Advanced Intelligent Systems
○ Wiley
Preprints posted in the last 30 days, ranked by how well they match Advanced Intelligent Systems's content profile, based on 11 papers previously published here. The average preprint has a 0.01% match score for this journal, so anything above that is already an above-average fit.
Cecchi, D.; Lasala, A.; Poli, F.; De Remigis, E.; Moccia, S.; Palagi, S.
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Fluorescence microscopy is routinely used in the quantitative analysis of Giant Unilamellar Vesicles (GUVs) because it makes GUVs visible and easy to detect. Nonetheless, it requires fluorescence-equipped microscopes and, especially, the addition of tagged molecules that may introduce undesired and unaccounted variations compared to the pristine formulation. Alternatively, phase-contrast microscopy can be used to observe GUVs without labelling, yet the automatic detection of GUVs in phase-contrast images by standard image processing techniques is difficult. To overcome this limitation, we developed a framework for the automated detection and size characterization of label-free GUVs based on YOLOv11 model, a state-of-art lightweight Convolutional Neural Network (CNN) for object detection. Trained on 164 phase-contrast microscope images containing 4666 manually annotated vesicles, the model enables robust detection under low contrast-to-noise ratio and heterogeneous illumination. The trained model is implemented in a stand-alone desktop application supporting multiple analysis in a single run, pixel-to-micrometer calibration, and automated size distribution analysis. This approach achieves high detection accuracy and competitive cross-microscope generalization using YOLOv11 nano on greyscale image. Retraining scripts compatible with standard hardware are also provided to enhance adaptability. This tool enables fast and reliable analysis of GUVs without altering their composition, supporting experimental reproducibility.
Chen, J.; Chen, W.
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Three-dimensional medical image segmentation is unusually heterogeneous in its computational needs: large homogeneous organ interiors are often resolved at coarse scale, whereas small organs, lesions, thin vessels, and ambiguous boundaries benefit from native-resolution processing. Yet modern 3D segmentation networks allocate nearly uniform computation across the entire volume. Existing dynamic-resolution or coarse-to fine methods reduce cost, but typically route regions using hand-crafted image complexity, prediction confidence, or a fixed crop, none of which directly estimates whether spending additional computation on a region will improve the segmentation. We introduce ERRORROUTE, a budgeted inference framework that learns the marginal refinement utility of candidate 3D blocks. A coarse branch first predicts global low-resolution semantics. During training, we estimate for each candidate block the loss reduction achievable by native resolution refinement and supervise a utility estimator to predict this gain before refinement. A differentiable budgeted selector then allocates a fixed compute budget to blocks with the highest expected utility per unit cost, while unselected regions remain on the efficient global path. Selected blocks are refined locally and merged through residual cross-scale fusion. Across six 3D CT/MRI benchmarks, ERRORROUTE achieves a mean Dice of 90.5% at 128 GFLOPs, improving the accuracy-compute frontier over dense 3D baselines and fixed coarse-to-fine systems. At a 30% routing budget, the learned utility estimator recovers 83% of the blocks selected by an oracle router, with the largest allocation concentrated on lesions, small structures, and complex boundaries. These results suggest that efficient medical segmentation benefits from learning where additional computation is useful, rather than merely where the model is uncertain.
Zhao, W.; Wang, N.; Han, Z.; Xu, X.; Ma, X.; Zhou, T.; Gui, J.; Liu, Y.; Liu, Q.; Qu, L.; Liu, W.; Ding, X.; Liu, X.; Ta, D.; Tan, J.; Chen, L.; Li, H.
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Capturing fast hemodynamics in deep organs is essential for understanding microvascular regulation of organ responses to physiological demands and pathological stress. However, non-invasive three-dimensional (3D) imaging of these microscale processes remains challenging due to trade-offs between spatial resolution, imaging speed, and penetration depth. Here, we present fluctuation-based high-order super-resolution acoustic microscope (FLAME), a tracking-free 3D ultrasound imaging technology capable of fast microvascular angiography and flow measurement. Using as few as 30 volumes, FLAME improves 3D ultrasound resolution by 8-fold (~50 m) and shortens data acquisition time by over two orders of magnitude, from tens of seconds to tens of milliseconds, compared with conventional tracking-based super-resolution approaches1-6. This high-throughput capability supports high-fidelity imaging of tissue functions under flexible experimental settings. FLAME achieves an unprecedented 3D super-resolution frame rate of ~40 Hz, capturing transient hemodynamic responses to various physiological and pathological stimuli in different mouse organs, from the microvascular level to the whole-body scale. With open-sourced implementation, FLAME provides an accessible platform for real-time, in-depth hemodynamic monitoring across diverse biomedical applications and clinical translation.
Rapilly, Q.; Badoual, A.; LIU, Y.-H.; Kervrann, C.
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In this paper, we introduce Nagini3D, a 3D biological image segmentation method that hybridizes two high-performing yet highly complementary segmentation paradigms: deep learning-based approaches and snake-based methods. Our approach leverages both the high efficiency of data-driven techniques and the convenient shape representation provided by active surfaces, which proves highly practical for incorporating a priori knowledge as well as for the post-analysis of segmented objects. By considering a complete 3D volume, our method ensures segmentation consistency across all three dimensions, unlike many approaches that handle volumes in a slice-by-slice manner. Additionally, it theoretically enables the representation of any object with spherical topology, overcoming the limitations of common 3D methods that rely on object convexity assumptions. Furthermore, the continuous representation of objects allows for a resolution-independent description, facilitating the extraction of various geometric features of interest (e.g., local curvature, principal deformations). We demonstrate both the quality of the segmentation produced by our method and the richness of the accessible geometric features through an extensive set of experiments conducted on multiple real and simulated datasets, including objects with challenging geometries.
Vasinkova, M.; Jochymek, L.; Gajdos, P.
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Machine learning benchmarks often treat image files as independent observations, although multiple files may represent repeated views or other related samples. Prior work has shown that non-independent partitioning can inflate model performance, but the sensitivity of a biomedical benchmark to progressively increasing train-test relatedness is less often quantified. Here, we quantified how apparent classification performance changes when recoverable sample groups are progressively allowed to cross training, validation, and test partitions. Using source structure and filenames, we reconstructed analysis groups in a microscopy corpus compiled from multiple sources and generated five class stratified partitioning conditions, ranging from an intact group reference to random assignment of individual files. We applied the same training recipe within each model family to 129,173 images spanning 14 classes of blood cells and hematopoietic cell states. For a pretrained Vision Transformer, random assignment of individual files increased the mean test macro F1 from 0.853 to 0.977 and accuracy from 0.851 to 0.990 relative to the intact group reference. Apparent performance increased progressively across the intermediate conditions. The endpoint pattern also appeared with a ResNet-50 backbone, and the largest apparent gains occurred in immature granulocytic and blast classes. A descriptive analysis within the random assignment condition further showed higher scores among test images that had another member of the same group in the training set, although the exposed and unexposed subsets were not exchangeable. The grouping workflow, exact split definitions, code, and compact result files are released as HSD-Bench to support reproducible evaluation of hidden sample dependence.
Vizcarra, J. C.; Kataria, B. C.; Chuah, C.-N.; Pearce, T. M.; Dugger, B. N.; Candano, C. A.; Rosado, A. M.; Cooper, L. A.; Gutman, D. A.
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Foundation models (FMs) have rapidly become dominant in artificial intelligence and are increasingly being adopted in computational pathology. Numerous pathology-specific FMs have been developed and evaluated for a variety of downstream tasks, most commonly using frozen image embeddings with linear probes. While a pathology FM, at least implicitly suggests broad reusability across tasks, the transferability of these models to neurodegenerative disease-related tasks remains largely unexplored. In this study, we evaluated fourteen frozen feature extractors, including general-vision and pathology FMs, across four pathology image classification datasets spanning two specialized neurodegenerative disease immuno-histochemistry (IHC) tasks (Tau neurofibrillary tangle (NFT) and amyloid-{beta} plaque classification) and two cancer-related hematoxylin and eosin (H&E) tasks (the breast tissue BACH dataset and the multi-tissue TIL dataset). The cancer-related H&E datasets represent tasks more closely aligned with the predominant pretraining domain of many current pathology FMs, whereas the neurodegenerative IHC datasets represent a more specialized domain with limited representation in existing FM pretraining cohorts. Frozen embedding linear probes were compared against a conventional supervised ResNet-50 convolutional neural network (CNN) trained directly on image tiles using identical train, validation, and test splits. Across both neurodegenerative IHC datasets, the supervised CNN substantially outperformed all frozen FM linear probes. In contrast, pathology FMs achieved performance comparable to, and in some cases exceeding, the supervised CNN across the two cancer-related H&E datasets, with UNI2-h achieving the highest performance on BACH and several pathology FMs performing on par with the CNN on the larger TIL dataset. Furthermore, a supervised CNN trained using only 1% of the Tau NFT training data (1,938 tiles) still exceeded the performance of the best frozen FM linear probe trained on the complete dataset. Together, these results suggest the transferability of frozen pathology FMs may depend strongly on how well the downstream task is represented by their pretraining domain. These findings demonstrate frozen pathology FMs transfer effectively to the cancer-related H&E tasks evaluated here but may be less effective for specialized neurodegenerative IHC tasks less well represented in current FM pretraining cohorts. Expanding pathology FM pretraining datasets to include a broader range of disease domains and staining modalities may therefore improve transferability to specialized pathology applications. Our findings also highlight the continued importance of conventional supervised learning and motivate future work investigating nonlinear probes and end-to-end FM fine-tuning.
Pelloux, M.; Tran, C. T.; Baig, J.; Khellaf, A.; Huang, S. L.; Nguyen, O. T.; Batardiere, M.; Diwan, S.-S.; Yang, Z.; Li, J.-L.; Vu, V.; Roy, S. F.; Soucy, G.; Nguyen, B. N.; Tessier-Cloutier, B.; Hosseini, M. S.; Knapp, D. J.; Valmary-Degano, S.; Trinh, V. Q.
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Current state of the art computational pathology foundation models attain satisfying accuracy most general pathology tasks including detection of pancreatic ductal adenocarcinoma (PDAC) but fail to provide sufficient explainability, operating as "black boxes". Some interpretability effort has been made, with models providing attention maps that project back on the slide the models high attended areas that pushed for the given prediction result, without further naming or reasoning explanation which can limit pathologists trust toward the prediction. We developed C3PRO, a cell-centric framework attempting to provide deeper understanding and explainability, by using embeddings extracted using a pathology foundation model, from cell centered tiles containing both the cell and its close neighborhood, which were then clustered into recurrent morphological phenotypes. A spatial graph was additionally constructed on top of the back-projected vocabulary to account for recurrent motifs. The evaluation of the model performance involved a cohort of 809 patients (825 diagnostic biopsies) using nested cross validation. C3PRO was benchmarked against four different multiple instance learning frameworks and achieved on-par performance for diagnostic (0.958 AUC), survival (0.617) and resectability (0.696) prediction, while providing additional insights compared to the strongest baseline models. Each clustered phenotype was probed and extracted as a representative tile mosaic and annotated by two expert pathologists, constituting a phenotype vocabulary available for statistical analysis. These phenotypes were scored by pathologists with physical brightfield microscopy and replicated the independent statistical significance impact on overall survival prediction from diagnostic biopsies.
Zhao, S.-C.; Mizuno, D.
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Scientific image sequences often feature recurring structures with unknown appearance and dynamics. While traditional methods using predefined templates, noise models, or motion classes work well when targets are known, such assumptions can hinder exploratory analysis where discovering that knowledge is the primary goal. This work presents a self-supervised framework that maps noisy video into a discrete vocabulary of local states and their spatiotemporal relationships. The method operates without clean targets, pretrained representations, semantic labels, feature templates, or prescribed trajectories, assuming only that informative structures recur for a finite duration within a bounded spatial neighborhood. A temporally masked vector-quantized network predicts the missing central frame from its neighbors, while a masked Transformer refines token assignments by weighing encoder evidence against contextual compatibility. Project-specific token sets define the feature family, while a support model quantifies the temporal evidence for individual occurrences, facilitating optional conservative suppression. The framework is first demonstrated on synthetic videos of moving particles. Without access to clean frames or particle coordinates during learning and selection, the learned states recover compact, point-spread-function-like structures. Tests on low-signal particle-tracking benchmarks yield frame-wise component precision between 87.0% and 94.5%, with recall decreasing as particle density rises. An experimental example using E. coli further shows that discrete states can separate biological structures from illumination and acquisition artifacts. In all cases, the primary outputs are interpretable state, activity, support, and component maps; rendered images serve as diagnostics rather than optimization targets.
Zhang, J.; Zheng, S. t.; Liu, X. f.; Zhang, W.; Gu, J.
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Text-driven medical image segmentation demands unified models based on multi-modal learning, capable of processing diverse imaging modalities while providing spatially reliable prediction estimates for clinical practice. Nevertheless, existing vision-language model solutions suffer from poor domain generalization and exhibit limited capability for uncertainty estimation to model the cumulative unreliability of cross-modal alignment across network layers. In this paper, we propose UniMedSeg, a unified vision-language framework for multi-modal medical image segmentation built upon frozen pre-trained vision-language backbones. Our framework incorporates modality-aware adaptation, cross-layer uncertainty-guided encoding, and a frequency-spatial dual-prompt decoder, wherein the progressive uncertainty propagation mechanism of the encoder suppresses misleading cross-modal features, whereas the decoder disentangles anatomical structure and boundary details in the frequency domain under dual textual prompts. Auxiliary losses further sharpen boundary representations and calibrate pixel-wise uncertainty maps. Extensive experiments demonstrate that UniMedSeg preserves competitive in-domain segmentation performance, improves domain generalization, and yields clinically useful spatial risk estimation.
Georvasilis, I.; Frias Nestares, M.; Badosa, C.; Musokhranova, U.; Oyarzabal, A.; Jimenez-Mallebrera, C.; Ruminski, J.; Roldan, M.
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Fluorescence microscopy is a central tool in biomedical image analysis, but its practical use is often constrained by incomplete and heterogeneous data. This is especially true in rare disease research, where patient cohorts are small, biological material is limited, and complex study designs frequently yield images with missing, degraded, or low-quality channels. We introduce Reliability-Mask-Conditioned DDPM (RM-DDPM), a conditional diffusion model that formulates multichannel image generation, missing-channel imputation, and degraded-channel restoration as a single reliability-guided inference problem. The model is trained on a curated subset of complete, high-quality multichannel images using a self-supervised masking objective to learn rich cross-channel dependencies from the available channels. At inference time, reliable observations condition the reverse diffusion process, allowing the same trained model to solve multiple reconstruction tasks without task-specific retraining. We evaluate RM-DDPM against task-specific baselines on one public and two in-house fluorescence microscopy datasets, comprising a rare-disease preclinical model study and a clinical patient cohort. Across these settings, RM-DDPM matches the specialized baselines in reconstruction quality, hallucinates fewer false structures, and remains more robust as degradation severity increases, while repeated sampling provides per-pixel uncertainty maps that flag unreliable reconstructions.
Duari, S.; Shome, R.; Raza, A.; Solanki, S.; Satija, S.; Sinha, S.; Kumar, S.; Chauhan, S.; Sharma, A.; Gautam, V.; Gupta, M.; Sengupta, D.; Ahuja, G.
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Interpretable computational pathology is constrained by a trade-off between scalable learned representations and biologically explicit phenotypes. Here we present NucXplore, a nuclear phenomics framework that converts each hematoxylin-and-eosin (H&E)-stained nucleus into 129 explicitly defined features spanning morphology, chromatin, intensity, texture, color, and spatial context. A Rust-based implementation accelerates extraction 17.6-fold over the Python reference implementation. Across three histopathology cohorts, NucXplore outperformed classical descriptors and larger deep-learning embeddings while retaining feature attribution. Applied to human skin, hierarchical multiple-instance models resolved eight cellular compartments and decoded age and sun-exposure states, revealing a largely shared aging program with age and cell-specific exposure-dependent recalibration. Conditional flow matching and neural ODE integration transformed cross-sectional phenotypes into model-implied trajectories, uncovering a midlife velocity minimum, late-life re-acceleration, and cell-specific changes in trajectory magnitude and direction. Source-free domain adaptation enabled preliminary transfer of age-associated predictions to an independent hospital cohort. NucXplore thus unifies interpretable representation, high-performance computing, and dynamical modeling for cellular phenotyping from routine histology.
Ye, E.; Wu, X.; Peng, R.; Hu, W.; Li, X.; Zhang, X.; Niu, M.; Guo, Y.; Sheng, X.; Wang, J.; Chen, L.; Sun, H.
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Microscopy-based cell profiling has broad applications in biological discovery, disease characterization, and phenotypic drug screening. Modern microscopy continues to push the limits of resolution, speed, depth and throughput, but better imaging does not automatically lead to better biomedical discovery and translation. A key bottleneck is feature representation: existing features are either handcrafted or learned as black-box embeddings and often lack explicit biological meaning. Here we propose biological grounding as a first principle for cell profiling and implement it through MorphAgent, an AI agent framework in which each quantitative feature originates from a biological hypothesis and is anchored to a cellular structure or process. By integrating biological knowledge, multimodal reasoning and automated validation, MorphAgent makes biologically grounded feature design systematic and scalable. We show biological grounding fundamentally changes the properties of cell profiling features. Across three microscopy modalities, MorphAgent produces compact, expressive and transferable features for drug screening, cell state characterization and disease morphology analysis. In wide-field Cell Painting, a common assay for phenotypic drug screening, it improves perturbation-retrieval mean average precision by 48% over CellProfiler and 20% over DeepProfiler while using substantially fewer dimensions. In confocal mitochondrial imaging, biologically grounded features transfer across independently acquired datasets and imaging resolutions, supporting accurate aging-state classification. In structured illumination microscopy (SIM) imaging of Tau-labeled samples, biologically grounded super resolution features reveal nanoscale morphologies inaccessible at conventional resolution, enhancing the discrimination of disease-associated mutations. Moreover, biologically grounded features provide a hierarchical organization of cellular morphology, enabling multilevel alignment with transcriptomics data and facilitating mechanistic understanding. Biologically grounded cell profiling thus brings AI reasoning closer to biological mechanisms, advancing biomedical discovery and translation.
Chen, J.; Chen, W.
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Synthetic medical data are commonly generated from manually chosen prompts, inverse frequency sampling, or fixed subgroup balancing rules. These strategies increase sample count but do not answer a more consequential question: which missing observations would most improve the current model? We introduce GAP DREAM, a closed-loop framework that treats a medical image generator as a queryable source of candidate evidence rather than an unlimited source of augmentation. At each round, GAP DREAM maps model failures into an evidence gap score that combines scarcity, predictive uncertainty, local representation density, and gradient novelty. A query policy translates high-value gaps into structured clinical requests, a modality specific generator proposes candidate imagetext pairs, and an independent selector retains only diagnostically consistent, diverse, non-duplicative candidates. The downstream model is then updated and the gap map recomputed. The central objective is therefore not image realism in isolation, but reduction of measurable evidence gaps under a fixed synthetic budget. Across a multi-modality protocol spanning long-tailed chest radiography, dermoscopy, clinical dermatology, and histopathology, GAPDREAM reaches stronger held-out real-data performance with substantially fewer generated samples than random,class-balanced, uncertainty-only, and static targeted augmentation. Results show gains in CXRLT tail mAP, HAM10000 macroAUROC, Came lyon17 out-of-domain accuracy, and worst group dermatology accuracy while reducing redundant synthetic samples. GAPDREAM reframes medical generation as active evidence acquisition: generate the examples the learner needs, not simply more examples of what is rare.
Lee, K.; Yoon, R. J.; Lee, R. S.; Lee, D.
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Cervical cytology screening relies on the visual assessment of nuclear and cellular morphology across thousands of cells per slide, a labor-intensive task that could benefit from quantitative computational support. Here we present CytoLens, a modular deep learning system that detects and counts nuclei, delineates nuclei and whole cells, measures the nuclear-to-cell area ratio (N:C), classifies cells from multifocal images and scores candidate abnormality. We trained and tested each module on a public dataset matched to its task, without pooling datasets. CytoLens detected nuclei with an F1 score of 0.931 (95% confidence interval (CI), 0.914-0.946) and counted 75 of 100 fields to within one nucleus. Point-prompted segmentation achieved mean Dice coefficients of 0.901 for nuclei and 0.847 for whole cells and, without retraining, transferred to a second dataset in which measured N:C increased with lesion grade in parallel with expert outlines. Multifocal hierarchical classification reached a mean accuracy of 0.947 across four Bethesda-aligned categories, and abnormality scores ranked annotated abnormal candidates with an area under the receiver operating characteristic curve (AUROC) of 0.865. These results support the evaluation of CytoLens on patient-linked clinical specimens.
Chen, W.; Chen, J.
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Coarse to fine segmentation is a natural strategy for three dimensional lesion delineation becausefull resolution processing of an entire scan is expensive while only a small fraction of the volume contains pathology. Its main failure mode, however, is structural: a lesion missed by the coarse stage is usually absent from the region of interest and therefore cannot be recovered by the fine stage. This problem is most severe for small, low contrast, and multifocal lesions. We introduce RECALLREFINE, a coverage constrained coarse to fine framework that treats candidate-region selection as a recall critical learning problem rather than a deterministic crop around the coarse mask.A full-volume network first predicts coarse lesion probabilities and multi-scale features. A candidate score network then combines coarse probability, predictive uncertainty, feature novelty, and spatial context to score overlapping 3D blocks. During training, a differentiable selector is optimized with a component-wise coverage objective that requires every annotated lesion instance to remain covered by at least one selected block under a fixed refinement budget. Selected blocks are processed at native resolution by a local refiner with a context-ring branch, and overlapping predictions are merged back into the full volume. Across four public 3D lesion benchmarks, RECALLREFINE improves mean tumor Dice from 76.5% for the strongest matched coarse-to-fine baseline to 78.4%, while increasing sensitivity for lesions below 10 mm from 60.9% to 70.8%.At a 20% candidate budget, the selector covers 94.4% of lesion instances, substantially reducing irreversible coarse-stage misses. The results suggest that the central problem in cascaded lesion segmentation is not only how to refine a detected lesion, but how to guarantee that difficult lesions are still represented in the refinement set.
Song, Z.; Shi, Z.; Sun, G.; Negi, S.; Fausther-Bovendo, H.; Zhao, H.
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Protein--Protein Interactions (PPIs) are fundamental to cellular processes and represent major targets for therapeutic development. Recent advances in machine learning for protein modeling, particularly protein language models, have enabled powerful representations of protein sequences and monomeric structures. However, compared with the increasingly generalizable representations available for individual proteins, transferable representations explicitly designed to capture protein-complex and interface contexts remain less developed. Here, we developed GraPPI, a unified graph-based deep learning framework for learning transferable representations of protein--protein complexes. At its core, PPIencoder represents protein complexes as heterogeneous residual graphs and uses self-supervised masked-edge prediction to learn interaction-aware representations that integrate sequence, three-dimensional geometry, and physicochemical properties. We then evaluated the transferability of these pretrained representations across complementary PPI tasks spanning binding-mode classification, binding-affinity prediction, and mutation-associated affinity prediction. GraPPI accurately distinguished plausible binding interfaces from perturbed or decoy binding interfaces, achieving AUROC values above 0.92 on an independent benchmark. For protein--protein binding-affinity prediction, GraPPI achieved Pearson correlation coefficients of 0.690 and 0.684 on the S90 and S79 benchmarks, respectively, with performance competitive with or exceeding that of recently reported affinity-prediction methods. GraPPI further transferred effectively to mutation-associated binding-affinity prediction, achieving a Pearson correlation of 0.84 and a mean absolute error of 1.20 kcal/mol on an independent SKEMPI benchmark. Together, these results establish GraPPI as a generalizable framework for protein-complex representation learning, extending pretrained protein representations from individual proteins toward the interaction context in which protein function is realized.
Zou, A.; Huang, E.; Wu, Q.; Barnard, M. E.; Zhang, C.
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Predicting molecular receptor status, including estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2), directly from routine hematoxylin and eosin (H&E)-stained whole-slide images (WSIs) could reduce reliance on costly immunohistochemistry, but image-only models have no access to the transcriptomic programs that define these subtypes. Here we show that coupling a whole-slide vision transformer (GigaPath) with bulk RNA-sequencing profiles through a training-time-only contrastive objective improves receptor-status classification from WSIs alone, without requiring any RNA-seq data at inference. Rather than training a dedicated bulk-transcriptomics encoder, we purpose the gene-sentence encoding and pretrained text encoder from BioBERT1 and apply them to bulk RNA-seq profiles from The Cancer Genome Atlas breast cancer cohort (TCGA-BRCA). Under five-fold cross-validation, this RNA-guided pre-training consistently improved AUROC for ER, PR, and HER2 relative to a contrastive-pre-training-free baseline and reproducibly introduced receptor-relevant structure into the learned slide representation. However, evaluation on independent external cohorts revealed a striking, receptor-dependent divergence: gains for ER and PR, both governed by broad, multi-gene luminal transcriptional programs, generalized robustly, whereas the apparent AUROC improvement for HER2, driven by focal ERBB2 amplification rather than a coordinated transcriptional signature, masked a severe loss of classification sensitivity at the operating threshold. These findings establish training-time transcriptomic supervision as a practical route to enhancing histology-only molecular subtyping, while showing that gene-selection strategy, and not model architecture alone, determines which molecular alterations such supervision can teach a vision model to recognize.
LE, L.; Zunino, A.; Slenders, E.; Bucci, A.; Donato, M.; Vicidomini, G.
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Image scanning microscopy (ISM) is emerging as the natural evolution of confocal laser-scanning microscopy, combining optical sectioning and a two-fold improvement in lateral resolution with high signal-to-noise ratio. Single-photon avalanche diode (SPAD) array detectors enable an effective implementation of ISM by recording a micro-image at each scan position with single-photon sensitivity, negligible readout noise, and without the fixed frame-readout constraints of conventional cameras. Nevertheless, SPAD array-based ISM remains largely confined to laboratories with expertise in microscope development. Here, we present BrightEyes-Scope, a reference implementation of a SPAD array-based ISM system. We provide step-by-step guidelines for optical design, system construction, alignment, calibration, data acquisition, and image reconstruction. Presented as a complete ISM microscope, BrightEyes-Scope also serves as a reference for upgrading existing laser-scanning microscopes. We anticipate that it will accelerate the adoption of ISM for quantitative super-resolution imaging.
CHEN, Z.; Yuan, Z.; Lu, Z.; Li, N.; Fu, C.; Sun, L.; Yao, H.; Liu, H.; Xian, Q.; Wang, T.; Li, W.; Liang, Z.; Li, B.; Zhang, Y.; Li, Y.; Wang, H.; Bian, W.-j.; Liu, X.; Chen, Y.; Li, B.; SUN, L.; Wang, Y.; Qiu, Z.
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Functional ultrasound (fUS) imaging provides deep, wide-field access to brain function, but instrument cost and complex acquisition and analysis workflows restrict its wider adoption. Here we introduce @fUS, an agentic transcranial imaging platform with an open architecture that integrates purpose-built hardware with executable experimental skills and persistent structured memory. The system combines 128-channel, 16-bit acquisition at 125 MHz with GPU-based processing and enables functional imaging through the intact scalp and skull of mice, with approximately 100-m spatial resolution and 10-Hz temporal sampling, providing a basis for longitudinal studies without cranial-window surgery. Its agent connects scientific objectives to experimental design, direct instrument control and data analysis, retaining context across interactions and supporting inspectable workflows through text and hands-free voice control. Neuroscience trainees with limited engineering experience independently configured the system within 30 min. Agent-assisted exploration of whisker-stimulation datasets revealed low-frequency vascular changes beyond conventional response mapping, supported by complementary two-photon measurements of single-vessel diameter. By combining transcranial imaging with accessible instrument control and analysis, @fUS provides a foundation for wider adoption and larger, more diverse neuroscience datasets. More broadly, it offers a framework for scientific instruments in which measurement, computation and experimental reasoning are developed as parts of the same system.
Royer, L. A.
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Volumetric microscopy produces datasets too large to share as files and too rich to convey as still images, yet exploring them interactively means installing desktop software or building web infrastructure. Here we present Luxar, an open-source Python framework that compiles multidimensional data into compact archives that any modern browser renders from static file hosting. Fluorescence volumes are represented as mixtures of anisotropic Gaussians (Gaussian splats): the image becomes geometry that the GPU draws directly, with no voxel grid to rebuild. Splats share one scene with points, trajectories and surfaces. Blind-spot cross-validation sets the number of Gaussians without noise-free references. Across 17 volumes from 12 datasets and four modalities, this yields a median 99-fold (6- to 340-fold) size reduction at 26 to 67 dB peak signal-to-noise ratio (PSNR), in minutes per volume on one GPU. We demonstrate Luxar on a 500-timepoint embryogenesis recording, super-resolution localisations, single-cell atlases and lineage tracking, each explorable through a shareable link.