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In-line digital holography is a simple yet powerful tool to image absorbing and/or phase objects. Nevertheless, the loss of the phase of the complex wavefront on the sensor can be critical in the reconstruction process. The simplicity of the setup must thus be counterbalanced by dedicated reconstruction algorithms, such as inverse approaches, in order to retrieve the object from its hologram. In the case of simple objects for which the diffraction pattern produced in the hologram plane can be modeled using few parameters, a model fitting algorithm is very effective. However, such an approach fails to reconstruct objects with more complex shapes, and an image reconstruction technique is then needed. The improved flexibility of these methods comes at the cost of a possible loss of reconstruction accuracy. In this work, we combine the two approaches (model fitting and regularized reconstruction) to benefit from their respective advantages. The sample to be reconstructed is modeled as the sum of simple parameterized objects and a complex-valued pixelated transmittance plane. These two components jointly scatter the incident illumination, and the resulting interferences contribute to the intensity on the sensor. The proposed hologram reconstruction algorithm is based on alternating a model fitting step and a regularized inversion step. We apply this algorithm in the context of fluid mechanics, where holograms of evaporating droplets are analyzed. In these holograms, the high contrast fringes produced by each droplet tend to mask the diffraction pattern produced by the surrounding vapor wake. With our method, the droplet and the vapor wake can be jointly reconstructed.
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This paper includes a tutorial on how to reconstruct in-line holograms using an inverse problems approach, starting with modeling the observations, selecting regularizations and constraints, and ending with the design of a reconstruction algorithm. A special focus is placed on the connections between the numerous alternating projections strategies derived from Fienup's phase retrieval technique and the inverse problems framework. In particular, an interpretation of Fienup's algorithm as iterates of a proximal gradient descent for a particular cost function is given. Reconstructions from simulated and experimental holograms of micrometric beads illustrate the theoretical developments. The results show that the transition from alternating projections techniques to the inverse problems formulation is straightforward and advantageous.
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Reconstruction of phase objects is a central problem in digital holography, whose various applications include microscopy, biomedical imaging, and fluid mechanics. Starting from a single in-line hologram, there is no direct way to recover the phase of the diffracted wave in the hologram plane. The reconstruction of absorbing and phase objects therefore requires the inversion of the non-linear hologram formation model. We propose a regularized reconstruction method that includes several physically-grounded constraints such as bounds on transmittance values, maximum/minimum phase, spatial smoothness or the absence of any object in parts of the field of view. To solve the non-convex and non-smooth optimization problem induced by our modeling, a variable splitting strategy is applied and the closed-form solution of the sub-problem (the so-called proximal operator) is derived. The resulting algorithm is efficient and is shown to lead to quantitative phase estimation on reconstructions of accurate simulations of in-line holograms based on the Mie theory. As our approach is adaptable to several in-line digital holography configurations, we present and discuss the promising results of reconstructions from experimental in-line holograms obtained in two different applications: the tracking of an evaporating droplet (size â¼ 100µm) and the microscopic imaging of bacteria (size â¼ 1µm).
Assuntos
Líquidos Corporais/fisiologia , Holografia/métodos , Aumento da Imagem/métodos , Interpretação de Imagem Assistida por Computador/métodos , Microbiologia , Microscopia/métodos , Algoritmos , Desenho de Equipamento , Escherichia coli/citologia , Fenômenos Físicos , Staphylococcus epidermidis/citologiaRESUMO
We propose a three-dimensional (3D) imaging platform based on lens-free microscopy to perform multiangle acquisitions on 3D cell cultures embedded in extracellular matrices. Lens-free microscopy acquisitions present some inherent issues such as the lack of phase information on the sensor plane and a limited angular coverage. We developed and compared three different algorithms based on the Fourier diffraction theorem to obtain fully 3D reconstructions. These algorithms present an increasing complexity associated with a better reconstruction quality. Two of them are based on a regularized inverse problem approach. To compare the reconstruction methods in terms of artefact reduction, signal-to-noise ratio, and computation time, we tested them on two experimental datasets: an endothelial cell culture and a prostate cell culture grown in a 3D extracellular matrix with large reconstructed volumes up to â¼5 mm3 with a resolution sufficient to resolve isolated single cells. The lens-free reconstructions compare well with standard microscopy.
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Tomographic iterative reconstruction methods need a very thorough modeling of data. This point becomes critical when the number of available projections is limited. At the core of this issue is the projector design, i.e., the numerical model relating the representation of the object of interest to the projections on the detector. Voxel driven and ray driven projection models are widely used for their short execution time in spite of their coarse approximations. Distance driven model has an improved accuracy but makes strong approximations to project voxel basis functions. Cubic voxel basis functions are anisotropic, accurately modeling their projection is, therefore, computationally expensive. Both smoother and more isotropic basis functions better represent the continuous functions and provide simpler projectors. These considerations have led to the development of spherically symmetric volume elements, called blobs. Set apart their isotropy, blobs are often considered too computationally expensive in practice. In this paper, we consider using separable B-splines as basis functions to represent the object, and we propose to approximate the projection of these basis functions by a 2D separable model. When the degree of the B-splines increases, their isotropy improves and projections can be computed regardless of their orientation. The degree and the sampling of the B-splines can be chosen according to a tradeoff between approximation quality and computational complexity. We quantitatively measure the good accuracy of our model and compare it with other projectors, such as the distance-driven and the model proposed by Long et al. From the numerical experiments, we demonstrate that our projector with an improved accuracy better preserves the quality of the reconstruction as the number of projections decreases. Our projector with cubic B-splines requires about twice as many operations as a model based on voxel basis functions. Higher accuracy projectors can be used to improve the resolution of the existing systems, or to reduce the number of projections required to reach a given resolution, potentially reducing the dose absorbed by the patient.