Open-cell recording of action potentials using active electrode arrays

Abstract

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.

Characterization of dorsal root ganglion neurons cultured on silicon micro-pillar substrates

“…Beside the classical electrophysiological techniques the activity of injured DRG neurons can be recorded by using micro-electrode arrays (MEAs), a technology that has been used by an increasing number of neuroscience laboratories456. With advanced MEAs based on integrated complementary metal oxide semiconductor (CMOS) any neuron grown over custom arrays can be recorded at high spatio-temporal resolution allowing us to better understand some aspects of its altered electrophysiological properties7.…”

Reflective lens-free imaging on high-density silicon microelectrode arrays for monitoring and evaluation of in vitro cardiac contractility

“…RCD was calculated in a region of 0.124 mm 2 around the electrodes of interest. In the first system, simultaneous intracellular action potential measurements were obtained by combining the RLFI setup with a CMOS-based microelectrode array with 16,384 individually addressable electrodes as previously described [7,39]. Briefly, intracellular access was achieved by electroporating the cell membrane locally through sub-cellular sized electrodes, achieving intracellular-like ('open-cell') recordings.…”

“…Primary neonatal rat ventricular cardiomyocytes were extracted from 2 day old Wistar rats and cultured in glass wells on both blank SiO 2 and microelectrode array chips, as described previously [7,35]. Cardiac culture medium (Ham F10 containing 5% FCS, 1% PenStrep, Hepes, 0.5% ITS, 0.1 mM Norepinephrine, and 2 µg/ml vitamin B-12) was refreshed on the first, third and fifth day.…”

“…These system address the limitations by enabling the recording of extracellular and intracellular [7][8][9] electrical activity from cellular monolayers for extended time periods [10]. Moreover, CMOS-based MEAs greatly enhance the throughput, spatial resolution and quality of these measurements compared to more simple glass-based MEAs [7,[11][12][13][14]. Another approach involves measuring the fluorescence of cardiac cells genetically altered to produce voltage sensitive dyes, but fluorescence is susceptible to photobleaching and phototoxicity [15].…”

“…A non-invasive alternative to record electrical activity are microelectrode arrays (MEAs). These system address the limitations by enabling the recording of extracellular and intracellular [7][8][9] electrical activity from cellular monolayers for extended time periods [10]. Moreover, CMOS-based MEAs greatly enhance the throughput, spatial resolution and quality of these measurements compared to more simple glass-based MEAs [7,[11][12][13][14].…”