<b>Dataset and reconstruction software for three-dimensional lattice structured illumination microscopy</b>
<p dir="ltr">Structured illumination microscopy (SIM) is widely used for live-cell super-resolution imaging. Compared with the two-dimensional implementation (2D-SIM), the three-dimensional implementation (3D-SIM) provides twofold improvement in both lateral and axial resolution. How...
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2025
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| Summary: | <p dir="ltr">Structured illumination microscopy (SIM) is widely used for live-cell super-resolution imaging. Compared with the two-dimensional implementation (2D-SIM), the three-dimensional implementation (3D-SIM) provides twofold improvement in both lateral and axial resolution. However, 3D-SIM requires more phase-shifted exposures at each axial plane and a higher photon budget per unit volume, which increases photobleaching and imposes heavy reconstruction overheads for volumetric imaging. Here we introduce 3D-Lattice-SIM, a gentle volumetric imaging strategy based on lattice illumination that encodes multiple spatial-frequency components per exposure, reducing redundant sampling and photon demand. We derive a reconstruction method for 3D-Lattice-SIM that enables accurate parameter estimation and robust physical reconstruction, and propose an ultrafast frequency-domain protocol that accelerates volumetric reconstruction by about three orders of magnitude. We further develop SIMPLEX, a self-supervised denoising network tailored to suppress noise-induced artifacts under photon-limited conditions. Within this framework, 3D-Lattice-SIM enables fast, high-fidelity 3D super-resolution reconstruction from about ten percent of the photon dose required by conventional 3D-SIM. We demonstrate live-cell imaging of endoplasmic reticulum remodeling and Lunapark redistribution at three-way junctions and volumetric visualization of multiple organelles from a single stain when combined with a pan-membrane labeling strategy. Overall, 3D-Lattice-SIM maintains volumetric three-dimensional super-resolution at a substantially reduced photon budget and provides a versatile platform for multicolor live-cell time-lapse imaging.<br>Please download and install the MATLAB runtime environment 2025a from https://www.mathworks.com/products/compiler/matlab-runtime.html<br></p> |
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