Three-dimensional (3D) structured illumination microscopy (SIM) improves spatial resolution by a factor of two in both lateral and axial directions. However, the adoption of 3D SIM is limited by low imaging speed, susceptibility to out-of-focus light, and likelihood of reconstruction errors. Here we present a novel approach for 3D SIM using a spinning disk. The disk generates a 3D lattice illumination pattern on the sample and optically reconstructs super-resolved images in real time. This technique achieves a 2-times resolution improvement with a speed up to 100 frames per second while physically rejecting 90% of the background signal.
© 2023 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement.