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URL: https://dx.doi.org/DOI:10.17504/protocols.io.be2ajgae

Authors: Dusan Velickovic, Jessica Lukowski, Guanshi Zhang, Theodore Alexandrov, Chris Anderton, Kumar Sharma

Group: KPMP

Summary: Mass spectrometry imaging is an exciting technologywhich enables a simultaneous analysis of multiple molecular components directly from single cells, tissues, and organs,. In combination with histological methods, this technique provides information about the spatial distribution of molecules in various biological tissues. Particularly, MALDI-MS imaging increases the coverage of metabolites by using different matrices and ion modes. In coordination with in situ analysis of proteins, transcripts and epigenetic marks, the complementary spatial information on metabolites will establish metabolic pathways that are dominant and characteristic of disease states. We have recently developed and optimized a spatial metabolomics approach to image small molecules in human kidneys and biopsy sized material. With our combined expertise at UTHSA, PNNL and EMBL and recent advances, we have established methods for identifying metabolites in human kidneys, employed ultra-high mass resolution MS imaging for tissue analysis, and developed a bioinformatics resource (METASPACE) to annotate metabolites for anatomical localization and 3-D reconstruction. Our integrated technology can easily connect with other TIS sites to provide biochemical readouts of genes/proteins in specific tissue and cellular compartments.Although METASPACE bioinformatics provides metabolite annotations of given reliability, there may be an inherent level of ambiguity due to the lack of untargeted MS/MS fragmentation in MALDI-MSI. Metabolite annotations from MALDI-MSI and METASPACE are structurally validated using orthogonal analytical techniques such as LC-MS/MS due to its high metabolome coverage, sensitivity, and throughput. The work flow for ‘bulk omics’ is shown in Figure 5.

Affiliations: Pacific Northwest National Laboratory, Pacific Northwest National Laboratory, University of Texas Health San Antonio, European Molecular Biology Laboratory, Pacific Northwest National Laboratory, University of Texas Health San Antonio

Version: 1

Publication Date: 2020

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