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Updated: Aug 15, 2026

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
Published on: May 30, 2016
Generalized low-pass filtering in structured illumination microscopy and spatial domain reconstructions with high
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Structured illumination microscopy (SIM) is a technique that employs non-uniform illumination to shift spatial frequencies that are normally unobservable into the non-zero region of the optical transfer function (OTF). Several differently illuminated images are combined to reconstruct these high spatial frequencies, yielding an up to two-fold increase in resolution beyond the diffraction limit. The reconstruction quality is affected by the noise level which depends on the specific procedure. In this work, we show that least-squares Fourier domain reconstruction results in the best possible theoretically achievable SSNR and thus can serve as a metric for reconstruction quality. We consider, in a general manner, how the choice of low-pass filtering in the reconstruction procedure affects the spectral signal-to-noise ratio (SSNR) in the reconstructed image. Finally, we present fast real-space reconstruction procedures that demonstrate near-optimal noise performance.
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