Spatial Molecular Imaging of the Glycome Using Mass Spectrometry

作者： Roberto A. Ribas1,2, Franca Bucco Paolasso1,2, Alison M. Ryan1,2, Manuel R. Sanchez1,2, Charles M. Soto1,2, Scarlett I. Caffee2, Reece C. Larson1,2, Derek B. Allison3,4, Matthew S. Gentry1,2, Ramon C. Sun1,2, Craig W. Vander Kooi1,2 1Center for Advanced Spatial Biomolecule Research (CASBR),University of Florida, Gainesville, 2Department of Biochemistry and Molecular Biology,University of Florida, Gainesville, 3Markey Cancer Center,University of Kentucky, 4Department of Pathology & Laboratory Medicine,University of Kentucky

简介：

Overview

This protocol details the key steps to enable rigorous and reproducible measurement of both glycogen and N-linked glycans using mass spectrometry imaging. The study addresses challenges in spatial biology, focusing on sample preparation and measurement sensitivity.

Key Study Components

Area of Science

• Mass Spectrometry Imaging
• Spatial Biology
• Glycobiology

Background

• Mass spectrometry-based technologies are essential for mapping biomolecules.
• Challenges include rigor and reproducibility in sample preparation.
• Understanding glycogen and glycan distribution is critical for biological research.
• Improved sensitivity and resolution are necessary for accurate measurements.

Purpose of Study

• To establish a protocol for measuring glycogen and N-linked glycans.
• To enhance reproducibility in mass spectrometry imaging.
• To provide insights into the spatial distribution of glycans in liver tissues.

Methods Used

• Preparation of isoamylase and PNGase F solutions.
• Use of humidity chambers for antigen retrieval.
• Application of dual enzyme spraying using HTX M5 sprayer.
• Matrix-assisted laser desorption ionization mass spectrometry imaging for detection.

Main Results

• Glycogen was localized throughout liver hepatocytes.
• N-linked glycans displayed distinct spatial distributions.
• Histological staining confirmed anatomical structures in liver tissues.
• Both glycogen and glycan chain length distributions were measurable.

Conclusions

• The protocol enables effective measurement of glycogen and N-linked glycans.
• Mass spectrometry imaging provides valuable spatial insights.
• Findings contribute to understanding glycobiology in liver tissues.

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