Time-encoded electrical detection of trace RNA biomarker by integrating programmable molecular amplifier on chip.

One of the serious challenges facing modern point-of-care (PoC) molecular diagnostic platforms relate to reliable detection of low concentration biomarkers such as nucleic acids or proteins in biological samples. Non-specific analyte-receptor interactions due to competitive binding in the presence of abundant molecules, inefficient mass transport and very low number of analyte molecules in sample volume, in general pose critical hurdles for successful implementation of such PoC platforms for clinical use. Focusing on these specific challenges, this work reports a unique PoC biosensor that combines the advantages of nanoscale biologically-sensitive field-effect transistor arrays (BioFET-arrays) realized in a wafer-scale top-down nanofabrication as high sensitivity electrical transducers with that of sophisticated molecular programs (MPs) customized for selective recognition of analyte miRNAs and amplification resulting in an overall augmentation of signal transduction strategy. The MPs realize a programmable universal molecular amplifier (PUMA) in fluidic matrix on chip and provide a biomarker-triggered exponential release of small nucleic acid sequences easily detected by receptor-modified BioFETs. A common miRNA biomarker LET7a was selected for successful demonstration of this novel biosensor, achieving limit of detection (LoD) down to 10 fM and wide dynamic ranges (10 pM-10 nM) in complex physiological solutions. As the determination of biomarker concentration is implemented by following the electrical signal related to analyte-triggered PUMA in time-domain instead of measuring the threshold shifts of BioFETs, and circumvents direct hybridization of biomarkers at transducer surface, this new strategy also allows for multiple usage (>3 times) of the biosensor platform suggesting exceptional cost-effectiveness for practical use.

Understanding Electrical Failure of Polyimide-Based Flexible Neural Implants: The Role of Thin Film Adhesion.

The lack of long-term stability of polymeric neural interfaces remains one of the most important and less tackled issues in this research field. To address this issue, we fabricated two test structures based on interdigitated electrodes (IDEs) encapsulated with polyimide (PI). One of the test samples was pretreated with barrel oxygen plasma prior to spin coating of the second PI layer. The second test structure was pretreated using a reactive ion etching (RIE) process. The test samples were immersed in an electrolyte solution at elevated temperatures to mimic the conditions inside the human brain. The samples were then electrically and mechanically stressed to accelerate their degradation. Real-time monitoring of the electrical insulation stability was used to compare the impact of the pretreatment on the long-term stability. Barrel-plasma-activated test samples showed a mean lifetime of 1.5 days, whereas RIE pretreatment increased the mean lifetime to 24 days. Therefore, RIE-pretreated test samples exhibited 16 times longer mean stability compared to purely chemically activated test samples. Furthermore, the electrical measurements were correlated with mechanical adhesion tests. Chemically activated test samples showed significant delamination, whereas RIE pretreatment enhanced the adhesion, and no delamination could be observed. The correlation of these investigations suggests that the adhesion between different layers is higher following RIE pretreatment compared to pretreatment with chemical barrel plasma. In conclusion, the adhesion between the two PI foils seems to play a key role in the long-term stability of such devices.

Delineating charge and capacitance transduction in system-integrated graphene-based BioFETs used as aptasensors for malaria detection.

Despite significant eradication efforts, malaria remains a persistent infectious disease with high mortality due to the lack of efficient point-of-care (PoC) screening solutions required to manage low-density asymptomatic parasitemia. In response, we demonstrate a quantitative electrical biosensor based on system-integrated two-dimensional field-effect transistors (2DBioFETs) of reduced graphene oxide (rGO) as transducer for high sensitivity screening of the main malaria biomarker, Plasmodium falciparum lactate dehydrogenase (PfLDH). The 2DBioFETs were biofunctionalized with pyrene-modified 2008s aptamers as specific PfLDH receptors. While we systematically optimize biosensor interface for optimal performance, aptamer-protein transduction at 2DBioFETs is elucidated based on delineation of charge and capacitance in an updated analytical model for two-dimensional rGO/biofunctional layer/electrolyte (2DiBLE) interfaces. Our 2DBioFET-aptasensors display a limit-of-detection down to 0.78 fM (0.11 pg/mL), dynamic ranges over 9 orders of magnitude (subfemto to submicromolar), high sensitivity, and selectivity in human serum validating their diagnostic potential as rapid PoC tests for malarial management.

Realization of a PEDOT:PSS/Graphene Oxide On-Chip Pseudo-Reference Electrode for Integrated ISFETs.

A stable reference electrode (RE) plays a crucial role in the performance of an ion-sensitive field-effect transistor (ISFET) for bio/chemical sensing applications. There is a strong demand for the miniaturization of the RE for integrated sensor systems such as lab-on-a-chip (LoC) or point-of-care (PoC) applications. Out of several approaches presented so far to integrate an on-chip electrode, there exist critical limitations such as the effect of analyte composition on the electrode potential and drifts during the measurements. In this paper, we present a micro-scale solid-state pseudo-reference electrode (pRE) based on poly(3,4-ethylene dioxythiophene): poly(styrene sulfonic acid) (PEDOT:PSS) coated with graphene oxide (GO) to deploy with an ion-sensitive field-effect transistor (ISFET)-based sensor platform. The PEDOT:PSS was electropolymerized from its monomer on a micro size gold (Au) electrode and, subsequently, a thin GO layer was deposited on top. The stability of the electrical potential and the cross-sensitivity to the ionic strength of the electrolyte were investigated. The presented pRE exhibits a highly stable open circuit potential (OCP) for up to 10 h with a minimal drift of ~0.65 mV/h and low cross-sensitivity to the ionic strength of the electrolyte. pH measurements were performed using silicon nanowire field-effect transistors (SiNW-FETs), using the developed pRE to ensure good gating performance of electrolyte-gated FETs. The impact of ionic strength was investigated by measuring the transfer characteristic of a SiNW-FET in two electrolytes with different ionic strengths (1 mM and 100 mM) but the same pH. The performance of the PEDOT:PSS/GO electrode is similar to a commercial electrochemical Ag/AgCl reference electrode.

Towards Long-Term Stable Polyimide-Based Flexible Electrical Insulation for Chronically Implanted Neural Electrodes.

For chronic applications of flexible neural implants, e.g., intracortical probes, the flexible substrate material has to encapsulate the electrical conductors with a long-term stability against the saline environment of the neural tissue. The biocompatible polymer polyimide is often used for this purpose. Due to its chemical inertness, the adhesion between two polyimide layers is, however, a challenge, which can lead to delamination and, finally, to short circuits. The state-of-the-art method to improve the adhesion strength is activating the polyimide surface using oxygen reactive ion etching (O2 RIE). However, the influence of the process variations (etching time, bias power) on the long-term stability is still unclear. Therefore, we establish a test method, where the aging of a gold interdigital structure embedded in two polyimide layers and immersed in saline solution is accelerated using an elevated temperature, mechanical stress and an electrical field. A continuous measurement of a leakage current is used to define the failure state. The results show that the variation of the O2 RIE plasma process has a significant effect on the long-term stability of the test samples. Comparing the two different plasma treatments 0.5 min at 25 W and 1 min at 50 W, the long-term stability could be increased from 20.9 ± 19.1 days to 44.9 ± 18.9 days. This corresponds to more than a doubled lifetime. An ideal solution for the delamination problem is still not available; however, the study shows that the fine-tuning of the fabrication processes can improve the long-term stability of chronically implanted neural electrodes.

Process Variability in Top-Down Fabrication of Silicon Nanowire-Based Biosensor Arrays.

Silicon nanowire field-effect transistors (SiNW-FET) have been studied as ultra-high sensitive sensors for the detection of biomolecules, metal ions, gas molecules and as an interface for biological systems due to their remarkable electronic properties. "Bottom-up" or "top-down" approaches that are used for the fabrication of SiNW-FET sensors have their respective limitations in terms of technology development. The "bottom-up" approach allows the synthesis of silicon nanowires (SiNW) in the range from a few nm to hundreds of nm in diameter. However, it is technologically challenging to realize reproducible bottom-up devices on a large scale for clinical biosensing applications. The top-down approach involves state-of-the-art lithography and nanofabrication techniques to cast SiNW down to a few 10s of nanometers in diameter out of high-quality Silicon-on-Insulator (SOI) wafers in a controlled environment, enabling the large-scale fabrication of sensors for a myriad of applications. The possibility of their wafer-scale integration in standard semiconductor processes makes SiNW-FETs one of the most promising candidates for the next generation of biosensor platforms for applications in healthcare and medicine. Although advanced fabrication techniques are employed for fabricating SiNW, the sensor-to-sensor variation in the fabrication processes is one of the limiting factors for a large-scale production towards commercial applications. To provide a detailed overview of the technical aspects responsible for this sensor-to-sensor variation, we critically review and discuss the fundamental aspects that could lead to such a sensor-to-sensor variation, focusing on fabrication parameters and processes described in the state-of-the-art literature. Furthermore, we discuss the impact of functionalization aspects, surface modification, and system integration of the SiNW-FET biosensors on post-fabrication-induced sensor-to-sensor variations for biosensing experiments.