Department of Chemistry─BMC, Uppsala University, 75123 Uppsala, Sweden.
Department of Medical Cell Biology, Uppsala University, 75123 Uppsala, Sweden.
J Am Soc Mass Spectrom. 2023 Nov 1;34(11):2518-2524. doi: 10.1021/jasms.3c00239. Epub 2023 Oct 13.
Single-cell metabolomics has the potential to reveal unique insights into intracellular mechanisms and biological processes. However, the detection of metabolites from individual cells is challenging due to their versatile chemical properties and concentrations. Here, we demonstrate a tapered probe for pneumatically assisted nanospray desorption electrospray ionization (PA nano-DESI) mass spectrometry that enables both chemical imaging of larger cells and global metabolomics of smaller 15 μm cells. Additionally, by depositing cells in predefined arrays, we show successful metabolomics from three individual INS-1 cells per minute, which enabled the acquisition of data from 479 individual cells. Several cells were used to optimize analytical conditions, and 93 or 97 cells were used to monitor metabolome alterations in INS-1 cells after exposure to a low or high glucose concentration, respectively. Our analytical approach offers insights into cellular heterogeneity and provides valuable information about cellular processes and responses in individual cells.
单细胞代谢组学有可能深入揭示细胞内机制和生物过程的独特见解。然而，由于代谢物具有多样的化学性质和浓度，因此从单个细胞中检测代谢物具有挑战性。在这里，我们展示了一种用于气动辅助纳喷雾解吸电喷雾电离（PA nano-DESI）质谱法的锥形探头，该探头能够对较大细胞进行化学成像，并对较小的 15μm 细胞进行全局代谢组学分析。此外，通过将细胞沉积在预定的阵列中，我们成功地实现了每分钟三个单个 INS-1 细胞的代谢组学分析，从而能够从 479 个单个细胞中获取数据。使用多个细胞来优化分析条件，并使用 93 或 97 个细胞来监测 INS-1 细胞在暴露于低或高葡萄糖浓度后的代谢组改变。我们的分析方法深入了解了细胞异质性，并为单个细胞中的细胞过程和反应提供了有价值的信息。
