In this two-part paper, we present a novel framework and methodology to analyze data from certain image-based biochemical assays, e.g., ELISPOT and Fluorospot assays. In this second part, we focus on our algorithmic contributions. We provide an algorithm for functional inverse diffusion that solves the variational problem we posed in Part I. As part of the derivation of this algorithm, we present the proximal operator for the non-negative group-sparsity regularizer, which is a novel result that is of interest in itself, also in comparison to previous results on the proximal operator of a sum of functions. We then present a discretized approximated implementation of our algorithm and evaluate it both in terms of operational cell-detection metrics and in terms of distributional optimal-transport metrics.
In this two-part paper, we present a novel framework and methodology to analyze data from certain imagebased biochemical assays, e.g., ELISPOT and Fluorospot assays. In this first part, we start by presenting a physical partial differential equations (PDE) model up to image acquisition for these biochemical assays. Then, we use the PDEs' Green function to derive a novel parametrization of the acquired images. This parametrization allows us to propose a functional optimization problem to address inverse diffusion. In particular, we propose a non-negative group-sparsity regularized optimization problem with the goal of localizing and characterizing the biological cells involved in the said assays. We continue by proposing a suitable discretization scheme that enables both the generation of synthetic data and implementable algorithms to address inverse diffusion. We end Part I by providing a preliminary comparison between the results of our methodology and an expert human labeler on real data. Part II is devoted to providing an accelerated proximal gradient algorithm to solve the proposed problem and to the empirical validation of our methodology.
We propose the use of Flat Metric to assess the performance of reconstruction methods for single-molecule localization microscopy (SMLM) in scenarios where the ground-truth is available. Flat Metric is intimately related to the concept of optimal transport between measures of different mass, providing solid mathematical foundations for SMLM evaluation and integrating both localization and detection performance. In this paper, we provide the foundations of Flat Metric and validate this measure by applying it to controlled synthetic examples and to data from the SMLM 2016 Challenge.
Accurate cell detection and counting in the image-based ELISpot and FluoroSpot immunoassays is a challenging task. Recently proposed methodology matches human accuracy by leveraging knowledge of the underlying physical process of these assays and using proximal optimization methods to solve an inverse problem. Nonetheless, thousands of computationally expensive iterations are often needed to reach a near-optimal solution. In this paper, we exploit the structure of the iterations to design a parameterized computation graph, SpotNet, that learns the patterns embedded within several training images and their respective cell information. Further, we compare SpotNet to a convolutional neural network layout customized for cell detection. We show empirical evidence that, while both designs obtain a detection performance on synthetic data far beyond that of a human expert, SpotNet is easier to train and obtains better estimates of particle secretion for each cell.
Robustness and stability of image-reconstruction algorithms have recently come under scrutiny. Their importance to medical imaging cannot be overstated. We review the known results for the topical variational regularization strategies ( 2 and 1 regularization) and present novel stability results for p -regularized linear inverse problems for p ∈ (1, ∞). Our results guarantee Lipschitz continuity for small p and Hölder continuity for larger p. They generalize well to the L p (Ω) function spaces.
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