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Tissue Glycome Mapping: Lectin Microarray-Based Differential Glycomic Analysis of Formalin-Fixed Paraffin-Embedded

Chiaki Nagai-Okatani1, Xia Zou2,3, Atsushi Matsuda2,4

  • 1Glycoscience and Glycotechnology Research Group, Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan. chiaki-okatani@aist.go.jp.

Methods in Molecular Biology (Clifton, N.J.)
|January 1, 2022
PubMed
Summary

This study introduces a laser microdissection-lectin microarray (LMD-LMA) method for detailed glycomic profiling of specific cells within formalin-fixed paraffin-embedded tissues. This technique enables reproducible tissue glycome mapping, aiding in understanding glycosylation in health and disease.

Keywords:
Cell type-selective analysisFormalin-fixed paraffin-embedded tissue sectionHistochemistryLaser microdissectionLectin microarrayTissue glycome mapping

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Area of Science:

  • Glycomics
  • Biochemistry
  • Histology

Background:

  • Lectin microarray (LMA) provides high-sensitivity glycan analysis for glycoproteins.
  • Analyzing specific cell populations within tissues requires advanced glycomic profiling methods.

Purpose of the Study:

  • To develop and standardize a laser microdissection-LMA (LMD-LMA) technique for spatial glycomic profiling of formalin-fixed paraffin-embedded (FFPE) tissues.
  • To enable detailed analysis of N- and O-glycans in specific cellular regions within FFPE samples.
  • To facilitate tissue glycome mapping for understanding region- and tissue-specific protein glycosylation.

Main Methods:

  • Integration of laser microdissection (LMD) for targeted tissue collection with lectin microarray (LMA) for glycan analysis.
  • Standardized protocol covering FFPE tissue preparation, staining, protein extraction, labeling, and LMA analysis.
  • Development of fluorescence LMD-LMA for cell type-selective glycomic profiling using lectin-antibody probes.

Main Results:

  • Establishment of a reproducible LMD-LMA protocol for obtaining comparable tissue glycomic profiles.
  • Demonstration of the feasibility of analyzing glycomic profiles from small, specific areas of FFPE tissues.
  • Validation of spatial tissue glycomic profiles for histochemical analysis and cell type-specific glycomic profiling.

Conclusions:

  • The standardized LMD-LMA method provides a powerful tool for tissue glycome mapping.
  • This approach enhances the utilization of FFPE specimens for discovering biological roles and disease-related alterations in protein glycosylation.
  • Spatial glycomic profiling advances the understanding of tissue-specific glycosylation patterns.