Simultaneous measurement of nonlinearity and electrochemical impedance for protein sensing using two-tone excitation.

Impedance biosensors detect the binding of a target to an immobilized probe by quantifying changes in the impedance of the electrode-electrolyte interface. The interface's I-V relationship is inherently nonlinear, varying with DC bias, and target binding can alter the degree of nonlinearity. We propose and demonstrate a method to simultaneously measure the nonlinearity and conventional small-signal impedance using intermodulation products from a two-tone input. Intermodulation amplitudes accurately reflect the impedance's manually-measured voltage dependence. We demonstrate that changes in nonlinearity can discriminate protein binding. Our measurements suggest that target binding can alter nonlinearity via the voltage dependence of the ionic double layer.

A System for Multiplexed Direct Electrical Detection of DNA Synthesis.

An electronic system for the multiplexed detection of DNA polymerization is designed and characterized. DNA polymerization is detected by the measurement of small transient currents arising from ion diffusion during polymerization. A transimpedance amplifier is used to detect these small currents; we implemented a twenty-four channel recording system on a single printed circuit board. Various contributions to the input-referred current noise are analyzed and characterized, as it limits the minimum detectable current and thus the biological limit of detection. We obtained 8.5 pA RMS mean noise current (averaged over all 24 channels) over the recording bandwidth (DC to 2 kHz). With digital filtering, the input-referred current noise of the acquisition system is reduced to 2.4 pA, which is much lower than the biological noise. Electrical crosstalk between channels is measured, and a model for the crosstalk is presented. Minimizing the crosstalk is critical because it can lead to erroneous microarray data. With proper precautions, crosstalk is reduced to a negligible value (less than 1.4%). Using a micro-fabricated array of 24 gold electrodes, we demonstrated system functionality by detecting the presence of a target DNA oligonucleotide which hybridized onto its corresponding target.

In vivo brain imaging using a portable 3.9 gram two-photon fluorescence microendoscope.

We introduce a compact two-photon fluorescence microendoscope based on a compound gradient refractive index endoscope probe, a DC micromotor for remote adjustment of the image plane, and a flexible photonic bandgap fiber for near distortion-free delivery of ultrashort excitation pulses. The imaging head has a mass of only 3.9 g and provides micrometer-scale resolution. We used portable two-photon microendoscopy to visualize hippocampal blood vessels in the brains of live mice.