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Fabrication and utility of a transparent graphene neural electrode array for electrophysiology, in vivo imaging, and optogenetics

Abstract

Transparent graphene-based neural electrode arrays provide unique opportunities for simultaneous investigation of electrophysiology, various neural imaging modalities, and optogenetics. Graphene electrodes have previously demonstrated greater broad-wavelength transmittance (90%) than other transparent materials such as indium tin oxide (80%) and ultrathin metals (60%). This protocol describes how to fabricate and implant a graphene-based microelectrocorticography (μECoG) electrode array and subsequently use this alongside electrophysiology, fluorescence microscopy, optical coherence tomography (OCT), and optogenetics. Further applications, such as transparent penetrating electrode arrays, multi-electrode electroretinography, and electromyography, are also viable with this technology. The procedures described herein, from the material characterization methods to the optogenetic experiments, can be completed within 3–4 weeks by an experienced graduate student. These protocols should help to expand the boundaries of neurophysiological experimentation, enabling analytical methods that were previously unachievable using opaque metal–based electrode arrays.

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Figure 1: Advantages of a transparent graphene neural electrode array.
Figure 2: Raman spectroscopy of four-layer graphene on Parylene C.
Figure 3: Optogenetic experiments in a Thy1::ChR2 mouse with varying optical stimulation power.
Figure 4: Surgery and electrophysiology equipment and setup.
Figure 5: Fabrication procedure for transparent graphene electrode array.
Figure 6: Graphene transfer and stacking processes.
Figure 7: Assembly of ZIF PCB and electrode arrays.
Figure 8: Steps for implantation of a graphene or platinum array into a rat model.
Figure 9: Steps for implantation of a graphene or platinum array into a mouse model.
Figure 10: Electrophysiology setup.
Figure 11: Anticipated results.

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Acknowledgements

This work was supported by grants from the National Institutes of Health (NIH NIBIB 1R01EB009103-01 and NIH NIBIB 2R01EB000856-06 to J.C.W.). This work was also sponsored by the Defense Advanced Research Projects Agency (DARPA) Biological Technology Office (BTO), under the auspices of D. Weber through Space and Naval Warfare Systems Command (SPAWAR) Systems Center (SSC) Pacific grants no. N66001-11-1-4013 and no. N66001-12-C-4025 to J.C.W. The work was also partly supported by the Army Research Office under grant W911NF-14-1-0652 to Z.M. The program manager is J.X. Qiu. D.-W.P. and S.M. were partly supported by the Office of Naval Research (ONR) under grant N00014-09-1-0803. D.-W.P. was partly supported by a grant from the National Institutes of Health (NIH DHHS/PHS R01HG000225). A.S. is supported by NIH NIBIB 1T32EB011434-01A1. S.M was partly supported by a Winslow Sargeant Fellowship. R.P. and F.A. were partially supported by National Science Foundation (NSF) Career Award grant 1454300 and Brain and Behavior Research Foundation (NARSAD) grant 20610. We thank K. Eliceiri for technical discussions related to electrode transparency measurements. We thank S. Kang for illustrations.

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D.-W.P., S.K.B., J.P.N., F.A., A.S., S.M., T.J.R., J.N., H.K., D.-H.B., J.B., S.T.F., and S.T. performed the experiments. D.-W.P., S.K.B., J.P.N., F.A., A.S., S.M., T.J.R., R.P., J.C.W., and Z.M. developed the protocol. D.-W.P., S.K.B., J.P.N., F.A., L.K.-H., K.I.S., W.L., J.C.W., and Z.M. wrote the manuscript.

Corresponding authors

Correspondence to Justin C Williams or Zhenqiang Ma.

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Competing interests

D.-W.P., A.S., S.M., Z.M. and J.C.W. declare competing financial interests in the form of a pending patent application (transparent and flexible neural electrode arrays).

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Park, DW., Brodnick, S., Ness, J. et al. Fabrication and utility of a transparent graphene neural electrode array for electrophysiology, in vivo imaging, and optogenetics. Nat Protoc 11, 2201–2222 (2016). https://doi.org/10.1038/nprot.2016.127

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