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Intracellular recording of action potentials by nanopillar electroporation

Abstract

Action potentials have a central role in the nervous system and in many cellular processes, notably those involving ion channels. The accurate measurement of action potentials requires efficient coupling between the cell membrane and the measuring electrodes. Intracellular recording methods such as patch clamping involve measuring the voltage or current across the cell membrane by accessing the cell interior with an electrode, allowing both the amplitude and shape of the action potentials to be recorded faithfully with high signal-to-noise ratios1. However, the invasive nature of intracellular methods usually limits the recording time to a few hours1, and their complexity makes it difficult to simultaneously record more than a few cells. Extracellular recording methods, such as multielectrode arrays2 and multitransistor arrays3, are non-invasive and allow long-term and multiplexed measurements. However, extracellular recording sacrifices the one-to-one correspondence between the cells and electrodes, and also suffers from significantly reduced signal strength and quality. Extracellular techniques are not, therefore, able to record action potentials with the accuracy needed to explore the properties of ion channels. As a result, the pharmacological screening of ion-channel drugs is usually performed by low-throughput intracellular recording methods4. The use of nanowire transistors5,6,7, nanotube-coupled transistors8 and micro gold-spine and related electrodes9,10,11,12 can significantly improve the signal strength of recorded action potentials. Here, we show that vertical nanopillar electrodes can record both the extracellular and intracellular action potentials of cultured cardiomyocytes over a long period of time with excellent signal strength and quality. Moreover, it is possible to repeatedly switch between extracellular and intracellular recording by nanoscale electroporation and resealing processes. Furthermore, vertical nanopillar electrodes can detect subtle changes in action potentials induced by drugs that target ion channels.

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Figure 1: Nanopillar electrode devices and their interactions with HL-1 cardiomyocytes.
Figure 2: Recording action potentials of a single HL-1 cell before and after electroporation.
Figure 3: Minimally invasive intracellular measurement of action potentials with high precision.
Figure 4: Parallel intracellular recording of multiple cells and the evolution of action potentials over consecutive days for single cells.
Figure 5: Effect of ion-channel blocking drugs on HL-1 cells.

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Acknowledgements

The HL-1 cardiac cell line was obtained from William C. Claycomb (Louisiana State University). This work was supported by the NSF (CAREER award no. 1055112), the NIH (grant no. NS057906), a Searle Scholar award, a Packard Science and Engineering Fellowship (to B.C.) and a National Defense Science and Engineering Graduate Fellowship (to Z.L.)

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All authors conceived the experiments. C.X., Z.L. and L.H. carried out experiments. All authors contributed to the scientific planning, discussions and writing of the manuscript.

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Correspondence to Yi Cui or Bianxiao Cui.

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Xie, C., Lin, Z., Hanson, L. et al. Intracellular recording of action potentials by nanopillar electroporation. Nature Nanotech 7, 185–190 (2012). https://doi.org/10.1038/nnano.2012.8

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