High-speed super-resolution structured illumination microscopy with a large field-of-view.

Structured illumination microscopy (SIM) has been extensively employed for observing subcellular structures and dynamics. However, achieving high-speed super-resolution SIM with a large field of view (FOV) remains challenging due to the trade-offs among spatial resolution, imaging speed and FOV under limited bandwidth constraints. Here, we report a novel SIM technique to address this issue. By utilizing a high-speed camera and a rolling image reconstruction strategy to accelerate super-resolution image acquisition, as well as using a deep resolution enhancement to further improve spatial resolution, this SIM technique achieves imaging with a spatial resolution of 94 nm, a FOV of 102 × 102 µm, and an imaging speed of 1333 frames per second. The exceptional imaging performance of this proposed SIM technique is experimentally demonstrated through the successful recording of the Brownian motion of fluorescent microspheres and the photobleaching of fluorescently labeled microtubules. This work offers a potential tool for the high-throughput observation of high-speed subcellular dynamics, which would bring significant applications in biomedical research.