Cryo-electron microscopy (cryo-EM) is a powerful technique for determining high-resolution 3D biomolecular structures from imaging data. Its unique ability to capture structural variability has spurred the development of heterogeneous reconstruction algorithms that can infer distributions of 3D structures from noisy, unlabeled imaging data. Despite the growing number of advanced methods, progress in the field is hindered by the lack of standardized benchmarks with ground truth information and reliable validation metrics. Here, we introduce CryoBench, a suite of datasets, metrics, and benchmarks for heterogeneous reconstruction in cryo-EM. CryoBench includes five datasets representing different sources of heterogeneity and degrees of difficulty. These include conformational heterogeneity generated from designed motions of antibody complexes or sampled from a molecular dynamics simulation, as well as {compositional heterogeneity from mixtures of ribosome assembly states or 100 common complexes present in cells. We then analyze state-of-the-art heterogeneous reconstruction tools, including neural and non-neural methods, assess their sensitivity to noise, and propose new metrics for quantitative evaluation. We hope that CryoBench will be a foundational resource for accelerating algorithmic development and evaluation in the cryo-EM and machine learning communities.

Calcium imaging permits optical measurement of neural activity. Since intracellular calcium concentration is an indirect measurement of neural activity, computational tools are necessary to infer the true underlying spiking activity from fluorescence measurements. Bayesian model inversion can be used to solve this problem, but typically requires either computationally expensive MCMC sampling, or faster but approximate maximum-a-posteriori optimization. Here, we introduce a flexible algorithmic framework for fast, efficient and accurate extraction of neural spikes from imaging data. Using the framework of variational autoencoders, we propose to amortize inference by training a deep neural network to perform model inversion efficiently.

In neuroscience, the similarity matrix of neural activity patterns in response to different sensory stimuli or under different cognitive states reflects the structure of neural representational space. Existing methods derive point estimations of neural activity patterns from noisy neural imaging data, and the similarity is calculated from these point estimations. We show that this approach translates structured noise from estimated patterns into spurious bias structure in the resulting similarity matrix, which is especially severe when signal-to-noise ratio is low and experimental conditions cannot be fully randomized in a cognitive task. We propose an alternative Bayesian framework for computing representational similarity in which we treat the covariance structure of neural activity patterns as a hyper-parameter in a generative model of the neural data, and directly estimate this covariance structure from imaging data while marginalizing over the unknown activity patterns. Converting the estimated covariance structure into a correlation matrix offers a much less biased estimate of neural representational similarity. Our method can also simultaneously estimate a signal-to-noise map that informs where the learned representational structure is supported more strongly, and the learned covariance matrix can be used as a structured prior to constrain Bayesian estimation of neural activity patterns.

Calcium imaging permits optical measurement of neural activity.

In neuroscience, the similarity matrix of neural activity patterns in response to different sensory stimuli or under different cognitive states reflects the structure of neural representational space. Existing methods derive point estimations of neural activity patterns from noisy neural imaging data, and the similarity is calculated from these point estimations. We show that this approach translates structured noise from estimated patterns into spurious bias structure in the resulting similarity matrix, which is especially severe when signal-to-noise ratio is low and experimental conditions cannot be fully randomized in a cognitive task. We propose an alternative Bayesian framework for computing representational similarity in which we treat the covariance structure of neural activity patterns as a hyper-parameter in a generative model of the neural data, and directly estimate this covariance structure from imaging data while marginalizing over the unknown activity patterns. Converting the estimated covariance structure into a correlation matrix offers a much less biased estimate of neural representational similarity. Our method can also simultaneously estimate a signal-to-noise map that informs where the learned representational structure is supported more strongly, and the learned covariance matrix can be used as a structured prior to constrain Bayesian estimation of neural activity patterns.

Temporal context in medicine is valuable in assessing key changes in patient health over time. We developed a machine learning framework to integrate diverse context from prior visits to improve health monitoring, especially when prior visits are limited and their frequency is variable. Our model first estimates initial risk of disease using medical data from the most recent patient visit, then refines this assessment using information digested from previously collected imaging and/or clinical biomarkers. We applied our framework to prostate cancer (PCa) risk prediction using data from a large population (28,342 patients, 39,013 magnetic resonance imaging scans, 68,931 blood tests) collected over nearly a decade. For predictions of the risk of clinically significant PCa at the time of the visit, integrating prior context directly converted false positives to true negatives, increasing overall specificity while preserving high sensitivity. False positive rates were reduced progressively from 51% to 33% when integrating information from up to three prior imaging examinations, as compared to using data from a single visit, and were further reduced to 24% when also including additional context from prior clinical data. For predicting the risk of PCa within five years of the visit, incorporating prior context reduced false positive rates still further (64% to 9%). Our findings show that information collected over time provides relevant context to enhance the specificity of medical risk prediction. For a wide range of progressive conditions, sufficient reduction of false positive rates using context could offer a pathway to expand longitudinal health monitoring programs to large populations with comparatively low baseline risk of disease, leading to earlier detection and improved health outcomes.

Identifying new disease-related patterns in medical imaging data with the help of machine learning enlarges the vocabulary of recognizable findings. This supports diagnostic and prognostic assessment. However, image appearance varies not only due to biological differences, but also due to imaging technology linked to vendors, scanning- or re- construction parameters. The resulting domain shifts impedes data representation learning strategies and the discovery of biologically meaningful cluster appearances. To address these challenges, we introduce an approach to actively learn the domain shift via post-hoc rotation of the data latent space, enabling disentanglement of biological and technical factors. Results on real-world heterogeneous clinical data showcase that the learned disentangled representation leads to stable clusters representing tissue-types across different acquisition settings. Cluster consistency is improved by +19.01% (ARI), +16.85% (NMI), and +12.39% (Dice) compared to the entangled representation, outperforming four state-of-the-art harmonization methods. When using the clusters to quantify tissue composition on idiopathic pulmonary fibrosis patients, the learned profiles enhance Cox survival prediction. This indicates that the proposed label-free framework facilitates biomarker discovery in multi-center routine imaging data. Code is available on GitHub https://github.com/cirmuw/latent-space-rotation-disentanglement.

Understanding the dynamic organization and homeostasis of living tissues requires high-resolution, time-resolved imaging coupled with methods capable of extracting interpretable, predictive insights from complex datasets. Here, we present the Vision Transformer Digital Twin Surrogate Network (VT-DTSN), a deep learning framework for predictive modeling of 3D+T imaging data from biological tissue. By leveraging Vision Transformers pretrained with DINO (Self-Distillation with NO Labels) and employing a multi-view fusion strategy, VT-DTSN learns to reconstruct high-fidelity, time-resolved dynamics of a Drosophila midgut while preserving morphological and feature-level integrity across imaging depths. The model is trained with a composite loss prioritizing pixel-level accuracy, perceptual structure, and feature-space alignment, ensuring biologically meaningful outputs suitable for in silico experimentation and hypothesis testing. Evaluation across layers and biological replicates demonstrates VT-DTSN's robustness and consistency, achieving low error rates and high structural similarity while maintaining efficient inference through model optimization. This work establishes VT-DTSN as a feasible, high-fidelity surrogate for cross-timepoint reconstruction and for studying tissue dynamics, enabling computational exploration of cellular behaviors and homeostasis to complement time-resolved imaging studies in biological research.

Integrating brain imaging data with clinical reports offers a valuable opportunity to leverage complementary multimodal information for more effective and timely diagnosis in practical clinical settings. This approach has gained significant attention in brain disorder research, yet a key challenge remains: how to effectively link objective imaging data with subjective text-based reports, such as doctors' notes. In this work, we propose a novel framework that aligns brain connectomes with clinical reports in a shared cross-modal latent space at both the subject and connectome levels, thereby enhancing representation learning. The key innovation of our approach is that we treat brain subnetworks as tokens of imaging data, rather than raw image patches, to align with word tokens in clinical reports. This enables a more efficient identification of system-level associations between neuroimaging findings and clinical observations, which is critical since brain disorders often manifest as network-level abnormalities rather than isolated regional alterations. We applied our method to mild cognitive impairment (MCI) using the ADNI dataset. Our approach not only achieves state-of-the-art predictive performance but also identifies clinically meaningful connectome-text pairs, offering new insights into the early mechanisms of Alzheimer's disease and supporting the development of clinically useful multimodal biomarkers.