Mass Spectrometry Imaging (MSI) is a powerful emerging technology opening a new frontier in the research field of Spatial ‘Omics. MSI maps the spatial and temporal distribution of 100’s to 1000’s of biomolecules (proteins, peptides, metabolites, drugs and chemicals) at once within biological tissues providing an enhanced understanding of underlying biological processes. Matrix assisted laser desorption ionization MS (MALDI-MS) is the most common MSI approach. Crucial to metabolite and peptide identification directly off tissues is the use of high mass resolution mass spectrometry, such as MALDI-FTMS. Several recent examples and strategies exemplifying the approach will be presented, including mapping and identification of peptide neurotoxins from the venom of sea anemone; and exploring the potential of measuring metabolite and peptide distributions within FFPE bio-banked tissues.