We developed 4Pi-SIM, which integrates 4Pi microscopy with structured illumination microscopy to achieve isotropic optical resolution through interference in both illumination and detection. We demonstrate its capabilities by time-lapse volumetric imaging of various subcellular structures at 100-nm resolution.
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References
Shao, L., Kner, P., Rego, E. H. & Gustafsson, M. G. Super-resolution 3D microscopy of live whole cells using structured illumination. Nat. Methods 8, 1044–1046 (2011). Seminal paper presenting 3D-SIM in living cells.
Shao, L. et al. I5S: wide-field light microscopy with 100-nm-scale resolution in three dimensions. Biophys. J. 94, 4971–4983 (2008). Seminal paper about combining 4Pi microscopy and SIM.
Zhang, Y. et al. Nanoscale subcellular architecture revealed by multicolor three-dimensional salvaged fluorescence imaging. Nat. Methods 17, 225–231 (2020). A study describing the implementation of a multicolor 4Pi-SMS microscope.
Huang, X. et al. Fast, long-term, super-resolution imaging with Hessian structured illumination microscopy. Nat. Biotechnol. 36, 451–459 (2018). A study introducing the Hessian algorithm for minimizing reconstruction artifacts in SIM.
Shao, L., Winoto, L., Agard, D. A., Gustafsson, M. G. & Sedat, J. W. Interferometer-based structured-illumination microscopy utilizing complementary phase relationship through constructive and destructive image detection by two cameras. J. Microsc. 246, 229–236 (2012). A study introducing the use of complementary phase images for estimating phase shifts in I5S.
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This is a summary of: Ouyang, Z. et al. Elucidating subcellular architecture and dynamics at isotropic 100-nm resolution with 4Pi-SIM. Nat. Methods https://doi.org/10.1038/s41592-024-02515-z (2024).
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Isotropic 100-nm resolution live-cell imaging with 4Pi-SIM. Nat Methods 22, 233–234 (2025). https://doi.org/10.1038/s41592-024-02514-0
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DOI: https://doi.org/10.1038/s41592-024-02514-0
