Method development for correlating lipid molecular information with anatomy in C. elegans

Caenorhabditis elegans (C. elegans) is an important model organism for studying fat storage and lipid metabolism. Mass-spectrometry imaging (MSI) is an emerging technology for mapping the spatial distribution of lipids. However, MSI analysis of C. elegans is limited by the lack of reproducible sample preparation methods. Here, we present a microfluidics-based workflow for preparing consecutive nematode sections while retaining their internal structures, such as the pharynx, intestine, and embryos. This method enables multimodal analysis of single nematodes by MSI and Oil Red O staining, revealing a number of lipids spatially distributed across different body parts. The feature-based image reconstruction technique enables the three-dimensional reconstruction of nematodes based on optical images and MSI-based lipid mapping. The present method can correlate MSI data with various imaging modalities to provide detailed correlations between anatomical features and lipid distribution in nematodes.