Analyzing cell type-specific dynamics of metabolism on kidney

Abstract Conventional single-cell metabolomics approaches such as matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) generate biochemical snapshots, neglecting the inherent dynamic nature of metabolism. Here we describe a platform based on isotope tracing and MALDI-MSI that allows in-situ dynamic measurements of cell type-specific metabolism at single-cell resolution, and thus unravel cell metabolism within its tissue architecture. We applied different 13C-isotope-labeled nutrients on vibratome slices of fresh mouse kidney. MALDI-MSI at single-cell resolution (i.e. pixel size of 5 × 5 μm2) was then applied to detect metabolites and lipids from the harvested tissues. Following MALDI-MSI analysis, post-MSI-analyzed sections were stained and subsequently imaged using multiplexed immunofluorescence (IF) microscopy for cell-type identification. This platform allows to achieve single-cell resolution in situ and hence interpret cell type-specific metabolic dynamics in the context of structure and metabolism of neighboring cells.