Nanomaterial based self-referencing microbiosensors for cell and tissue physiology research.

Physiological studies require sensitive tools to directly quantify transport kinetics in the cell/tissue spatial domain under physiological conditions. Although biosensors are capable of measuring concentration, their applications in physiological studies are limited due to the relatively low sensitivity, excessive drift/noise, and inability to quantify analyte transport. Nanomaterials significantly improve the electrochemical transduction of microelectrodes, and make the construction of highly sensitive microbiosensors possible. Furthermore, a novel biosensor modality, self-referencing (SR), enables direct measurement of real-time flux and drift/noise subtraction. SR microbiosensors based on nanomaterials have been used to measure the real-time analyte transport in several cell/tissue studies coupled with various stimulators/inhibitors. These studies include: glucose uptake in pancreatic ß cells, cancer cells, muscle tissues, intestinal tissues and P. Aeruginosa biofilms; glutamate flux near neuronal cells; and endogenous indole-3-acetic acid flux near the surface of Zea mays roots. Results from the SR studies provide important insights into cancer, diabetes, nutrition, neurophysiology, environmental and plant physiology studies under dynamic physiological conditions, demonstrating that the SR microbiosensors are an extremely valuable tool for physiology research.

An aqueous media based approach for the preparation of a biosensor platform composed of graphene oxide and Pt-black.

The combination of Pt nanoparticles and graphene was more effective in enhancing biosensing than either nanomaterial alone according to previous reports. Based on the structural similarities between water soluble graphene oxide (GrO(x)) and graphene, we report the fabrication of an aqueous media based GrO(x)/Pt-black nanocomposite for biosensing enhancement. In this approach GrO(x) acted as a nanoscale molecular template for the electrodeposition of Pt-black, an amorphously nanopatterned isoform of platinum metal. Scanning electron microscopy (SEM) images and energy-dispersive X-ray spectroscopy (EDS) showed that Pt-black was growing along GrO(x). The effective surface area and electrocatalytic activity towards H(2)O(2) oxidation of GrO(x)/Pt-black microelectrodes were significantly higher than for Pt-black microelectrodes. When used to prepare a bio-nanocomposite based on protein functionalization with the enzyme glucose oxidase (GOx), the GrO(x)/Pt-black microbiosensors exhibited improved sensitivity over the Pt-black microbiosensors. This suggested that the GrO(x)/Pt-black nanocomposite facilitated an increase in electron transfer, and/or minimized mass transport limitations as compared to Pt-black used alone. Glucose microbiosensors based on GrO(x)/Pt-black exhibited high sensitivity (465.9 ± 48.0 nA/mM), a low detection limit of 1 µM, a linear response range of 1 µM-2mM, and response time of ≈ 4s. Additionally the sensor was stable and highly selective over potential interferents.

Electrical characterization of a single cell electroporation biochip with the 2-D scanning vibrating electrode technology.

Advancements in microfabrication technology have lead to the development of planar micro-pore electroporation technology. This technology has been shown to provide greater control in single cell manipulation, and electroporation which is independent from cell size. In this work we report direct and spatially resolved characterization of electric currents within a planar micropore electroporation biochip to better understand this phenomenon at the cellular level. This work was performed using a two-dimensional (2-D) vibrating probe (VP). Analysis of the spatial patterns of current density yielded a 4th order polynomial profile in the planes parallel to the biochip's surface and a three parameter hyperbolic decay profile in the planes perpendicular to the chip surface. A finite element model was developed which correlates with actual measurements on the micropore. Preliminary VP current density measurements of electroporated HepG2 cells revealed a significantly high current density minutes after electroporation even with non-electroporative pulses. These results indicate that cells take a considerable amount of time for complete electrophysiological recovery and indicate the use of the VP as a cell viability indicator for optimized electroporation.