Electrical coupling of mammalian neurons to microelectrodes with 3D nanoprotrusions

Substrate integrated planar microelectrode arrays is the “gold standard” method for millisecond-resolution, long-term, large-scale, cell-noninvasive electrophysiological recordings from mammalian neuronal networks. Nevertheless, these devices suffer from drawbacks that are solved by spike-detecting, spike-sorting and signal-averaging techniques which rely on estimated parameters that require user supervision to correct errors, merge clusters and remove outliers. Here we show that primary rat hippocampal neurons grown on micrometer sized gold mushroom-shaped microelectrodes (gMμE) functionalized simply by poly-ethylene-imine/laminin undergo self-assembly processes to form loose patch-like hybrid structures. More than 90% of the hybrids formed in this way record monophasic positive action potentials (APs). Of these, 34.5% record APs with amplitudes above 300 μV and up to 5,085 μV. This self-assembled neuron-gMμE configuration improves the recording quality as compared to planar MEA. This study characterizes and analyzes the electrophysiological signaling repertoire generated by the neurons-gMμE configuration, and discusses prospects to further improve the technology.

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“…4a,b)262728293441. The circuits illustrate two operational modes of the membrane facing the gMμE (the junctional membranes).…”

Section: Resultsmentioning

confidence: 99%

Multisite electrophysiological recordings by self-assembled loose-patch-like junctions between cultured hippocampal neurons and mushroom-shaped microelectrodes

Shmoel

Rabieh

Ojovan

et al. 2016

Sci Rep

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“…This, together with increased conductance of the neuronal membrane that faces the electrode (the junctional membrane - jm ), makes it possible to record action potentials and subthreshold synaptic potentials with qualities and biophysics similar to perforated patch recordings 31 . Ultrastructural studies have revealed that various cell types including NIH/3T3, CHO, PC-12, H9C2, HL-1 cell lines as well as primary cultured rat hippocampal neurons also engulf mushroom shaped electrodes 25 26 27 30 32 33 34 , thus suggesting that the cell-biological mechanisms leading to gMμE engulfment may be ubiquitous.…”

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confidence: 99%

“…Although no studies have attempted to optimize IN-CELL recordings by increasing the size of gMμEs, several laboratories have explored the potential use of nanometric size mushroom shaped electrodes 32 34 36 . These studies are based on the reasoning that nanometric sized mushroom-shaped microelectrodes may be more suitable for interfacing with small (10–20 μm diameter) mammalian neurons than the very large Aplysia neurons (50–80 μm diameter) used in our proof-of-concept studies.…”

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confidence: 99%

A feasibility study of multi-site,intracellular recordings from mammalian neurons by extracellular gold mushroom-shaped microelectrodes

Ojovan

Rabieh

Shmoel

et al. 2015

Sci Rep

103

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The development of multi-electrode array platforms for large scale recording of neurons is at the forefront of neuro-engineering research efforts. Recently we demonstrated, at the proof-of-concept level, a breakthrough neuron-microelectrode interface in which cultured Aplysia neurons tightly engulf gold mushroom-shaped microelectrodes (gMμEs). While maintaining their extracellular position, the gMμEs record synaptic- and action-potentials with characteristic features of intracellular recordings. Here we examined the feasibility of using gMμEs for intracellular recordings from mammalian neurons. To that end we experimentally examined the innate size limits of cultured rat hippocampal neurons to engulf gMμEs and measured the width of the “extracellular” cleft formed between the neurons and the gold surface. Using the experimental results we next analyzed the expected range of gMμEs-neuron electrical coupling coefficients. We estimated that sufficient electrical coupling levels to record attenuated synaptic- and action-potentials can be reached using the gMμE-neuron configuration. The definition of the engulfment limits of the gMμEs caps diameter at ≤2–2.5 μm and the estimated electrical coupling coefficients from the simulations pave the way for rational development and application of the gMμE based concept for in-cell recordings from mammalian neurons.

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“…In this method micrometer-sized, extracellular gold mushroom-shaped microelectrodes (gMμEs) record attenuated synaptic and action potentials (APs) with characteristic features of intracellular recordings while the electrode maintains an extracellular position2829303132333435. These studies show that a range of cell types tightly engulf gMμEs to form high seal resistance ( R s )2930363738. This, together with the increased conductance of the neuronal membrane that faces the electrode (the junctional membrane – jm ), makes it possible to record from cultured Aplysia neurons APs and subthreshold synaptic potentials with qualities and biophysics similar to perforated patch recordings3940.…”

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confidence: 99%

On-chip, multisite extracellular and intracellular recordings from primary cultured skeletal myotubes

Rabieh

Ojovan

Shmoel

et al. 2016

Sci Rep

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In contrast to the extensive use of microelectrode array (MEA) technology in electrophysiological studies of cultured neurons and cardiac muscles, the vast field of skeletal muscle research has yet to adopt the technology. Here we demonstrate an empowering MEA technology for high quality, multisite, long-term electrophysiological recordings from cultured skeletal myotubes. Individual rat skeletal myotubes cultured on micrometer sized gold mushroom-shaped microelectrode (gMμE) based MEA tightly engulf the gMμEs, forming a high seal resistance between the myotubes and the gMμEs. As a consequence, spontaneous action potentials generated by the contracting myotubes are recorded as extracellular field potentials with amplitudes of up to 10 mV for over 14 days. Application of a 10 ms, 0.5–0.9 V voltage pulse through the gMμEs electroporated the myotube membrane, and transiently converted the extracellular to intracellular recording mode for 10–30 min. In a fraction of the cultures stable attenuated intracellular recordings were spontaneously produced. In these cases or after electroporation, subthreshold spontaneous potentials were also recorded. The introduction of the gMμE-MEA as a simple-to-use, high-quality electrophysiological tool together with the progress made in the use of cultured human myotubes opens up new venues for basic and clinical skeletal muscle research, preclinical drug screening, and personalized medicine.

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Electrical coupling of mammalian neurons to microelectrodes with 3D nanoprotrusions

Cited by 24 publications

References 17 publications

Multisite electrophysiological recordings by self-assembled loose-patch-like junctions between cultured hippocampal neurons and mushroom-shaped microelectrodes

Multisite electrophysiological recordings by self-assembled loose-patch-like junctions between cultured hippocampal neurons and mushroom-shaped microelectrodes

A feasibility study of multi-site,intracellular recordings from mammalian neurons by extracellular gold mushroom-shaped microelectrodes

On-chip, multisite extracellular and intracellular recordings from primary cultured skeletal myotubes

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