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Abstract

A Low-Cost Testbed for Neural Microelectrodes †

Department of Biosystems Science and Engineering, ETH Zurich, 4058 Basel, Switzerland
*
Author to whom correspondence should be addressed.
Presented at the XXXV EUROSENSORS Conference, Lecce, Italy, 10–13 September 2023.
Proceedings 2024, 97(1), 62; https://doi.org/10.3390/proceedings2024097062
Published: 21 March 2024

Abstract

:
The performances of microelectrode arrays for neural interfaces strongly depend on electrode design. Due to a lack of simulation tools, electrode engineers often have to refine new designs empirically. This process requires setups of electrical and electrophysiological hardware that are not specific to electrode testing and unnecessarily costly. We propose a low-cost testbed for specifically targeting metrics relevant to electrode performance and functions, which relies on an off-the-shelf measurement tool and only on components necessary for such testing. We experimentally demonstrate the platform by characterizing microelectrodes by means of impedance spectroscopy and recording the extracellular action potentials from in vitro primary rat neurons.

1. Introduction

Electrode design is crucial for the performance of microelectrode arrays (MEAs) used for neural interfaces and cellular electrophysiology [1]. New electrode developments often involve iterative experimentation using certain sets of performance metrics [1,2]. Setups assembled using standard instruments and MEA recording systems are, however, neither cost-optimized nor readily configurable for electrode characterizations. Therefore, we propose a testbed that provides performance and functional testing for electrode development featuring low costs of ~EUR 500, ease of assembly, and compactness. The proposed testbed includes two important tests for electrodes, namely, electrochemical impedance spectroscopy (EIS) and the capability to record action potentials (APs) of electrogenic cells. We experimentally demonstrate the testbed on in-house made MEAs.

2. Materials and Methods

The testbed was built using an off-the-shelf system-on-chip (STEMLab 125-10, Red Pitaya, Solkan, Slovenia) and a custom-printed circuit board featuring a set of multiplexers (MUX) and two amplifiers. MEAs were fabricated on glass wafers (150 nm Pt metal and 1-μm SiO2 passivation layer), featuring 60 working electrodes (WEs) and a reference electrode (RE). Chips were packaged to form a bath chamber. Pt-black was electro-deposited on the WEs in selected chips. The testbed was operated in two modes: (1) EIS and (2) electrophysiological recording (Figure 1a).
For EIS, the MEA bath was filled with phosphate-buffered saline. A sinusoidal voltage excitation signal was swept from 10 Hz to 10 kHz, and the current was recorded from each WE. The impedances were calculated by a digital demodulation technique. For electrophysiological recordings, E18 rat cortical neurons were plated on the arrays. On 31 days in vitro (DIV), the voltage at each WE was amplified (gain = 430), recorded at 15.26 kS/s, and filtered (300 Hz–5 kHz). Continuous recordings of 1.07 s duration were performed repeatedly.

3. Discussion

We used the testbed to obtain impedance spectra for electrodes of 4 and 8 μm diameter, with and without Pt-black (Figure 1b), showing the effects of size and Pt-black on impedance reduction.
We tested 8 μm Pt-black electrodes for the AP recording of rat cortical neurons. Figure 1c shows a representative voltage recording, where extracellular APs are clearly visible.

Author Contributions

Conceptualization, C.-V.H.B. and F.C.; methodology, C.-V.H.B. and F.C.; software, C.-V.H.B., N.M. and F.C.; investigation, C.-V.H.B., N.M. and F.C.; writing—original draft, C.-V.H.B.; writing— review and editing, H.U., A.H. and F.C.; supervision, A.H.; funding acquisition, H.U., A.H. and F.C. All authors have read and agreed to the published version of the manuscript.

Funding

This work was supported by the European Research Council Advanced Grant 694829 ‘neuroXscales’ and the Swiss National Science Foundation project 205320_188910/1.

Institutional Review Board Statement

All experimental protocols involving animals were approved by the Basel-Stadt veterinary office according to Swiss federal laws on animal welfare, and were carried out in accordance with the approved guidelines.

Informed Consent Statement

Not applicable.

Data Availability Statement

The raw data supporting the conclusions of this article will be made available by the authors on request.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Viswam, V.; Obien, M.E.J.; Franke, F.; Frey, U.; Hierlemann, A. Optimal Electrode Size for Multi-Scale Extracellular-Potential Recording from Neuronal Assemblies. Front. Neurosci. 2019, 13, 385. [Google Scholar] [CrossRef] [PubMed]
  2. Boehler, C.; Carli, S.; Fadiga, L.; Stieglitz, T.; Asplund, M. Tutorial: Guidelines for Standardized Performance Tests for Electrodes Intended for Neural Interfaces and Bioelectronics. Nat. Protoc. 2020, 15, 3557–3578. [Google Scholar] [CrossRef] [PubMed]
Figure 1. Testbed schematic and results. (a) An MEA was connected to a 61:4 MUX bank. Two of the outputs were connected to a low-noise amplifier (LNA) and a transimpedance amplifier (TIA). The STEMLab 125-10 hosts a system-on-chip, analog–digital and digital–analog converters (ADC/DAC), and digital outputs (I/Os) to operate the MUXs. Switches S1–5 set up either EIS or recording mode. For EIS, a frequency-swept voltage signal (VA) is applied to the RE, and the current through the WE (IA) is measured by the TIA. For electrophysiological recordings, the RE is grounded, and the voltage (VB) is recorded from a WE after amplification by the LNA. (b) Spectra of mean impedance magnitudes (|Z|) of electrodes of 4 and 8 μm diameter, with and without Pt-black (n = 15 each). (c) Representative voltage recording of extracellular APs (marked with red triangles) from 31 DIV rat neurons.
Figure 1. Testbed schematic and results. (a) An MEA was connected to a 61:4 MUX bank. Two of the outputs were connected to a low-noise amplifier (LNA) and a transimpedance amplifier (TIA). The STEMLab 125-10 hosts a system-on-chip, analog–digital and digital–analog converters (ADC/DAC), and digital outputs (I/Os) to operate the MUXs. Switches S1–5 set up either EIS or recording mode. For EIS, a frequency-swept voltage signal (VA) is applied to the RE, and the current through the WE (IA) is measured by the TIA. For electrophysiological recordings, the RE is grounded, and the voltage (VB) is recorded from a WE after amplification by the LNA. (b) Spectra of mean impedance magnitudes (|Z|) of electrodes of 4 and 8 μm diameter, with and without Pt-black (n = 15 each). (c) Representative voltage recording of extracellular APs (marked with red triangles) from 31 DIV rat neurons.
Proceedings 97 00062 g001
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MDPI and ACS Style

Bui, C.-V.H.; Maliakal, N.; Ulusan, H.; Hierlemann, A.; Cardes, F. A Low-Cost Testbed for Neural Microelectrodes. Proceedings 2024, 97, 62. https://doi.org/10.3390/proceedings2024097062

AMA Style

Bui C-VH, Maliakal N, Ulusan H, Hierlemann A, Cardes F. A Low-Cost Testbed for Neural Microelectrodes. Proceedings. 2024; 97(1):62. https://doi.org/10.3390/proceedings2024097062

Chicago/Turabian Style

Bui, Cat-Vu H., Neethu Maliakal, Hasan Ulusan, Andreas Hierlemann, and Fernando Cardes. 2024. "A Low-Cost Testbed for Neural Microelectrodes" Proceedings 97, no. 1: 62. https://doi.org/10.3390/proceedings2024097062

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