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Abstract: FR-PO0752

Super-Resolved Three-Dimensional (3D) Imaging and Analysis of Whole Mouse Glomeruli via GloMAP

Session Information

Category: Glomerular Diseases

  • 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology

Authors

  • Vaughan, Joshua C., University of Washington, Seattle, Washington, United States
  • Ali, Adilijiang, University of Washington, Seattle, Washington, United States
  • Poudel, Chetan, University of Washington, Seattle, Washington, United States
  • Eng, Diana G., University of Washington, Seattle, Washington, United States
  • Pippin, Jeffrey W., University of Washington, Seattle, Washington, United States
  • Shankland, Stuart J., University of Washington, Seattle, Washington, United States
Background

Glomerular structure is central to kidney function, yet traditional imaging relies on thin sections or modest spatial resolution, limiting the ability to study whole glomeruli as 3D functional units. To address this challenge, we developed GloMAP, a pipeline combining super-resolution optical microscopy and human-assisted computational segmentation to reconstruct whole mouse glomeruli at 100nm resolution.

Methods

We used a novel fluorescent tricolor stain to label tissue sections and we imaged whole hydrogel-expanded glomeruli by confocal microscopy. Manual and machine-learning-assisted segmentation defined six structural compartments (e.g., GBM, Bowman’s capsule) and enabled classification of cell types. These data were used to analyze volume fractions, surface areas, basement membrane thicknesses, and other properties.

Results

24 glomeruli from young adult, aged, and experimental FSGS mice were reconstructed, allowing analysis of glomerular compartments and structures and classification of the 4 principal glomerular cell types with >90% accuracy. This revealed age- and disease-associated changes, including segmental basement membrane thickening, altered mesangial structure, and altered cell type proportions. Global maps of GBM and BCBM thickness revealed distinct remodeling patterns across conditions. In experimental FSGS glomeruli, we observed multilayered BCBM, podocyte hypertrophy, and parietal epithelial cell migration (Fig. 1).

Conclusion

GloMAP enables high-resolution, 3D reconstruction of whole glomeruli using accessible methods. It is a powerful platform for studying whole glomerular architecture and it provides a foundation for integrative modeling and therapeutic evaluation.

Fig. 1. Representative segmental pathological patterns from an experimental FSGS mouse glomerulus.

Funding

  • NIDDK Support

Digital Object Identifier (DOI)