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Technology

2014
Journal of Modern Optics

Real-time super-resolution imaging by high-speed fluorescence emission difference microscopy

Authors:

Rong, Z., Li, S., Kuang, C., Xu, Y., & Liu, X.

Keywords:

FED, diffraction barrier, real-time imaging, high speed, galvo mirror

Abstract:

The recently proposed fluorescence emission difference (FED) microscopy has been demonstrated to be capable of breaking the diffraction barrier, which restricts the spatial resolution of far-field fluorescence microscopy. In this paper, we report a novel high-speed FED system that can realize real-time super-resolution imaging. By replacing the conventional nanopositioning stage with a galvo mirror, the temporal resolution of FED is improved to nearly one frame per second, which is 100 times faster than that of best existing FED, while the system maintains a super-high spatial resolution of 150 nm, which is far beyond the diffraction barrier. Therefore, the high-speed FED is suitable for large-area observations while avoiding photobleaching. Detailed theoretical analysis, simulations, and experimental real-time resolution tests on 100 nm nanoparticles and biological cells are reported.

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