  - Mhlanga, Musa M.
AU  - Henriques, Ricardo
TI  - Super-Beacons: open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy
AID  - 10.1101/2020.01.20.912311
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.01.20.912311
4099  - http://biorxiv.org/content/early/2020/02/04/2020.01.20.912311.short
4100  - http://biorxiv.org/content/early/2020/02/04/2020.01.20.912311.full
AB  - 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.