| The main goal of this research is to design, fabricate, and test a nanomaterial-based platform adequate for the measurement of physiological changes in living cells. The two primary objectives toward this end are (1) the synthesis and selection of a suitable nanomaterial and (2) the demonstration of cellular response to a direct stimulus. Determining a useful nanomaterial morphology and behavior within a sensor configuration presented challenges based on cellular integration and access to electrochemical characterization. The prospect for feasible optimization and eventual scale-up in technology were also significant. Constraining criteria are that the nanomaterial detector must (a) be cheap and relatively easy to fabricate controllably, (b) encourage cell attachment, (c) exhibit consistent wettability over time, and (d) facilitate electrochemical processes. The ultimate goal would be to transfer a proof-of-principle and proof-of-design for a whole-cell sensor technology that is cost effective and has a potential for hand-held packaging.;Initial tasks were to determine an effective and highly-functional nanomaterial for biosensors by assessing wettability, morphology and conductivity behavior of several candidate materials: gallium nitride nanowires, silicon dioxide nanosprings and nanowires, and titania nanofibers. Electrospinning poly(vinyl pyrrolidone)-coated titania nano- and microfibers (O20 nm--2 microm) into a pseudo-random network is controllable to a uniformity of 1--2° in contact angle. The final electrode can be prepared with a precise wettability ranging from partial wetting to ultrahydrophobic (170°) on a variety of substrates: glass, indium tin oxide, silicon, and aluminum. Fiber mats exhibit excellent mechanical stability against rinsing, and support the incubation of epithelial (skin) and pancreatic cells. Impedance spectroscopy on the whole-cell sensor shows resistive changes attributed to cell growth as well as complex frequency-dependent behavior that can be interpreted as simple RCL circuit behavior with changing component parameters. Upon addition of lactic acid, some cell death is evident but complex impedance measurements indicate competing cell growth with adjustment to media pH. The surface impedance of the PVP-titania fiber-ITO electrode has been used in a novel measurement method to reveal significant qualitative and quantitative materials response characteristics associated with changes in solution environment, fiber mat morphology, and the state of the cells' attachment, proliferation and death. |
