MALDI-Mass Spectrometric Imaging for the Investigation of Metabolites in Medicago truncatula Root Nodules
MALDI-Mass Spectrometric Imaging for the Investigation of Metabolites in Medicago truncatula Root Nodules
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DOI：
10.3791/51434
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发表时间：
2014-03-01
影响因子：
1.2
通讯作者：
Li, Lingjun
中科院分区：
文献类型：
--
作者：
Gemperline, Erin;Li, Lingjun
用于研究小分子（如药物或内源性代谢物）的大多数技术采用组织提取物，需要对可能导致所研究代谢途径变化的目标组织进行均质化1。质谱成像（MSI）是一种强大的分析工具，可以提供生物组织样本完整切片内分析物的空间信息1 -5。该技术已广泛用于研究各种类型的化合物，包括蛋白质、肽、脂质和小分子如内源性代谢物。基质辅助激光解吸/电离（MALDI）-MSI可以同时检测多种代谢物的空间分布。在此，提出了一种专门用于对豆科植物根和根瘤进行非靶向代谢组学MSI实验的方法，该方法可以揭示发生的生物过程。这里介绍的方法显示了一个典型的MSI工作流程，从样品制备到图像采集，并重点介绍了矩阵应用步骤，展示了几种可用于检测小分子的矩阵应用技术。一旦生成MS图像，将讨论和演示感兴趣的代谢物的分析和鉴定。这里介绍的标准工作流程可以很容易地针对不同的组织类型、分子种类和仪器进行修改。
Most techniques used to study small molecules, such as pharmaceutical drugs or endogenous metabolites, employ tissue extracts which require the homogenization of the tissue of interest that could potentially cause changes in the metabolic pathways being studied1. Mass spectrometric imaging (MSI) is a powerful analytical tool that can provide spatial information of analytes within intact slices of biological tissue samples1-5. This technique has been used extensively to study various types of compounds including proteins, peptides, lipids, and small molecules such as endogenous metabolites. With matrix-assisted laser desorption/ionization (MALDI)-MSI, spatial distributions of multiple metabolites can be simultaneously detected. Herein, a method developed specifically for conducting untargeted metabolomics MSI experiments on legume roots and root nodules is presented which could reveal insights into the biological processes taking place. The method presented here shows a typical MSI workflow, from sample preparation to image acquisition, and focuses on the matrix application step, demonstrating several matrix application techniques that are useful for detecting small molecules. Once the MS images are generated, the analysis and identification of metabolites of interest is discussed and demonstrated. The standard workflow presented here can be easily modified for different tissue types, molecular species, and instrumentation.