Masuda Keiko, Abouleila Yasmine, Ali Ahmed, Yanagida Toshio, Masujima Tsutomu
Laboratory for Single-Cell Mass Spectrometry, Quantitative Biology Center, RIKEN, Osaka, Japan.
Methods Mol Biol. 2018;1778:269-282. doi: 10.1007/978-1-4939-7819-9_19.
Live single-cell mass spectrometry (LSC-MS) allows for the detection of hundreds to thousands of metabolite peaks acquired from a single plant cell within a few minutes. Plant cells are first observed under a stereomicroscope, a cell of interest is chosen, and then sampled using a metal-coated glass microcapillary for subsequent analysis. A few microliters of ionization solvent is then added to the rear end of the capillary followed by the introduction of the capillary's content directly into the mass spectrometer. High voltage is applied between the capillary and the mass spectrometer inlet to induce nanospray ionization. Metabolite structural confirmation is performed using tandem mass spectrometry analysis (MS/MS) and fragments are matched with MS/MS databases to predict metabolic pathways. This method enables swift and direct molecular detection and identification of specific metabolites from a single plant cell along with their localization within the cell, which will allow for comprehensive understanding of plant metabolomics on a single cell level.
实时单细胞质谱分析（LSC-MS）能够在几分钟内检测从单个植物细胞中获取的数百到数千个代谢物峰。首先在立体显微镜下观察植物细胞，选择感兴趣的细胞，然后使用金属涂层玻璃微毛细管进行采样，以便后续分析。然后向毛细管后端加入几微升电离溶剂，接着将毛细管中的物质直接引入质谱仪。在毛细管和质谱仪入口之间施加高电压以诱导纳米喷雾电离。使用串联质谱分析（MS/MS）进行代谢物结构确认，并将碎片与MS/MS数据库进行匹配以预测代谢途径。该方法能够快速、直接地从单个植物细胞中检测和鉴定特定代谢物及其在细胞内的定位，从而有助于在单细胞水平上全面了解植物代谢组学。
