Implementation of a fluorescence spatiotemporal modulation super-resolution microscope

Super-resolution microscopy (SRM) unveils details of subcellular organelles and provides a technical foundation for cellular biology research. Long-term, non-invasive live-cell super-resolution imaging requires low-intensity illumination and high image quality. Here, we present a new, to the best of our knowledge, method based on time-resolved detection termed fluorescence spatiotemporal modulation, in which highly spatially resolved photons in the beam center are extracted by taking the difference of the photons in the beam periphery with a weighted coefficient. The experimental results show a sub-100 nm resolution at tens of microwatts of laser power. Our proposed method requires only one laser, laying a foundation for a lower-cost multi-color super-resolution imaging system.