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Super-resolution imaging using the spatial-frequency filtered intensity fluctuation correlation

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 Added by Jane Sprigg
 Publication date 2014
  fields Physics
and research's language is English




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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 intensity fluctuation correlation of thermal light, the demonstrated camera recovered the image of the resolution testing gauge. This method could be adapted to long distance imaging, such as satellite imaging, which requires large diameter camera lenses to achieve high image resolution.



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Developing a chip-based super-resolution imaging technique with large field-of-view (FOV), deep subwavelength resolution, and compatibility for both fluorescent and non-fluorescent samples is desired for material science, biomedicine, and life researches, etc. Previous on-chip super-resolution methods focus on either fluorescent or non-fluorescent imaging, putting an urgent requirement on the general imaging technique compatible with both of them. Here, we introduce a universal super-resolution imaging method based on tunable virtual-wavevector spatial frequency shift (TVSFS), realizing both labeled and label-free super-resolution imaging on a single delicately fabricated scalable photonic chip. Theoretically, with TVSFS, the diffraction limit of a linear optical system can be overcome, and the resolution can be improved more than three times, which is the limitation for most super-resolution imaging based on spatial frequency engineering. Diffractive units were fabricated on the chips surface to provide a wavevector-variable evanescent wave illumination and induce tunable deep SFS in the samples Fourier space. A resolution of {lambda}/4.7 for the label-free sample and {lambda}/7.1 for the labeled sample with a large FOV could be achieved with a CMOS-compatible process on a GaP chip. The large FOV, high-compatibility, and high-integration TVSFS chip may advance the fields like cell engineering, precision inspection in the industry, chemical research, etc.
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, nonclassical properties of the illumination beam, or near-field probing. Thus, they are not applicable whenever such interaction is not possible, for example, in astronomy or non-invasive biological imaging. Far-field, linear-optical super-resolution techniques based on passive analysis of light coming from the object would cover these gaps. In this paper, we present the first proof-of-principle demonstration of such a technique. It works by accessing information about spatial correlations of the image optical field and, hence, about the object itself via measuring projections onto Hermite-Gaussian transverse spatial modes. With a basis of 21 spatial modes in both transverse dimensions, we perform two-dimensional imaging with twofold resolution enhancement beyond the diffraction limit.
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.
Despite super-resolution fluorescence blinking microscopes break the diffraction limit, the intense phototoxic illumination and long-term image sequences thus far still pose to major challenges in visualizing live-organisms. Here, we proposed a super-resolution method based on auto-correlation two-step deconvolution (SACD) to enhance the temporal resolution at lower signal intensity levels. Unlike conventional techniques, such as super-resolution optical fluctuation imaging (SOFI) or stochastic optical reconstruction microscopy (STORM), our model allows 16 frames to generate super-resolution images, without noticeable degradation in recording quality. We demonstrate SACD both in simulated predictions and experimental validations, with the resulting spatial resolution of 64 nm and 2-10 fold speed improvements. The use of low signal-to-noise ratio acquisition of image sequences, our SACD enables fast, background-free, super-resolution imaging that maybe advance implemented as a suitable tool for rapid live-cells imaging.
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 lens. However, strong debating happens on the introduced drain problem at the imaging position. In this work, we for the first time find that a solid immersion Maxwells fish-eye lens could be used for super-resolution imaging. We find that it is due to the perfect focusing and total reflection at the outer interface, such that a super-resolution image is formed at the required position in the air background. This simple mechanism will also be valid for other absolute instruments and more versatile super-imaging systems will be anticipated.
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