We report two 3D printed devices that can be used for electrochemical detection. In both cases, the electrode is housed in commercially available, polymer-based fittings so that the various electrode materials (platinum, platinum black, carbon, gold, silver) can be easily added to a threaded receiving port printed on the device; this enables a module-like approach to the experimental design, where the electrodes are removable and can be easily repolished for reuse after exposure to biological samples. The first printed device represents a microfluidic platform with a 500 × 500 μm channel and a threaded receiving port to allow integration of either polyetheretherketone (PEEK) nut-encased glassy carbon or platinum black (Pt-black) electrodes for dopamine and nitric oxide (NO) detection, respectively. The embedded 1 mm glassy carbon electrode had a limit of detection (LOD) of 500 nM for dopamine and a linear response (R2= 0.99) for concentrations between 25-500 μM. When the glassy carbon electrode was coated with 0.05% Nafion, significant exclusion of nitrite was observed when compared to signal obtained from equimolar injections of dopamine. When using flow injection analysis with a Pt/Pt-black electrode and standards derived from NO gas, a linear correlation (R2 = 0.99) over a wide range of concentrations (7.6 - 190 μM) was obtained, with the LOD for NO being 1 μM. The second application showcases a 3D printed fluidic device that allows collection of the biologically relevant analyte adenosine triphosphate (ATP) while simultaneously measuring the release stimulus (reduced oxygen concentration). The hypoxic sample (4.76 ± 0.53 ppm oxygen) released 2.37 ± 0.37 times more ATP than the normoxic sample (8.22 ± 0.60 ppm oxygen). Importantly, the results reported here verify the reproducible and transferable nature of using 3D printing as a fabrication technique, as devices and electrodes were moved between labs multiple times during completion of the study.

The combination of hydrodynamic focusing with embedded capillaries in a microfluidic device is shown to enable both surface enhanced Raman scattering (SERS) and electrochemical characterization of analytes at nanomolar concentrations in flow. The approach utilizes a versatile polystyrene device that contains an encapsulated microelectrode and fluidic tubing, which is shown to enable straightforward hydrodynamic focusing onto the electrode surface to improve detection. A polydimethyslsiloxane (PDMS) microchannel positioned over both the embedded tubing and SERS active electrode (aligned ∼200 μm from each other) generates a sheath flow that confines the analyte molecules eluting from the embedded tubing over the SERS electrode, increasing the interaction between the Riboflavin (vitamin B2) and the SERS active electrode. The microfluidic device was characterized using finite element simulations, amperometry, and Raman experiments. This device shows a SERS and amperometric detection limit near 1 and 100 nM, respectively. This combination of SERS and amperometry in a single device provides an improved method to identify and quantify electroactive analytes over either technique independently.

A new method of fabricating electrodes for microchip devices that involves the use of Teflon molds and a commercially available epoxy to embed electrodes of various size and composition is described. The resulting epoxy base can be polished to generate a fresh electrode and sealed against PDMS-based fluidic structures. Microchip-based flow injection analysis was used to characterize the epoxy-embedded electrodes. It was shown that gold electrodes can be amalgamated with liquid mercury and the resulting mercury/gold electrode used to selectively detect glutathione from lysed red blood cells. The ability to encapsulate multiple electrode materials of differing composition enabled the integration of microchip electrophoresis with electrochemical detection. Finally, a unique feature of this approach is that the electrode connection is made from the bottom of the epoxy base. This enables the creation of three-dimensional gold pillar electrodes (65 µm in diameter and 27 µm in height) that can be integrated within a fluidic network. As compared to the use of a flat electrode of a similar diameter, the use of the pillar electrode led to improvements in both the sensitivity (72.1 pA/µM for the pillar vs. 4.2 pA/µM for the flat electrode) and limit of detection (20 nM for the pillar vs. 600 nM for the flat electrode), with catechol being the test analyte. These epoxy-embedded electrodes hold promise for the creation of inexpensive microfluidic devices that can be used to electrochemically detect biologically important analytes in a manner where the electrodes can be polished and a fresh electrode surface generated as desired.

In this paper, we present two new methodologies of improving the performance of microchip-based electrochemical detection in microfluidic devices. The first part describes the fabrication and characterization of epoxy-embedded gold microelectrode arrays that are evenly spaced and easily modified. Electrodepositions using a gold plating solution can be performed on the electrodes to result in a 3-dimenional pillar array that, when used with microchip-based flow injection analysis, leads to an 8-fold increase in signal (when compared to a single electrode), with the limit of detection (LOD) for catechol being 4 nM. For detecting analytically challenging molecules such as nitric oxide (NO), platinization of electrodes is commonly used to increase the sensitivity. It is shown here that microchip devices containing either the pillar arrays or more traditional glassy carbon electrodes can be modified with platinum black for NO detection. In the case of using glassy carbon electrodes for NO detection, integration of the resulting platinized electrode with microchip-based flow analysis resulted in a 10 times signal increase relative to use of a bare glassy carbon electrode. In addition, it is demonstrated that these electrodes can be coated with Nafion to impart selectivity towards NO over interfering species such as nitrite. The LOD for NO when using the platinum black/Nafion-coated glassy carbon electrode was 9 nM. These electrodes can also be embedded in a polystyrene substrate, with the applicability of these sensitive and selective electrodes being demonstrated by monitoring the ATP-mediated release of NO from endothelial cells immobilized in a microfluidic network without any adhesion factor.

This paper describes the use of epoxy-encapsulated electrodes to integrate microchip-based electrophoresis with electrochemical detection. Devices with various electrode combinations can easily be developed. This includes a palladium decoupler with a downstream working electrode material of either gold, mercury/gold, platinum, glassy carbon, or a carbon fiber bundle. Additional device components such as the platinum wires for the electrophoresis separation and the counter electrode for detection can also be integrated into the epoxy base. The effect of the decoupler configuration was studied in terms of the separation performance, detector noise, and the ability to analyze samples of a high ionic strength. The ability of both glassy carbon and carbon fiber bundle electrodes to analyze a complex mixture was demonstrated. It was also shown that a PDMS-based valving microchip can be used along with the epoxy embedded electrodes to integrate microdialysis sampling with microchip electrophoresis and electrochemical detection, with the microdialysis tubing also being embedded in the epoxy substrate. This approach enables one to vary the detection electrode material as desired in a manner where the electrodes can be polished and modified in a similar fashion to electrochemical flow cells used in liquid chromatography.