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Updated: Sep 14, 2025

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
Published on: May 30, 2016
Synthesis of Arbitrary Interference Patterns Using a Single Galvanometric Mirror and Its Application to Structured
Ke Guo1, Abderrahim Boualam1, James D Manton2
1Department of Bioengineering, Imperial College London, Exhibition Road, London SW7 2AZ, U.K.
None:
Structured illumination microscopy (SIM) overcomes the diffraction limit of optical microscopy by projecting finely spaced interference fringes with different orientations and phases onto the sample and imaging the result. A major challenge of SIM is to generate the different illumination patterns with a high contrast and switching speed, which commonly requires expensive devices and the sacrifice of illumination power efficiency. We present a new way of generating interference patterns for 2D and 3D SIM achromatically, with high speed and high power efficiency, using only one moving part. The interference patterns are created by a common-path interferometer, with the orientation, polarization, and phase of interference patterns controlled by a single galvanometric mirror. We characterize the contrast and switching speed of the interference patterns and demonstrate their utility by performing high-speed (980 raw frames per second) 2D SIM imaging on fluorescent nanoparticles and 3D SIM on fixed iFluor 488 phalloidin-stained U-2 OS cells.
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