The shape of objects is an important source of visual information in a wide range of applications. One of the core challenges of shape quantification is to ensure that the extracted measurements remain invariant to transformations that preserve an object’s intrinsic geometry, such as changing its size, orientation, and position in the image. In this work, we introduce ShapeEmbed, a self-supervised representation learning framework designed to encode the contour of objects in 2D images, represented as a Euclidean distance matrix, into a shape descriptor that is invariant to translation, scaling, rotation, reflection, and point indexing. Our approach overcomes the limitations of traditional shape descriptors while improving upon existing state-of-the-art autoencoder-based approaches. We demonstrate that the descriptors learned by our framework outperform their competitors in shape classification tasks on natural and biological images. We envision our approach to be of particular relevance to biological imaging applications.

SUMMARY: We introduce BioImage Archive Binder (bia-binder), an open-source, cloud-architectured, and web-based coding environment tailored to bioimage analysis that is freely accessible to all researchers. The service generates easy-to-use Jupyter Notebook coding environments hosted on EMBL-EBI's Embassy Cloud, an academically hosted compute service which provides significant computational resources. The bia-binder architecture is free, open-source and publicly available for deployment. It features fast and direct access to images in the BioImage Archive, the Image Data Resource, and the BioStudies databases. We believe that this service can play a role in mitigating the current inequalities in access to scientific resources across academia. As bia-binder produces permanent links to compiled coding environments, we foresee the service to become widely used within the community and enable exploratory research. AVAILABILITY AND IMPLEMENTATION: bia-binder is built and deployed using helmsman and helm and released under the MIT licence. It can be accessed at binder.bioimagearchive.org and runs on any standard web browser.

High-resolution posture tracking of C. elegans has applications in genetics, neuroscience, and drug screening. While classic methods can reliably track isolated worms on uniform backgrounds, they fail when worms overlap, coil, or move in complex environments. Model-based tracking and deep learning approaches have addressed these issues to an extent, but there is still significant room for improvement in tracking crawling worms. Here we train a version of the DeepTangle algorithm developed for swimming worms using a combination of data derived from Tierpsy tracker and hand-annotated data for more difficult cases. DeepTangleCrawl (DTC) outperforms existing methods, reducing failure rates and producing more continuous, gap-free worm trajectories that are less likely to be interrupted by collisions between worms or self-intersecting postures (coils). We show that DTC enables the analysis of previously inaccessible behaviours and increases the signal-to-noise ratio in phenotypic screens, even for data that was specifically collected to be compatible with legacy trackers including low worm density and thin bacterial lawns. DTC broadens the applicability of high-throughput worm imaging to more complex behaviours that involve worm-worm interactions and more naturalistic environments including thicker bacterial lawns.

MOTIVATION: The nuclear pore complex (NPC) is the only passageway for macromolecules between nucleus and cytoplasm, and an important reference standard in microscopy: it is massive and stereotypically arranged. The average architecture of NPC proteins has been resolved with pseudoatomic precision, however observed NPC heterogeneities evidence a high degree of divergence from this average. Single-molecule localization microscopy (SMLM) images NPCs at protein-level resolution, whereupon image analysis software studies NPC variability. However, the true picture of this variability is unknown. In quantitative image analysis experiments, it is thus difficult to distinguish intrinsically high SMLM noise from variability of the underlying structure. RESULTS: We introduce CIR4MICS ('ceramics', Configurable, Irregular Rings FOR MICroscopy Simulations), a pipeline that synthesizes ground truth datasets of structurally variable NPCs based on architectural models of the true NPC. Users can select one or more N- or C-terminally tagged NPC proteins, and simulate a wide range of geometric variations. We also represent the NPC as a spring-model such that arbitrary deforming forces, of user-defined magnitudes, simulate irregularly shaped variations. Further, we provide annotated reference datasets of simulated human NPCs, which facilitate a side-by-side comparison with real data. To demonstrate, we synthetically replicate a geometric analysis of real NPC radii and reveal that a range of simulated variability parameters can lead to observed results. Our simulator is therefore valuable to test the capabilities of image analysis methods, as well as to inform experimentalists about the requirements of hypothesis-driven imaging studies. AVAILABILITY AND IMPLEMENTATION: Code: https://github.com/uhlmanngroup/cir4mics. Simulated data: BioStudies S-BSST1058.

MOTIVATION: Rotated template matching is an efficient and versatile algorithm to analyze microscopy images, as it automates the detection of stereotypical structures, such as organelles that can appear at any orientation. Its performance however quickly degrades in noisy image data. RESULTS: We introduce Steer'n'Detect, an ImageJ plugin implementing a recently published algorithm to detect patterns of interest at any orientation with high accuracy from a single template in 2D images. Steer'n'Detect provides a faster and more robust substitute to template matching. By adapting to the statistics of the image background, it guarantees accurate results even in the presence of noise. The plugin comes with an intuitive user interface facilitating results analysis and further post-processing. AVAILABILITY AND IMPLEMENTATION: https://github.com/Biomedical-Imaging-Group/Steer-n-Detect. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

We provide a complete pipeline for the detection of patterns of interest in an image. In our approach, the patterns are assumed to be adequately modeled by a known template, and are located at unknown positions and orientations that we aim at retrieving. We propose a continuous-domain additive image model, where the analyzed image is the sum of the patterns to localize and a background with self-similar isotropic power-spectrum. We are then able to compute the optimal filter fulfilling the SNR criterion based on one single template and background pair: it strongly responds to the template while being optimally decoupled from the background model. In addition, we constrain our filter to be steerable, which allows for a fast template detection together with orientation estimation. In practice, the implementation requires to discretize a continuous-domain formulation on polar grids, which is performed using quadratic radial B-splines. We demonstrate the practical usefulness of our method on a variety of template approximation and pattern detection experiments. We show that the detection performance drastically improves when we exploit the statistics of the background via its power-spectrum decay, which we refer to as spectral-shaping. The proposed scheme outperforms state-of-the-art steerable methods by up to 50% of absolute detection performance.

We propose a novel sparse dictionary learning method for planar shapes in the sense of Kendall, namely configurations of landmarks in the plane considered up to similitudes. Our shape dictionary method provides a good trade-off between algorithmic simplicity and faithfulness with respect to the nonlinear geometric structure of Kendall's shape space. Remarkably, it boils down to a classical dictionary learning formulation modified using complex weights. Existing dictionary learning methods extended to nonlinear spaces map the manifold either to a reproducing kernel Hilbert space or to a tangent space. The first approach is unnecessarily heavy in the case of Kendall's shape space and causes the geometrical understanding of shapes to be lost, while the second one induces distortions and theoretical complexity. Our approach does not suffer from these drawbacks. Instead of embedding the shape space into a linear space, we rely on the hyperplane of centered configurations, including preshapes from which shapes are defined as rotation orbits. In this linear space, the dictionary atoms are scaled and rotated using complex weights before summation. Furthermore, our formulation is more general than Kendall's original one: it applies to discretely defined configurations of landmarks as well as continuously defined interpolating curves. We implemented our algorithm by adapting the method of optimal directions combined to a Cholesky-optimized order recursive matching pursuit. An interesting feature of our shape dictionary is that it produces visually realistic atoms, while guaranteeing reconstruction accuracy. Its efficiency can mostly be attributed to a clear formulation of the framework with complex numbers. We illustrate the strong potential of our approach for the characterization of datasets of shapes up to similitudes and the analysis of patterns in deforming two-dimensional shapes.

Motivation: We introduce a formulation for the general task of finding diverse shortest paths between two end-points. Our approach is not linked to a specific biological problem and can be applied to a large variety of images thanks to its generic implementation as a user-friendly ImageJ/Fiji plugin. It relies on the introduction of additional layers in a Viterbi path graph, which requires slight modifications to the standard Viterbi algorithm rules. This layered graph construction allows for the specification of various constraints imposing diversity between solutions. Results: The software allows obtaining a collection of diverse shortest paths under some user-defined constraints through a convenient and user-friendly interface. It can be used alone or be integrated into larger image analysis pipelines. Availability and implementation: http://bigwww.epfl.ch/algorithms/diversepathsj. Contact: [email protected] or [email protected]. Supplementary information: Supplementary data are available at Bioinformatics online.

We present a new family of snakes that satisfy the property of multiresolution by exploiting subdivision schemes. We show in a generic way how to construct such snakes based on an admissible subdivision mask. We derive the necessary energy formulations and provide the formulas for their efficient computation. Depending on the choice of the mask, such models have the ability to reproduce trigonometric or polynomial curves. They can also be designed to be interpolating, a property that is useful in user-interactive applications. We provide explicit examples of subdivision snakes and illustrate their use for the segmentation of bioimages. We show that they are robust in the presence of noise and provide a multiresolution algorithm to enlarge their basin of attraction, which decreases their dependence on initialization compared to singleresolution snakes. We show the advantages of the proposed model in terms of computation and segmentation of structures with different sizes.

BACKGROUND: Automated classification using machine learning often relies on features derived from segmenting individual objects, which can be difficult to automate. WND-CHARM is a previously developed classification algorithm in which features are computed on the whole image, thereby avoiding the need for segmentation. The algorithm obtained encouraging results but requires considerable computational expertise to execute. Furthermore, some benchmark sets have been shown to be subject to confounding artifacts that overestimate classification accuracy. RESULTS: We developed CP-CHARM, a user-friendly image-based classification algorithm inspired by WND-CHARM in (i) its ability to capture a wide variety of morphological aspects of the image, and (ii) the absence of requirement for segmentation. In order to make such an image-based classification method easily accessible to the biological research community, CP-CHARM relies on the widely-used open-source image analysis software CellProfiler for feature extraction. To validate our method, we reproduced WND-CHARM's results and ensured that CP-CHARM obtained comparable performance. We then successfully applied our approach on cell-based assay data and on tissue images. We designed these new training and test sets to reduce the effect of batch-related artifacts. CONCLUSIONS: The proposed method preserves the strengths of WND-CHARM - it extracts a wide variety of morphological features directly on whole images thereby avoiding the need for cell segmentation, but additionally, it makes the methods easily accessible for researchers without computational expertise by implementing them as a CellProfiler pipeline. It has been demonstrated to perform well on a wide range of bioimage classification problems, including on new datasets that have been carefully selected and annotated to minimize batch effects. This provides for the first time a realistic and reliable assessment of the whole image classification strategy.

A crucial component of steerable wavelets is the radial profile of the generating function in the frequency domain. In this paper, we present an infinite-dimensional optimization scheme that helps us find the optimal profile for a given criterion over the space of tight frames. We consider two classes of criteria that measure the localization of the wavelet. The first class specifies the spatial localization of the wavelet profile, and the second that of the resulting wavelet coefficients. From these metrics and the proposed algorithm, we construct tight wavelet frames that are optimally localized and provide their analytical expression. In particular, one of the considered criterion helps us finding back the popular Simoncelli wavelet profile. Finally, the investigation of local orientation estimation, image reconstruction from detected contours in the wavelet domain, and denoising indicate that optimizing wavelet localization improves the performance of steerable wavelets, since our new wavelets outperform the traditional ones.

We propose a framework for the detection of junctions in images. Although the detection of edges and key points is a well examined and described area, the multiscale detection of junction centers, especially for odd orders, poses a challenge in pattern analysis. The goal of this paper is to build optimal junction detectors based on 2D steerable wavelets that are polar-separable in the Fourier domain. The approaches we develop are general and can be used for the detection of arbitrary symmetric and asymmetric junctions. The backbone of our construction is a multiscale pyramid with a radial wavelet function where the directional components are represented by circular harmonics and encoded in a shaping matrix. We are able to detect M -fold junctions in different scales and orientations. We provide experimental results on both simulated and real data to demonstrate the effectiveness of the algorithm.

We introduce a new model of parametric contours defined in a continuous fashion. Our curve model relies on Hermite spline interpolation and can easily generate curves with sharp discontinuities; it also grants direct access to the tangent at each location. With these two features, the Hermite snake distinguishes itself from classical spline-snake models and allows one to address certain bioimaging problems in a more efficient way. More precisely, the Hermite snake construction allows introducing sharp corners in the snake curve and designing directional energy functionals relying on local orientation information in the input image. Using the formalism of spline theory, the model is shown to meet practical requirements such as invariance to affine transformations and good approximation properties. Finally, the dependence on initial conditions and the robustness to the noise is studied on synthetic data in order to validate our Hermite snake model, and its usefulness is illustrated on real biological images acquired using brightfield, phase-contrast, differential-interference-contrast, and scanning-electron microscopy.

Parametric active contours are an attractive approach for image segmentation, thanks to their computational efficiency. They are driven by application-dependent energies that reflect the prior knowledge on the object to be segmented. We propose an energy involving shape priors acting in a regularization-like manner. Thereby, the shape of the snake is orthogonally projected onto the space that spans the affine transformations of a given shape prior. The formulation of the curves is continuous, which provides computational benefits when compared with landmark-based (discrete) methods. We show that this approach improves the robustness and quality of spline-based segmentation algorithms, while its computational overhead is negligible. An interactive and ready-to-use implementation of the proposed algorithm is available and was successfully tested on real data in order to segment Drosophila flies and yeast cells in microscopic images.

We present a new exponential B-spline basis that enables the construction of active contours for the analysis of biomedical images. Our functions generalize the well-known polynomial Hermite B-splines and provide us with a direct control over the tangents of the parameterized contour, which is absent in traditional spline-based active contours. Our basis functions have been designed to perfectly reproduce elliptical and circular shapes. Moreover, they can approximate any closed curve up to arbitrary precision by increasing the number of anchor points. They are therefore well-suited to the segmentation of the roundish objects that are commonly encountered in the analysis of bioimages. We illustrate the performance of an active contour built using our functions on some examples of real biological data.

We study the issue of localization in the context of isotropic wavelet frames. We define a variance-type measure of localization and propose an algorithm based on calculus of variations to minimize this criterion under the constraint of a tight wavelet frame. Based on these calculations, we design the variance-optimal wavelet (VOW). Finally, we demonstrate the advantage of better localization in a practical image-processing task.