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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The authors declare that all data supporting the findings of this study are available within the paper and its Supplementary Information.
Sasaki, T., Minamisawa, G., Takahashi, N., Matsuki, N. & Ikegaya, Y. Reverse optical trawling for synaptic connections in situ. J. Neurophysiol. 102, 636–643 (2009).
Petreanu, L., Huber, D., Sobczyk, A. & Svoboda, K. Channelrhodopsin-2-assisted circuit mapping of long-range callosal projections. Nat. Neurosci. 10, 663–668 (2007).
Shemesh, O. A. et al. Temporally precise single-cell-resolution optogenetics. Nat. Neurosci. 20, 1796–1806 (2017).
Jäckel, D. et al. Combination of high-density microelectrode array and patch clamp recordings to enable studies of multisynaptic integration. Sci. Rep. 7, 978 (2017).
Spira, M. E. & Hai, A. Multi-electrode array technologies for neuroscience and cardiology. Nat. Nanotech. 8, 83–94 (2013).
Perin, R., Berger, T. K. & Markram, H. A synaptic organizing principle for cortical neuronal groups. Proc. Natl Acad. Sci. USA 108, 5419–5424 (2011).
Robinson, J. T. et al. Vertical nanowire electrode arrays as a scalable platform for intracellular interfacing to neuronal circuits. Nat. Nanotech. 7, 180–184 (2012).
Abbott, J. et al. CMOS nanoelectrode array for all-electrical intracellular electrophysiological imaging. Nat. Nanotech. 12, 460–466 (2017).
Eversmann, B. et al. A 128 × 128 CMOS Biosensor array for extracellular recording of neural activity. IEEE J. Solid-State Circuits 38, 2306–2317 (2003).
Berdondini, L. et al. Active pixel sensor array for high spatio-temporal resolution electrophysiological recordings from single cell to large scale neuronal networks. Lab Chip 9, 2644–2651 (2009).
Frey, U. et al. Switch-matrix-based high-density microelectrode array in CMOS technology. IEEE J. Solid-State Circuits 45, 467–482 (2010).
Tsai, D., Sawyer, D., Bradd, A., Yuste, R. & Shepard, K. L. A very large-scale microelectrode array for cellular-resolution electrophysiology. Nat. Commun. 8, 1802 (2017).
Lopez, C. M. et al. A 16,384-electrode 1,024-channel multimodal CMOS MEA for high-throughput intracellular action potential measurements and impedance spectroscopy in drug-screening applications. In IEEE International Solid-State Circuits Conference (eds Anderson, J. H. et al.) 61, 464–466 (IEEE, 2018).
Fertig, N., Blick, R. H. & Behrends, J. C. Whole cell patch clamp recording performed on a planar glass chip. Biophys. J. 82, 3056–3062 (2002).
Lau, A. Y., Hung, P. J., Wu, A. R. & Lee, L. P. Open-access microfluidic patch-clamp array with raised lateral cell trapping sites. Lab Chip 6, 1510–1515 (2006).
Dunlop, J., Bowlby, M., Peri, R., Vasilyev, D. & Arias, R. High-throughput electrophysiology: an emerging paradigm for ion-channel screening and physiology. Nat. Rev. Drug Discov. 7, 358–368 (2008).
Hochbaum, D. R. et al. All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins. Nat. Methods 11, 825–833 (2014).
Kim, C. K., Adhikari, A. & Deisseroth, K. Integration of optogenetics with complementary methodologies in systems neuroscience. Nat. Rev. Neurosci. 18, 222–235 (2017).
Ikegaya, Y. et al. Synfire chains and cortical songs: temporal modules of cortical activity. Science 304, 559–564 (2004).
Stetter, O., Battaglia, D., Soriano, J. & Geisel, T. Model-free reconstruction of excitatory neuronal connectivity from calcium imaging signals. PLoS Comput. Biol. 8, e1002653 (2012).
Ahrens, M. B. et al. Brain-wide neuronal dynamics during motor adaptation in zebrafish. Nature 485, 471–477 (2012).
Akerboom, J. et al. Optimization of a GCaMP calcium indicator for neural activity imaging. J. Neurosci. 32, 13819–13840 (2012).
Gong, Y. et al. High-speed recording of neural spikes in awake mice and flies with a fluorescent voltage sensor. Science 350, 1361–1366 (2015).
Woodford, C. R. et al. Improved PeT molecules for optically sensing voltage in neurons. J. Am. Chem. Soc. 137, 1817–1824 (2015).
Lou, S. et al. Genetically targeted all-optical electrophysiology with a transgenic cre-dependent optopatch mouse. J. Neurosci. 36, 11059–11073 (2016).
Li, L.-L. et al. Morphological control of platinum nanostructures for highly efficient dye-sensitized solar cells. J. Mater. Chem. 22, 6267 (2012).
Dipalo, M. et al. Intracellular and extracellular recording of spontaneous action potentials in mammalian neurons and cardiac cells with 3D plasmonic nanoelectrodes. Nano Lett. 17, 3932–3939 (2017).
Lin, Z. C., Xie, C., Osakada, Y., Cui, Y. & Cui, B. Iridium oxide nanotube electrodes for sensitive and prolonged intracellular measurement of action potentials. Nat. Commun. 5, 3206 (2014).
Hai, A., Shappir, J. & Spira, M. E. In-cell recordings by extracellular microelectrodes. Nat. Methods 7, 200–202 (2010).
Liu, R. et al. High density individually addressable nanowire arrays record intracellular activity from primary rodent and human stem cell derived neurons. Nano Lett. 17, 2757–2764 (2017).
Fromherz, P. Self-gating of ion channels in cell adhesion. Phys. Rev. Lett. 78, 4131–4134 (1997).
Obien, M. E. J., Deligkaris, K., Bullmann, T., Bakkum, D. J. & Frey, U. Revealing neuronal function through microelectrode array recordings. Front. Neurosci. 9, 423 (2015).
Massobrio, P., Tessadori, J., Chiappalone, M. & Ghirardi, M. In vitro studies of neuronal networks and synaptic plasticity in invertebrates and in mammals using multielectrode arrays. Neural Plast. 2015, 1–18 (2015).
Froemke, R. C., Debanne, D. & Bi, G.-Q. Temporal modulation of spike-timing-dependent plasticity. Front. Syn. Neurosci. 2, 19 (2010).
Hai, A. & Spira, M. E. On-chip electroporation, membrane repair dynamics and transient in-cell recordings by arrays of gold mushroom-shaped microelectrodes. Lab Chip 12, 2865–2873 (2012).
Xie, C., Lin, Z., Hanson, L., Cui, Y. & Cui, B. Intracellular recording of action potentials by nanopillar electroporation. Nat. Nanotech. 7, 185–190 (2012).
Duan, X. et al. Intracellular recordings of action potentials by an extracellular nanoscale field-effect transistor. Nat. Nanotechnol. 7, 174–179 (2012).
Lee, K.-Y. et al. Vertical nanowire probes for intracellular signaling of living cells. Nanoscale Res. Lett. 9, 56 (2014).
Tian, B. et al. Three-dimensional, flexible nanoscale field-effect transistors as localized bioprobes. Science 329, 830–834 (2010).
Hai, A., Shappir, J. & Spira, M. E. Long-term, multisite, parallel, in-cell recording and stimulation by an array of extracellular microelectrodes. J. Neurophysiol. 104, 559–568 (2010).
Shmoel, N. et al. Multisite electrophysiological recordings by self-assembled loose-patch-like junctions between cultured hippocampal neurons and mushroom-shaped microelectrodes. Sci. Rep. 6, 27110 (2016).
Fitzsimonds, R. M., Song, H. J. & Poo, M. M. Propagation of activity-dependent synaptic depression in simple neural networks. Nature 388, 439–448 (1997).
Kullmann, D. M. & Nicoll, R. A. Long-term potentiation is associated with increases in quantal content and quantal amplitude. Nature 357, 240–244 (1992).
Hardingham, N. R. et al. Quantal analysis reveals a functional correlation between presynaptic and postsynaptic efficacy in excitatory connections from rat neocortex. J. Neurosci. 30, 1441–1451 (2010).
Shein-Idelson, M., Pammer, L., Hemberger, M. & Laurent, G. Large-scale mapping of cortical synaptic projections with extracellular electrode arrays. Nat. Methods 14, 882–890 (2017).
Sabatini, B. L. & Regehr, W. G. Timing of neurotransmission at fast synapses in the mammalian brain. Nature 384, 170–172 (1996).
Barthó, P. et al. Characterization of neocortical principal cells and interneurons by network interactions and extracellular features. J. Neurophysiol. 92, 600–608 (2004).
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.).
The authors declare no competing interests.
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Supplementary figures, tables, discussion, references and video captions.
Intracellular recordings of neuronal action potentials across a connected network. Large network bursts involving 1,837 pixels.
Intracellular recordings of neuronal action potentials across a connected network. Large network bursts involving 1,882 pixels.
Intracellular stimulation across a neuronal network.
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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