Controlled Humidity Levels for Fine Spatial Detail Information in Enzyme-Assisted N-Glycan MALDI MSI

Dušan Veličković, Kumar Sharma, Theodore Alexandrov, Jeffrey B. Hodgin, Christopher R. Anderton

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


Investigation of the spatial distribution of N-glycans in tissue specimens has emerged as a powerful tool in clinical research, in part, because altered N-glycans are often a hallmark of disease progression. Mass spectrometry imaging of N-glycans relies on peptide N-glycanase spraying and tissue incubation for efficient in situ release of N-glycans from their carrier proteins. Unstandardized and uncontrolled incubation steps often cause significant delocalization of released N-glycans, resulting in the inability to link given N-glycan composition to a specific microanatomical region in the tissue. Herein, we optimized the incubation step to provide accurate and sensitive MALDI-MSI of N-glycans. Specifically, we tested saturated solutions of various salts that maintain constant relative humidity in the incubation chamber. We showed that the best performance was achieved using a saturated solution of KNO3that maintains an 89% RH. Under these conditions, near maximal sensitivity was achieved with the minutest ion delocalization, which we demonstrated at a 35 μm spatial resolution, where we observed six distinct spatial patterns that colocalize to distinct microanatomical compartments in a kidney nephrectomy tissue section.

Original languageEnglish (US)
Pages (from-to)1577-1580
Number of pages4
JournalJournal of the American Society for Mass Spectrometry
Issue number8
StatePublished - Aug 3 2022

ASJC Scopus subject areas

  • Structural Biology
  • Spectroscopy


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