Simultaneous multicolor fluorescence imaging using PSF splitting

Van den Eynde, Robin; Hertel, Fabian; Abakumov, Sergey; Krajnik, Bartosz; Hugelier, Siewert; Auer, Alexander; Hellmeier, Joschka; Schlichthaerle, Thomas; Grattan, Rachel M.; Lidke, Diane S.; Jungmann, Ralf; Leutenegger, Marcel; Vandenberg, Wim; Dedecker, Peter

doi:10.1038/s41592-024-02383-7

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Simultaneous multicolor fluorescence imaging using PSF splitting

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Leutenegger, Marcel
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Citation

Van den Eynde, R., Hertel, F., Abakumov, S., Krajnik, B., Hugelier, S., Auer, A., et al. (2024). Simultaneous multicolor fluorescence imaging using PSF splitting. Nature Methods. doi:10.1038/s41592-024-02383-7.

Cite as: https://hdl.handle.net/21.11116/0000-000F-DDBA-7

Abstract

We present a way to encode more information in fluorescence imaging by splitting the original point spread function (PSF), which offers broadband operation and compatibility with other PSF engineering modalities and existing analysis tools. We demonstrate the approach using the ‘Circulator’, an add-on that encodes the fluorophore emission band into the PSF, enabling simultaneous multicolor super-resolution and single-molecule microscopy using essentially the full field of view.