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A nanoelectrode array for obtaining intracellular recordings from thousands of connected neurons

Abstract

Current electrophysiological or optical techniques cannot reliably perform simultaneous intracellular recordings from more than a few tens of neurons. Here we report a nanoelectrode array that can simultaneously obtain intracellular recordings from thousands of connected mammalian neurons in vitro. The array consists of 4,096 platinum-black electrodes with nanoscale roughness fabricated on top of a silicon chip that monolithically integrates 4,096 microscale amplifiers, configurable into pseudocurrent-clamp mode (for concurrent current injection and voltage recording) or into pseudovoltage-clamp mode (for concurrent voltage application and current recording). We used the array in pseudovoltage-clamp mode to measure the effects of drugs on ion-channel currents. In pseudocurrent-clamp mode, the array intracellularly recorded action potentials and postsynaptic potentials from thousands of neurons. In addition, we mapped over 300 excitatory and inhibitory synaptic connections from more than 1,700 neurons that were intracellularly recorded for 19 min. This high-throughput intracellular-recording technology could benefit functional connectome mapping, electrophysiological screening and other functional interrogations of neuronal networks.

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Fig. 1: The CNEI.
Fig. 2: Intracellular recording and stimulation of disassociated rat neurons using the pCC and pVC configurations.
Fig. 3: Network-wide intracellular measurements of dissociated rat neurons in the pCC configuration.
Fig. 4: Measurement of chemical synapse characteristics and network-wide mapping of synaptic connectivity with the pCC configuration.

Data availability

The authors declare that all data supporting the findings of this study are available within the paper and its Supplementary Information.

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Acknowledgements

Post-fabrication and characterization were performed, in part, at the Center for Nanoscale Systems at Harvard University. The authors are grateful for the support of this research by Samsung Advanced Institute of Technology, Samsung Electronics (A37734 to H.P. and D.H.), Catalyst Foundation (J.A., H.P. and D.H.), the Army Research Office (W911NF-15-1-0565 to D.H.), the Army Research Office (W911NF-17-1-0425 to D.H.), the National Science Foundation Graduate Research Fellowship Program (DGE1745303 to K.K.), the National Institutes of Health (1-U01-MH105960-01 to H.P.), the Gordon and Betty Moore Foundation (to H.P.), and the US Army Research Laboratory and the US Army Research Office (W911NF1510548 to H.P.).

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Contributions

H.P., D.H., J.A., T.Y. and K.K. conceived and designed the experiments. J.A. and L.Q. designed the CMOS IC, J.A., Y.K. and W.W. designed the interface electronics and T.Y., S.B. and K.K. performed post-fabrication and device packaging. J.A., T.Y., K.K. and R.S.G. performed the experiments and J.A., T.Y., K.K., H.P. and D.H. analysed the data. H.P. and D.H. supervised the project. J.A., T.Y., K.K., D.H. and H.P. wrote the manuscript, and all authors read and discussed it.

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Correspondence to Hongkun Park or Donhee Ham.

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Supplementary information

Supplementary Information

Supplementary figures, tables, discussion, references and video captions.

Reporting Summary

Supplementary Video 1

Intracellular recordings of neuronal action potentials across a connected network. Large network bursts involving 1,837 pixels.

Supplementary Video 2

Intracellular recordings of neuronal action potentials across a connected network. Large network bursts involving 1,882 pixels.

Supplementary Video 3

Intracellular stimulation across a neuronal network.

Supplementary Video 4

Intracellular mapping of depolarization potential propagations on the application of a drug.

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Abbott, J., Ye, T., Krenek, K. et al. A nanoelectrode array for obtaining intracellular recordings from thousands of connected neurons. Nat Biomed Eng 4, 232–241 (2020). https://doi.org/10.1038/s41551-019-0455-7

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