Abstract
Super-resolution microscopy surpasses the diffraction limit to enable the observation of the fine details in sub-cellular structures and their dynamics in diverse biological processes within living cells. Structured illumination microscopy (SIM) uses a relatively low illumination light power compared with other super-resolution microscopies and has great potential to meet the demands of live-cell imaging. However, the imaging acquisition and reconstruction speeds limit its further applications. In this article, recent developments all targeted at improving the overall speed of SIM are reviewed. These comprise both hardware and software improvements, which include a reduction in the number of raw images, GPU acceleration, deep learning and the spatial domain reconstruction. We also discuss the application of these developments in live-cell imaging.
Highlights
Fluorescence microscopy is a powerful tool for visualizing biological processes of molecules and intracellular structures in living cells
3-frame structured-illumination microscopy (SIM) and Deep learning methods perform the best and receive the most stars. This makes sense because less raw data are needed for SR image reconstruction and this results in better ways to track sample movement
Deep learning and SDR methods have a faster postprocessing speed, which has the benefit of displaying reconstructed SR images of live-cells in real-time
Summary
Fluorescence microscopy is a powerful tool for visualizing biological processes of molecules and intracellular structures in living cells. Based on this setup, we introduce several improvements for fast SIM . Whereas the camera used by fast structured illumination systems usually reads the imaging data out in an asynchronous way as a result of rolling shutter mode This leads to synchronization problems limiting the acquisition speed of fast SIM. By presenting different SIM patterns during the start exposure and readout line, the SLM maintains the readout line of the camera inside the dark region when the camera runs in continuous rolling shutter mode This approach reached an acquisition rate of 79 fps with the raw frame rate of 714 fps, the field of view at 16.5 μm × 16.5 μm and the laser illuminating time of 0.5 s.
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