gSTED Microscopy with an OPSL: Cutting Edge Super-Resolution Breaking the diffraction limit with a CW OPSL

In order to probe cell biology at the molecular level and correlate single molecule activity with macroscopic cellular behavior in real-time, biologists need farfield microscopy techniques with the highest possible spatial as well as temporal resolution. Driven by this need, the use of lasers in confocal microscopy has taken fluorescence microscopy to the diffraction limit, which was long considered the theoretical and practical limit on spatial resolution of this technique. In the 1990’s novel super-resolution far-field microscopy (or nanoscopy) approaches based on the reversible modulation of the fluorescence signal have evolved which pushed the spatial resolution to the nanoscale [1]. And Stimulated Emission Depletion (STED) nanoscopy has been developed [2] as a broadly applicable method and can be used with both fixed and live tissue samples [1]. The combination of a continuous-wave (CW) laser [3] with a gated detection (gSTED) [4] and the availability of a yellow (577 nm) low-noise laser has enabled further optimization.