Dataset Information

Long-term Intracellular Recording of Optogenetically-induced Electrical Activities using Vertical Nanowire Multi Electrode Array.

ABSTRACT: Continuous recording of intracellular activities in single cells is required for deciphering rare, dynamic and heterogeneous cell responses, which are missed by population or brief single-cell recording. Even if the field of intracellular recording is constantly proceeding, several technical challenges are still remained to conquer this important approach. Here, we demonstrate long-term intracellular recording by combining a vertical nanowire multi electrode array (VNMEA) with optogenetic stimulation to minimally disrupt cell survival and functions during intracellular access and measurement. We synthesized small-diameter and high-aspect-ratio silicon nanowires to spontaneously penetrate into single cells, and used light to modulate the cell's responsiveness. The light-induced intra- and extracellular activities of individual optogenetically-modified cells were measured simultaneously, and each cell showed distinctly different measurement characteristics according to the cell-electrode configuration. Intracellular recordings were achieved continuously and reliably without signal interference and attenuation over 24?hours. The integration of two controllable techniques, vertically grown nanowire electrodes and optogenetics, expands the strategies for discovering the mechanisms for crucial physiological and dynamic processes in various types of cells.

SUBMITTER: Yoo J

PROVIDER: S-EPMC7062878 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Long-term Intracellular Recording of Optogenetically-induced Electrical Activities using Vertical Nanowire Multi Electrode Array.

Yoo Jisoo J Kwak Hankyul H Kwon Juyoung J Ha Go Eun GE Lee Elliot H EH Song Seungwoo S Na Jukwan J Lee Hyo-Jung HJ Lee Jaejun J Hwangbo Areum A Cha Eunkyung E Chae Youngcheol Y Cheong Eunji E Choi Heon-Jin HJ

Scientific reports 20200309 1

Continuous recording of intracellular activities in single cells is required for deciphering rare, dynamic and heterogeneous cell responses, which are missed by population or brief single-cell recording. Even if the field of intracellular recording is constantly proceeding, several technical challenges are still remained to conquer this important approach. Here, we demonstrate long-term intracellular recording by combining a vertical nanowire multi electrode array (VNMEA) with optogenetic stimul ...[more]

PMID: 32152369

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Project description:Multi-electrode arrays, both active or passive, emerged as ideal technologies to unveil intricated electrophysiological dynamics of cells and tissues. Active MEAs, designed using complementary metal oxide semiconductor technology (CMOS), stand over passive devices thanks to the possibility of achieving single-cell resolution, the reduced electrode size, the reduced crosstalk and the higher functionality and portability. Nevertheless, most of the reported CMOS MEA systems mainly rely on a single operational modality, which strongly hampers the applicability range of a single device. This can be a limiting factor considering that most biological and electrophysiological dynamics are often based on the synergy of multiple and complex mechanisms acting from different angles on the same phenomena. Here, we designed a CMOS MEA chip with 16,384 titanium nitride electrodes, 6 independent operational modalities and 1,024 parallel recording channels for neuro-electrophysiological studies. Sixteen independent active areas are patterned on the chip surface forming a 4 × 4 matrix, each one including 1,024 electrodes. Electrodes of four different sizes are present on the chip surface, ranging from 2.5 × 3.5 μm2 up to 11 × 11.0 μm2, with 15 μm pitch. In this paper, we exploited the impedance monitoring and voltage recording modalities not only to monitor the growth and development of primary rat hippocampal neurons, but also to assess their electrophysiological activity over time showing a mean spike amplitude of 144.8 ± 84.6 μV. Fixed frequency (1 kHz) and high sampling rate (30 kHz) impedance measurements were used to evaluate the cellular adhesion and growth on the chip surface. Thanks to the high-density configuration of the electrodes, as well as their dimension and pitch, the chip can appreciate the evolutions of the cell culture morphology starting from the moment of the seeding up to mature culture conditions. The measurements were confirmed by fluorescent staining. The effect of the different electrode sizes on the spike amplitudes and noise were also discussed. The multi-modality of the presented CMOS MEA allows for the simultaneous assessment of different physiological properties of the cultured neurons. Therefore, it can pave the way both to answer complex fundamental neuroscience questions as well as to aid the current drug-development paradigm.

Project description:Nanowire field-effect transistors (NW-FETs) have been shown to be powerful building blocks for nanoscale bioelectronic interfaces with cells and tissue due to their excellent sensitivity and their capability to form strongly coupled interfaces with cell membranes. Graphene has also been shown to be an attractive building block for nanoscale electronic devices, although little is known about its interfaces with cells and tissue. Here we report the first studies of graphene field effect transistors (Gra-FETs) as well as combined Gra- and NW-FETs interfaced to electrogenic cells. Gra-FET conductance signals recorded from spontaneously beating embryonic chicken cardiomyocytes yield well-defined extracellular signals with signal-to-noise ratio routinely >4. The conductance signal amplitude was tuned by varying the Gra-FET working region through changes in water gate potential, V(wg). Signals recorded from cardiomyocytes for different V(wg) result in constant calibrated extracellular voltage, indicating a robust graphene/cell interface. Significantly, variations in V(wg) across the Dirac point demonstrate the expected signal polarity flip, thus allowing, for the first time, both n- and p-type recording to be achieved from the same Gra-FET simply by offsetting V(wg). In addition, comparisons of peak-to-peak recorded signal widths made as a function of Gra-FET device sizes and versus NW-FETs allowed an assessment of relative resolution in extracellular recording. Specifically, peak-to-peak widths increased with the area of Gra-FET devices, indicating an averaged signal from different points across the outer membrane of the beating cells. One-dimensional silicon NW- FETs incorporated side by side with the two-dimensional Gra-FET devices further highlighted limits in both temporal resolution and multiplexed measurements from the same cell for the different types of devices. The distinct and complementary capabilities of Gra- and NW-FETs could open up unique opportunities in the field of bioelectronics in the future.

Dataset Information

Long-term Intracellular Recording of Optogenetically-induced Electrical Activities using Vertical Nanowire Multi Electrode Array.

Publications

Long-term Intracellular Recording of Optogenetically-induced Electrical Activities using Vertical Nanowire Multi Electrode Array.

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