Wei Zhenwei, Zhang Xiaochao, Si Xingyu, Gong Xiaoyun, Zhang Sichun, Zhang Xinrong
Department of Chemistry, Tsinghua University, Beijing, China.
Methods Mol Biol. 2020;2064:31-59. doi: 10.1007/978-1-4939-9831-9_4.
In this chapter, we introduced a Pico-ESI strategy for metabolomics analysis with picoliter-level samples. This Pico-ESI strategy was technically achieved by pulsed direct current electrospray ionization source (Pulsed-DC-ESI). This source could collect MS signals for a few minutes from a cell, enabling us to obtain large-scale MS data of metabolite IDs in single-cell analysis. Further identification of the single-cell metabolome such as the database match and chemical modification to metabolome was thereby achieved. Technically, this source could ionize sample with no need of sample and electrode contact, which can be potentially applied for high-throughput analysis. We also introduced several strategies related to Pico-ESI to reduce the matrix interference especially for extremely small samples developed in our group, including step-voltage nanoelectrospray, picoliter sample desalting method, droplet-based microextraction method, and probe-ESI, etc. All these strategies have been successfully applied to single-cell analysis.
在本章中，我们介绍了一种用于皮升级样本代谢组学分析的皮升电喷雾电离（Pico-ESI）策略。这种Pico-ESI策略通过脉冲直流电喷雾电离源（Pulsed-DC-ESI）在技术上得以实现。该源能够从单个细胞收集数分钟的质谱信号，使我们能够在单细胞分析中获得代谢物ID的大规模质谱数据。从而实现了对单细胞代谢组的进一步鉴定，如数据库匹配和代谢组的化学修饰。从技术上讲，该源能够在无需样品与电极接触的情况下使样品离子化，这有可能应用于高通量分析。我们还介绍了几种与Pico-ESI相关的策略，以减少基质干扰，特别是针对我们团队开发的极少量样品，包括步进电压纳米电喷雾、皮升样品脱盐方法、基于液滴的微萃取方法和探针电喷雾电离等。所有这些策略都已成功应用于单细胞分析。
