MALDI mass spectrometry imaging of N-glycans was established 4 years ago and is now routinely used by 3 groups around the World. Since our group’s proof-of-principle paper in 2015, we have published several other papers that spatially investigate the N-glycome of formalin-fixed paraffin-embedded (FFPE) tissues that have been extracted from osteoarthritis and ovarian cancer patients. However, further work is required by our group to (i) improve and optimize this workflow, and (ii) verify whether this technique can be used to discover potential diagnostic markers for early-stage diseases. This presentation will provide (i) detailed instructions on our established method for groups new to MALDI mass spectrometry imaging and (ii) a vision for the future of this new glycomic technology.