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

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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.
Funding Information
  • European Research Council (758752)
  • European Research Council (101069260)
  • European Innovation Council (101046928)
  • European Innovation Council (101259077)
  • European Commission (Marie Curie Skłodowska 847548)

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