ArticleAnalytical chemistry2026
Filtration-Based Rapid ECM-Hydrogel Elimination Enables Robust MALDI MSI of Intestinal Organoids.
Article in Analytical chemistry, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
The rapid development of matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) offers a powerful approach for the 2D metabolic profiling of intestinal organoids. However, its application remains constrained by challenges in sample preparation. Conventional extracellular matrix (ECM)-based hydrogel removal methods, such as repeated centrifugation or enzymatic digestion, are labor-intensive and time-consuming, often causing structural disruption of organoids and degradation of metabolites, thereby compromising the spectral quality and metabolite coverage. In this study, we established a filtration-based rapid ECM-hydrogel elimination (FREE) method using a 70 μm filter, which reduced treatment time from 75 to 25 min while preserving organoid structural integrity, cell viability, and metabolic stability and, more importantly, improved data quality, with a 26.6% increase in metabolite coverage and enhanced signal intensity. Applying this workflow to an LPS-induced inflammatory intestinal organoid model, we observed spatial metabolic remodeling associated with epithelial polarity disruption. Notably, phosphatidylserine (PS) became enriched at the apical membrane, while phosphatidylethanolamine (PE) exhibited a basolateral side, suggesting a link between loss of polarity and early apoptotic signaling. Overall, FREE provides an efficient pretreatment strategy that enables MSI research on intestinal organoids to dissect the mechanisms of inflammation-associated metabolic remodeling.
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