Volume electron microscopy (EM) now enables nanometric-scale 3D reconstructions of neural tissue, opening the door to quantitative, morphology-driven neuroscience beyond connectivity alone. While previous studies relied on handcrafted descriptors and classical machine learning for morphology analysis, recent progress in deep learning for 3D shape understanding offers new opportunities to learn robust, task-specific representations directly from geometric data. In this paper we present NeMoCo , a geometry learning framework that targets the key practical bottleneck in connectomics and ultrastructural analysis: the scarcity and cost of dense expert annotations for the long tail of neurite and organelle phenotypes. NeMoCo formulates representation learning for EM-derived neurite meshes in a self-supervised Momentum Contrast (MoCo) style. We use DiffusionNet (Sharp et al., 2022) as a mesh encoder with intrinsic spectral descriptors (HKS) and train with a momentum-updated teacher encoder and a large memory bank of negatives. To learn invariances that are essential in practice, we generate paired geometric views via controlled affine transformations and resolution changes (including mesh decimation), encouraging embeddings to be stable under nuisance variability while remaining discriminative. We provide an extensive study of augmentation strength and temperature, and evaluate learned representations through frozen retrieval and non-parametric classification (frozen kNN), as well as downstream supervised fine-tuning under limited labels. NeMoCo demonstrates that MoCo-style self-supervision yields robust neurite morphology embeddings on EM meshes, improving label-efficiency and offering a scalable foundation for retrieval, clustering, and phenotype discovery in ultrastructural neuroscience. All the data and the code used for NeMoCo are available at https://github.com/Uzshah/NeMoCo .

This systematic review and meta-analysis comprehensively analyzes deep learning approaches for brain electron microscopy (EM) segmentation, addressing the critical challenge of extracting neuroanatomical information at nanometer resolution. Following PRISMA guidelines, we identified 60 studies through structured database searches, with quantitative meta-analysis of 27 studies (46 experiments) across 10 datasets providing the first unified benchmark comparison in this domain. Our analysis reveals a field transitioning from traditional CNN approaches toward foundation models and hybrid architectures. The meta-analysis demonstrates that foundation models outperform traditional CNNs by 13%–35% across key metrics, with the 3D Transformer + U-Net achieving the highest composite score (0.954) across five datasets. Meta-analysis confirms significant advantages for foundation models in instance-based metrics (Cohen’s d = − 6 . 44 ), while only 26% of experiments validate across multiple datasets. Four key evolutionary trends emerge: (1) transition from 2D to 3D architectures optimized for ultrastructural complexity; (2) development of topology-preserving loss functions and evaluation metrics (clDice, ERL) that prioritize neural connectivity over pixel-wise accuracy; (3) emergence of self-supervised and foundation model adaptation techniques reducing annotation dependency; and (4) evolution toward specialized architectures capturing long-range dependencies critical for neural structures. Performance analysis reveals that mitochondria segmentation achieves highest accuracy (Jaccard scores 87.2–90.5%), while computational requirements vary from single-GPU implementations to distributed systems with 48 GPUs for teravoxel-scale volumes. Despite progress, reproducibility challenges persist with only 54% of studies providing public code repositories. These advances drive innovation in 3D computer vision, establish new benchmarks for volumetric instance segmentation, and address fundamental challenges in processing massive biological datasets. Our unified benchmarks and comprehensive analysis provide a foundation for systematic progress tracking and evidence-based method selection, positioning brain EM segmentation to enable large-scale connectomics studies and detailed neuroanatomical mapping across scales.

We introduce NeuroVerse, a framework for exploring 3D nanometric-scale reconstructions of neural and glial cellular processes in the central nervous system. Using image stacks from volume electron microscopy, NeuroVerse generates 3D mesh models through a SAM2-based segmentation pipeline and integrates absorption signals for deployment in a Metaverse environment. The framework includes a SAM2 adapter optimized for biological microscopy imaging, adapted with feature enhancement blocks and dual decoders to improve the segmentation of complex cellular structures. An interactive virtual AI agent, powered by Heygen and OpenAI models with domain-specific knowledge, provides semi-real-time assistance. NeuroVerse supports education and collaborative analysis for neuroanatomy and neuroscience. It includes a pipeline for the creation of 3D models, automated segmentation, mesh reconstruction, and heatmap computation, optimized for the Spatial.io ecosystem. Contributions include a virtual anatomy lab for neuroanatomy education and collaborative sessions on spatial morphology correlation and neuroenergetic absorption models. Evaluations show that the SAM2 adapter preserves fine cellular details and manages irregular boundaries. Preliminary sessions indicate potential to enhance neuroscience education, improve remote collaboration among scientists, and provide access to advanced neuroscientific data and tools. Evaluation of the virtual AI agent confirms its ability to provide context-aware support, interpret complex cellular structures, and facilitate understanding through semi-real-time assistance for students analyzing neural and glial reconstructions. NeuroVerse combines imaging, segmentation, and AI technologies within an immersive Metaverse platform for neuroscience education and research. • Development of digital twin of the Human Anatomy Institute of the University of Turin. • AI-Based pipeline for EM image segmentation and Data interpretation. • Case study for multiple usage for Data Analysis and Education in the Metaverse.

Ultraliser is a neuroscience-specific software framework capable of creating accurate and biologically realistic 3D models of complex neuroscientific structures at intracellular (e.g. mitochondria and endoplasmic reticula), cellular (e.g. neurons and glia) and even multicellular scales of resolution (e.g. cerebral vasculature and minicolumns). Resulting models are exported as triangulated surface meshes and annotated volumes for multiple applications in in silico neuroscience, allowing scalable supercomputer simulations that can unravel intricate cellular structure-function relationships. Ultraliser implements a high-performance and unconditionally robust voxelization engine adapted to create optimized watertight surface meshes and annotated voxel grids from arbitrary non-watertight triangular soups, digitized morphological skeletons or binary volumetric masks. The framework represents a major leap forward in simulation-based neuroscience, making it possible to employ high-resolution 3D structural models for quantification of surface areas and volumes, which are of the utmost importance for cellular and system simulations. The power of Ultraliser is demonstrated with several use cases in which hundreds of models are created for potential application in diverse types of simulations. Ultraliser is publicly released under the GNU GPL3 license on GitHub (BlueBrain/Ultraliser). SIGNIFICANCE: There is crystal clear evidence on the impact of cell shape on its signaling mechanisms. Structural models can therefore be insightful to realize the function; the more realistic the structure can be, the further we get insights into the function. Creating realistic structural models from existing ones is challenging, particularly when needed for detailed subcellular simulations. We present Ultraliser, a neuroscience-dedicated framework capable of building these structural models with realistic and detailed cellular geometries that can be used for simulations.

We present a shape processing framework for visual exploration of cellular nuclear envelopes extracted from microscopic images arising in histology and neuroscience. The framework is based on a novel shape descriptor of closed contours in 2D and 3D. In 2D, it relies on a geodesically uniform resampling of discrete curves to compute unsigned curvatures at vertices and edges based on discrete differential geometry. Our descriptor is, by design, invariant under translation, rotation, and parameterization. We achieve the latter invariance under parameterization shifts by using elliptic Fourier analysis on the resulting curvature vectors. Uniform scale-invariance is optional and is a result of scaling curvature features to z-scores. We further augment the proposed descriptor with feature coefficients obtained through sparse coding of the extracted cellular structures using K-sparse autoencoders. For the analysis of 3D shapes, we compute mean curvatures based on the Laplace-Beltrami operator on triangular meshes, followed by computing a spherical parameterization through mean curvature flow. Finally, we compute the Spherical Harmonics decomposition to obtain invariant energy coefficients. Our invariant descriptors provide an embedding into a fixed-dimensional feature space that can be used for various applications, e.g., as input features for deep and shallow learning techniques or as input for dimension reduction schemes to provide a visual reference for clustering shape collections. We demonstrate the capabilities of our framework in the context of visual analysis and unsupervised classification of 2D histology images and 3D nuclear envelopes extracted from serial section electron microscopy stacks.

We present a novel virtual reality (VR) based framework for the exploratory analysis of nanoscale 3D reconstructions of cellular structures acquired from rodent brain samples through serial electron microscopy. The system is specifically targeted on medial axis representations (skeletons) of branched and tubular structures of cellular shapes, and it is designed for providing to domain scientists: i) effective and fast semi-automatic interfaces for tracing skeletons directly on surface-based representations of cells and structures, ii) fast tools for proofreading, i.e., correcting and editing of semi-automatically constructed skeleton representations, and iii) natural methods for interactive exploration, i.e., measuring, comparing, and analyzing geometric features related to cellular structures based on medial axis representations. Neuroscientists currently use the system for performing morphology studies on sparse reconstructions of glial cells and neurons extracted from a sample of the somatosensory cortex of a juvenile rat. The framework runs in a standard PC and has been tested on two different display and interaction setups: PC-tethered stereoscopic head-mounted display (HMD) with 3D controllers and tracking sensors, and a large display wall with a standard gamepad controller. We report on a user study that we carried out for analyzing user performance on different tasks using these two setups.

We present a novel immersive environment for the interactive analysis of nanoscale cellular reconstructions of rodent brain samples acquired through electron microscopy. The system is focused on medial axis representations (skeletons) of branched and tubular structures of brain cells, and it is specifically designed for: i) effective semi-automatic creation of skeletons from surface-based representations of cells and structures ii) fast proofreading, i.e., correcting and editing of semi-automatically constructed skeleton representations, and iii) useful exploration, i.e., measuring, comparing, and analyzing geometric features related to cellular structures based on medial axis representations. The application runs in a standard PC-tethered virtual reality (VR) setup with a head mounted display (HMD), controllers, and tracking sensors. The system is currently used by neuroscientists for performing morphology studies on sparse reconstructions of glial cells and neurons extracted from a sample of the somatosensory cortex of a juvenile rat.

Abstract Digital acquisition and processing techniques are changing the way neuroscience investigation is carried out. Emerging applications range from statistical analysis on image stacks to complex connectomics visual analysis tools targeted to develop and test hypotheses of brain development and activity. In this work, we focus on neuroenergetics, a field where neuroscientists analyze nanoscale brain morphology and relate energy consumption to glucose storage in form of glycogen granules. In order to facilitate the understanding of neuroenergetic mechanisms, we propose a novel customized pipeline for the visual analysis of nanometric‐level reconstructions based on electron microscopy image data. Our framework supports analysis tasks by combining i) a scalable volume visualization architecture able to selectively render image stacks and corresponding labelled data, ii) a method for highlighting distance‐based energy absorption probabilities in form of glow maps, and iii) a hybrid connectivitybased and absorption‐based interactive layout representation able to support queries for selective analysis of areas of interest and potential activity within the segmented datasets. This working pipeline is currently used in a variety of studies in the neuroenergetics domain. Here, we discuss a test case in which the framework was successfully used by domain scientists for the analysis of aging effects on glycogen metabolism, extracting knowledge from a series of nanoscale brain stacks of rodents somatosensory cortex.

This paper presents Abstractocyte, a system for the visual analysis of astrocytes and their relation to neurons, in nanoscale volumes of brain tissue. Astrocytes are glial cells, i.e., non-neuronal cells that support neurons and the nervous system. The study of astrocytes has immense potential for understanding brain function. However, their complex and widely-branching structure requires high-resolution electron microscopy imaging and makes visualization and analysis challenging. Furthermore, the structure and function of astrocytes is very different from neurons, and therefore requires the development of new visualization and analysis tools. With Abstractocyte, biologists can explore the morphology of astrocytes using various visual abstraction levels, while simultaneously analyzing neighboring neurons and their connectivity. We define a novel, conceptual 2D abstraction space for jointly visualizing astrocytes and neurons. Neuroscientists can choose a specific joint visualization as a point in this space. Interactively moving this point allows them to smoothly transition between different abstraction levels in an intuitive manner. In contrast to simply switching between different visualizations, this preserves the visual context and correlations throughout the transition. Users can smoothly navigate from concrete, highly-detailed 3D views to simplified and abstracted 2D views. In addition to investigating astrocytes, neurons, and their relationships, we enable the interactive analysis of the distribution of glycogen, which is of high importance to neuroscientists. We describe the design of Abstractocyte, and present three case studies in which neuroscientists have successfully used our system to assess astrocytic coverage of synapses, glycogen distribution in relation to synapses, and astrocytic-mitochondria coverage.

Glucose is the main energy substrate in the adult brain under normal conditions. Accumulating evidence, however, indicates that lactate produced in astrocytes (a type of glial cell) can also fuel neuronal activity. The quantitative aspects of this so-called astrocyte-neuron lactate shuttle (ANLS) are still debated. To address this question, we developed a detailed biophysical model of the brain's metabolic interactions. Our model integrates three modeling approaches, the Buxton-Wang model of vascular dynamics, the Hodgkin-Huxley formulation of neuronal membrane excitability and a biophysical model of metabolic pathways. This approach provides a template for large-scale simulations of the neuron-glia-vasculature (NGV) ensemble, and for the first time integrates the respective timescales at which energy metabolism and neuronal excitability occur. The model is constrained by relative neuronal and astrocytic oxygen and glucose utilization, by the concentration of metabolites at rest and by the temporal dynamics of NADH upon activation. These constraints produced four observations. First, a transfer of lactate from astrocytes to neurons emerged in response to activity. Second, constrained by activity-dependent NADH transients, neuronal oxidative metabolism increased first upon activation with a subsequent delayed astrocytic glycolysis increase. Third, the model correctly predicted the dynamics of extracellular lactate and oxygen as observed in vivo in rats. Fourth, the model correctly predicted the temporal dynamics of tissue lactate, of tissue glucose and oxygen consumption, and of the BOLD signal as reported in human studies. These findings not only support the ANLS hypothesis but also provide a quantitative mathematical description of the metabolic activation in neurons and glial cells, as well as of the macroscopic measurements obtained during brain imaging.

Sodium entry during an action potential determines the energy efficiency of a neuron. The classic Hodgkin-Huxley model of action potential generation is notoriously inefficient in that regard with about 4 times more charges flowing through the membrane than the theoretical minimum required to achieve the observed depolarization. Yet, recent experimental results show that mammalian neurons are close to the optimal metabolic efficiency and that the dynamics of their voltage-gated channels is significantly different than the one exhibited by the classic Hodgkin-Huxley model during the action potential. Nevertheless, the original Hodgkin-Huxley model is still widely used and rarely to model the squid giant axon from which it was extracted. Here, we introduce a novel family of Hodgkin-Huxley models that correctly account for sodium entry, action potential width and whose voltage-gated channels display a dynamics very similar to the most recent experimental observations in mammalian neurons. We speak here about a family of models because the model is parameterized by a unique parameter the variations of which allow to reproduce the entire range of experimental observations from cortical pyramidal neurons to Purkinje cells, yielding a very economical framework to model a wide range of different central neurons. The present paper demonstrates the performances and discuss the properties of this new family of models.