Super-resolution structured illumination microscopy of cellular dynamics with fluorescence lifetime contrast

Raphaël Marchand; Henning Ortkrass; Daniel Aziz; Franz Pfanner; Eman Abbas; Silvio O. Rizzoli; Wolfgang Hübner; Adam J. Bowman; Thomas Huser; Thomas Juffmann

doi:10.1364/OPTICA.589837

Super-resolution structured illumination microscopy of cellular dynamics with fluorescence lifetime contrast

Raphaël Marchand
, Henning Ortkrass
, Daniel Aziz
, Franz Pfanner
, Eman Abbas
, Silvio O. Rizzoli
, Wolfgang Hübner
, Adam J. Bowman
, Thomas Huser
, Thomas Juffmann (Corresponding author)

Publications: Contribution to journal › Article › Peer Reviewed

Abstract

Super-resolution structured illumination microscopy (SR-SIM) enables fluorescence microscopy beyond the diffraction limit at high frame rates. Compared to other super-resolution microscopy techniques, the low photon fluence used in SR-SIM makes it readily compatible with live-cell imaging. Here, we combine SR-SIM with electro-optic fluorescence lifetime imaging (EOFLIM), demonstrating super-resolved multiplexed imaging of spectrally overlapping fluorophores, enhanced contrast due to optical sectioning, and environmental sensing at a spatial resolution of 156 nm. The high photon detection throughput of EOFLIM enables a combination of lifetime precision, size of the field of view, spatial resolution, and speed, which, taken together, is unprecedented, enabling the super-resolved imaging of cellular dynamics.

Original language	English
Pages (from-to)	1065-1074
Number of pages	10
Journal	Optica
Volume	13
Issue number	6
DOIs	https://doi.org/10.1364/OPTICA.589837
Publication status	Published - 20 Jun 2026

Funding

European Research Council, Horizon 2020 research and innovation program (758752, 101069260); European Innovation Council (101046928, 101259077); European Union (Marie Curie Skłodowska 847548). We acknowledge useful discussions with Francisco Balzarotti, Lukas Hille, Nele Jarnot, Clara Conrad-Billroth, Philipp Weber, Jörg Enderlein, and Ivana Matoušová Víšová for help with the acquisition of the isogyre patterns. Marcel Müller is acknowledged for software support. We thank the biooptics facility of the Max Perutz Labs for support. We also acknowledge PicoQuant GmbH for the loan of the laser source free of charge. We also acknowledge the Büllesbach family, the Wegscheid family, and flatmates from August-Bebel-Straße, Bielefeld, for their kind accommodation offer free of charge.

Funders	Funder number
European Research Council	758752, 101069260
European Commission	101046928, 101259077, 847548

Austrian Fields of Science 2012

103021 Optics

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10.1364/OPTICA.589837Licence: Other

Cite this

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title = "Super-resolution structured illumination microscopy of cellular dynamics with fluorescence lifetime contrast",

abstract = "Super-resolution structured illumination microscopy (SR-SIM) enables fluorescence microscopy beyond the diffraction limit at high frame rates. Compared to other super-resolution microscopy techniques, the low photon fluence used in SR-SIM makes it readily compatible with live-cell imaging. Here, we combine SR-SIM with electro-optic fluorescence lifetime imaging (EOFLIM), demonstrating super-resolved multiplexed imaging of spectrally overlapping fluorophores, enhanced contrast due to optical sectioning, and environmental sensing at a spatial resolution of 156 nm. The high photon detection throughput of EOFLIM enables a combination of lifetime precision, size of the field of view, spatial resolution, and speed, which, taken together, is unprecedented, enabling the super-resolved imaging of cellular dynamics.",

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AU - Rizzoli, Silvio O.

AU - Hübner, Wolfgang

AU - Bowman, Adam J.

AU - Huser, Thomas

AU - Juffmann, Thomas

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AB - Super-resolution structured illumination microscopy (SR-SIM) enables fluorescence microscopy beyond the diffraction limit at high frame rates. Compared to other super-resolution microscopy techniques, the low photon fluence used in SR-SIM makes it readily compatible with live-cell imaging. Here, we combine SR-SIM with electro-optic fluorescence lifetime imaging (EOFLIM), demonstrating super-resolved multiplexed imaging of spectrally overlapping fluorophores, enhanced contrast due to optical sectioning, and environmental sensing at a spatial resolution of 156 nm. The high photon detection throughput of EOFLIM enables a combination of lifetime precision, size of the field of view, spatial resolution, and speed, which, taken together, is unprecedented, enabling the super-resolved imaging of cellular dynamics.

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Super-resolution structured illumination microscopy of cellular dynamics with fluorescence lifetime contrast

Abstract

Funding

Austrian Fields of Science 2012

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Other files and links

HIGH-RESOLUTION FUNCTIONAL OPTICAL IMAGING OF CHEMORESISTANCE (CHEM-SCAN)

EOFLIM: Fast Electro-Optic Fluorescence Lifetime Microscopy (EOFLIM) for super-resolution microscopy and brain surgery.

Integration of solid-state nanopores into a functional device designed for electrical and optical cross-monitoring

Cite this

Super-resolution structured illumination microscopy of cellular dynamics with fluorescence lifetime contrast

Abstract

Funding

Austrian Fields of Science 2012

Access to Document

Other files and links

Projects

HIGH-RESOLUTION FUNCTIONAL OPTICAL IMAGING OF CHEMORESISTANCE (CHEM-SCAN)

EOFLIM: Fast Electro-Optic Fluorescence Lifetime Microscopy (EOFLIM) for super-resolution microscopy and brain surgery.

Integration of solid-state nanopores into a functional device designed for electrical and optical cross-monitoring

Cite this