Fast structured illumination microscopy using rolling shutter cameras

Spatial light modulators (SLM) update in a synchronous manner, whereas the data readoutprocess in fast structured illumination systems is usually done using a rolling shutter camerawith asynchronous readout. In structured illumination microscopy (SIM), this leads tosynchronization problems causing a speed limit for fast acquisition. In this paper we presenta configuration to overcome this limit by exploiting the extremely fast SLM display anddividing it into several segments along the direction of the rolling shutter of the sCMOScamera and displaying multiple SLM frames per camera acquisition. The sCMOS runs incontinuous rolling shutter mode and the SLM keeps the readout-line always inside a darkregion presenting different SIM patterns before and after the readout/start-exposure line.Using this approach, we reached a raw frame rate of 714 frames per second (fps) resulting ina two-beam SIM acquisition rate of 79 fps with a region of interest (ROI) of 16.5 × 16.5 μm2.