Simultaneous multicolor fluorescence imaging using PSF splitting

Authors:

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

Keywords:

Fluorescence imaging; Super-resolution microscopy; point spread function; PSF; multicolor super-resolution; single-molecule microscopy

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.

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