title: Integrated Droplet-Based Microextraction with ESI-MS for Removal of Matrix Interference in Single-Cell Analysis.
abstract: 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.
authors.affiliation: Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
authors.forename: Xiao-Chao
authors.initials: XC
authors.lastname: Zhang
authors.affiliation: Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
authors.forename: Zhen-Wei
authors.initials: ZW
authors.lastname: Wei
authors.affiliation: National Institute of Metrology, Beijing 100013, China.
authors.forename: Xiao-Yun
authors.initials: XY
authors.lastname: Gong
authors.affiliation: Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
authors.forename: Xing-Yu
authors.initials: XY
authors.lastname: Si
authors.affiliation: Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
authors.forename: Yao-Yao
authors.initials: YY
authors.lastname: Zhao
authors.affiliation: Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
authors.forename: Cheng-Dui
authors.initials: CD
authors.lastname: Yang
authors.affiliation: Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
authors.forename: Si-Chun
authors.initials: SC
authors.lastname: Zhang
authors.affiliation: Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing 100084, China.
authors.forename: Xin-Rong
authors.initials: XR
authors.lastname: Zhang
mesh_terms: D000249:Adenosine Monophosphate / Q000032:analysis / Q000302:isolation & purification; D005944:Glucosamine / Q000031:analogs & derivatives / Q000032:analysis / Q000302:isolation & purification; D005978:Glutathione / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D066292:Lipid Droplets / Q000737:chemistry*; D059627:Liquid Phase Microextraction / Q000379:methods*; D061986:MCF-7 Cells; D044085:Microfluidics; D059010:Single-Cell Analysis / Q000379:methods*; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D012997:Solvents; C026712:UDP-glucosamine; D000249:Adenosine Monophosphate; D005978:Glutathione; D005944:Glucosamine