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Doubling the resolution of light-sheet fluorescence microscopy

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A combination of light-sheet fluorescence microscopy (LSFM) with structured illumination doubles resolving power over LSFM alone. We show a practical implementation using a single objective for illumination and fluorescence detection and demonstrate its use for rapid volumetric imaging.

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Fig. 1: Light-sheet fluorescence microscopy (LSFM) with structured illumination (SIM).

References

  1. Stelzer, E. H. K. et al. Light sheet fluorescence microscopy. Nat. Rev. Methods Primers 1, 73 (2021). A primer of the current state of the art in light-sheet fluorescence microscopy (LSFM).

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  2. Gustafsson, M. G. L. Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. J. Microsc. 198, 82–87 (2000). Seminal paper for structured illumination microscopy, which was used here to improve the resolution in LSFM.

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  3. Dunsby, C. Optically sectioned imaging by oblique plane microscopy. Opt. Express 16, 20306–20316 (2008). Seminal paper for oblique plane microscopy, which was used here as a starting point for combining LSFM with structured illumination.

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  4. Chang, B.-J. et al. Real-time multi-angle projection imaging of biological dynamics. Nat. Methods 18, 829–834 (2021). A study describing a method to form projections from a 3D volume, which conceptually can be applied to the work presented here.

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  5. Chang, B.-J., Meza, V. D. P. & Stelzer, E. H. K. csiLSFM combines light-sheet fluorescence microscopy and coherent structured illumination for a lateral resolution below 100 nm. Proc. Natl Acad. Sci. USA 114, 4869–4874 (2017). A previous effort to combine structured illumination with LSFM using two illumination and one detection objective.

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This is a summary of: Chen, B. et al. Resolution doubling in light-sheet microscopy via oblique plane structured illumination. Nat. Methods https://doi.org/10.1038/s41592-022-01635-8 (2022).

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Doubling the resolution of light-sheet fluorescence microscopy. Nat Methods 19, 1355–1356 (2022). https://doi.org/10.1038/s41592-022-01636-7

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