Title : Beyond plaques: 10-nanometer-scale pathoconnectomics of Alzheimer’s disease
Abstract:
Alzheimer’s neuropathology is commonly described by lesion burden, yet cognitive decline emerges from the disruption of neural circuits. Amyloid-β (Aβ) plaques are associated with local dendritic spine loss, excitatory synapse depletion, impaired axonal transport, and dystrophic neurites [2–5]. In the 5xFAD model, plaque-associated axonal spheroids (PAAS) can impede action-potential propagation and disrupt long-range connectivity [6]. Studies in mouse and human Alzheimer’s tissue further implicate endolysosomal, cytoskeletal, lipid-transport, and mTOR-associated pathways in spheroid pathology [7]. These findings identify the plaque–PAAS microenvironment as a critical unit of disease, but one that remains difficult to interrogate: fluorescence microscopy (FM) provides molecular specificity without continuous nanoscale architecture, whereas electron microscopy (EM) reveals ultrastructure but is difficult to molecularly multiplex and scale across translational cohorts.
To address this gap, Panluminate developed, to our knowledge, the first integrated, scalable, all-optical pathoconnectomics platform that co-registers continuous 3D ultrastructure with multiplexed molecular information in a single tissue volume. Panluminate’s proprietary expansion chemistry and dense pan-protein labeling reveal neurites, synapses, organelles, and neuropil, while multiplexed immunolabeling identifies the cell types, disease proteins, and signaling compartments embedded within that architecture. The underlying method achieves approximately 14 ± 3 nm effective lateral resolution under ~20-fold expansion conditions [8]. Automated sample processing and high-NA volumetric light-sheet imaging extend this capability beyond selected microscopic fields toward reproducible, cohort-scale studies.
In collaboration with translational Alzheimer’s researchers, Panluminate is applying this workflow to age-matched 5xFAD and control mouse brain tissue. Initial 8-plex datasets resolve Aβ deposits together with NF-H-positive axons, MBP-positive myelin, endolysosomal compartments, nuclear architecture, and pan-protein neuropil. We are augmenting a panel of 35 validated brain-tissue antibodies with 15 Alzheimer’s-focused targets to quantify plaque-distance-dependent changes in PAAS burden, axonal caliber and continuity, organelle accumulation, myelin organization, and synaptic architecture. In parallel, we are adapting the workflow to postmortem human brain tissue to establish a translational bridge between animal models and human neuropathology.
Because disease-specific molecular panels can be exchanged without sacrificing ultrastructural context, the same platform can be applied to other neurodegenerative and neurological disorders in which protein pathology, cellular identity, and circuit remodeling must be measured together. Panluminate is now extending this new pathoconnectomics paradigm through collaborations with neuropathologists, translational researchers, and therapeutic-development teams.
