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基质辅助激光解吸/电离质谱成像技术对细菌脂肽抗生素的原位分析

In Situ Analysis of Bacterial Lipopeptide Antibiotics by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging.

作者信息

Debois Delphine, Ongena Marc, Cawoy Hélène, De Pauw Edwin

机构信息

Mass Spectrometry Laboratory (LSM-GIGA-R), Chemistry Department, University of Liege - Allee du 6 aout, 11 - B6c - Chimie Licence et Recherche - Quartier Agora, Liege-1 (Sart Tilman), B-4000, Belgium.

Gembloux Agro-Bio Tech, University of Liège, Gembloux, Belgium.

出版信息

Methods Mol Biol. 2016;1401:161-73. doi: 10.1007/978-1-4939-3375-4_11.

Abstract

Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) is a technique developed in the late 1990s enabling the two-dimensional mapping of a broad variety of biomolecules present at the surface of a sample. In many applications including pharmaceutical studies or biomarker discovery, the distribution of proteins, lipids or drugs, and metabolites may be visualized within tissue sections. More recently, MALDI MSI has become increasingly applied in microbiology where the versatility of the technique is perfectly suited to monitor the metabolic dynamics of bacterial colonies. The work described here is focused on the application of MALDI MSI to map secondary metabolites produced by Bacilli, especially lipopeptides, produced by bacterial cells during their interaction with their environment (bacteria, fungi, plant roots, etc.). This chapter addresses the advantages and challenges that the implementation of MALDI MSI to microbiological samples entails, including detailed protocols on sample preparation (from both microbiologist and mass spectrometrist points of view), matrix deposition, and data acquisition and interpretation. Lipopeptide images recorded from confrontation plates are also presented.

摘要

基质辅助激光解吸/电离质谱成像(MALDI MSI)是20世纪90年代末开发的一项技术,能够对样品表面存在的多种生物分子进行二维映射。在包括药物研究或生物标志物发现在内的许多应用中,蛋白质、脂质、药物和代谢物的分布可以在组织切片中可视化。最近,MALDI MSI在微生物学中的应用越来越多,该技术的多功能性非常适合监测细菌菌落的代谢动态。本文所述工作聚焦于应用MALDI MSI绘制芽孢杆菌产生的次级代谢产物,特别是细菌细胞在与环境(细菌、真菌、植物根等)相互作用过程中产生的脂肽。本章阐述了将MALDI MSI应用于微生物样品所带来的优势和挑战,包括详细的样品制备方案(从微生物学家和质谱学家的角度)、基质沉积以及数据采集和解释。还展示了从对峙平板记录的脂肽图像。

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