RESUMEN
A potentiostat circuit for the application of bipolar electrode voltages and detection of bidirectional currents using a microelectrode array is presented. The potentiostat operates as a regulated-cascode amplifier for positive input currents, and as an active-input regulated-cascode mirror for negative input currents. This topology enables constant-potential amperometry and fast-scan cyclic voltammetry (FSCV) at microelectrode arrays for parallel recording of quantal release events, electrode impedance characterization, and high-throughput drug screening. A 64-channel FSCV detector array, fabricated in a 0.5-$\mu$m, 5-V CMOS process, is also demonstrated. Each detector occupies an area of 45 $\mu$m $\times$ 30 $\mu$m and consists of only 14 transistors and a 50-fF integrating capacitor. The system was validated using prerecorded input stimuli from actual FSCV measurements at a carbon-fiber microelectrode.
Asunto(s)
Técnicas Electroquímicas/métodos , Microelectrodos , Animales , Técnicas Biosensibles/métodos , Membrana Celular/metabolismo , Exocitosis/fisiología , Humanos , Neuronas/metabolismo , RuidoRESUMEN
Amperometry is a powerful method to record quantal release events from chromaffin cells and is widely used to assess how specific drugs modify quantal size, kinetics of release, and early fusion pore properties. Surface-modified CMOS-based electrochemical sensor arrays allow simultaneous recordings from multiple cells. A reliable, low-cost technique is presented here for efficient targeting of single cells specifically to the electrode sites. An SU-8 microwell structure is patterned on the chip surface to provide insulation for the circuitry as well as cell trapping at the electrode sites. A shifted electrode design is also incorporated to increase the flexibility of the dimension and shape of the microwells. The sensitivity of the electrodes is validated by a dopamine injection experiment. Microwells with dimensions slightly larger than the cells to be trapped ensure excellent single-cell targeting efficiency, increasing the reliability and efficiency for on-chip single-cell amperometry measurements. The surface-modified device was validated with parallel recordings of live chromaffin cells trapped in the microwells. Rapid amperometric spikes with no diffusional broadening were observed, indicating that the trapped and recorded cells were in very close contact with the electrodes. The live cell recording confirms in a single experiment that spike parameters vary significantly from cell to cell but the large number of cells recorded simultaneously provides the statistical significance.