bioRxiv Science⌕ Search

Biology subjects

Gomez-de-Mariscal, E.

Publications and source records attributed to Gomez-de-Mariscal, E..

13 recordsLinked to original sources

PhenoMe: An interactive AI framework for interpretable, microscopy-driven phenotyping

Extracting meaningful biological patterns from microscopy image datasets requires tools that quantitatively connect visual similarities and experimental context. Deep learning models, and in particular novel foundation models, have the capacity to translate images into high-dimensional embeddings that can better represent complex visual patterns. Using these complex representations analytically and linking them to experimental information, it is possible to build advanced exploratory and phenotyping capabilities. Here, we present PhenoMe, an open-source, model-agnostic framework to handle pretrained foundation model-derived image representations alongside quantitative image properties and experimental metadata. PhenoMe integrates multiple scriptable and interactive workflows for embedding space visualisation, dimensionality reduction and clustering, similarity-based comparison and retrieval of representative images for each phenotype, among others. It also supports correlation analyses that link the embedding structure to interpretable image properties. We demonstrate PhenoMe's capabilities on seven public microscopy datasets, ranging from cell painting assays, nanoscale synaptic images, drug-response analyses and dynamic cell-phenotyping. These case studies show how readily available image representations can support data contextualisation, curation and analysis. Through guided notebooks, reproducible configurations, extensible model wrappers and a napari plugin, PhenoMe makes representation-based microscopy exploration accessible to researchers without dedicated technical expertise. PhenoMe lays the groundwork for applying foundation models and representation learning to microscopy data analysis.

bioinformatics↗

AI4Life Open Calls and Public Challenges: why, how, and what we have learned.

Within AI4Life, we ran three Open Calls and three Public Challenges (2023-2025), supporting 22 bioimage analysis projects from 151 applications and engaging 225 challenge participants, with the aim of applying FAIR deep learning in the life sciences. Our experience offers a view of the current state of bioimage analysis, the landscape of available tools, as well as the existing gaps between method developers, tool producers and potential users. It highlights that even after careful selection for AI-ready projects, most still require substantial effort to apply deep learning, and that the field still relies heavily on established, well-rounded methods to solve common problems. We come to the conclusion that for scientific AI in biology, the rate-limiting step is not methods and models but data, annotations, and shared infrastructure underneath them.

bioinformatics↗

ReScale4DL: Balancing Pixel and Contextual Information for Enhanced Bioimage Segmentation

Deep learning has established itself as the state-of-the-art approach for segmentation in bioimage analysis. However, these powerful algorithms present an intriguing paradox regarding image resolution: contrary to intuition, lower-resolution images can yield superior performance for specific image analysis carried out by deep learning. This phenomenon is particularly significant in microscopy imaging, where high-resolution acquisitions come with substantial costs in throughput, data storage requirements, and potential photodamage to specimens. Through systematic experimentation, we evaluate how varying image resolution impacts deep learning performance in cellular image segmentation tasks. We trained popular architectures on datasets downsampled to 6-50% of their original resolution, mimicking acquisitions at lower image magnification, and compared their performance against models trained on native-resolution images. Our results show that segmentation accuracy either improves (by up to 25% of mean Intersection over Union (IoU)) or experiences only minimal degradation (< 5% of mean IoU) when using images downsampled by up to a factor of 4 (25% of the original resolution). This downsampling proportionally increases information throughput while reducing data storage requirements and inference time. With these findings, we contribute systematic guidelines to deep learning practitioners in creating efficient experimental pipelines for image-driven discoveries. This approach improves the sustainability and cost-effectiveness of bioimaging studies by reducing data and computing needs while optimising microscopy techniques.

biophysics↗

DINOSim: Zero-Shot Object Detection and Semantic Segmentation on Electron Microscopy Images

We present DINOSim, a novel method for detecting and segmenting objects in microscopy images without the need for large annotated datasets or additional training. DINOSim builds on the pretrained DINOv2 image encoder, which captures semantic information from images. By comparing the encoders features of images patches to those of a user-selected reference, DINOSim generates pseudo-labels that guide object detection and segmentation. Subsequently, a k-nearest neighbors framework is then used to refine predictions across new images. Our experiments show that DINOSim can effectively identify and segment previously unseen objects in diverse microscopy datasets, offering performance comparable to supervised approaches while avoiding the need for costly manual labeling. We also investigate how different choices of user prompts selection and model size affect accuracy and generalization. To make the method widely accessible, we provide an open-source Napari plugin (github.com/AAitorG/napari-DINOSim), enabling researchers to easily apply DINOSim to their own data. Overall, DINOSim offers a fast, flexible and practical solution for bioimage analysis, particularly valuable in resource-constrained settings.

bioinformatics↗

PhotoFiTT: A Quantitative Framework for Assessing Phototoxicity in Live-Cell Microscopy Experiments

Phototoxicity in live-cell fluorescence microscopy can compromise experimental outcomes, yet quantitative methods to assess its impact remain limited. Here we present PhotoFiTT (Phototoxicity Fitness Time Trial), an integrated framework combining a standardised experimental protocol with advanced image analysis to quantify light-induced cellular stress in label-free settings. PhotoFiTT leverages machine learning and cell cycle dynamics to analyse mitotic timing, cell size changes, and overall cellular activity in response to controlled light exposure. Using adherent mammalian cells, we demonstrate PhotoFiTTs ability to detect wavelength- and dose-dependent effects, showcasing that near-UV light induces significant mitotic delays at doses as low as 0.6 J/cm2, while longer wavelengths require higher doses for comparable effects. PhotoFiTT enables researchers to establish quantitative benchmarks for acceptable levels of photodamage, facilitating the optimisation of imaging protocols that balance image quality with sample health.

cell biology↗

Gut Analysis Toolbox: Automating quantitative analysis of enteric neurons

The enteric nervous system (ENS) plays an important role in coordinating gut function. The ENS consists of an extensive network of neurons and glial cells within the wall of the gastrointestinal tract. Alterations in neuronal distribution, function, and type are strongly associated with enteric neuropathies and gastrointestinal (GI) dysfunction and can serve as biomarkers for disease. However, current methods for assessing neuronal counts and distribution suffer from undersampling. This is partly due to challenges associated with imaging and analyzing large tissue areas, and operator bias due to manual analysis. Here, we present the Gut Analysis Toolbox (GAT), an image analysis tool designed for characterization of enteric neurons and their neurochemical coding using 2D images of GI wholemount preparations. GAT is developed for the Fiji distribution of ImageJ. It has a user-friendly interface and offers rapid and accurate cell segmentation. Custom deep learning (DL) based cell segmentation models were developed using StarDist. GAT also includes a ganglion segmentation model which was developed using deepImageJ. In addition, GAT allows importing of segmentation generated by other software. DL models have been trained using ZeroCostDL4Mic on diverse datasets sourced from different laboratories. This captures the variability associated with differences in animal species, image acquisition parameters, and sample preparation across research groups. We demonstrate the robustness of the cell segmentation DL models by comparing them against the state-of-the-art cell segmentation software, Cellpose. To quantify neuronal distribution GAT applies proximal neighbor-based spatial analysis. We demonstrate how the proximal neighbor analysis can reveal differences in cellular distribution across gut regions using a published dataset. In summary, GAT provides an easy-to-use toolbox to streamline routine image analysis tasks in ENS research. GAT enhances throughput allowing unbiased analysis of larger tissue areas, multiple neuronal markers and numerous samples rapidly.

neuroscience↗

Bridging the Gap: Integrating Cutting-edge Techniques into Biological Imaging with deepImageJ

This manuscript showcases the latest advancements in deepImageJ, a pivotal Fiji/ImageJ plugin for bioimage analysis in the life sciences. The plugin, known for its user-friendly interface, facilitates the application of diverse pre-trained neural networks to custom data. The manuscript demonstrates a number of deepImageJ capabilities, particularly in executing complex pipelines, 3D analysis, and processing large images. A key development is the integration of the Java Deep Learning Library (JDLL), expanding deepImageJs compatibility with various deep learning frameworks, including TensorFlow, PyTorch, and ONNX. This allows for running multiple engines within a single Fiji/ImageJ instance, streamlining complex bioimage analysis tasks. The manuscript details three case studies to demonstrate these capabilities. The first explores integrated image-to image translation and nuclei segmentation. The second focuses on 3D nuclei segmentation. The third case study deals with large image segmentation. These studies underscore deepImageJs versatility and power in bioimage analysis, emphasizing its role as a critical tool for life scientists and researchers. The advancements in deepImageJ bridge the gap between deep learning model developers and end-users, enabling a more accessible and efficient approach to biological image analysis. The advancements in deepImageJ, detailed in this paper, represent a significant leap in bioimage analysis, crucial for life sciences. By enhancing this Fiji/ImageJ plugin, the research bridges the gap between complex deep learning models and practical applications, making advanced bioimage analysis accessible to a broader audience. This integration of the Java Deep Learning Library (JDLL) within deepImageJ is particularly noteworthy, as it expands compatibility with leading deep learning frameworks. This allows for the seamless execution of multiple models in a single instance, simplifying the construction of complex image analysis pipelines. The implications of this research are far-reaching, extending beyond academic circles to potentially impact various sectors, including healthcare, pharmaceuticals, and biotechnology. The enhanced capabilities of deepImageJ in handling intricate pipelines, 3D analysis, and large images facilitate detailed and efficient analysis of biological data. Such advancements are vital for accelerating research and development in medical imaging, drug discovery, and understanding complex biological processes. This manuscript contribution to the field of bioimage analysis is significant, offering a tool that empowers researchers, irrespective of their computational expertise, to leverage advanced technologies in their work. The wide applicability and ease of use of deepImageJ have the potential to foster interdisciplinary collaborations, drive innovation, and facilitate discoveries across various scientific and industrial sectors.

bioinformatics↗

DL4MicEverywhere: Deep learning for microscopy made flexible, shareable, and reproducible

Deep learning has revolutionised the analysis of extensive microscopy datasets, yet challenges persist in the widespread adoption of these techniques. Many lack access to training data, computing resources, and expertise to develop complex models. We introduce DL4MicEverywhere, advancing our previous ZeroCostDL4Mic platform, to make deep learning more accessible. DL4MicEverywhere uniquely allows flexible training and deployment across diverse computational environments by encapsulating methods in interactive Jupyter notebooks within Docker containers -a standalone virtualisation of required packages and code to reproduce a computational environment-. This enhances reproducibility and convenience. The platform includes twice as many techniques as originally provided by ZeroCostDL4Mic and enables community contributions via automated build pipelines. DL4MicEverywhere empowers participatory innovation and aims to democratise deep learning for bioimage analysis.

bioinformatics↗

Transertion and cell geometry organize the Escherichia coli nucleoid during rapid growth

AO_SCPLOWBSTRACTC_SCPLOWBacterial chromosomes are spatiotemporally organized and sensitive to environmental changes. However, the mechanisms underlying chromosome configuration and reorganization are currently not fully understood. Using 3D single-molecule localization microscopy and live-cell imaging, we show that the Escherichia coli nucleoid adopts a condensed, membrane-associated configuration during rapid growth. To study the influence of different biosynthetic processes on nucleoid morphology and positioning, we recorded multi-colour super-resolution images during drug treatment. After developing analysis routines for confocal and super-resolution images, we captured highly resolved snapshots which revealed the complete loss of the membrane-bound state of the nucleoid within 10 minutes of halting transcription and translation. This indicates an active role of transertion (coupled transcription, translation and membrane insertion) in nucleoid organization. In contrast, cell wall synthesis inhibition only affects nucleoid organization during morphological changes. Further, we provide evidence that the E. coli nucleoid spatially correlates with MreB in unperturbed E. coli cells, while this correlation diminishes in cells with changed cell geometry or upon inhibition of protein biosynthesis. Replication inhibition experiments, as well as multi-drug treatments highlight the role of entropic effects and transcription in nucleoid condensation and positioning. In summary, we provide experimental evidence for transertion as a principal organiser of the bacterial nucleoid, and show that an altered metabolic state and antibiotic treatment both lead to major changes in the degree of transertion and overall spatial organization of the nucleoid. Our high-resolution characterization reveals dynamics of antibiotic action and provides tools to quantify bacterial chromosome organization. This does not only provide valuable insights into the role of transertion, but can also be applied to study other cell-biological processes.

microbiology↗

Small extracellular vesicles promote stiffness-mediated metastasis

Tissue stiffness is a critical prognostic factor in breast cancer and is associated with metastatic progression. Here we show an alternative and complementary hypothesis of tumor progression whereby physiological matrix stiffness affects the quantity and protein cargo of small EVs produced by cancer cells, which in turn drive their metastasis. Primary patient breast tissue produces significantly more EVs from stiff tumor tissue than soft tumor adjacent tissue. EVs released by cancer cells on matrices that model human breast tumors (25 kPa; stiff EVs) feature increased adhesion molecule presentation (ITG2{beta}1, ITG6{beta}4, ITG6{beta}1, CD44) compared to EVs from softer normal tissue (0.5 kPa; soft EVs), which facilitates their binding to extracellular matrix (ECM) protein collagen IV, and a 3-fold increase in homing ability to distant organs in mice. In a zebrafish xenograft model, stiff EVs aid cancer cell dissemination through enhanced chemotaxis. Moreover, normal, resident lung fibroblasts treated with stiff and soft EVs change their gene expression profiles to adopt a cancer associated fibroblast (CAF) phenotype. These findings show that EV quantity, cargo, and function depend heavily on the mechanical properties of the extracellular microenvironment. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/545937v3_ufig1.gif" ALT="Figure 1"> View larger version (42K): org.highwire.dtl.DTLVardef@1be1dbdorg.highwire.dtl.DTLVardef@928710org.highwire.dtl.DTLVardef@1e14e9borg.highwire.dtl.DTLVardef@efdb7f_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

MLL1 regulates cytokine-driven cell migration and metastasis

Cell migration is a critical requirement for cancer metastasis. Cytokine production and its role in cancer cell migration have been traditionally associated with immune cells in the tumor microenvironment. MLL1 is a histone methyltransferase that controls 3D cell migration via the secretion of cytokines, IL-6 and TGF-{beta}1, by the cancer cells themselves. In vivo, MLL1 depletion reduced metastatic burden and prolonged survival. MLL1 exerts its effects with its scaffold protein, Menin. Mechanistically, the MLL1-Menin interaction controls actin filament assembly via the IL-6/pSTAT3/Arp3 axis and acto-myosin contractility via the TGF-{beta}1/Gli2/ROCK1/2/pMLC2 axis, which regulate dynamic protrusion generation and 3D cell migration. MLL1 also regulates cell proliferation via mitosis-based and cell cycle-related pathways. Combining an MLL1-Menin inhibitor with Paclitaxel, a standard chemotherapeutic, abrogated tumor growth and metastasis in a syngeneic model. These results highlight the potential of targeting the MLL1 in metastasis prevention and its potential to be combined with currently administered chemotherapeutics. Statement of SignificanceWe identify MLL1 as being vital to metastasis, which causes the vast majority of cancer-related deaths. MLL1 controls cell migration, a requirement for metastasis, by regulating the secretion of cytokines. MLL1 inhibition lowers metastatic burden independent of its impact on primary tumor growth, highlighting its anti-metastatic potential in TNBC.

cancer biology↗

BioImage Model Zoo: A Community-Driven Resource for Accessible Deep Learning in BioImage Analysis

Deep learning-based approaches are revolutionizing imaging-driven scientific research. However, the accessibility and reproducibility of deep learning-based workflows for imaging scientists remain far from sufficient. Several tools have recently risen to the challenge of democratizing deep learning by providing user-friendly interfaces to analyze new data with pre-trained or fine-tuned models. Still, few of the existing pre-trained models are interoperable between these tools, critically restricting a models overall utility and the possibility of validating and reproducing scientific analyses. Here, we present the BioImage Model Zoo (https://bioimage.io): a community-driven, fully open resource where standardized pre-trained models can be shared, explored, tested, and downloaded for further adaptation or direct deployment in multiple end user-facing tools (e.g., ilastik, deepImageJ, QuPath, StarDist, ImJoy, ZeroCostDL4Mic, CSBDeep). To enable everyone to contribute and consume the Zoo resources, we provide a model standard to enable cross-compatibility, a rich list of example models and practical use-cases, developer tools, documentation, and the accompanying infrastructure for model upload, download and testing. Our contribution aims to lay the groundwork to make deep learning methods for microscopy imaging findable, accessible, interoperable, and reusable (FAIR) across software tools and platforms.

bioinformatics↗

DeepBacs: Bacterial image analysis using open-source deep learning approaches

Deep Learning (DL) is rapidly changing the field of microscopy, allowing for efficient analysis of complex data while often out-performing classical algorithms. This revolution has led to a significant effort to create user-friendly tools allowing biomedical researchers with little background in computer sciences to use this technology effectively. Thus far, these approaches have mainly focused on analysing microscopy images from eukaryotic samples and are still underused in microbiology. In this work, we demonstrate how to use a range of state-of-the-art artificial neural-networks particularly suited for the analysis of bacterial microscopy images, using our recently developed ZeroCostDL4Mic platform. We showcase different DL approaches for segmenting bright field and fluorescence images of different bacterial species, use object detection to classify different growth stages in time-lapse imaging data, and carry out DL-assisted phenotypic profiling of antibiotic-treated cells. To also demonstrate the DL capacity to enhance low-phototoxicity live-cell microscopy, we showcase how image denoising can allow researchers to attain high-fidelity data in faster and longer imaging. Finally, artificial labelling of cell membranes and predictions of super-resolution images allow for accurate mapping of cell shape and intracellular targets. To aid in the training of novice users, we provide a purposefully-built database of training and testing data, enabling bacteriologists to quickly explore how to analyse their data through DL. We hope this lays a fertile ground for the efficient application of DL in microbiology and fosters the creation of novel tools for bacterial cell biology and antibiotic research.

microbiology↗