The sensitivity of zinc (Zn(II)) detection using fast-scan cyclic voltammetry (FSCV) with carbon fiber microelectrodes (CFMEs) is low compared to other neurochemicals. We have shown previously that Zn(II) plates to the surface of CFME's and we speculate that it is because of the abundance of oxide functionality on the surface. Plating reduces sensitivity over time and causes significant disruption to detection stability. This limited sensitivity and stability hinders Zn(II) detection, especially in complex matrices like the brain. To address this, we developed plasma-treated gold fiber microelectrodes (AuMEs) which enable sensitive and stable Zn(II) detection with FSCV. Typically, gold fibers are treated using corrosive acids to clean the surface and this step is important for preparing the surface for electrochemistry. Likewise, because FSCV is an adsorption-based technique, it is also important for Zn(II) to adsorb and desorb to prevent irreversible plating. Because of these requirements, careful optimization of the electrode surface was necessary to render the surface for Zn(II) adsorption yet strike a balance between attraction to the surface vs. irreversible interactions. In this study, we employed oxygen plasma treatment to activate the gold fiber surface without inducing significant morphological changes. This treatment effectively removes the organic layer while functionalizing the surface with oxygen, enabling Zn(II) detection that is not possible on untreated gold surfaces. Our results demonstrate significantly improved Zn(II) detection sensitivity and stability on AuME compared to CFME's. Overall, this work provides an advance in our understanding of Zn(II) electrochemistry and a new tool for improved metallotransmitter detection in the brain.

17β-Estradiol (E2) is a ubiquitously expressed hormone that is active in a wide range of neuroprotective and regenerative roles throughout the brain. In particular, it is a well-known dopamine (DA) regulator and is responsible for modulating the expression of dopaminergic receptors and transporters. Recent studies point to E2 release occurring on a rapid time scale and having impacts on DA activity within seconds to minutes. As such, tools capable of monitoring the release of both E2 and DA in real time are essential for developing an accurate understanding of their interactive roles in neurotransmission and regulation. Currently, no analytical techniques capable of codetection of both analytes with high sensitivity, spatiotemporal resolution, extended monitoring, and minimal tissue damage exist. We describe a modified waveform using fast-scan cyclic voltammetry that is capable of low nanomolar detection of both DA and E2 on a subsecond time scale. Both analytes have limits of detection at or below 30 nM and high sensitivity: 11.31 ± 0.55 nA/μM for DA and 9.47 ± 0.36 nA/μM for E2. The waveform is validated in a tissue matrix, confirming its viability for measurement in a biologically relevant setting. This is the first method capable of codetection of fluctuations in DA and E2 with the temporal, spatial, and sensitivity requirements necessary for studying real-time neurochemical signaling.

Carbon-based sensors have remained critical materials for electrochemical detection of neurochemicals, rooted in their inherent biocompatibility and broad potential window. Real-time monitoring using fast-scan cyclic voltammetry has resulted in the rise of minimally invasive carbon fiber microelectrodes as the material of choice for making measurements in tissue, but challenges with carbon fiber's innate properties have limited its applicability to understudied neurochemicals. Here, we provide a critical review of the state of carbon-based real-time neurochemical detection and offer insight into ways we envision addressing these limitations in the future. This piece focuses on three main hinderances of traditional carbon fiber based materials: diminished temporal resolution due to geometric properties and adsorption/desorption properties of the material, poor selectivity/specificity to most neurochemicals, and the inability to tune amorphous carbon surfaces for specific interfacial interactions. Routes to addressing these challenges could lie in methods like computational modeling of single-molecule interfacial interactions, expansion to tunable carbon-based materials, and novel approaches to synthesizing these materials. We hope this critical piece does justice to describing the novel carbon-based materials that have preceded this work, and we hope this review provides useful solutions to innovate carbon-based material development in the future for individualized neurochemical structures.

A facile platform derived from deposition of ethynyl linkers on carbon fiber microelectrodes has been developed for sensitive and selective sensing of Cu(ii). This study is the first to demonstrate the successful anodic deposition of ethynyl linkers, specifically 1,4-diethynylbenzene, onto carbon fiber microelectrodes. Multi-scan deposition of DEB on these microelectrodes resulted in an increased sensitivity and selectivity towards Cu(ii) that persists amidst other divalent interferents and displays sustained performance over four days while stored at room temperature. This method can be extended to other ethynyl terminal moieties, thereby creating a versatile chemical platform that will enable improved sensitivity and selectivity for a new frontier of biomarker measurement.

Carbon fiber microelectrodes (CFMEs) have been used to detect neurotransmitters and other biomolecules using fast-scan cyclic voltammetry (FSCV) for the past few decades. This technique measures neurotransmitters such as dopamine and, more recently, physiologically relevant neuropeptides. Oxytocin, a pleiotropic peptide hormone, is physiologically important for adaptation, development, reproduction, and social behavior. This neuropeptide functions as a stress-coping molecule, an anti-inflammatory agent, and serves as an antioxidant with protective effects especially during adversity or trauma. Here, we measure tyrosine using the Modified Sawhorse Waveform (MSW), enabling enhanced electrode sensitivity for the amino acid and oxytocin peptide. Applying the MSW, decreased surface fouling and enabled codetection with other monoamines. As oxytocin contains tyrosine, the MSW was also used to detect oxytocin. The sensitivity of oxytocin detection was found to be 3.99 ± 0.49 nA/μM, (n=5). Additionally, we demonstrate that applying the MSW on CFMEs allows for real time measurements of exogenously applied oxytocin on rat brain slices. These studies may serve as novel assays for oxytocin detection in a fast, sub-second timescale with possible implications for in vivo measurements and further understanding of the physiological role of oxytocin.