Improving glutamate microsensors by optimizing the composition of the redox hydrogel

Amperometric hydrogel-coated glutamate microsensors form a promising concept to detect glutamate levels directly in brain tissue. These microsensors are constructed by coating a carbon fiber electrode (CFE) (10 mu m diameter; 300-500 mu m long) with a five-component redox-hydrogel, in which L-glutamate oxidase, horseradish peroxidase, and ascorbate oxidase are wired via poly(ethylene glycol) diglycidyl ether to an osmium-containing redox polymer. Coating with a thin Nafion film completes the construction. Prior to use in vivo, a reliable and reproducible construction of microsensors with a high performance is required. For an optimal microsensor performance, the balance between the five individual hydrogel components is critical. However, due to their small size, hydrogel application to CFE's need to be performed by dip-coating. Dip-coating is a difficult procedure to control and does not allow individual application of hydrogel constituents. To improve the microsensor construction and to better control the dip-coating procedure, we have recently developed an automated device. Throughout this study, automatic dip-coating was performed with premixed solutions, in which the amount of a single component was varied. This allowed us to optimize the hydrogel composition, which resulted in a significant improvement of the microsensor properties in terms of sensitivity, current density, linearity, detection limit, and interference by ascorbic acid.