Plasmonic meta-electrodes allow intracellular recordings at network level on high-density CMOS-multi-electrode arrays

Abstract

The ability to monitor electrogenic cells accurately plays a pivotal role in neuroscience, cardiology and cell biology. Despite pioneering research and long-lasting efforts, the existing methods for intracellular recording of action potentials on the large network scale suffer limitations that prevent their widespread use. Here, we introduce the concept of a meta-electrode, a planar porous electrode that mimics the optical and biological behaviour of three-dimensional plasmonic antennas but also preserves the ability to work as an electrode. Its synergistic combination with plasmonic optoacoustic poration allows commercial complementary metal–oxide semiconductor multi-electrode arrays to record intracellular action potentials in large cellular networks. We apply this approach to measure signals from human-induced pluripotent stem cell-derived cardiac cells, rodent primary cardiomyocytes and immortalized cell types and demonstrate the possibility of non-invasively testing a variety of relevant drugs. Due to its robustness and easiness of use, we expect the method will be rapidly adopted by the scientific community and by pharmaceutical companies.

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Fig. 1: Overview of the optoacoustic poration mechanism.
Fig. 2: Intracellular recordings of hiPSCs, rat primary and HL-1 cardiomyocytes on CMOS-MEA.
Fig. 3: Massive poration on CMOS-MEA and stability of the intracellular coupling.
Fig. 4: Drug detection on planar meta-electrodes.

Change history

  • 28 August 2018

    In the version of this Article originally published, the affiliation for the author Francesca Santoro was incorrectly given; it should have been ‘Center for Advanced Biomaterials for Healthcare, Istituto Italiano di Tecnologia, Napoli, Italy’. This has now been corrected in all versions of the Article.

References

  1. 1.

    Wu, A. H. Cardiotoxic drugs: clinical monitoring and decision making. Heart 94, 1503–1509 (2008).

    CAS  Article  Google Scholar 

  2. 2.

    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).

    Google Scholar 

  3. 3.

    Müller, J. et al. High-resolution CMOS MEA platform to study neurons at subcellular, cellular, and network levels. Lab Chip 15, 2767–2780 (2015).

    Article  Google Scholar 

  4. 4.

    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).

    CAS  Article  Google Scholar 

  5. 5.

    Braeken, D. et al. Open-cell recording of action potentials using active electrode arrays. Lab Chip 12, 4397–4402 (2012).

    CAS  Article  Google Scholar 

  6. 6.

    Huys, R. et al. Single-cell recording and stimulation with a 16k micro-nail electrode array integrated on a 0.18 æm CMOS chip. Lab Chip 12, 1274–1280 (2012).

    CAS  Article  Google Scholar 

  7. 7.

    Xie, C., Lin, Z., Hanson, L., Cui, Y. & Cui, B. Intracellular recording of action potentials by nanopillar electroporation. Nat. Nanotech. 7, 185–190 (2012).

    CAS  Article  Google Scholar 

  8. 8.

    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).

    CAS  Article  Google Scholar 

  9. 9.

    Spira, M. E. & Hai, A. Multi-electrode array technologies for neuroscience and cardiology. Nat. Nanotech. 8, 83–94 (2013).

    CAS  Article  Google Scholar 

  10. 10.

    Abbott, J. et al. CMOS nanoelectrode array for all-electrical intracellular electrophysiological imaging. Nat. Nanotech. 12, 460–466 (2017).

    CAS  Article  Google Scholar 

  11. 11.

    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).

    CAS  Article  Google Scholar 

  12. 12.

    Zhao, W. et al. Nanoscale manipulation of membrane curvature for probing endocytosis in live cells. Nat. Nanotech. 12, 750–756 (2017).

    CAS  Article  Google Scholar 

  13. 13.

    Yu, N. & Capasso, F. Flat optics with designer metasurfaces. Nat. Mater. 13, 139–150 (2014).

    CAS  Article  Google Scholar 

  14. 14.

    Messina, G. C. et al. Spatially, temporally, and quantitatively controlled delivery of broad range of molecules into selected cells through plasmonic nanotubes. Adv. Mater. 27, 7145–7149 (2015).

    CAS  Article  Google Scholar 

  15. 15.

    Zilio, P., Dipalo, M., Tantussi, F., Messina, G. C. & de Angelis, F. Hot electrons in water: injection and ponderomotive acceleration by means of plasmonic nanoelectrodes. Light Sci. Appl. 6, e17002 (2017).

    CAS  Article  Google Scholar 

  16. 16.

    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).

    CAS  Article  Google Scholar 

  17. 17.

    Meinzer, N., Barnes, W. L. & Hooper, I. R. Plasmonic meta-atoms and metasurfaces. Nat. Photon. 8, 889–898 (2014).

    CAS  Article  Google Scholar 

  18. 18.

    Qian, L. H., Yan, X. Q., Fujita, T., Inoue, A. & Chen, M. W. Surface enhanced Raman scattering of nanoporous gold: smaller pore sizes stronger enhancements. Appl. Phys. Lett. 90, 153120 (2007).

    Article  Google Scholar 

  19. 19.

    Garoli, D. et al. Boosting infrared energy transfer in 3D nanoporous gold antennas. Nanoscale 9, 915–922 (2017).

    CAS  Article  Google Scholar 

  20. 20.

    Fu, Y., Zhang, J., Nowaczyk, K. & Lakowicz, J. R. Enhanced single molecule fluorescence and reduced observation volumes on nanoporous gold (NPG) films. Chem. Commun. 49, 10874–10876 (2013).

    CAS  Article  Google Scholar 

  21. 21.

    Recum, A. F. et al. Surface roughness, porosity, and texture as modifiers of cellular adhesion. Tissue Eng. 2, 241–253 (1996).

    CAS  Article  Google Scholar 

  22. 22.

    Hallab, N. J., Bundy, K. J., O’Connor, K., Moses, R. L. & Jacobs, J. J. Evaluation of metallic and polymeric biomaterial surface energy and surface roughness characteristics for directed cell adhesion. Tissue Eng. 7, 55–71 (2001).

    CAS  Article  Google Scholar 

  23. 23.

    Imfeld, K. et al. Large-scale, high-resolution data acquisition system for extracellular recording of electrophysiological activity. IEEE Trans. Biomed. Eng. 55, 2064–2073 (2008).

    Article  Google Scholar 

  24. 24.

    Hilgen, G. et al. Pan-retinal characterisation of light responses from ganglion cells in the developing mouse retina. Sci. Rep. 7, 42330 (2017).

    CAS  Article  Google Scholar 

  25. 25.

    DeBusschere, B. D. & Kovacs, G. T. Portable cell-based biosensor system using integrated CMOS cell-cartridges. Biosens. Bioelectron. 16, 543–556 (2001).

    CAS  Article  Google Scholar 

  26. 26.

    Lin, Z. C. et al. Accurate nanoelectrode recording of human pluripotent stem cell-derived cardiomyocytes for assaying drugs and modeling disease. Microsyst. Nanoeng. 3, 16080 (2017).

    CAS  Article  Google Scholar 

  27. 27.

    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).

    Article  Google Scholar 

  28. 28.

    Camm, J. Antiarrhythmic drugs for the maintenance of sinus rhythm: risks and benefits. Int. J. Cardiol. 155, 362–371 (2012).

    Article  Google Scholar 

  29. 29.

    Virginio, C., Ugolini, A., Ballini, E., Carignani, C. & Corsi, M. Spontaneous cardiac action potentials recordings from human cardiomyocytes derived from pluripotent stem cells: an experimental paradigm in agreement with CiPA recommendation. J. Pharmacol. Toxicol. Methods 75, 200 (2015).

    Article  Google Scholar 

  30. 30.

    Shryock, J. C., Song, Y., Rajamani, S., Antzelevitch, C. & Belardinelli, L. The arrhythmogenic consequences of increasing late INa in the cardiomyocyte. Cardiovasc. Res. 99, 600–611 (2013).

    CAS  Article  Google Scholar 

  31. 31.

    Santoro, F. et al. Revealing the cell–material interface with nanometer resolution by focused ion beam/scanning electron microscopy. ACS Nano 11, 8320–8328 (2017).

    CAS  Article  Google Scholar 

  32. 32.

    Belu, A. et al. Ultra-thin resin embedding method for scanning electron microscopy of individual cells on high and low aspect ratio 3D nanostructures. J. Microsc. 263, 78–86 (2016).

    CAS  Article  Google Scholar 

  33. 33.

    Dipalo, M. et al. 3D plasmonic nanoantennas integrated with MEA biosensors. Nanoscale 7, 3703–3711 (2015).

    CAS  Article  Google Scholar 

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Acknowledgements

We thank M. Gandolfo and A. Maccione for discussions and for the impedance spectroscopy data of the CMOS-MEA electrodes. The research that led to these results received funding from the European Research Council under the European Union’s Seventh Framework Programme (FP/2007-2013)/ERC Grant Agreement no. [616213], CoG: Neuro-Plasmonics.

Author information

Affiliations

Authors

Contributions

F.D.A. and M.D. conceived and designed the experiments. M.D., G.M. and L.L. performed the electrophysiology experiments. F.S. and V.C. performed the focused ion beam cross-sections and SEM imaging. A.J. and M.D. analysed the data. G.B., A.J. and D.G. characterized the porous meta-electrodes. G.B. and M.D. fabricated the passive MEA devices. M.D. and F.T. designed the experimental set-up. A.A. and A.S. performed the electromagnetic and thermal simulations. F.D.A. supervised the work. All the authors discussed the results and wrote the manuscript.

Corresponding author

Correspondence to Francesco De Angelis.

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The authors declare no competing interests.

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Supplementary information

41565_2018_222_MOESM3_ESM.avi

Whole CMOS-MEA recording during optoacoustic poration on one electrode. The optoacoustic poration occurs at t = 5 s.

41565_2018_222_MOESM4_ESM.avi

Propagation wave in extracellular mode of human-induced pluripotent stem cells-derived cardiomyocytes on CMOS-MEA. The pixel colours represent the signals’ amplitude.

41565_2018_222_MOESM5_ESM.avi

Propagation wave in intracellular mode of human-induced pluripotent stem cells-derived cardiomyocytes on CMOS-MEA. The pixel colours represent the signals’ amplitude.

41565_2018_222_MOESM6_ESM.avi

Propagation wave in extracellular mode of human-induced pluripotent stem cells-derived cardiomyocytes on CMOS-MEA, including example extracellular waveforms from few electrodes. The brown vertical stripes in the graphs represent the time-bins used for calculating the amplitude for the pixel colouring in the map. The time-traces shown in the videos are highlighted by black square contours in the colour maps.

41565_2018_222_MOESM7_ESM.avi

Propagation wave in intracellular mode of human-induced pluripotent stem cells-derived cardiomyocytes on CMOS-MEA, including example intracellular waveforms from few electrodes. The brown vertical stripes in the graphs represent the time-bins used for calculating the amplitude for the pixel colouring in the map. The time-traces shown in the videos are highlighted by black square contours in the colour maps.

Supplementary Information

Supplementary Methods, Supplementary Figures 1–15, Supplementary References

Reporting Summary

Supplementary Video 1

Whole CMOS-MEA recording during optoacoustic poration on one electrode. The optoacoustic poration occurs at t = 5 s.

Supplementary Video 2

Propagation wave in extracellular mode of human-induced pluripotent stem cells-derived cardiomyocytes on CMOS-MEA. The pixel colours represent the signals’ amplitude.

Supplementary Video 3

Propagation wave in intracellular mode of human-induced pluripotent stem cells-derived cardiomyocytes on CMOS-MEA. The pixel colours represent the signals’ amplitude.

Supplementary Video 4

Propagation wave in extracellular mode of human-induced pluripotent stem cells-derived cardiomyocytes on CMOS-MEA, including example extracellular waveforms from few electrodes. The brown vertical stripes in the graphs represent the time-bins used for calculating the amplitude for the pixel colouring in the map. The time-traces shown in the videos are highlighted by black square contours in the colour maps.

Supplementary Video 5

Propagation wave in intracellular mode of human-induced pluripotent stem cells-derived cardiomyocytes on CMOS-MEA, including example intracellular waveforms from few electrodes. The brown vertical stripes in the graphs represent the time-bins used for calculating the amplitude for the pixel colouring in the map. The time-traces shown in the videos are highlighted by black square contours in the colour maps.

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Dipalo, M., Melle, G., Lovato, L. et al. Plasmonic meta-electrodes allow intracellular recordings at network level on high-density CMOS-multi-electrode arrays. Nature Nanotech 13, 965–971 (2018). https://doi.org/10.1038/s41565-018-0222-z

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