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Integrated Droplet-Based Microextraction with ESI-MS for Removal of Matrix Interference in Single-Cell Analysis

机译:基于ESI-MS的基于液滴的集成微萃取技术可消除单细胞分析中的基质干扰

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摘要

Integrating droplet-based microfluidics with mass spectrometry is essential to high-throughput and multiple analysis of single cells. Nevertheless, matrix effects such as the interference of culture medium and intracellular components influence the sensitivity and the accuracy of results in single-cell analysis. To resolve this problem, we developed a method that integrated droplet-based microextraction with single-cell mass spectrometry. Specific extraction solvent was used to selectively obtain intracellular components of interest and remove interference of other components. Using this method, UDP-Glc-NAc, GSH, GSSG, AMP, ADP and ATP were successfully detected in single MCF-7 cells. We also applied the method to study the change of unicellular metabolites in the biological process of dysfunctional oxidative phosphorylation. The method could not only realize matrix-free, selective and sensitive detection of metabolites in single cells, but also have the capability for reliable and high-throughput single-cell analysis.
机译:将基于液滴的微流控技术与质谱相集成对于单个细胞的高通量和多重分析至关重要。尽管如此,诸如培养基和细胞内成分的干扰之类的基质效应仍会影响单细胞分析结果的敏感性和准确性。为解决此问题,我们开发了一种将基于液滴的微萃取与单细胞质谱相结合的方法。使用特定的提取溶剂可选择性地获得目标细胞内组分并消除其他组分的干扰。使用此方法,可以在单个MCF-7细胞中成功检测到UDP-Glc-NAc,GSH,GSSG,AMP,ADP和ATP。我们还应用该方法研究功能失调的氧化磷酸化的生物过程中单细胞代谢产物的变化。该方法不仅可以实现无细胞,无选择性,灵敏的单细胞代谢物检测,而且具有可靠,高通量的单细胞分析能力。

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