Super-resolved Optical Imaging, Reconstruction, and Spatial Analysis of Whole Mouse Renal Glomeruli via GloMAP

Abstract

The glomerulus plays a crucial role in blood filtration and is made up of several key components that function collaboratively. Traditionally, optical microscopy has provided insights into general physiology and molecular distributions, and electron microscopy has been used to reveal ultrastructural details of the glomerular structures. While past studies have extensively examined local changes in glomerular diseases and aging, the global relationships and coordination among glomerular structures remain poorly understood due to the limitations of two-dimensional and partial analyses. To address these limitations, I present a novel pipeline that employs super-resolution fluorescence microscopy to achieve holistic three-dimensional imaging, reconstruction, and analysis of whole mouse glomeruli at 100 nm resolution. This workflow integrates advanced tissue labeling techniques and super-resolution imaging to capture entire mouse glomeruli, and combines manual segmentation with machine learning methods to reconstruct all glomerular compartments in 3D. I further demonstrate the versatility of this approach by applying it to various glomerular types, including cortical and juxtamedullary, as well as different conditions such as aging and focal segmental glomerulosclerosis. I also show that the detailed spatial analyses of the resulting models reveal new insights into the spatial correlations among glomerular components in both aged and diseased mice. Once published, the unique datasets generated by this approach will serve as a valuable resource for the nephrology community.