ArticleBiomedical optics express2026
Super-resolution imaging using fluorescence-lifetime emission-difference.
Article in Biomedical optics express, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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6 authors.
Funding
Abstract
The diffraction of light limits the ability of light microscopes to resolve biological structures and intracellular processes with nanoscale spatial information. Fluorescence emission difference (FED) improves image contrast and effective spatial resolution of confocal microscopy by alternating the excitation of the sample with a Gaussian and donut-shaped laser beam. Improvement in image resolution is obtained by subtracting images captured with both excitation modes. The combination of this technique with fluorescence lifetime imaging microscopy (FLIM) has remained limited. Here, we propose a super-resolution fluorescence-lifetime emission-difference (FLED) microscopy approach obtained through a multi-image weighted-subtraction algorithm that combines images reconstructed from multiple temporal bins. A high-throughput implementation was demonstrated using a commercial time-resolved stimulated-emission depletion microscope (STED). Through simulations of fluorescent beads and experimental validation on biological samples, we demonstrated a 1.6× resolution enhancement corresponding to 0.22
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