Label-free DNA sensing platform with low-voltage electrolyte-gated transistors.

We report a method to measure DNA hybridization potentiometrically in a manner conducive to portable or hand-held biosensors. An electrolyte-gated transistor (EGT) based on poly(3-hexylthiophene) (P3HT) and an ion-gel serves as a transducer for surface hybridization of DNA. The key aspect of the design is the use of a floating-gate electrode functionalized with ssDNA whose potential is determined by both capacitive coupling with a primary, addressable gate electrode and the presence of adsorbed molecules. When DNA is hybridized at the floating gate, it offsets the primary gate voltage felt by the P3HT semiconductor; the offset is directly measurable and quantitatively related to the number density of dsDNA molecules. The presented sensing strategy can be readily adapted to other biomolecules of interest and integrated into a microfluidic system for field applications of biosensors.

Detection and Sourcing of Gluten in Grain with Multiple Floating-Gate Transistor Biosensors.

We report a chemically tunable electronic sensor for quantitation of gluten based on a floating-gate transistor (FGT) architecture. The FGTs are fabricated in parallel and each one is functionalized with a different chemical moiety designed to preferentially bind a specific grain source of gluten. The resulting set of FGT sensors can detect both wheat and barley gluten below the gluten-free limit of 20 ppm (w/w) while providing a source-dependent signature for improved accuracy. This label-free transduction method does not require any secondary binding events, resulting in a ca. 45 min reduction in analysis time relative to state-of-the-art ELISA kits with a simple and easily implemented workflow.

Interfacial Charge Contributions to Chemical Sensing by Electrolyte-Gated Transistors with Floating Gates.

The floating gate, electrolyte-gated transistor (FGT) is a chemical sensing device utilizing a floating gate electrode to physically separate and electronically couple the active sensing area with the transistor. The FGT platform has yielded promising results for the detection of DNA and proteins, but questions remain regarding its fundamental operating mechanism. Using carboxylic acid-terminated self-assembled monolayers (SAMs) exposed to solutions of different pH, we create a charged surface and hence characterize the role that interfacial charge concentration plays relative to capacitance changes. The results agree with theoretical predictions from conventional double-layer theory, rationalizing nonlinear responses obtained at high analyte concentrations in previous work using the FGT architecture. Our study elucidates an important effect in the sensing mechanism of FGTs, expanding opportunities for the rational optimization of these devices for chemical and biochemical detection.

Neural pathfinding on uni- and multidirectional photopolymerized micropatterns.

Overcoming signal resolution barriers of neural prostheses, such as the commercially available cochlear impant (CI) or the developing retinal implant, will likely require spatial control of regenerative neural elements. To rationally design materials that direct nerve growth, it is first necessary to determine pathfinding behavior of de novo neurite growth from prosthesis-relevant cells such as spiral ganglion neurons (SGNs) in the inner ear. Accordingly, in this work, repeating 90° turns were fabricated as multidirectional micropatterns to determine SGN neurite turning capability and pathfinding. Unidirectional micropatterns and unpatterned substrates are used as comparisons. Spiral ganglion Schwann cell alignment (SGSC) is also examined on each surface type. Micropatterns are fabricated using the spatial reaction control inherent to photopolymerization with photomasks that have either parallel line spacing gratings for unidirectional patterns or repeating 90° angle steps for multidirectional patterns. Feature depth is controlled by modulating UV exposure time by shuttering the light source at given time increments. Substrate topography is characterized by white light interferometry and scanning electron microscopy (SEM). Both pattern types exhibit features that are 25 µm in width and 7.4 ± 0.7 µm in depth. SGN neurites orient randomly on unpatterned photopolymer controls, align and consistently track unidirectional patterns, and are substantially influenced by, but do not consistently track, multidirectional turning cues. Neurite lengths are 20% shorter on multidirectional substrates compared to unidirectional patterns while neurite branching and microfeature crossing events are significantly higher. For both pattern types, the majority of the neurite length is located in depressed surface features. Developing methods to understand neural pathfinding and to guide de novo neurite growth to specific stimulatory elements will enable design of innovative biomaterials that improve functional outcomes of devices that interface with the nervous system.

Photopolymerized microfeatures for directed spiral ganglion neurite and Schwann cell growth.

Cochlear implants (CIs) provide auditory perception to individuals with severe hearing impairment. However, their ability to encode complex auditory stimuli is limited due, in part, to poor spatial resolution caused by electrical current spread in the inner ear. Directing nerve cell processes towards target electrodes may reduce the problematic current spread and improve stimulatory specificity. In this work, photopolymerization was used to fabricate micro- and nano-patterned methacrylate polymers to probe the extent of spiral ganglion neuron (SGN) neurite and Schwann cell (SGSC) contact guidance based on variations in substrate topographical cues. Micropatterned substrates are formed in a rapid, single-step reaction by selectively blocking light with photomasks which have parallel line-space gratings with periodicities of 10-100 µm. Channel amplitudes of 250 nm-10 µm are generated by modulating UV exposure time, light intensity, and photoinitiator concentration. Gradual transitions are observed between ridges and grooves using scanning electron and atomic force microscopy. The transitions stand in contrast to vertical features generated via etching lithographic techniques. Alignment of neural elements increases significantly with increasing feature amplitude and constant periodicity, as well as with decreasing periodicity and constant amplitude. SGN neurite alignment strongly correlates (r = 0.93) with maximum feature slope. Multiple neuronal and glial types orient to the patterns with varying degrees of alignment. This work presents a method to fabricate gradually-sloping micropatterns for cellular contact guidance studies and demonstrates spatial control of inner ear neural elements in response to micro- and nano-scale surface topography.