Deciphering the neuronal code—the rules by which neuronal circuits store and process information—is a major scientific challenge1,2. Currently, these efforts are impeded by a lack of experimental tools that are sensitive enough to quantify the strength of individual synaptic connections and also scalable enough to simultaneously measure and control a large number of mammalian neurons with single-cell resolution3,4. Here, we report a scalable intracellular electrode platform based on vertical nanowires that allows parallel electrical interfacing to multiple mammalian neurons. Specifically, we show that our vertical nanowire electrode arrays can intracellularly record and stimulate neuronal activity in dissociated cultures of rat cortical neurons and can also be used to map multiple individual synaptic connections. The scalability of this platform, combined with its compatibility with silicon nanofabrication techniques, provides a clear path towards simultaneous, high-fidelity interfacing with hundreds of individual neurons.
Subscribe to Journal
Get full journal access for 1 year
only $15.58 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Yuste, R. Circuit neuroscience: the road ahead. Front. Neurosci. 2, 6–9 (2008).
Bock, D. D. et al. Network anatomy and in vivo physiology of visual cortical neurons. Nature 471, 177–182 (2011).
Pine, J. A history of MEA development, in Advances in Network Electrophysiology (eds, Taketani, M. & Baudry, M.) 3–23 (Springer, 2006).
Molleman, A. Patch Clamping (Wiley, 2003).
Rolston, J. D., Gross, R. E. & Potter, S. M. A low-cost multielectrode system for data acquisition enabling real-time closed-loop processing with rapid recovery from stimulation artifacts. Front. Neuroeng. 2, 12–12 (2009).
Voelker, M. & Fromherz, P. Signal transmission from individual mammalian nerve cell to field-effect transistor. Small 1, 206–210 (2005).
Kim, D-H. et al. Dissolvable films of silk fibroin for ultrathin conformal bio-integrated electronics. Nature Mater. 9, 511–517 (2010).
Viventi, J. et al. A conformal, bio-interfaced class of silicon electronics for mapping cardiac electrophysiology. Sci. Trans. Med. 2, 24ra22 (2010).
Patolsky, F. et al. Detection, stimulation, and inhibition of neuronal signals with high-density nanowire transistor arrays. Science 313, 1100–1104 (2006).
Eschermann, J. F. et al. Action potentials of HL-1 cells recorded with silicon nanowire transistors. Appl. Phys. Lett. 95, 083703 (2009).
Wang, K., Fishman, H. A., Dai, H. & Harris, J. S. Neural stimulation with a carbon nanotube microelectrode array. Nano Lett. 6, 2043–2048 (2006).
McKnight, T. E. et al. Resident neuroelectrochemical interfacing using carbon nanofiber arrays. J. Phys. Chem. B 110, 15317–15327 (2006).
Hai, A. et al. Spine-shaped gold protrusions improve the adherence and electrical coupling of neurons with the surface of micro-electronic devices. J. R. Soc. Interface 6, 1153–1165 (2009).
Hai, A., Shappir, J. & Spira, M. E. In-cell recordings by extracellular microelectrodes. Nature Methods 7, 200–202 (2010).
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).
Li, X., Klemic, K. G., Reed, M. A. & Sigworth, F. J. Microfluidic system for planar patch clamp electrode arrays. Nano Lett. 6, 815–819 (2006).
Sigworth, F. J. & Klemic, K. G. Microchip technology in ion-channel research. IEEE Trans. Nanobiosci. 4, 121–127 (2005).
Tian, B. et al. Three-dimensional, flexible nanoscale field-effect transistors as localized bioprobes. Science 329, 830–834 (2010).
Nikolenko, V., Poskanzer, K. E. & Yuste, R. Two-photon photostimulation and imaging of neural circuits. Nature Methods 4, 943–950 (2007).
Peterka, D. S., Takahashi, H. & Yuste, R. Imaging voltage in neurons. Neuron 69, 9–21 (2011).
Zhang, F. et al. Optogenetic interrogation of neural circuits: technology for probing mammalian brain structures. Nature Protoc. 5, 439–456 (2010).
Shalek, A. K. et al. Vertical silicon nanowires as a universal platform for delivering biomolecules into living cells. Proc. Natl Acad. Sci. USA 107, 1870–1875 (2010).
Thomas, P. & Smart, T. G. HEK293 cell line: a vehicle for the expression of recombinant proteins. J. Pharmacol. Toxicol. Methods 51, 187–200 (2005).
Rols, M. P. & Teissié, J. Electropermeabilization of mammalian cells. Quantitative analysis of the phenomenon. Biophys. J. 58, 1089–1098 (1990).
Moulton, S. E. et al. Studies of double layer capacitance and electron transfer at a gold electrode exposed to protein solutions. Electrochim. Acta 49, 4223–4230 (2004).
Dichter, M. A. Rat cortical neurons in cell culture: culture methods, cell morphology, electrophysiology, and synapse formation. Brain Res. 149, 279–293 (1978).
Romijn, H. J., Mud, M. T., Habets, A. M. & Wolters, P. S. A quantitative electron microscopic study on synapse formation in dissociated fetal rat cerebral cortex in vitro. Brain Res. 227, 591–605 (1981).
Kole, M. H. P. & Stuart, G. J. Is action potential threshold lowest in the axon? Nature Neurosci. 11, 1253–1255 (2008).
Dan, Y. & Poo, M-M. Spike timing-dependent plasticity: from synapse to perception. Physiol. Rev. 86, 1033–1048 (2006).
Mason, A., Nicoll, A. & Stratford, K. Synaptic transmission between individual pyramidal neurons of the rat visual cortex in vitro. J. Neurosci. 11, 72–84 (1991).
Nicolelis, M. A. L. Brain–machine interfaces to restore motor function and probe neural circuits. Nature Rev. Neurosci. 4, 417–422 (2003).
Arancio, O., Kandel, E. R. & Hawkins, R. D. Activity-dependent long-term enhancement of transmitter release by presynaptic 3′,5′-cyclic GMP in cultured hippocampal neurons. Nature 376, 74–80 (1995).
The authors thank J. MacArthur, E. Soucy, J. Greenwood, L. DeFeo, N. Sanjana, A. Dibos, G. Lau, B. Ilic, M. Metzler, L. Xie and E. Macomber for scientific discussions and technical assistance. The VNEA fabrication and characterization were performed in part at the Center for Nanoscale Systems at Harvard University. This work was supported by an NIH Pioneer award (5DP1OD003893-03) and an NSF EFRI award (EFRI-0835947).
The authors declare no competing financial interests.
About this article
Cite this article
Robinson, J., Jorgolli, M., Shalek, A. et al. Vertical nanowire electrode arrays as a scalable platform for intracellular interfacing to neuronal circuits. Nature Nanotech 7, 180–184 (2012) doi:10.1038/nnano.2011.249
ACS Nano (2019)
Nature Nanotechnology (2019)
Cell Reports (2019)
ACS Applied Materials & Interfaces (2019)
Scientific Reports (2019)