Transfer-free Fabrication and Assessment of Transparent Multilayer CVD Graphene MEAs for in-vitro Optogenetic Studies

Abstract

Electronic interfaces, particularly microelectrode arrays (MEAs), are crucial for studying electrophysiological processes in the body, with applications ranging from implants to deep brain simulators. In neuroscience, they play a vital role in exploring neuronal cell distribution and behaviour, as well as disorders like epilepsy and Alzheimer’s disease. However, electrophysiological recordings have limitations, that have led to the exploration of optical approaches like calcium imaging. To address the shortcomings, a promising strategy involves integrating electrophysiology and optical methods for simultaneous cellular activity measurement, capitalising on their combined temporal and spatial resolution. The challenge lies in developing fully transparent MEAs to overcome the limitations of traditional opaque electrodes.
Graphene’s versatile properties, spanning from electrical conductivity to mechanical flexibility, position it as an ideal material for transparent and flexible electronics, particularly in neural recording and stimulation. Due to these properties, graphene MEAs (gMEAs) allow integration with various optical techniques, overcoming limitations associated with traditional opaque MEAs.
In this project, we designed and fabricated a transparent gMEA, intended to perform electrical signal recordings and optical voltage mappings simultaneously from photostimulated optogenetic cell lines. The design allows for photostimulation from a source beneath the gMEA, while enabling unobstructed optical measurements from above. The electrodes were crafted from multilayer chemical vapour deposition (CVD) graphene, chosen for its transparency and favourable electrical properties. Quartz and sapphire were evaluated as potential substrates for the device. After demonstrating the synthesis of multilayer graphene was possible on both substrates, quartz was selected as the preferred material due to its resistance to graphene delamination.
Characterisation of the gMEAs was done using various techniques, including optical transmittance (OT), electrochemical impedance spectroscopy (EIS), and measurements of the signal-to-noise ratio (SNR). The stability of the gMEAs was also assessed by immersing the devices in cell culture medium and with ageing tests performed in PBS. Initial electrochemical characterisation of the gMEAs exhibited promising signal detection despite a relatively high baseline noise of ∼ 23 μV . In comparison, commercially available MultiChannel Systems MEA (60MEA200/30iR-Ti), showed a lower baseline noise (∼ 4 μV ), but gMEAs achieved comparable signal sensitivity. EIS of gMEAs revealed an impedance at 1 kHz ranging from 3.2 to 9.89 MΩ, largely surpassing values in other studies. However, when area normalised, the impedance remained comparable to reported values. Stability tests identified issues related to the permeability of the encapsulation layer and degradation of molybdenum structures, causing large variations in the SNR and EIS measurements after exposure to liquid media.