Biology subjects

Wu, W.

Publications and source records attributed to Wu, W..

At least 19 recordsLinked to original sources

bioRxiv2026

Sequential vaccination matures VRC01-class HIV-1 neutralizing antibodies in humans

Inducing a broadly neutralizing antibody (bnAb) response is a key goal of HIV-1 vaccine development. Despite significant progress, it is unknown whether vaccine-primed B cells can be reactivated and matured to secrete bona fide bnAbs. In the IAVI C107/C110 single-arm, open-label human trial, VRC01-class memory B cells previously primed by the GT1.1 germline-targeting envelope glycoprotein (Env) trimer were reactivated by a BG505 SOSIP.664 trimer boost. Somatic hypermutation in the activated B cells drove higher antibody affinity for the CD4-binding site on fully glycosylated, heterologous Env trimers. Three VRC01-class monoclonal antibodies derived from independent B cell lineages neutralized 18-31% of 119 viruses in a globally diverse panel. This work advances the HIV-1 germline-targeting vaccination concept from proof-of-priming towards proof-of-breadth and defines a benchmark for bnAb induction.

immunology↗

bioRxiv2026

Metabolite-centric Knowledge Graph Unveils Predictive and Mechanistic Insights through Self-supervised Multimodal Learning

Metabolomics research is advancing toward a comprehensive view of metabolites and their biological functions, yet conventional, dictionary-like databases fall short of presenting these complexities. To overcome this limitation, we developed MetaKG, a metabolite-centric knowledge graph that integrates diverse chemical, biological, and semantic data, coupled with a global self-supervised multimodal learning framework that generates advanced embeddings (MetaKGE) for both predictive and mechanistic analyses. MetaKG outperforms existing models in standard representation learning metrics and across 12 external datasets. Furthermore, MetaKG refines existing database information by detecting misannotated labels and uncovering previously overlooked biochemical relationships. Its predictive capability naturally yields testable biochemical hypotheses, as demonstrated in studies of G6PD deficiency and colorectal cancer. By transforming passive, database-scale information into extractable knowledge and actionable insights, MetaKG paves the way for hypothesis-driven discovery in metabolomics and holds promise for informing future research in clinical biomarker identification and therapeutic development.

bioinformatics↗

bioRxiv2026

Cortex-wide single-cell activity mapping in freely moving mice

How the cortex supports complex, naturalistic behavior can be revealed only in freely moving animals. Although head-mounted microscopes promise access to the neural dynamics underlying such behavior, fundamental trade-offs among field of view, spatial resolution, device weight, and background contamination have prevented cortex-wide single-neuron recording, leaving distributed cortical coordination largely inaccessible. To meet these challenges, we introduce COMET (Cortex-wide Observational Miniature Epifluorescence Technique), a 3.75-g head-mounted mesoscope that integrates an aberration-controlled wide-field optical design, uniform high-efficiency excitation and single-coaxial power-and-data transmission with an end-to-end denoising and signal-extraction pipeline. This enables cortex-wide calcium imaging at single-cell resolution in freely moving mice. COMET captures activity from >15,000 layer 2/3 neurons per 5-min recording over fields of view up to 10 mm in diameter while preserving natural behavior. Benchmarking against benchtop two-photon microscopy demonstrates high spatial concordance of detected neurons and matched event-level calcium dynamics. We apply COMET to social interaction to quantify region-specific engagement and to an auditory trace fear-conditioning task that requires natural movement and behavioral expression, revealing time-ordered changes in regional firing-rate structure and functional connectivity preceding a global conditioned response. COMET provides an open, lightweight hardware-software framework for linking single-neuron activity to cortex-scale network dynamics during freely expressed behaviors under changing internal or external conditions.

neuroscience↗

bioRxiv2026

Scalable localization of clustering instability for spatial transcriptomics

Standard spatial transcriptomics workflows rely on clustering methods to define regions of interest for downstream analyses, such as differential expression and pathway enrichment. Although researchers are presented with a single partition, most clustering methods are stochastic, meaning their output depends on random algorithmic choices, typically controlled by a seed; thus, repeating the analysis changing only the seed can yield a different result. Spatial transcriptomics data may be especially sensitive to algorithmic perturbations due to high dimensionality, sparsity, and mixed expression profiles at tissue boundaries and transitional zones; however, the extent of seed-dependent variation and its downstream impact have yet to be systematically evaluated. To bridge this gap, we present InSTability, a framework that localizes disagreement among partitions at the observation level. InSTability is built on (1) an interpretable, per-spot score that quantifies the variability of a spot's cluster assignment across partitions, and (2) an algorithm with time complexity linear in the number of spots for a specified number of runs for computing the instability score at all spots. Together, these advances enable spatially-resolved clustering instability maps for large, high-resolution datasets. Applying InSTability to repeated runs of seven established methods (BayesSpace, GraphST, Leiden, Louvain, SEDR, STAGATE, and SpiceMix) on 18 benchmark samples reveals that clustering instability varies across tissue sections, with shared spatial patterns emerging among different methods applied to the same sample. Furthermore, clustering instability correlates with poor agreement against reference annotations and propagates downstream into differential expression analyses, resulting in inconsistent gene sets across seeds. Thus, InSTability can strengthen spatial transcriptomics studies by highlighting regions where clustering is seed-sensitive and further validation is warranted. InSTability is open-source and available at https://github.com/molloy-lab/InSTability.

bioinformatics↗

bioRxiv2026

SEEKER: A genome-scale library-on-library screening platform for deciphering T cell recognition of antigen

Decoding which antigens activate a given T cell receptor (TCR) is a central challenge in immunology. We present SEEKER, a functional genome-scale library-on-library screening platform built on three elements: a Jurkat-derived T-APC cell (TAPCell) that combinatorially expresses one TCR and one peptide-MHC drawn from separate libraries together with an NFAT activation reporter; encapsulation of single TAPCells in a thermos-reversible hydrogel for isolated clonal expansion, so recognition is read out as intraclonal activation; and dual-asymmetric PCR that links TCR- and pMHC-encoding sequences into a heritable unit, enabling progressive hit enrichment over screening rounds. SEEKER interrogates ~10^8 TCR-pMHC combinations per run without prior knowledge. Screening joint-infiltrating CD8+ T cells from HLA-B*27+ ankylosing spondylitis patients against proteome-wide libraries validated 53 TCR-peptide pairs, uncovered extensive cross-recognition of self and common viral epitopes, and revealed highly polyreactive T cells recognizing up to 80 autoantigens, implicating virus-triggered poly-autoreactivity in multi-organ autoimmune diseases.

immunology↗

bioRxiv2026

Hypomagnetic field inhibits seed germination via ROS signaling and reprogramming transcriptome in Arabidopsis thaliana

The geomagnetic field (GMF) is a ubiquitous yet poorly understood environmental factor that affects plant growth and development. It has been suggested that the blue-light photoreceptor cryptochrome (CRY) functions as a critical magnetic-field sensor in birds and insects through the radical-pair mechanism (RPM). However, whether CRY acts as a bona fide transducer of GMF signals and what are the downstream cascades remain largely elusive in plants. To address these key issues, we generated hypomagnetic field (HMF) via Helmholtz coils and passive magnetic shielding to investigate the GMF deprivation effects in Arabidopsis thaliana. HMF significantly delays the germination speed without altering the final germination rate, suggesting that HMF specifically modulates germination kinetics rather than seed viability. Time-resolved transcriptomic profiling revealed a coordinated transcriptional shift characterized by downregulation of growth-promoting genes and concurrent upregulation of defense-related genes under HMF. This biphasic transcriptional reprogramming coincides with a significant increase of reactive oxygen species (ROS), linking redox perturbation to the germination delay. Consistently, supplementation with ROS-scavenging antioxidants (e.g., reduced glutathione and ascorbic acid) rescues both germination speed and the misregulated expression of a subset of HMF-responsive genes, suggesting the central role of ROS in mediating HMF bioeffects. Genetic analysis using cry1 cry2 double mutants further indicated that HMF-induced germination delay operates through both CRY-dependent and -independent signaling pathways, suggesting the involvement of additional magnetosensitive modules beyond the canonical CRY-based RPM. These findings suggest that the GMF acts as a positive environmental cue that fine tunes the growth-defense trade-off through a redox-dependent signaling.

plant biology↗

bioRxiv2026

A self-limiting, TREM2-dependent anabolic program confers microglial resilience in Alzheimer's disease

Microglia are key drivers of Alzheimer's disease (AD), and TREM2, one of the strongest genetic risk factors, enables the disease-associated microglia (DAM) state required for plaque engagement. How TREM2 sustains this protection, and why it falters as disease advances, remain unclear. Using proteomics, single-cell transcriptomics, and in vivo metabolic labeling, we show that plaque-associated microglia mount a TREM2-dependent anabolic program coupling nascent protein synthesis to mitochondrial biogenesis. This program, not the DAM signature, marks phagocytically competent microglia; its loss is associated with proteostatic overload and mitochondrial dysfunction. Unexpectedly, this anabolic program is self-limiting, peaking at low amyloid burden and declining as burden rises. This decline is conserved in humans: across 523 donors, anabolic capacity tracks TREM2 but falls at advanced Braak stage even as DAM signature keeps rising. Thus, anabolic capacity, rather than activation state, marks microglial resilience in AD, and sustained stimulation of phagocytosis by anti-amyloid antibodies risks biosynthetic exhaustion and self-limiting efficacy.

neuroscience↗

bioRxiv2026

A multi-tissue epigenomic atlas links the sheep non-coding genome to domestication and complex traits

Non-coding regulatory variation drives complex traits, domestication, and evolutionary adaptation, yet the sheep genome lacks high-resolution functional annotation. Here we present SheepEpimap, a multi-tissue regulatory atlas harmonizing 516 CUT&Tag histone modifications, ATAC-seq, and RNA-seq datasets across 43 adult tissues in sheep. We annotated 2.93 million cis-regulatory elements, yielding 557,441 enhancer-gene pairs and 145,407 variants with allele-specific effects. By training a sequence-to-function deep-learning model, we decoded the base-pair syntax of chromatin accessibility, annotated transcription factor motif instances genome-wide, and constructed 12,210 tissue-specific gene regulatory networks (GRNs). Integrating this resource with multi-tissue expression quantitative trait loci, selection sweeps, and genome-wide association studies prioritized non-coding variants driving domestication and complex traits. Finally, cross-species analysis revealed that sequence-conserved, tissue-matched enhancers were significantly enriched in the heritability of complex traits and diseases in humans. In summary, SheepEpimap (https://genome.ucsc.edu/s/mengzhu/SheepEpimap) provides an open-access foundational ecosystem for sheep functional genomics, precision breeding, and comparative biology.

genetics↗

bioRxiv2026

Pto evolved from malectin-like RLKs phosphorylates AvrPtoB to promote Prf-mediated immunity in Solanum pimpinellifolium

Bacterial speck, caused by Pseudomonas syringae pv. tomato (Pst), is a devastating disease of tomato that severely limits global tomato productivity. Understanding the molecular mechanisms underlying Pst and tomato is essential for developing disease resistant varieties. Here, we demonstrate that Pto, the first disease-resistance gene conferring recognition of a specific pathogen, phosphorylates Pst type III effector AvrPtoB at serine 335 site. This post-translational modification triggers the dissociation of the Prf immune complex, enhancing immune signaling and reducing bacterial pathogenicity. Furthermore, evolutionary analyses indicate that Pto-associated proteins originated from malectin-like receptor kinases (MLRs) through loss of the extracellular domain. Crucially, we identified two key amino acid substitutions, Arg158 and Glu258 in Pto, which replace the ancestral lysine residues in MLRs (SpHREK1-1, SpHERK1-2 and SpHERK1-3). These substitutions stabilize Pto by preventing degradation mediated by AvrPtoB's E3 ubiquitin ligase activity. Our findings reveal a novel mechanism, by which Pto phosphorylates a bacterial effector to trigger enhanced immunity and elucidate the key evolutionary adaptations that have shaped Pto into a stable resistance protein.

plant biology↗

bioRxiv2026

Immunometabolic Reprogramming by Thyroid-Stimulating Antibodies Drives Orbital Adipogenesis via Histone Lactylation in Thyroid Eye Disease

Thyroid eye disease (TED) is conventionally viewed as an autoimmune inflammatory disorder. However, the metabolic determinants driving orbital tissue remodeling remain largely undefined. Here, we uncover a paradigm-shifting mechanism whereby thyroid-stimulating antibodies (TSAbs) instigate profound metabolic reprogramming in orbital fibroblasts (OFs), driving a switch toward glycolysis and markedly elevating lactate production via the CREB/LDHA axis. Critically, we demonstrate that this TSAbs-induced metabolic perturbation is not a mere byproduct but the principal driver of pathology. Mechanistically, lactate functions as a signaling metabolite that directly potentiates adipogenesis via histone lactylation-dependent transcriptional activation of the master adipogenic regulators PPAR{gamma} and C/EBP. Collectively, our findings underscore metabolic reprogramming as a central pathogenic mechanism in TED, whereby autoimmune cues drive disease progression through metabolic-epigenetic crosstalk. This work substantiates the paradigm of TED as an immune- metabolic disorder.

immunology↗

bioRxiv2026

Language Dysfunction Associated with Pathological Brain Connectivity in Children with Epilepsy

Language impairment is common in childhood epilepsy and may arise in part from disruption of the distributed brain networks that support language. In self-limited epilepsy with centrotemporal spikes (SeLECTS)--the most common focal epilepsy of childhood--hyperconnectivity has been linked to poor language outcomes, but the specific network patterns associated with language dysfunction remain unclear, limiting the ability to target neurostimulation rationally. We recorded high-density EEG from 27 children with SeLECTS and 29 age-matched controls during verb generation and rest, and quantified functional connectivity across multiple frequency bands and bilateral frontal, temporal, occipital, and motor regions. Using multivariate pattern analysis, we identified connectivity patterns that predicted language ability, distinguished patterns shared across groups from those specific to SeLECTS and tested whether spatially specific connectivity provided information beyond whole-brain or hemispheric averages and conventional clinical variables. Frontal and occipitotemporal connectivity, particularly within the left hemisphere, predicted language ability across groups, whereas motor-network connectivity emerged as the dominant SeLECTS-specific predictor, linking the epileptogenic network to language dysfunction. Connectivity between specific regions outperformed averaged connectivity measures and predicted language beyond epilepsy diagnosis and antiseizure medication use. Task-based connectivity also outperformed resting-state connectivity. These findings show that language ability is associated with distributed yet spatially specific patterns of brain connectivity, while epilepsy introduces distinct alterations centered on the epileptogenic network. Identifying these disease-specific network patterns provides mechanistic insight into language dysfunction and a rational basis for spatially targeted neuromodulation.

neuroscience↗

bioRxiv2026

Mechanism of fatty acid uptake and inhibition in human FATP2

Fatty Acid Transport Protein 2 (FATP2) couples fatty acid uptake to intracellular activation and is associated with pathological lipid accumulation in cancer and nonalcoholic fatty liver disease. Here, we present cryo-electron microscopy structures of human FATP2 across its reaction cycle. Our structures suggest that FATP2 recruits fatty acids directly from the membrane interface through a hydrophobic tunnel. Catalysis involves a [~]130{degrees} rotation of the C-terminal domain, a transition trapped by the antihypertensive drugs isradipine and benidipine. Both drugs lock the enzyme in a thioester-forming state, but benidipine exhibits superior efficacy by extending a bulky moiety into the primary catalytic tunnel to sterically block substrate entry. Furthermore, we identify a product inhibition mechanism where excess acyl-CoA traps the enzyme, potentially limiting metabolic overload. These findings provide a structural framework for understanding vectorial fatty acid channeling and a scaffold for developing modulators of metabolic flux. HighlightsO_LICryo-EM structures of human FATP2 reveal a membrane-anchored lollipop topology C_LIO_LIEndogenous fatty acids within a hydrophobic tunnel delineate the fatty acid uptake pathway C_LIO_LIIsradipine and benidipine displace fatty acids to trap a non-productive conformation C_LIO_LIAcyl-CoA product inhibition may provide negative feedback via steric occlusion C_LI

biophysics↗

bioRxiv2026

5-HT4 receptor ligand RS67333 modulates striatal acetylcholine and dopamine release via inhibition of acetylcholinesterase

Serotonin 5-HT4 receptors (5-HT4Rs) have emerged as potential therapeutic targets in neuropsychiatric and neurodegenerative disorders by modulating circuits that shape mood, cognition, and motor function. Ligands for 5-HT4Rs can modify dopamine (DA) and acetylcholine (ACh) transmission but mechanisms and circuits have not been fully resolved. Some 5-HT4R agonists have been suggested to have effects that include inhibition of acetylcholinesterase (AChE), raising speculation that 5-HT4R ligands might modulate ACh and/or DA through this action. Here, we investigated the impact of RS67333, a partial 5-HT4R agonist, on DA and ACh release dynamics in the striatum detected ex vivo in mouse brain slices using fast-scan cyclic voltammetry and genetically encoded ACh sensor GRABACh3.0 respectively. We found that RS67333 significantly modulated electrically evoked DA release in dorsolateral striatum and nucleus accumbens core, effects that were abolished by a nicotinic receptor (nAChR) antagonist. In parallel, RS67333 altered evoked ACh signals by extending extracellular ACh lifetime, and correspondingly, RS67333 was found to inhibit striatal AChE enzymatic activity. By contrast, BIMU8, an alternative 5-HT4R ligand that did not inhibit striatal AChE, had no effect on evoked striatal ACh or DA release. These findings indicate that RS67333 modulates striatal ACh transmission, which shapes downstream regulation of DA release by nAChRs, not through 5-HT4Rs but through AChE inhibition. These findings emphasize the caution due in attributing functions to 5-HT4Rs, but also highlight an alternative pharmacological profile of some purported 5-HT4R ligands as AChE inhibitors of potential utility for treating ACh/DA disorders.

neuroscience↗

bioRxiv2026

SOLiD-MaP: A Photoproximity Labeling Platform for Small Molecule Binding Site Mapping on RNA

RNA-targeting small molecules are emerging as promising therapeutic modalities, but their development requires methods that define binding sites and evaluate RNA target selectivity. Existing approaches for detecting ligand-RNA interactions have provided powerful foundations, yet many rely on direct crosslinking or covalent-capture chemistries whose performance depends on ligand-specific probe design, warhead compatibility, and local reaction geometry. Here, we report Singlet Oxygen footprinting on RNA in a Ligand-Directed manner for Mutational Profiling (SOLiD-MaP), a photoproximity labelling platform for small molecule-RNA interaction analysis. Using the Mango-II aptamer and thiazole orange derivatives as a model system, we establish aniline as an efficient nucleophile for singlet oxygen-mediated RNA labelling and demonstrate target-selective labelling driven by ligand-localized photosensitization. We further show that labelling selectivity can be tuned by chemically constraining the singlet oxygen diffusion with a quencher. Finally, we develop a pairwise reverse transcription stop assay and a mutational profiling with next-generation sequencing readouts to infer ligand-proximal regions and unambiguously map binding sites. SOLiD-MaP provides a new, orthogonal strategy for studying small molecule-RNA recognition and should support RNA-focused mechanism-of-action studies.

biochemistry↗

bioRxiv2026

GYS2 Promotes the Progression of Aortic Dissection and Aortic Aneurysm via C5a/NF-κB-Mediated Pro-Inflammatory Macrophage Polarization

BackgroundAortic aneurysm and aortic dissection (AAD) are lethal cardiovascular emergencies characterized by sudden onset and extremely high early mortality. The pro-inflammatory polarization of macrophages is one of the core factors driving the pathogenesis of AAD, but its underlying mechanism remains unclear. This study focuses on the aberrant expression of glycogen synthase 2 (GYS2) in the AAD microenvironment and its role in driving macrophage polarization toward the pro-inflammatory M1 phenotype. MethodsProteomic analysis was conducted to identify protein heterogeneity associated with AAD. Clinical and animal samples were used to evaluate the correlation between GYS2 expression and AAD progression. Whole-body GYS2 knockout mice and adeno-associated virus (AAV)-mediated macrophage-specific gain- and loss-of-function models were utilized to investigate the regulatory role of GYS2 in macrophage polarization and complement activation. Downstream molecular pathways were identified and validated through in vitro stimulation and in vivo exogenous C5a rescue experiments. ResultsGYS2 expression was significantly upregulated in AAD tissues and primarily localized in macrophages. Activation of GYS2 by LiCl or macrophage-specific overexpression of GYS2 exacerbated aortic dilation and extracellular matrix degradation, and increased mortality in AAD mice. Conversely, whole-body GYS2 knockout or macrophage-specific GYS2 knockdown suppressed inflammatory factors, significantly reduced the incidence of AAD, and attenuated vascular injury. Mechanistically, excessive GYS2 in macrophages specifically triggered the complement-coagulation cascade, promoting the generation of the potent anaphylatoxin C5a. C5a further bound to its receptor C5AR1, activating the downstream PLC{beta}3/NF-{kappa}B signaling pathway, thereby inducing M1 macrophage polarization and matrix metalloproteinase-mediated extracellular matrix degradation. In vivo exogenous C5a rescue completely reversed the vascular protective effects conferred by GYS2 deficiency. ConclusionsThis study demonstrates that highly expressed GYS2 regulates the pro-inflammatory polarization of macrophages and extracellular matrix degradation via the complement C5a/PLC{beta}3/NF-{kappa}B signaling axis, which is a key mechanism driving AAD progression. Specific inhibition of macrophage GYS2 can effectively alleviate aortic vascular inflammation and prevent AAD progression, providing a promising novel strategy for the clinical conservative treatment of AAD.

Molecular Biology↗

bioRxiv2026

Coordinated immune-epithelial dynamics in the nasal epithelium protect against respiratory virus infection

Epithelial remodeling is a hallmark of host antiviral responses that strengthen barrier defenses against infection. Our current understanding of viral pathophysiology within the nasal epithelium, the initial site of most respiratory viral infections, remains incomplete due to limited understanding of the native tissue architecture and protective epithelial remodeling during immune events. Leveraging coronavirus disease 2019 (COVID-19) as a model, we applied spatial multi-omics on nasal cross-sectional tissues to characterize the immune-epithelial landscape during infection, identifying a coordinated increase in goblet cells and suppressive macrophages associated with elevated IL13 in the immune compartment. Our results further reveal that goblet cell and suppressive macrophage enrichment are spatially linked in proximity to IL13-expressing CD4 T cells, consistent with IL13-driven remodeling in situ. Using a primary human nasal air-liquid interface model, we demonstrate that IL13 alone is sufficient to remodel epithelial composition and morphology, subsequently restricting viral infection by reshaping the apical mucus barrier of the nasal epithelium. Our findings uncover a spatially organized, IL13-driven circuit for immune-epithelial remodeling as a protective barrier against respiratory viral infections.

immunology↗

bioRxiv2026

Cell type-centric interaction networks define spatial architecture of intrahepatic cholangiocarcinoma

Tumor spatial organization critically shapes disease progression and therapeutic response, yet remains poorly defined. Intrahepatic cholangiocarcinoma (iCCA), a rare and aggressive liver malignancy with extensive stromal and immune remodeling, provides a compelling model to study tumor architecture. We generated a single-cell spatial atlas of 1 million cells from 131 iCCA patients using 53-plex spatial proteomics. To systemically characterize tumor spatial organization, we developed a graph-based deep learning framework to define cell type-centric interaction networks, identifying 41 distinct multicellular spatial patterns. Integration of these networks revealed higher-order tumor- and immune-enriched microenvironments associated with patient outcomes. Notably, neutrophil-associated tumor-enriched and tumor-desert microenvironments delineated patient groups with opposing clinical outcomes and distinct neutrophil states. These findings were validated by single-cell spatial transcriptomic profiling of 6 million cells from 162 iCCA patients. Together, this study defines the spatial architecture of iCCA and provides a comprehensive resource for exploring tumor spatial organization.

cancer biology↗

bioRxiv2026

An open-source stereotaxic container with an integrated cutting guide for human brain fixation during magnetic resonance imaging and sectioning for histology

IntroductionPostmortem imaging at ultrahigh field strengths, such as 7 Tesla (7T) magnetic resonance imaging (MRI), enables unprecedented visualization of fine brain structures and pathology. However, precise registration between MRI and histology is often compromised by tissue deformation and lack of standardized tools for preserving anatomical orientation. A reliable, high-resolution MRI-to-histology workflow is essential for MRI-guided sampling to investigate neurodegenerative diseases. MethodsWe developed a human postmortem brain container composed of stereotaxic enclosure and cutting guide system using iterative 3D printing. The final design is constructed from Victrex AM 200, a polyaryletherketone material engineered with MRI compatibility, high mechanical strength, and high resistance to fixatives. Human brains were embedded in an agar/sucrose mixture within the container and scanned at 7T using high-contrast and high-resolution imaging sequences. The cutting guide enables reproducible sectioning at defined tissue coronal slab thicknesses (9.2 {+/-} 0.6 mm). Block face photographs of each coronal slab were then co-registered with MRI for targeted tissue sampling. ResultsTo date, 215 human postmortem brains have been imaged ex vivo using this container. The container maintains consistent spatial orientation from imaging through sectioning, supporting correlation between MR-visible features and histological findings. Imaging-visible abnormalities, such as white matter hyperintensities and cerebral microbleeds, were localized and sampled with minimal deformation. The container enables single-scan workflows, avoiding custom cutting templates or repeat imaging. It supports deep learning-based lesion segmentation and MRI-guided histology registration, facilitating large-scale neuropathological investigations. ConclusionThe stereotaxic cutting guide system addresses a critical gap in postmortem imaging by integrating MRI-compatible materials, precision-guided sectioning, and high-throughput imaging. It has enabled multimodal studies of aging and neurodegeneration and is being adopted in neuropathological workflows. This platform provides a reproducible, scalable, and anatomically precise solution for aligning ultrahigh field MRI with histology, enhancing both clinical research and digital pathology efforts.

bioengineering↗