Comprehensive MSI-based protocols for the spatial lipidomics characterization of microscale organisms
Spatial omics technologies are essential for characterizing molecular disruptions induced by pathological conditions or environmental stressors while preserving the native spatial context of biomolecules. Among these, mass spectrometry imaging (MSI) has become a central platform for spatial lipidomi...
| Autores: | , , , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2025 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/398218 |
| Acceso en línea: | http://hdl.handle.net/10261/398218 https://api.elsevier.com/content/abstract/scopus_id/105013571573 |
| Access Level: | acceso abierto |
| Palabra clave: | Zebrafish embryos MALDI MALDI-2 Mass Spectrometry Imaging SALDI Sample embedding Spatial Lipidomics http://metadata.un.org/sdg/11 http://metadata.un.org/sdg/3 http://metadata.un.org/sdg/6 http://metadata.un.org/sdg/9 Ensure healthy lives and promote well-being for all at all ages Ensure availability and sustainable management of water and sanitation for all Build resilient infrastructure, promote inclusive and sustainable industrialization and foster innovation Make cities and human settlements inclusive, safe, resilient and sustainable |
| Sumario: | Spatial omics technologies are essential for characterizing molecular disruptions induced by pathological conditions or environmental stressors while preserving the native spatial context of biomolecules. Among these, mass spectrometry imaging (MSI) has become a central platform for spatial lipidomics. However, applying MSI to small and fragile organisms, such as zebrafish embryos (ZFEs), presents significant challenges. In this study, we introduce an integrative MSI-based workflow for spatial lipidomics, optimized for ZFEs but broadly adaptable to other delicate biological microsystems. The workflow integrates a refined sample preparation protocol that enabled high-quality consecutive tissue sections with a systematic comparison of ionization strategies, including matrix-assisted laser desorption/ionization (MALDI), surface-assisted laser desorption/ionization (SALDI), and advanced laser post-ionization (MALDI-2), achieving reproducible multimodal MSI analysis. Beyond comparative assessment, our results provide new insights into the ionization yields for each MSI platform. MALDI-2 stands out as a technique for spatial lipidomics, demonstrating superior ionization efficiency and enabling high-resolution imaging down to a pixel size of 5 μm. This pixel size enhancement improves visualization of anatomical structures and detailed lipid mapping, which are critical for understanding lipid biology in these small models. While MALDI proved highly effective for glycerophospholipids and sphingolipids, SALDI showed better performance for low-molecular-weight and neutral lipids. These results establish a conceptual model linking experimental conditions and analyte properties to optimize ionization across lipid classes, providing a robust framework for MSI-based spatial lipidomics in ZFEs. Therefore, this advancement contributes to the broader application of spatial omics using novel biological systems on biomedical, pharmacological and toxicological research. |
|---|