Convex Total Variation Denoising of Poisson Fluorescence Confocal Images With Anisotropic Filtering

Fluorescence confocal microscopy (FCM) is now one of the most important tools in biomedicine research. In fact, it makes it possible to accurately study the dynamic processes occurring inside the cell and its nucleus by following the motion of fluorescent molecules over time. Due to the small amount of acquired radiation and the huge optical and electronics amplification, the FCM images are usually corrupted by a severe type of Poisson noise. This noise may be even more damaging when very low intensity incident radiation is used to avoid phototoxicity. In this paper, a Bayesian algorithm is proposed to remove the Poisson intensity dependent noise corrupting the FCM image sequences. The observations are organized in a 3-D tensor where each plane is one of the images acquired along the time of a cell nucleus using the fluorescence loss in photobleaching (FLIP) technique. The method removes simultaneously the noise by considering different spatial and temporal correlations. This is accomplished by using an anisotropic 3-D filter that may be separately tuned in space and in time dimensions. Tests using synthetic and real data are described and presented to illustrate the application of the algorithm. A comparison with several state-of-the-art algorithms is also presented

Feature-preserving image restoration and its application in biological fluorescence microscopy

This thesis presents a new investigation of image restoration and its application to fluorescence cell microscopy. The first part of the work is to develop advanced image denoising algorithms to restore images from noisy observations by using a novel featurepreserving diffusion approach. I have applied these algorithms to different types of images, including biometric, biological and natural images, and demonstrated their superior performance for noise removal and feature preservation, compared to several state of the art methods. In the second part of my work, I explore a novel, simple and inexpensive super-resolution restoration method for quantitative microscopy in cell biology. In this method, a super-resolution image is restored, through an inverse process, by using multiple diffraction-limited (low) resolution observations, which are acquired from conventional microscopes whilst translating the sample parallel to the image plane, so referred to as translation microscopy (TRAM). A key to this new development is the integration of a robust feature detector, developed in the first part, to the inverse process to restore high resolution images well above the diffraction limit in the presence of strong noise. TRAM is a post-image acquisition computational method and can be implemented with any microscope. Experiments show a nearly 7-fold increase in lateral spatial resolution in noisy biological environments, delivering multi-colour image resolution of ~30 nm

SPITFIR(e): A supermaneuverable algorithm for fast denoising and deconvolution of 3D fluorescence microscopy images and videos

International audienceModern fluorescent microscopy imaging is still limited by the optical aberrations and the photon budget available in the specimen. A direct consequence is the necessity to develop flexible and "off-road" algorithms in order to recover structural details and improve spatial resolution, which is critical when restraining the illumination to low levels in order to limit photo-damages. Here, we report SPITFIR(e) a flexible method designed to accurately and quickly restore 2D-3D fluorescence microscopy images and videos (4D images). We designed a generic sparse-promoting regularizer to subtract undesirable out-of-focus background and we developed a primal-dual algorithm for fast optimization. SPITFIR(e) is a "swiss-knife" method for practitioners as it adapts to any microscopy techniques, to various sources of signal degradation (noise, blur), to variable image contents, aswell as to low signal-to-noise ratios. Our method outperforms existing state-of-the-art algorithms, and is more flexible than supervised deep-learning methods requiring ground truth datasets. The performance, the flexibility, and the ability to push the spatiotemporal resolution limit of sub-diffracted fluorescence microscopy techniques are demonstrated on experimental datasets acquired with various microscopy techniques from 3D spinning-disk confocal up to lattice light sheet microscopy

Mathematical Methods for the Quantification of Actin-Filaments in Microscopic Images

In cell biology confocal laser scanning microscopic images of the actin filament of human osteoblasts are produced to assess the cell development. This thesis aims at an advanced approach for accurate quantitative measurements about the morphology of the bright-ridge set of these microscopic images and thus about the actin filament. Therefore automatic preprocessing, tagging and quantification interplay to approximate the capabilities of the human observer to intuitively recognize the filaments correctly. Numerical experiments with random models confirm the accuracy of this approach

Non-parametric regression for patch-based fluorescence microscopy image sequence denoising

We present a non-parametric regression method for denoising 3D image sequences acquired in fluorescence microscopy. The proposed method exploits 3D+time information to improve the signal-to-noise ratio of images corrupted by mixed Poisson-Gaussian noise. A variance stabilization transform is first applied to the image-data to introduce independence between the mean and variance. This pre-processing requires the knowledge of parameters related to the acquisition system, also estimated in our approach. In a second step, we propose an original statistical patch-based framework for noise reduction and preservation of space-time discontinuities. In our study, discontinuities are related to small moving spots with high velocity observed in fluorescence video-microscopy. The idea is to minimize an objective nonlocal energy functional involving spatio-temporal image patches. The minimizer has a simple form and is defined as the weighted average of input data taken in spatially-varying neighborhoods. The size of each neighborhood is optimized to improve the performance of the pointwise estimator. The performance of the algorithm which requires no motion estimation, is then demonstrated on both synthetic and real image sequences using qualitative and quantitative criteria

Coupling Image Restoration and Segmentation: A Generalized Linear Model/Bregman Perspective

We introduce a new class of data-fitting energies that couple image segmentation with image restoration. These functionals model the image intensity using the statistical framework of generalized linear models. By duality, we establish an information-theoretic interpretation using Bregman divergences. We demonstrate how this formulation couples in a principled way image restoration tasks such as denoising, deblurring (deconvolution), and inpainting with segmentation. We present an alternating minimization algorithm to solve the resulting composite photometric/geometric inverse problem.We use Fisher scoring to solve the photometric problem and to provide asymptotic uncertainty estimates. We derive the shape gradient of our data-fitting energy and investigate convex relaxation for the geometric problem. We introduce a new alternating split- Bregman strategy to solve the resulting convex problem and present experiments and comparisons on both synthetic and real-world images