Random illumination microscopy (RIM) could surpass the diffraction barrier in fluorescence microscopy by illuminating an object with unknown speckle patterns. It has been demonstrated that the resolution in RIM using second-order statistics is as good as that of conventional structured illumination microscopy (SIM) from the asymptotic point of view. Compared to classical SIM, RIM is more robust to optical aberrations and scattering introduced by thick samples. In this work, I show that the quantum correlations could further improve the resolution in random illumination microscopy due to the photon antibunching property of fluorophore emitters. In theory, the super-resolution capacity of this quantum-enhanced version of RIM corresponds to the fourth power of the point spread function under the general epi-illumination geometry.
Random illumination microscopy (RIM) could surpass the diffraction barrier in fluorescence microscopy by illuminating an object with unknown speckle patterns. It has been demonstrated that the resolution in RIM using second-order statistics is as good as that of conventional structured illumination microscopy (SIM) from the asymptotic point of view. Compared to classical SIM, RIM is more robust to optical aberrations and scattering introduced by thick samples. In this work, I show that the quantum correlations could further improve the resolution in random illumination microscopy due to the photon antibunching property of fluorophore emitters. In theory, the super-resolution capacity of this quantum-enhanced version of RIM corresponds to the fourth power of the point spread function under the general epi-illumination geometry.
Far-field optical microscopy beyond the Abbe diffraction limit, making use of nonlinear excitation (e.g. STED), or temporal fluctuations in fluorescence (PALM, STORM, SOFI) is already a reality. In contrast, overcoming the diffraction limit using non-classical properties of light is very difficult to achieve due to the fragility of quantum states of light. Here, we experimentally demonstrate superresolution microscopy based on quantum properties of light naturally emitted by fluorophores used as markers in fluorescence microscopy. Our approach is based on photon antibunching, the tendency of fluorophores to emit photons one by one rather than in bursts. Although a distinctively quantum phenomenon, antibunching is readily observed in most common fluorophores even at room temperature. This nonclassical resource can be utilized directly to enhance the imaging resolution, since the non-classical far-field intensity correlations induced by antibunching carry high spatial frequency information on the spatial distribution of emitters. Detecting photon statistics simultaneously in the entire field of view, we were able to detect non-classical correlations of the second and third order, and reconstructed images with resolution significantly beyond the diffraction limit. Alternatively, we demonstrate the utilization of antibunching for augmenting the capabilities of localization-based superresolution imaging in the presence of multiple emitters, using a novel detector comprised of an array of single photon detectors connected to a densely packed fiber bundle. These features allow us to enhance the spatial and temporal resolution with which multiple emitters can be imaged compared with other techniques that rely on CCD cameras.
We propose to use intensity correlation microscopy in combination with structured illumination to image quantum emitters that exhibit antibunching with a spatial resolution reaching far beyond the Rayleigh limit. Combining intensity measurements and intensity auto correlations up to order m creates an effective PSF with FWHM shrunk by the factor m. Structured Illumination microscopy on the other hand introduces a resolution improvement of factor 2 by use of the principle of moiré fringes. Here, we show that for linear low-intensity excitation and linear optical detection the simultaneous use of both techniques leads to an in theory unlimited resolution power with the improvement scaling favorably as m+m in dependence of the correlation order m. Hence, yielding this technique to be of interest in microscopy for imaging a variety of samples including biological ones. We present the underlying theory and simulations demonstrating the highly increased spatial superresolution, and point out requirements for an experimental implementation.
We propose to use intensity correlation microscopy in combination with structured illumination to image quantum emitters that exhibit antibunching with a spatial resolution reaching far into the sub-classical regime. Combining intensity measurements and intensity auto correlations up to order m creates an effective PSF with FWHM shrunk by the factor p m. Structured Illumination microscopy on the other hand introduces a resolution improvement of factor 2 by use of the principle of moiré fringes. Here, we show that for linear low-intensity excitation and linear optical detection the simultaneous use of both techniques leads to an in theory unlimited resolution power with the improvement scaling favorably as m+sqrt m in dependence of the correlation order m. Hence, yielding this technique to be of the utmost interest for biological imaging. We present the underlying theory and simulations demonstrating the highly increased spatial superresolution, and point out requirements for an experimental implementation.
The classical optical diffraction limit can be overcome by exploiting the quantum properties of light in several theoretical studies; however, they mostly rely on an entangled light source. Recent experiments have demonstrated that quantum properties are preserved in many fluorophores, which makes it possible to add a new dimension of information for super-resolution fluorescence imaging. Here, we developed a statistical quantum coherence model for fluorescence emitters and proposed a new super-resolution method using fluorescence quantum coherence in fluorescence microscopy. In this study, by exploiting a single-photon avalanche detector (SPAD) array with a time-correlated single-photon-counting technique to perform spatial-temporal photon statistics of fluorescence coherence, the subdiffraction-limited spatial separation of emitters is obtained from the determined coherence. We numerically demonstrate an example of two-photon interference from two common fluorophores using an achievable experimental procedure. Our model provides a bridge between the macroscopic partial coherence theory and the microscopic dephasing and spectral diffusion mechanics of emitters. By fully taking advantage of the spatial-temporal fluctuations of the emitted photons as well as coherence, our quantum-enhanced imaging method has the significant potential to improve the resolution of fluorescence microscopy even when the detected signals are weak.
Reaching high-resolution imaging of delicate samples at a low-intensity level is a major challenge in microscopy. Here, we present and experimentally implement an advanced quantum super-resolution imaging technique based on photon statistics measurement. Our reconstruction algorithm adapts to any kind of non-Poissonian emitters, outperforming the corresponding classical SOFI method, in particular in low-light conditions. It offers sub-diffraction resolution improvement that scales with the j , where j is the highest-order central moments of the photocounts. More remarkably, we show that in combination with structured illumination, an improvement of j + j can be reached, providing a promising avenue for non-invasive super-resolution microscopy.
Intensity correlation microscopy (ICM), which is prominently known through antibunching microscopy or super-resolution optical fluctuation imaging (SOFI), provides super-resolution through a correlation analysis of antibunching of independent quantum emitters or temporal fluctuations of blinking fluorophores. For correlation order m the PSF in the signal is effectively taken to the mth power, and is thus directly shrunk by the factor m. Combined with deconvolution, a close to linear resolution improvement of factor m can be obtained. Yet, analysis of high correlation orders is challenging, which limits the achievable resolutions. Here we propose to use three dimensional structured illumination along with mth-order correlation analysis to obtain an enhanced scaling of up to m + m = 2m. Including the stokes shift or plasmonic sub-wavelength illumination enhancements beyond 2m can be achieved. Hence, resolutions far below the diffraction limit in full 3D imaging and with already low correlation orders, can potentially be achieved. Since ICM operates in the linear regime our approach may be particularly promising for enhancing the resolution in biological imaging at low illumination levels.