Single-Cell Metabolic Profiling: Metabolite Formulas from Isotopic Fine Structures in Heterogeneous Plant Cell Populations
Single-Cell Metabolic Profiling: Metabolite Formulas from Isotopic Fine Structures in Heterogeneous Plant Cell Populations
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DOI：
10.1021/acs.analchem.0c00936
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发表时间：
2020-05-19
影响因子：
7.4
通讯作者：
Vertes, Akos
中科院分区：
文献类型：
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作者：
Samarah, Laith Z.;Khattar, Rikkita;Vertes, Akos
细胞群中代谢异质性的表征需要对单细胞进行分析。目前大多数单细胞分析方法依赖于细胞操作，可能会改变单个细胞中代谢物的丰度。小的样品体积和代谢物的化学多样性是单细胞代谢组学中的额外挑战。在这里，我们描述了基于光纤的激光烧蚀电喷雾电离（f-LAESI）与21 T傅立叶变换离子回旋共振质谱（21 TFTICR-MS）的组合，用于植物组织中的原位单细胞代谢分析。选择被细菌感染的单个植物细胞，并直接从组织中取样，而不进行细胞操作，通过用细光纤尖端进行中红外消融，以通过f-LAESI进行电离。超高性能21 T-FTICR-MS能够同时捕获47种已知化合物和11种未知化合物的同位素精细结构（IFS），从而阐明它们在单细胞中的元素组成，并提供细胞群中代谢异质性的信息。
Characterization of the metabolic heterogeneity in cell populations requires the analysis of single cells. Most current methods in single-cell analysis rely on cell manipulation, potentially altering the abundance of metabolites in individual cells. A small sample volume and the chemical diversity of metabolites are additional challenges in single-cell metabolomics. Here, we describe the combination of fiber-based laser ablation electrospray ionization (f-LAESI) with 21 T Fourier transform ion cyclotron resonance mass spectrometry (21TFTICR-MS) for in situ single-cell metabolic profiling in plant tissue. Single plant cells infected by bacteria were selected and sampled directly from the tissue without cell manipulation through mid-infrared ablation with a fine optical fiber tip for ionization by f-LAESI. Ultrahigh performance 21T-FTICR-MS enabled the simultaneous capture of isotopic fine structures (IFSs) for 47 known and 11 unknown compounds, thus elucidating their elemental compositions from single cells and providing information on metabolic heterogeneity in the cell population.