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Related Experiment Video

Updated: May 14, 2026

Spatial Molecular Imaging of the Glycome Using Mass Spectrometry
08:52

Spatial Molecular Imaging of the Glycome Using Mass Spectrometry

Published on: November 28, 2025

Resolving Sialylated N-Glycans and Immune Cell Landscapes Using a Unified Same-Section IMC-MSI Workflow.

Jake Griner1, Reto Gerber2, Mark D Robinson2

  • 1Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC, USA.

Biorxiv : the Preprint Server for Biology
|May 13, 2026
PubMed
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The IMC-first approach is superior for multimodal spatial analysis on FFPE tissues. This workflow preserves antibody signals and glycan data integrity, enabling robust integration of imaging mass cytometry and mass spectrometry imaging.

Area of Science:

  • Spatial biology
  • Biomolecular imaging
  • Analytical chemistry

Background:

  • Multimodal spatial analysis on formalin-fixed paraffin-embedded (FFPE) tissues combines antibody imaging and mass spectrometry imaging (MSI).
  • Chemically intensive derivatization for isomer-resolved glycan MSI can interfere with antibody detection.
  • Optimizing acquisition order is crucial for reliable multimodal data integration.

Purpose of the Study:

  • To systematically evaluate the analytical compatibility and optimal acquisition order of imaging mass cytometry (IMC) and N-glycan MALDI-MSI on the same FFPE section.
  • To assess the impact of Amidation-Activation-X-Linkage (AAXL) derivatization on antibody detection and glycan analysis.
  • To establish a preferred workflow for integrating IMC and MSI data.

Main Methods:

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Last Updated: May 14, 2026

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Published on: November 28, 2025

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  • Comparison of two same-section workflows: AAXL-MALDI MSI followed by IMC (MALDI-first) and IMC followed by AAXL-MALDI MSI (IMC-first).
  • Systematic evaluation of antibody signal preservation and glycan data integrity using specific markers and derivatization strategies.
  • High-precision co-registration for multimodal data integration at domain and pixel levels.

Main Results:

  • The MALDI-first workflow resulted in significant loss of antibody signal, compromising downstream analysis.
  • The IMC-first workflow successfully preserved quantitative antibody performance.
  • IMC-first acquisition maintained spatial glycan distributions, relative abundance, and isomer-specific signal integrity in subsequent AAXL-MALDI MSI.
  • Analytically robust integration of IMC and MSI data was achieved with the IMC-first strategy.

Conclusions:

  • IMC-first acquisition is the preferred strategy for combining antibody imaging with chemically intensive, isomer-resolved glycan MSI on FFPE sections.
  • This workflow enables reliable multimodal spatial analysis, preserving data integrity from both techniques.
  • Provides generalizable guidance for designing future multimodal spatial mass spectrometry experiments.