Microscopy-Directed Imaging Mass Spectrometry for Rapid High Spatial Resolution Molecular Imaging of Glomeruli

Journal Article (2023)
Authors

Allison B. Esselman (VanderBilt University)

Nathan Heath Patterson (VanderBilt University)

L.G. Migas (TU Delft - Team Raf Van de Plas, VanderBilt University)

Martin Dufresne (VanderBilt University)

Katerina V. Djambazova (VanderBilt University)

Madeline E. Colley (VanderBilt University)

R Van de Plas (TU Delft - Team Raf Van de Plas, VanderBilt University)

Jeffrey M. Spraggins (VanderBilt University)

Research Group
Team Raf Van de Plas
Copyright
© 2023 Allison B. Esselman, Nathan Heath Patterson, L.G. Migas, Martin Dufresne, Katerina V. Djambazova, Madeline E. Colley, Raf Van de Plas, Jeffrey M. Spraggins
To reference this document use:
https://doi.org/10.1021/jasms.3c00033
More Info
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Publication Year
2023
Language
English
Copyright
© 2023 Allison B. Esselman, Nathan Heath Patterson, L.G. Migas, Martin Dufresne, Katerina V. Djambazova, Madeline E. Colley, Raf Van de Plas, Jeffrey M. Spraggins
Research Group
Team Raf Van de Plas
Issue number
7
Volume number
34
Pages (from-to)
1305-1314
DOI:
https://doi.org/10.1021/jasms.3c00033
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Abstract

The glomerulus is a multicellular functional tissue unit (FTU) of the nephron that is responsible for blood filtration. Each glomerulus contains multiple substructures and cell types that are crucial for their function. To understand normal aging and disease in kidneys, methods for high spatial resolution molecular imaging within these FTUs across whole slide images is required. Here we demonstrate a workflow using microscopy-driven selected sampling to enable 5 μm pixel size matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) of all glomeruli within whole slide human kidney tissues. Such high spatial resolution imaging entails large numbers of pixels, increasing the data acquisition times. Automating FTU-specific tissue sampling enables high-resolution analysis of critical tissue structures, while concurrently maintaining throughput. Glomeruli were automatically segmented using coregistered autofluorescence microscopy data, and these segmentations were translated into MALDI IMS measurement regions. This allowed high-throughput acquisition of 268 glomeruli from a single whole slide human kidney tissue section. Unsupervised machine learning methods were used to discover molecular profiles of glomerular subregions and differentiate between healthy and diseased glomeruli. Average spectra for each glomerulus were analyzed using Uniform Manifold Approximation and Projection (UMAP) and k-means clustering, yielding 7 distinct groups of differentiated healthy and diseased glomeruli. Pixel-wise k-means clustering was applied to all glomeruli, showing unique molecular profiles localized to subregions within each glomerulus. Automated microscopy-driven, FTU-targeted acquisition for high spatial resolution molecular imaging maintains high-throughput and enables rapid assessment of whole slide images at cellular resolution and identification of tissue features associated with normal aging and disease.

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