Fig. 4

Demonstration of the high image frame rates achieved by high-speed SR-SIM. a Cutouts of eight consecutive SR-SIM frames taken at 57.8 SIM frames (520 raw frames) per second, with 0.5 ms illumination time for each of the nine raw frames per SR-SIM image. The sample consists of 0.2 µm TetraSpeck Microspheres in a 50/50 mixture of stock solution and glycerol. The excitation wavelength was 647 nm, laser power was about 3 mW before entering the objective lens. The short acquisition time allows the acquisition of images with low motion blur, even though the sample is diffusing quickly, and the high SR-SIM frame rate leads to a high temporal resolution. b Movement of the diffusing microspheres along the x-axis over time displayed as a kymograph. It spans over 107 consecutive SIM frames, or ~1.85 s, including the images shown in (a). Note that the diffusing microspheres could also be observed in real time and super-resolution by the system operator. Fixed microspheres were also imaged to demonstrate the general capabilities of the SIM system, see Supplementary Fig. 7