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Explicitly Normalized ImaGing MAss Spectrometry

Final Report Summary - ENIGMAS (Explicitly Normalized ImaGing MAss Spectrometry)

The Explicitly Normalized ImaGing MAss Spectrometry (ENIGMAS) project aimed at developing methodologies for imaging mass spectrometry (MS) to enable its application to the study of pre-clinical animal models of neurological disease.

Main developments/achievements include:

- Reproducible on-tissue digestion protocols for protein analysis in imaging MS.
- Reproducible sample preparation protocols for metabolite, neuropeptide and intact protein analysis by imaging MS.
- Comprehensive database of peptides and proteins identified by imaging MS in mouse brain tissue sections.
- Bio informatics pipelines for automated and anatomical driven analysis of imaging MS datasets from large animal cohorts.
- Application of imaging MS to pre-clinical models of neurological disease.
- Novel biomolecular insights regarding the changes produced in mouse brain by cortical spreading depression, the electrophysiological equivalent of migraine aura in animals.

The work developed during the grant period was enthusiastically disseminated across the imaging MS and neuroscience communities through the publication of peer reviewed manuscripts (2 published manuscripts, 1 submitted manuscript, 1 manuscript in preparation), participation in scientific conferences (oral and poster communications), participation in workshops and advanced training across Europe through the COST programme BM1104 – Imaging Mass Spectrometry: New Tools For Healthcare Research. The technological improvements will be made publically available through the imaging MS website ( for widespread application across different imaging MS laboratories.

The innovative technologies/methodologies developed will have a profound impact on the imaging MS and on the neurosciences field. Most important, they will allow:

- Facile protein identification through the developed imaging MS methods and protein databases;
- Automatic annotation of imaging MS datasets with the tissue histology/anatomy;
- Facile comparison of animal cohorts in pre-clinical models of neurological disease, whether by extracting data from specific regions in the mouse brain or comparing distributions in different animals;
- Alignment of imaging MS datasets to the Allen Brain Atlas, a genome-wide repository of data from the mouse brain, allows the comparison of imaging MS data with genomic information, thereby increasing the impact of imaging MS data and biological information retrieved;
- Widespread application of imaging MS to other neurological diseases of present day importance, such as stroke, seizure, Alzheimer’s, Parkinson’s.