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Updated: Feb 19, 2026

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
Published on: May 30, 2016
Super-Resolution Structured Illumination Microscopy
Rainer Heintzmann1,2, Thomas Huser3,4
1Leibniz Institute of Photonic Technology , Albert-Einstein Straße 9, 07745 Jena, Germany.
Abstract:
Super-resolved structured illumination microscopy (SR-SIM) is among the most rapidly growing fluorescence microscopy techniques that can surpass the optical diffraction limit. The strength of SR-SIM is that it can be readily applied to samples prepared for conventional fluorescence microscopy, requiring no sophisticated sample preparation protocols. As an extension of wide-field fluorescence microscopy, it is inherently capable of multicolor imaging and optical sectioning and, with sufficiently fast implementations, permits live cell imaging. Image reconstruction, however, currently relies on sophisticated computational procedures, susceptible to reconstruction artifacts, requiring trained users to recognize and avoid them. Here, we review the latest developments in SR-SIM research. Starting from a historical overview of the development of SR-SIM, we review how this method can be implemented in various experimental schemes, we provide an overview of the important parameters involved in successful image reconstruction, we summarize recent biological applications, and we provide a brief outlook of the directions in which we believe SR-SIM is headed in the future.
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