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. To demonstrate this principle, we developed a new class of DNA-based open-source superresolution probes named super-beacons, with photoswitching kinetics that can be tuned structurally, thermally and chemically. The potential of these probes for livecell compatible super-resolution microscopy without high-illumination or toxic imaging buffers is revealed by imaging interferon inducible transmembrane proteins (IFITMs) at sub-100 nm resolutions. K E Y W O R D S DNA, fluorescence microscopy, fluorescence quenching, molecular-beacons, live-cell, superresolution SYNOPSIS We developed a new class of DNA-based self-quenching open-source super-resolution probes named super-beacons, with photoswitching kinetics that can be tuned structurally, thermally and chemically. These probes alleviate the need for high-intensity illumination and toxic buffers usually needed in single-molecule localization super-resolution approaches. We showcase their applicability in fixed-and live-cell imaging with sub-100 nm resolutions to explore the organisation of interferon inducible transmembrane proteins (IFTM) at the cell surface. Ab link; Link to antibody of interest. mW.cm-2; Live-cell compatible illumination. Pedro M. Pereira and Nils Gustafsson equally contributed to this study.