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Confocal laser scanning microscopy (CLSM) stands out as one of the most widely used microscopy techniques, thanks to its three-dimensional imaging capability and its sub-diffraction spatial resolution, achieved through the closure of a pinhole in front of a single-element detector. However, the pinhole also rejects useful photons and beating the diffraction limit comes at the price of irremediably compromising the signal-to-noise ratio (SNR) of the data. Image scanning microscopy (ISM) emerged as the natural evolution of CLSM, exploiting a small array detector in place of the pinhole and the single-element detector. Each sensitive element is small enough to achieve sub-diffraction resolution through the confocal effect, but the size of the whole detector is large enough to guarantee excellent collection efficiency and SNR. However, the raw data produced by an ISM setup consists of a 4D dataset which can be seen as a set of confocal-like images. Thus, fusing the dataset into a single super-resolved image requires a dedicated reconstruction algorithm. Conventional methods are multi-image deconvolution, which requires prior knowledge of the system point spread functions (PSF), or adaptive pixel reassignment (APR), which is effective only on a limited range of experimental conditions. In this work, we describe and validate a novel concept for ISM image reconstruction based on autocorrelation inversion. We leverage unique properties of the autocorrelation to discard low-frequency components and maximize the resolution of the reconstructed image, without any assumption on the image or any knowledge of the PSF. Our results push the quality of the ISM reconstruction beyond the level provided by APR and open new perspectives for multi-dimensional image processing., Comment: 21 pages, 5 figures

In the field of disordered photonics, a common objective is to characterize optically opaque materials for controlling light delivery or performing imaging. Among various complex devices, multi-mode optical fibers stand out as cost-effective and easy-to-handle tools, making them attractive for several tasks. In this context, we leverage the reservoir computing paradigm to recast these fibers into random hardware projectors, transforming an input dataset into a higher dimensional speckled image set. The goal of our study is to demonstrate that using such randomized data for classification by training a single logistic regression layer improves accuracy compared to training on direct raw images. Interestingly, we found that the classification accuracy achieved using the reservoir is also higher than that obtained with the standard transmission matrix model, a widely accepted tool for describing light transmission through disordered devices. We find that the reason for such improved performance could be due to the fact that the hardware classifier operates in a flatter region of the loss landscape when trained on fiber data, which aligns with the current theory of deep neural networks. These findings strongly suggest that multi-mode fibers possess robust generalization properties, positioning them as promising tools for optically-assisted neural networks. With this study, in fact, we want to contribute to advancing the knowledge and practical utilization of these versatile instruments, which may play a significant role in shaping the future of machine learning.

Recovering the transmission matrix of a disordered medium is a challenging problem in disordered photonics. Usually, its reconstruction relies on a complex inversion that aims at connecting a fully-controlled input to the deterministic interference of the light field scrambled by the device. At the moment, iterative phase-retrieval protocols provide the fastest reconstructing frameworks, converging in a few tens of iterations. Exploiting the knowledge of speckle correlations, we construct a new phase retrieval algorithm that reduces the computational cost to a single iteration. Besides being faster, our method is effective also using less measurements than state-of-the-art protocols. Thanks to reducing computation time by one order of magnitude, our result can be groundbreaking for real-time optical operations in medical imaging., Comment: 9 pages, 7 figures

The aberrations induced by the sample and/or by the sample holder limit the resolution of optical microscopes. Wavefront correction can be achieved using a deformable mirror with wavefront sensorless optimization algorithms but, despite the complexity of these systems, the level of correction is often limited to a small area in the field of view of the microscope. In this work, we present a plug and play module for aberration measurement and correction. The wavefront correction is performed through direct wavefront reconstruction using the spinning-pupil aberration measurement and controlling a deformable lens in closed loop. The lens corrects the aberrations in the center of the field of view, leaving residual aberrations at the margins, that are removed by anisoplanatic deconvolution. We present experimental results obtained in fluorescence microscopy, with a wide field and a light sheet fluorescence microscope. These results indicate that detection and correction over the full field of view can be achieved with a compact transmissive module placed in the detection path of the fluorescence microscope., Comment: 7 pages, 5 figures

The aberrations in an optical microscope are commonly measured and corrected at one location in the field of view, within the so-called isoplanatic patch. Full-field correction is desirable for high-resolution imaging of large specimens. Here we present a novel wavefront detector, based on pupil sampling with sub-apertures, which measures the aberrated wavefront phase at each position of the specimen. Based on this measurement, we propose a region-wise deconvolution that provides an anisoplanatic reconstruction of the sample image. Our results indicate that the measurement and correction of the aberrations can be performed in a wide-field fluorescence microscope over its entire field of view., Comment: 5 pages, 4 figures

In multi-view fluorescence microscopy, each angular acquisition needs to be aligned with care to obtain an optimal volumetric reconstruction. Here, instead, we propose a neat protocol based on auto-correlation inversion, that leads directly to the formation of inherently aligned tomographies. Our method generates sharp reconstructions, with the same accuracy reachable after sub-pixel alignment but with improved point-spread-function. The procedure can be performed simultaneously with deconvolution further increasing the reconstruction resolution.

Recovering a signal from auto-correlations or, equivalently, retrieving the phase linked to a given Fourier modulus, is a wide-spread problem in imaging. This problem has been tackled in a number of experimental situations, from optical microscopy to adaptive astronomy, making use of assumptions based on constraints and prior information about the recovered object. In a similar fashion, deconvolution is another common problem in imaging, in particular within the optical community, allowing high-resolution reconstruction of blurred images. Here we address the mixed problem of performing the auto-correlation inversion while, at the same time, deconvolving its current estimation. To this end, we propose an I-divergence optimization, driving our formalism into a widely used iterative scheme, inspired by Bayesian-based approaches. We demonstrate the method recovering the signal from blurred auto-correlations, further analysing the cases of blurred objects and band-limited Fourier measurements., Comment: 8 pages, 5 figures

One of the biggest challenges in the field of biomedical imaging is the comprehension and the exploitation of the photon scattering through disordered media. Many studies have pursued the solution to this puzzle, achieving light-focusing control or reconstructing images in complex media. In the present work, we investigate how statistical inference helps the calculation of the transmission matrix in a complex scrambling environment, enabling its usage like a normal optical element. We convert a linear input-output transmission problem into a statistical formulation based on pseudolikelihood maximization, learning the coupling matrix via random sampling of intensity realizations. Our aim is to uncover insights from the scattering problem, encouraging the development of novel imaging techniques for better medical investigations, borrowing a number of statistical tools from spin-glass theory.

Linear problems appear in a variety of disciplines and their application for the transmission matrix recovery is one of the most stimulating challenges in biomedical imaging. Its knowledge turns any random media into an optical tool that can focus or transmit an image through disorder. Here, converting an input-output problem into a statistical mechanical formulation, we investigate how inference protocols can learn the transmission couplings by pseudolikelihood maximization. Bridging linear regression and thermodynamics let us propose an innovative framework to pursue the solution of the scattering-riddle.

Dementia disorders are increasingly becoming sources of a broad range of problems, strongly interfering with normal daily tasks of a growing number of individuals. Such neurodegenerative diseases are often accompanied with progressive brain atrophy that, at late stages, leads to drastically reduced brain dimensions. At the moment, this structural involution can be followed with XCT or MRI measurements that share numerous disadvantages in terms of usability, invasiveness and costs. In this work, we aim to retrieve information concerning the brain atrophy stage and its evolution, proposing a novel approach based on non-invasive time-resolved Near Infra-Red (tr-NIR) measurements. For this purpose, we created a set of human-head atlases, in which we eroded the brain as it would happen in a clinical brain-atrophy progression. With these realistic meshes, we reproduced a longitudinal tr-NIR study exploiting a Monte-Carlo photon propagation algorithm to model the varying cerebral spinal fluid (CSF). The study of the time-resolved reflectance curve at late photon arrival times exhibited peculiar slope-changes upon CSF layer increase that were confirmed under several measurement conditions. The performance of the technique suggests good sensitivity to CSF variation, useful for a fast and non-invasive observation of the dementia progression., Comment: 32 pages, double spaced, 11 figures

Hyperuniform structures possess the ability to confine and drive light, although their fabrication is extremely challenging. Here we demonstrate that speckle patters obtained by a superposition of randomly arranged sources of Bessel beams can be used to generate hyperunifrom scalar fields. By exploiting laser light tailored with a spatial filter, we experimentally produce (without requiring any computational power) a speckle pattern possessing maxima at locations corresponding to a hyperuniform distribution. By properly filtering out intensity fluctuation from the same speckle pattern, it is possible to retrieve an intensity profile satisfying the hyperuniformity requirements. Our findings are supported by extensive numerical simulations., Comment: 13 pages, 7 figures

The field of biomedical imaging has undergone a rapid growth in recent years, mostly due to the implementation of ad-hoc designed experimental setups, theoretical support methods and numerical reconstructions. Especially for biological samples, the high number of scattering events occurring during the photon propagation process limit the penetration depth and the possibility to outperform direct imaging in thicker and not transparent samples. In this thesis, we will examine theoretically and experimentally the scattering process from two opposite points of view, focusing also on the continuous stimulus offered by the will to tackle some specific challenges in the emerging optical imaging science. Firstly, we will discuss the light propagation in diffusive biological tissues considering the particular case of the presence of optically transparent regions enclosed in a highly scattering environment. The correct inclusion of this information, can ultimately lead to higher resolution reconstruction, especially in neuroimaging. On the other hand, we will examine the extreme case of the three-dimensional imaging of a totally hidden sample, in which the phase has been scrambled by a random scattering layer. By making use of appropriate numerical methods, we will prove how it is possible to outperform such hidden reconstruction in a very efficient way, opening the path toward the unexplored field of three-dimensional hidden imaging. Finally, we will present how, the properties noticed while addressing these problems, leaded us to the development of a novel alignment-free three-dimensional tomographic technique that we refer to as Phase-Retrieved Tomography. Ultimately, we used this technique for the study of the fluorescence distribution in a three-dimensional spherical tumor model, the cancer cell spheroid, one of the most important biological model for the study of such disease., Comment: Original title: "Development of novel light propagation algorithms in turbid media with varying optical heterogeneity"

Optical tomographic imaging of biological specimen bases its reliability on the combination of both accurate experimental measures and advanced computational techniques. In general, due to high scattering and absorption in most of the tissues, multi view geometries are required to reduce diffuse halo and blurring in the reconstructions. Scanning processes are used to acquire the data but they inevitably introduces perturbation, negating the assumption of aligned measures. Here we propose an innovative, registration free, imaging protocol implemented to image a human tumor spheroid at mesoscopic regime. The technique relies on the calculation of autocorrelation sinogram and object autocorrelation, finalizing the tomographic reconstruction via a three dimensional Gerchberg Saxton algorithm that retrieves the missing phase information. Our method is conceptually simple and focuses on single image acquisition, regardless of the specimen position in the camera plane. We demonstrate increased deep resolution abilities, not achievable with the current approaches, rendering the data alignment process obsolete., Comment: 21 pages, 5 figures

Wavefront shaping has revolutionized the concepts of optical imaging and focusing. Contrary to what was believed, strong scattering in the optical paths can be exploited in favor of light focusing through turbid media and ultimately improve optical imaging and light manipulation capabilities. The use of light shapers and appropriately engineered scattering structures, i.e. opaque lenses enables the production of nano-scale confined foci and of extended fields of view. Exploiting this concept we fabricate configurable scattering structures by direct femtosecond laser writing. The properly shaped light trespassing the customized structure, a photonic lattice of parallel rods, forms a light-sheet at user defined positions. We demonstrate that our technique enables light-sheets with sub-micron resolution and extended depth of focus, a significant advantage when compared to the existing free space systems. Moreover, our approach permits to focus light of different wavelengths onto the same defined position without moving any optical element and correcting for chromatic aberrations. Furthermore, we demonstrate how an opaque cylindrical lens can be created using a biological structure, hence unveiling the potential advantages for optical microscopic and mesoscopic imaging., Comment: 22 pages, 5 figures

Bessel beams are non-diffracting light structures, which can be produced with simple tabletop optical elements such as axicon lenses or ring spatial filters and coherent laser beams. One of their main characteristic is that Bessel beams maintain their spatial characteristics after meters of propagation. In this paper we demonstrate a system and method for generating Bessel beams from amorphous speckle patterns, exploiting adaptive optimization by a spatial light modulator. These speckles are generated by light modes transmitted through a scattering curtain and selected by a ring shaped filter. With the proposed strategy it is possible to produce at user defined positions, reconfigurable, non-diffracting Bessel beams through a disordered medium.