Department of Chemistry-BMC, Uppsala University, Sweden.
Analyst. 2017 Oct 7;142(19):3639-3647. doi: 10.1039/c7an00885f. Epub 2017 Aug 24.
Molecular profiling of single cells has the potential to significantly advance our understanding of cell function and cellular processes of importance to health and disease. In particular, small molecules with rapid turn-over rates can reveal activated metabolic pathways resulting from an altered chemical environment or cellular events such as differentiation. Consequently, techniques for quantitative metabolite detection acquired in a higher throughput manner are needed to characterize the biological variability between seemingly homogenous cells. Here, we show that nanospray desorption electrospray ionization (nano-DESI) mass spectrometry (MS) enables sensitive molecular profiling and quantification of endogenous species in single cells in a higher throughput manner. Specifically, we show a large number of detected amino acids and phospholipids, including plasmalogens, readily detected from single cheek cells. Further, by incorporating a phosphatidylcholine (PC) internal standard into the nano-DESI solvent, we determined the total amount of PC in one cell to be 1.2 pmoles. Finally, we describe a higher throughput approach where molecules in single cells are automatically profiled. These developments in single cell analysis provide a basis for future studies to understand cellular processes related to drug effects, cell differentiation and altered chemical microenvironments.
单细胞的分子特征分析有可能极大地增进我们对细胞功能和重要健康与疾病相关细胞过程的理解。特别是，具有快速周转率的小分子可以揭示由于化学环境改变或细胞分化等事件而导致的激活代谢途径。因此，需要采用高通量方式获取用于定量检测代谢物的技术，以对看似同质的细胞之间的生物学变异性进行特征描述。在这里，我们展示了纳喷雾解吸电喷雾电离（nano-DESI）质谱（MS）能够以高通量方式灵敏地对单细胞中的内源性物质进行分子特征分析和定量。具体而言，我们显示了大量的检测到的氨基酸和磷脂，包括脑磷脂，很容易从单个脸颊细胞中检测到。此外，通过将磷脂酰胆碱（PC）内标物纳入 nano-DESI 溶剂中，我们确定一个细胞中的 PC 总量为 1.2 皮摩尔。最后，我们描述了一种高通量方法，其中可以自动对单细胞中的分子进行分析。这些单细胞分析方面的进展为未来的研究提供了基础，这些研究旨在了解与药物作用、细胞分化和改变的化学微环境相关的细胞过程。
