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Sci Rep ; 6: 24730, 2016 04 29.
Artigo em Inglês | MEDLINE | ID: mdl-27126222

RESUMO

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.


Assuntos
Gotículas Lipídicas/química , Microextração em Fase Líquida/métodos , Análise de Célula Única/métodos , Espectrometria de Massas por Ionização por Electrospray , Monofosfato de Adenosina/análise , Monofosfato de Adenosina/isolamento & purificação , Glucosamina/análogos & derivados , Glucosamina/análise , Glucosamina/isolamento & purificação , Glutationa/análise , Glutationa/isolamento & purificação , Humanos , Células MCF-7 , Microfluídica , Solventes/química
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