Glassy carbon (GC) microelectrodes have been successfully used for the detection of electroactive neurotransmitters such as
dopamine and
serotonin through voltammetry. However, non-electroactive neurotransmitters such as
glutamate,
lactate, and
gamma-aminobutyric acid (
GABA) are inherently unsuitable for detection through voltammetry
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Glassy carbon (GC) microelectrodes have been successfully used for the detection of electroactive neurotransmitters such as
dopamine and
serotonin through voltammetry. However, non-electroactive neurotransmitters such as
glutamate,
lactate, and
gamma-aminobutyric acid (
GABA) are inherently unsuitable for detection through voltammetry techniques without functionalizing the surface of the microelectrodes. To this end, we present here the immobilization of the
L-glutamate oxidase (
GluOx) enzyme on the surface of GC microelectrodes to enable the catalysis of a chemical reaction between
L-glutamate, oxygen, and water to produce H
2O
2, an electroactive byproduct that is readily detectable through voltammetry. This immobilization of
GluOx on the surface of bare GC microelectrodes and the subsequent catalytic reduction in H
2O
2 through fast-scan cyclic voltammetry (FSCV) helped demonstrate the indirect in vitro detection of
glutamate, a non-electroactive molecule, at concentrations as low as 10 nM. The functionalized microelectrodes formed part of a four-channel array of microelectrodes (30 μm × 60 μm) on a 1.6 cm long neural probe that was supported on a flexible polymer, with potential for in vivo applications. The types and strengths of the bond between the GC microelectrode surface and its functional groups, on one hand, and
glutamate and the immobilized functionalization matrix, on the other hand, were investigated through molecular dynamic (MD) modeling and Fourier transform infrared spectroscopy (FTIR). Both MD modeling and FTIR demonstrated the presence of several covalent bonds in the form of
C-O (carbon–oxygen polar covalent bond),
C=O (carbonyl),
C-H (alkenyl),
N-H (hydrogen bond),
C-N (carbon–nitrogen single bond), and
C≡N (
triple carbon–nitrogen bond). Further, penetration tests on an agarose hydrogel model confirmed that the probes are mechanically robust, with their penetrating forces being much lower than the fracture force of the probe material.
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