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Compressed Raman spectroscopy is a promising technique for fast chemical analysis. In particular, classification between species with known spectra can be performed with measures acquired through a few binary filters. Moreover, it is possible to reconstruct spectra by using enough filters. As classification and reconstruction are competing, designing filters allowing one to perform both tasks is challenging. To tackle this problem, we propose to build optimal trade-off filters, i.e., filters so that there exist no filters achieving better performance in both classification and reconstruction. With this approach, users get an overview of reachable performance and can choose the trade-off most fitting their application.
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3D-scanning fluorescence imaging of living tissue is in demand for less phototoxic acquisition process. For the imaging of biological surfaces, adaptive and sparse scanning schemes have been proven to efficiently reduce the light dose by concentrating acquisitions around the surface. In this paper, we focus on optimizing the scanning scheme at a constant photon budget, when the problem is to estimate the position of a biological surface whose intensity profile is modeled as a Gaussian shape. We propose an approach based on the Cramér-Rao bound to optimize the positions and number of scanning points, assuming signal-dependant Gaussian noise. We show that, in the case of regular sampling, the optimization problem can be reduced to a few parameters, allowing us to define quasi-optimal acquisition strategies, first when no prior knowledge of the surface location is available and then when the user has a prior on this location.
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Imageamento Tridimensional , Microscopia ConfocalRESUMO
Compressed Raman methods allow classification between known chemical species with only a few measurements through binary filters. We propose a methodology for binary filter optimization, in which filters are modified at each pixel so that classification can still be achieved pixel by pixel with a few measurements acquired in parallel, while retaining the ability to reconstruct a full spectrum when combining measurements from several pixels. This approach is robust to intensity variations between pixels. It relies on a generalized Bhattacharyya bound and on the Cramér-Rao bound to tailor filters with optimized performance.
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Scanning fluorescence microscopes are now able to image large biological samples at high spatial and temporal resolution. This comes at the expense of an increased light dose which is detrimental to fluorophore stability and cell physiology. To highly reduce the light dose, we designed an adaptive scanning fluorescence microscope with a scanning scheme optimized for the unsupervised imaging of cell sheets, which underly the shape of many embryos and organs. The surface of the tissue is first delineated from the acquisition of a very small subset (~0.1%) of sample space, using a robust estimation strategy. Two alternative scanning strategies are then proposed to image the tissue with an improved photon budget, without loss in resolution. The first strategy consists in scanning only a thin shell around the estimated surface of interest, allowing high reduction of light dose when the tissue is curved. The second strategy applies when structures of interest lie at the cell periphery (e.g. adherens junctions). An iterative approach is then used to propagate scanning along cell contours. We demonstrate the benefit of our approach imaging live epithelia from Drosophila melanogaster. On the examples shown, both approaches yield more than a 20-fold reduction in light dose -and up to more than 80-fold- compared to a full scan of the volume. These smart-scanning strategies can be easily implemented on most scanning fluorescent imaging modality. The dramatic reduction in light exposure of the sample should allow prolonged imaging of the live processes under investigation.
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Classification of different species with Raman measurements is analyzed when a total of exactly $ N $N photons are detected with binary filtered Raman spectra instead of fixing the measuring time. The optimal classification method for this problem leads to classification error probabilities upper-bounded by the Bhattacharyya bound and that are invariant to the multiplication of the spectrum intensities by an unknown factor. Furthermore, it is shown that this approach can be implemented with a number of binary filters smaller than the number of species to discriminate.
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Bhattacharyya bounds of classification error probability between two species with Raman and binary compressed Raman measurements limited by Poisson photon noise are analyzed. They exhibit the relevant physical parameters and lead to a simple expression of a minimal number of photons necessary to upper bound the optimal classification error probability.
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The precision of proportion estimation with binary filtering of a Raman spectrum mixture is analyzed when the number of binary filters is equal to the number of present species and when the measurements are corrupted with Poisson photon noise. It is shown that the Cramer-Rao bound provides a useful methodology to analyze the performance of such an approach, in particular when the binary filters are orthogonal. It is demonstrated that a simple linear mean square error estimation method is efficient (i.e., has a variance equal to the Cramer-Rao bound). Evolutions of the Cramer-Rao bound are analyzed when the measuring times are optimized or when the considered proportion for binary filter synthesis is not optimized. Two strategies for the appropriate choice of this considered proportion are also analyzed for the binary filter synthesis.
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We report a simple add-on for broadband stimulated Raman scattering (SRS) microscopes to enable fast and programmable spectroscopy acquisition. It comprises a conventional dispersive spectrometer layout incorporating a fast digital micromirror device (DMD). The approach is validated by acquiring SRS spectra of standard chemicals. We demonstrate a DMD's advantage in broadband SRS by showing higher signal-to-noise ratio using a multiplexed Hadamard spectral basis and compressive sensing detection. Our results apply to a variety of frequency-domain pump-probe spectroscopy.
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Second-harmonic generation microscopy can provide estimation of some local molecule distribution properties. However, in order not to get erroneous conclusions, it is important to detect measurements with insufficient precision. Such a detection technique is developed considering an approximation of the ultimate precision provided by the Cramer-Rao bound. This method is characterized and a simple approximation of its detection and false alarm probabilities is developed.
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We address the detection of manufactured objects in different types of environments with active polarimetric imaging. Using an original, fully adaptive imager, we compare several imaging modes having different numbers of polarimetric degrees of freedom. We demonstrate the efficiency of active polarimetric imaging for decamouflage and hazardous object detection, and underline the characteristics that a polarimetric imager aimed at this type of application should possess. We show that in most encountered scenarios the Mueller matrices are nearly diagonal, and sufficient detection performance can be obtained with simple polarimetric imaging systems having reduced degrees of freedom. Moreover, intensity normalization of images is of paramount importance to better reveal polarimetric contrast.
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We designed and built an active polarimetric imager with laser illumination at 1.5 µm wavelength for adaptive polarimetric contrast optimization. It can generate and analyze any polarization state on the Poincaré sphere in order to best adapt to the polarimetric properties of the scene. Polarimetric contrast optimization is performed by analyzing the scene with an ultrafast active-contour-based segmentation algorithm. This device is, to the best of our knowledge, the first fully adaptive imager controlled by image processing algorithms for polarimetric contrast enhancement. Its capabilities are illustrated in some examples of real-world decamouflage applications.
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Single-cell dry mass measurement is used in biology to follow cell cycle, to address effects of drugs, or to investigate cell metabolism. Quantitative phase imaging technique with quadriwave lateral shearing interferometry (QWLSI) allows measuring cell dry mass. The technique is very simple to set up, as it is integrated in a camera-like instrument. It simply plugs onto a standard microscope and uses a white light illumination source. Its working principle is first explained, from image acquisition to automated segmentation algorithm and dry mass quantification. Metrology of the whole process, including its sensitivity, repeatability, reliability, sources of error, over different kinds of samples and under different experimental conditions, is developed. We show that there is no influence of magnification or spatial light coherence on dry mass measurement; effect of defocus is more critical but can be calibrated. As a consequence, QWLSI is a well-suited technique for fast, simple, and reliable cell dry mass study, especially for live cells.
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Interferometria/métodos , Análise de Célula Única/métodos , Algoritmos , Animais , Artefatos , Automação , Células COS , Calibragem , Chlorocebus aethiops , Desenho de Equipamento , Eritrócitos/citologia , Processamento de Imagem Assistida por Computador , Luz , Microscopia/métodos , Mitose , Distribuição Normal , Óptica e Fotônica , Reprodutibilidade dos TestesRESUMO
Underwater images often suffer from poor visibility due to photon scattering. However, in some cases, optical polarization filtering techniques can decrease the contribution of the scattered light and improve the visual image quality. In this Letter, the influence of these techniques for underwater image registration is analyzed, particularly when backscattered light is the main perturbation induced by the submarine environment. This analysis is performed using the Cramer-Rao bound and relies on a standard image formation model, taking into account various kinds of noises.
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Underwater optical image simulation is a valuable tool for oceanic science, especially for the characterization of image processing techniques such as color restoration. In this context, simulating images with a correct color rendering is crucial. This paper presents an extension of existing image simulation models to RGB imaging. The influence of the spectral discretization of the model parameters on the color rendering of the simulated images is studied. It is especially shown that, if only RGB data of the scene chosen for simulations are available, a spectral reconstruction step prior to the simulations improves the image color rendering.
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We present a method for automatic target detection based on the iterative interplay between an active polarimetric imager with adaptive capabilities and a snake-based image segmentation algorithm. It successfully addresses the difficult situations where the target and the background differ only by their polarimetric properties. This method illustrates the benefits of integrating digital processing algorithms at the heart of the image acquisition process rather than using them only for postprocessing.
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Algoritmos , Interpretação de Imagem Assistida por Computador/métodos , Reconhecimento Automatizado de Padrão/métodos , Refratometria/métodos , Polarimetria de Varredura a Laser/métodos , Aumento da Imagem/métodos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
This paper deals with point target detection in infrared images of the sky for which there are local variations of the gray level mean value. We show that considering a simple image model with the gray level mean value varying as a linear or a quadratic function of the pixel coordinates can improve mixed segmentation-detection performance in comparison to homogeneous model-based approaches.
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We propose a new method based on the minimization of the stochastic complexity for fast and efficient tracking adapted to video images with a static camera. The obtained criterion combines the advantages of background-subtraction-based techniques and those of using measures of similarities to a target model without requiring any tuning of a weighting parameter. It is then demonstrated that this approach can be implemented with a fast integral image technique to estimate the location and the rectangular shape of the target in a few milliseconds.
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A new snake-based segmentation technique of a single object (simply connected) in the presence of inhomogeneous Gaussian noise is proposed, in which the mean in each region is modeled as a polynomial function of the coordinates and which is thus adapted to inhomogeneous illumination. It is shown that the minimization of the stochastic complexity of the image, which can be implemented efficiently, allows one to automatically estimate not only the number and the position of the nodes of the polygonal contour used to describe the object but also the degree of the polynomials that model the variations of the mean.
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We present a generalization of a new statistical technique of image partitioning into homogeneous regions to cases where the family of the probability laws of the gray-level fluctuations is a priori unknown. For that purpose, the probability laws are described with step functions whose parameters are estimated. This approach is based on a polygonal grid which can have an arbitrary topology and whose number of regions and regularity of its boundaries are obtained by minimizing the stochastic complexity of the image. We demonstrate that efficient homogeneous image partitioning can be obtained when no parametric model of the probability laws of the gray levels is used and that this approach leads to a criterion without parameter to be tuned by the user. The efficiency of this technique is compared to a statistical parametric technique on a synthetic image and is compared to a standard unsupervised segmentation method on real optical images.
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Algoritmos , Artefatos , Aumento da Imagem/métodos , Interpretação de Imagem Assistida por Computador/métodos , Armazenamento e Recuperação da Informação/métodos , Modelos Estatísticos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Processos EstocásticosRESUMO
We propose a nonparametric statistical snake technique that is based on the minimization of the stochastic complexity (minimum description length principle). The probability distributions of the gray levels in the different regions of the image are described with step functions with parameters that are estimated. The segmentation is thus obtained by minimizing a criterion that does not include any parameter to be tuned by the user. We illustrate the robustness of this technique on various types of images with level set and polygonal contour models. The efficiency of this approach is also analyzed in comparison with parametric statistical techniques.