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Wireless nanopore electrodes for analysis of single entities

Nature Protocols volume 14, pages 2015–2035 (2019)Cite this article

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Abstract

Measurements of a single entity underpin knowledge of the heterogeneity and stochastics in the behavior of molecules, nanoparticles, and cells. Electrochemistry provides a direct and fast method to analyze single entities as it probes electron/charge-transfer processes. However, a highly reproducible electrochemical-sensing nanointerface is often hard to fabricate because of a lack of control of the fabrication processes at the nanoscale. In comparison with conventional micro/nanoelectrodes with a metal wire inside, we present a general and easily implemented protocol that describes how to fabricate and use a wireless nanopore electrode (WNE). Nanoscale metal deposition occurs at the tip of the nanopipette, providing an electroactive sensing interface. The WNEs utilize a dynamic ionic flow instead of a metal wire to sense the interfacial redox process. WNEs provide a highly controllable interface with a 30- to 200-nm diameter. This protocol presents the construction and characterization of two types of WNEs—the open-type WNE and closed-type WNE—which can be used to achieve reproducible electrochemical measurements of single entities. Combined with the related signal amplification mechanisms, we also describe how WNEs can be used to detect single redox molecules/ions, analyze the metabolism of single cells, and discriminate single nanoparticles in a mixture. This protocol is broadly applicable to studies of living cells, nanomaterials, and sensors at the single-entity level. The total time required to complete the protocol is ~10–18 h. Each WNE costs ~$1–$3.

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Fig. 1: Schematic illustration of the concept of the WNE and its applications.
Fig. 2: Fabrication procedure and characterization of the closed-type and open-type WNEs.
Fig. 3: Schematic of the adapted setup for WNE experiments.
Fig. 4: Electrical and optical monitoring of the closed-type WNE fabrication process.
Fig. 5: Detection of Hg2+ at different concentrations using open-type WNEs.
Fig. 6: Electrical monitoring of the redox target in living cells using WNE.
Fig. 7: Electrical recording of single-nanoparticle collision with closed-type WNEs.

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The authors declare that the main data supporting the findings of this study are available within the article and its Supplementary Information files.

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Acknowledgements

This research was supported by the National Natural Science Foundation of China (21834001 and 61871183), the Innovation Program of the Shanghai Municipal Education Commission (2017-01-07-00-02-E00023), the National Ten Thousand Talent Program for young top-notch talent, and the Shanghai Rising-Star Program (19QA1402300).

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  1. These authors contributed equally: Rui Gao, Yao Lin.

Authors and Affiliations

  1. School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, China

    Rui Gao, Yao Lin, Yi-Lun Ying, Yong-Xu Hu, Su-Wen Xu, Lin-Qi Ruan, Ru-Jia Yu, Yuan-Jie Li, Hao-Wen Li, Ling-Fei Cui & Yi-Tao Long

  2. School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, China

    Yi-Lun Ying & Yi-Tao Long

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Contributions

R.G., Y.-L.Y., and Y.-T.L. conceived and designed the research; R.G., Y.L., Y.-X.H., S.-W.X., L.-Q.R., R.-J.Y, Y.-J.L., and L.-F.C. performed the WNE fabrication and characterization experiments; R.G. and Y.-X.H. implemented the applications by using WNEs and analyzed the data; Y.-X.H. and H.-W.L. cultured the cells. R.G., Y.L., Y.-X.H., Y.-L.Y., and Y.-T.L. organized and summarized the figures; R.G., Y.L., Y.-L.Y., and Y.-T.L. wrote the manuscript. All authors were involved in the discussions of the manuscript at all stages.

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Correspondence to Yi-Lun Ying.

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Key references using this protocol

Ying, Y.-L. et al. J. Am. Chem. Soc. 140, 5385–5392 (2018): https://pubs.acs.org/doi/abs/10.1021/jacs.7b12106

Gao, R. et al. Angew. Chem. Int. Ed. 57, 1011–1015 (2018): https://onlinelibrary.wiley.com/doi/10.1002/anie.201710201

Key data used in this protocol

Gao, R., Ying, Y.-L., Hu, Y.-X., Li, Y.-J. & Long, Y.-T. Anal. Chem. 89, 7382–7387 (2017): https://pubs.acs.org/doi/abs/10.1021/acs.analchem.7b00729

Ying, Y.-L. et al. J. Am. Chem. Soc. 140, 5385–5392 (2018): https://pubs.acs.org/doi/abs/10.1021/jacs.7b12106

Gao, R. et al. Angew. Chem. Int. Ed. 57, 1011–1015 (2018): https://onlinelibrary.wiley.com/doi/10.1002/anie.201710201

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Gao, R., Lin, Y., Ying, YL. et al. Wireless nanopore electrodes for analysis of single entities. Nat Protoc 14, 2015–2035 (2019). https://doi.org/10.1038/s41596-019-0171-5

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