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.
Highlights
Droplet-based microfluidics could manipulate tiny droplets to perform fast[1,2], low-consuming[3,4,5], and high-throughput assays[6,7,8,9], which is suitable for single-cell analysis
We have detected UDP-Glc-NAc, GSH, GSSG, Adenosine 5′ - monophosphate (AMP), Adenosine 5′ - diphosphate (ADP) and Adenosine 5′ -triphosphate (ATP) in single MCF-7 cells, and we applied this method to study the biological process of dysfunctional oxidative phosphorylation
Each dish of MCF-7 cells was added in 1 mL solvent and the cellular compounds was extracted for 20 min
Summary
Droplet-based microfluidics could manipulate tiny droplets to perform fast[1,2], low-consuming[3,4,5], and high-throughput assays[6,7,8,9], which is suitable for single-cell analysis. It is essential to integrate droplet-based microfluidics with mass spectrometry for multiple analysis at single-cell levels. Some methods based on ESI-MS have been successfully applied to single-cell metabolite analysis[19,20,21]. We proposed a method to integrate droplet-based microextraction with ESI mass spectrometry (see Fig. 1). The extract was sucked back to the capillary’s tip, and detected by ESI-MS after evaporating and redissolving with a small-volume assisted solvent (20 ~ 100 pL). We have detected UDP-Glc-NAc, GSH, GSSG, AMP, ADP and ATP in single MCF-7 cells, and we applied this method to study the biological process of dysfunctional oxidative phosphorylation
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