Catalytic activity of glucose oxidase after dielectrophoretic immobilization on nanoelectrodes.

Dielectrophoresis (DEP) is an AC electrokinetic effect that is proven to be effective for the immobilization of not only cells, but also of macromolecules, for example, antibodies and enzyme molecules. In our previous work, we have already demonstrated the high catalytic activity of immobilized horseradish peroxidase after DEP. To evaluate the suitability of the immobilization method for sensing or research in general, we want to test it for other enzymes, too. In this study, glucose oxidase (GOX) from Aspergillus niger was immobilized on TiN nanoelectrode arrays by DEP. Fluorescence microscopy showed the intrinsic fluorescence of the immobilized enzymes flavin cofactor on the electrodes. The catalytic activity of immobilized GOX was detectable, but a fraction of less than 1.3% of the maximum activity that was expected for a full monolayer of immobilized enzymes on all electrodes was stable for multiple measurement cycles. Therefore, the effect of DEP immobilization on the catalytic activity strongly depends on the used enzyme.

AC electrokinetic immobilization of influenza virus.

The use of alternating current (AC) electrokinetic forces, like dielectrophoresis and AC electroosmosis, as a simple and fast method to immobilize sub-micrometer objects onto nanoelectrode arrays is presented. Due to its medical relevance, the influenza virus is chosen as a model organism. One of the outstanding features is that the immobilization of viral material to the electrodes can be achieved permanently, allowing subsequent handling independently from the electrical setup. Thus, by using merely electric fields, we demonstrate that the need of prior chemical surface modification could become obsolete. The accumulation of viral material over time is observed by fluorescence microscopy. The influences of side effects like electrothermal fluid flow, causing a fluid motion above the electrodes and causing an intensity gradient within the electrode array, are discussed. Due to the improved resolution by combining fluorescence microscopy with deconvolution, it is shown that the viral material is mainly drawn to the electrode edge and to a lesser extent to the electrode surface. Finally, areas of application for this functionalization technique are presented.

Fluid streaming above interdigitated electrodes in dielectrophoresis experiments.

For the investigation of alternating current electrokinetic effects, a system is presented that allows for the simultaneous observation of fluid flow above and around microelectrodes in all three directions in space. Beside the usual microscopical view from top, lateral observation through the same objective is made possible by two small mirrors that are placed next to the electrodes. Fluid flow and movement of fluorescent nanoparticles above interdigitated electrodes are monitored by fluorescence microscopy and digital imaging and are further analysed by image processing. Field frequencies are varied from 10 Hz to 1 GHz at up to 10V(rms) . Electrical conductivity of the fluid is monitored in situ in the actual measuring chamber.