Lab-on-a-chip devices that combine, e.g. chemical synthesis with integrated on-chip analytics and multi-compartment organ-on-a-chip approaches, are a fast and attractive evolving research area. While integration of appropriate cell models in microfluidic setups for monitoring the biological activity of synthesis products or test compounds is already in focus, the integration of label-free bioelectronic analysis techniques is still poorly realized. In this context, we investigated the capabilities of impedance spectroscopy as a non-destructive real-time monitoring technique for adherent cell models in a microfluidic setup. While an initial adaptation of a microelectrode array (MEA) layout from a static setup revealed clear restrictions in the application of impedance spectroscopy in a microfluidic chip, we could demonstrate the advantage of a FEM simulation based rational MEA layout optimization for an optimum electrical field distribution within microfluidic structures. Furthermore, FEM simulation based analysis of shear stress and time-dependent test compound distribution led to identification of an optimal flow rate. Based on the simulation derived optimized microfluidic MEA, comparable impedance spectra characteristics were achieved for HEK293A cells cultured under microfluidic and static conditions. Furthermore, HEK293A cells expressing Y1 receptors were used to successfully demonstrate the capabilities of impedimetric monitoring of cellular alterations in the microfluidic setup. More strikingly, the maximum impedimetric signal for the receptor activation was significantly increased by a factor of 2.8. Detailed investigations of cell morphology and motility led to the conclusion that cultivation under microfluidic conditions could lead to an extended and stabilized cell–electrode interface.
El desarrollo de biosensores para identificar marcadores moleculares es fundamental para la implementación de nuevas técnicas que permitan la detección de mutaciones genéticas de manera rápida, económica y simple. En este estudio se presenta el desarrollo de un biosensor de ADN complementario (cADN) basado en mediciones de espectroscopía de bioimpedancia eléctrica, y se evalúa su potencial utilidad en la detección de un gen característico de obesidad. Los resultados indican la factibilidad técnica de desarrollar un biosensor de marcadores moleculares o genes específicos a través de la inmovilización de cADN y su detección con mediciones de espectroscopía de bioimpedancia eléctrica.
For miniaturization and integration of chemical synthesis and analytics on small length scales, the development of complex lab-on-chip (LOC) systems is in the focus of many current research projects. While application specific synthesis and analytic modules and LOC devices are widely described, the combination and integration of different modules is intensively investigated. Problems for in-line processes such as solvent incompatibilities, e.g., for a multistep synthesis or the combination of an organic drug synthesis with a cell-based biological activity testing system, require a solvent exchange between serialized modules. Here, we present a continuously operating microfluidic solvent exchanger based on the principle of free-flow electrophoresis for miscible organic/aqueous fluids. We highlight a proof-of-principle and describe the working principle for the model compound fluorescein, where the organic solvent DMSO is exchanged against an aqueous buffer. The DMSO removal performance could be significantly increased to 95% by optimization of the microfluidic layout. Moreover, the optimization of the inlet flow ratio resulted in a minimized dilution factor of 5, and we were able to demonstrate that a reduction of the supporting instrumentation is possible without a significant decrease of the DMSO removal performance. Finally, the compatibility of the developed solvent exchanger for cell based downstream applications was proven. The impedimetric monitoring of HEK293A cells in a continuously operating microfluidic setup revealed no adverse effects of the residual DMSO after the solvent replacement. Our solvent exchanger device demonstrates the power of micro-free-flow electrophoresis not only as a powerful technique for separation and purification of compound mixtures but also for solvent replacement.
Microfluidic platforms capable of complex on-chip processing and liquid handling enable a wide variety of sensing, cellular, and material-related applications across a spectrum of disciplines in engineering and biology. However, there is a general lack of available microscale sensors capable of non-optically monitoring and quantifying on-chip fluid motion. Hence, many microfluidic systems are confined to the laboratory because their use requires optical microscopy. Here, we present a method for dynamically tracking laminar interfacial flows in microfluidic channels non-optically using impedance spectroscopy. Using a microfluidic T-channel, we generate a liquid interface by co-flowing two different electrolyte streams side-by-side. The interface is driven through an array of “displacement” electrodes where it is electrokinetically deflected across the microchannel. The interfacial flow is monitored downstream using an array of interdigitated “impedance” electrodes which dynamically measure the electrochemical impedance near the surface of the flow channel. We demonstrate that the impedance spectrum is sensitively influenced by the position of the deflected interface. While laminar fluid interfaces are ubiquitous to microfluidic flows and used extensively in rheology and biomolecular detection, it is currently difficult to measure their position non-optically. The sensing method presented here enables the interfacial position to be dynamically determined without a microscope and provides a new tool for lowering the barriers to operating microfluidic devices outside the confines of a traditional laboratory.