In Situ Measurement of ATP in Single Cells by an Amphiphilic Aptamer-Assisted Electrochemical Nano-Biosensor

In situ and quantitative measurements of adenosine 5 '- triphosphate (ATP) in single living cells are highly desired for understanding several sorts of necessary physiological and pathological processes. Due to its small size and high sensitivity, an ultramicroelectrode can be used for single-cell analysis. However, ATP is difficult to detect in single cells because it is nonelectroactive and low in content. Herein, we introduced an electrochemical nano-biosensor based on an amphiphilic aptamer-assisted carbon fiber nanoelectrode (aptCFNE) with high signal-to-noise ratio. The low current (e.g., 60 pA) and the tiny diameter of the tip (ca. 400 nm) of the nanosensor made it noninvasive to living cells. The amphiphilic aptamer has good biocompatibility and can be stably modified to the surface of functionalized electrodes. CFNE, which was modified with ferrocene-labeled aptamer, could quickly and selectively detect ATP content in the nucleus, cytoplasm, and extracellular space of single HeLa cells. The results showed that the ATP contents in the nucleus, cytoplasm, and extracellular space were 568 +/- 9, 461 +/- 20, and 312 +/- 4 mu M, respectively. The anticancer drug treatment effects on the cellular level were further recorded, which was of great significance for understanding ATP-related biological processes and drug screenings. This strategy is universally applicable to detect other targets by changing the aptamer sequence, which will greatly improve our understanding of cell heterogeneity and provide a more reliable scientific basis for exploring major diseases at the single-cell level.

Automated summary

The study introduces an electrochemical nano-biosensor based on a carbon fiber nanoelectrode assisted by an amphiphilic aptamer, which can quickly and selectively detect ATP content in the nucleus, cytoplasm, and extracellular space of single HeLa cells. The effects of anticancer drugs on the cellular level were recorded, providing insights into ATP-related biological processes and drug screenings.