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Diffraction unlimited super-resolution imaging critically depends on the switching of fluorophores between at least two states, often induced using intense laser light and special buffers. The high illumination power or UV light required for appropriate blinking kinetics is currently hindering live-cell experiments. Recently, so-called self-blinking dyes that switch spontaneously between an open, fluorescent on-state and a closed colorless off-state were introduced. Here we exploit the synergy between super-resolution optical fluctuation imaging (SOFI) and spontaneously switching fluorophores for 2D functional and for volumetric imaging. SOFI tolerates high labeling densities, on-time ratios, and low signal-to-noise by analyzing higher-order statistics of a few hundred to thousand frames of stochastically blinking fluorophores. We demonstrate 2D imaging of fixed cells with a uniform resolution up to 50-60 nm in 6th order SOFI and characterize changing experimental conditions. We extend multiplane cross-correlation analysis to 4th order using biplane and 8-plane volumetric imaging achieving up to 29 (virtual) planes. The low laser excitation intensities needed for self-blinking SOFI are ideal for live-cell imaging. We show proof-of-principal time-resolved imaging by observing slow membrane movements in cells. Self-blinking SOFI provides a route for easy-to-use 2D and 3D high-resolution functional imaging that is robust against artefacts and suitable for live-cell imaging.
We report an experimental demonstration of a nonclassical imaging mechanism with super-resolving power beyond the Rayleigh limit. When the classical image is completely blurred out due to the use of a small imaging lens, by taking advantage of the in
Beam self-cleaning (BSC) in graded-index (GRIN) multimode fibres (MMFs) has been recently reported by different research groups. Driven by the interplay between Kerr effect and beam self-imaging, BSC counteracts random mode coupling, and forces laser
Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the universal application of supe
The resolution of optical imaging devices is ultimately limited by the diffraction of light. To circumvent this limit, modern super-resolution microscopy techniques employ active interaction with the object by exploiting its optical nonlinearities, n
It has been shown that negative refraction makes a perfect lens. However, with little loss, the imaging functionality will be strongly compromised. Later on, it was proved that positive refraction from Maxwells fish-eye lens can also makes a perfect