Open-cell recording of action potentials using active electrode arrays.

The investigation of complex communication in cellular networks requires superior measurement tools than those available to date. Electrode arrays integrated onto silicon electronics are increasingly used to measure the electrical activity of cells in an automated and highly parallelized fashion, but they are restricted to recording extracellular potentials. Here, we report on an array of TiN electrodes built using standard silicon electronics for intracellular action potential recording. Intracellular access, possible at each of the 16 384 electrodes on the chip, was accomplished by local membrane electroporation using electrical stimulation with subcellular, micrometer-sized electrodes. Access to the cell interior was transient and could be tuned in duration by adapting the electroporation protocol. Intracellular sensing was found to be minimally invasive in the short and long-term, allowing consecutive intracellular recordings from the same cell over the course of days. Finally, we applied this method to investigate the effect of an ion channel blocker on cardiac electrical activity. This technique opens the door to massively parallel, long-term intracellular recording for fundamental electrophysiology and drug screening.

[1]  A. Lambacher,et al.  Identifying firing mammalian neurons in networks with high-resolution multi-transistor array (MTA) , 2011 .

[2]  P. Fromherz,et al.  Signal transmission from individual mammalian nerve cell to field-effect transistor. , 2005, Small.

[3]  B. Cui,et al.  Intracellular Recording of Action Potentials by Nanopillar Electroporation , 2012, Nature nanotechnology.

[4]  E. Neher,et al.  Single Na+ channel currents observed in cultured rat muscle cells , 1980, Nature.

[5]  B. Sakmann,et al.  Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches , 1981, Pflügers Archiv.

[6]  J. Camm.  Antiarrhythmic drugs for the maintenance of sinus rhythm: risks and benefits. , 2012, International journal of cardiology.

[7]  Bo Yu,et al.  Nanochannel electroporation delivers precise amounts of biomolecules into living cells. , 2011, Nature nanotechnology.

[8]  Andreas Hierlemann,et al.  Growing Cells Atop Microelectronic Chips: Interfacing Electrogenic Cells In Vitro With CMOS-Based Microelectrode Arrays , 2011, Proceedings of the IEEE.

[9]  Charles M. Lieber,et al.  Three-Dimensional, Flexible Nanoscale Field-Effect Transistors as Localized Bioprobes , 2010, Science.

[10]  Enrico Marani,et al.  Geometry-based finite-element modeling of the electrical contact between a cultured neuron and a microelectrode , 2003, IEEE Transactions on Biomedical Engineering.

[11]  Jan Wouters,et al.  Single-cell recording and stimulation with a 16k micro-nail electrode array integrated on a 0.18 μm CMOS chip. , 2012, Lab on a chip.

[12]  Andrei G. Pakhomov,et al.  Analysis of Plasma Membrane Integrity by Fluorescent Detection of Tl+ Uptake , 2010, The Journal of Membrane Biology.

[13]  J. Weaver,et al.  Theory of electroporation: A review , 1996 .

[14]  Jacob T. Robinson,et al.  Vertical nanowire electrode arrays as a scalable platform for intracellular interfacing to neuronal circuits. , 2012, Nature nanotechnology.

[15]  Bozhi Tian,et al.  Intracellular recordings of action potentials by an extracellular nanoscale field-effect transistor , 2011, Nature nanotechnology.

[16]  Isabelle Fonteille,et al.  The development of high quality seals for silicon patch-clamp chips. , 2010, Biomaterials.

[17]  Luca Berdondini,et al.  Active pixel sensor array for high spatio-temporal resolution electrophysiological recordings from single cell to large scale neuronal networks. , 2009, Lab on a chip.

[18]  N J Izzo,et al.  HL-1 cells: a cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[19]  J. Shappir,et al.  In-cell recordings by extracellular microelectrodes , 2010, Nature Methods.

[20]  Yusuf Leblebici,et al.  Electrical modeling of the cell-electrode interface for recording neural activity from high-density microelectrode arrays , 2009, Neurocomputing.