Direct, rapid, and label-free detection of enzyme-substrate interactions in physiological buffers using CMOS-compatible nanoribbon sensors.

We demonstrate the versatility of Al2O3-passivated Si nanowire devices ("nanoribbons") in the analysis of enzyme-substrate interactions via the monitoring of pH change. Our approach is shown to be effective through the detection of urea in phosphate buffered saline (PBS), and penicillinase in PBS and urine, at limits of detection of <200 µM and 0.02 units/mL, respectively. The ability to extract accurate enzyme kinetics and the Michaelis-Menten constant (Km) from the acetylcholine-acetylcholinesterase reaction is also demonstrated.

Record high efficiency single-walled carbon nanotube/silicon p-n junction solar cells.

Carrier transport characteristics in high-efficiency single-walled carbon nanotubes (SWNTs)/silicon (Si) hybrid solar cells are presented. The solar cells were fabricated by depositing intrinsic p-type SWNT thin-films on n-type Si wafers without involving any high-temperature process for p-n junction formation. The optimized cells showed a device ideality factor close to unity and a record-high power-conversion-efficiency of >11%. By investigating the dark forward current density characteristics with varying temperature, we have identified that the temperature-dependent current rectification originates from the thermally activated band-to-band transition of carriers in Si, and the role of the SWNT thin films is to establish a built-in potential for carrier separation/collection. We have also established that the dominant carrier transport mechanism is diffusion, with minimal interface recombination. This is further supported by the observation of a long minority carrier lifetime of ~34 µs, determined by the transient recovery method. This study suggests that these hybrid solar cells operate in the same manner as single crystalline p-n homojunction Si solar cells.

Determination of molecular configuration by debye length modulation.

Silicon nanowire field effect transistors (FETs) have emerged as ultrasensitive, label-free biodetectors that operate by sensing bound surface charge. However, the ionic strength of the environment (i.e., the Debye length of the solution) dictates the effective magnitude of the surface charge. Here, we show that control of the Debye length determines the spatial extent of sensed bound surface charge on the sensor. We apply this technique to different methods of antibody immobilization, demonstrating different effective distances of induced charge from the sensor surface.

Novel electronic biosensor for automated inoculum preparation to accelerate antimicrobial susceptibility testing.

A key predictor of morbidity and mortality for patients with a bloodstream infection is time to appropriate antimicrobial therapy. Accelerating antimicrobial susceptibility testing from positive blood cultures is therefore key to improving patient outcomes, yet traditional laboratory approaches can require 2-4 days for actionable results. The eQUANT-a novel instrument utilizing electrical biosensors-produces a standardized inoculum equivalent to a 0.5 McFarland directly from positive blood cultures. This proof-of-concept study demonstrates that eQUANT inocula prepared from clinically significant species of Enterobacterales were comparable to 0.5 McF inocula generated from bacterial colonies in both CFU/ml concentration and performance in antimicrobial susceptibility testing, with ≥ 95% essential and categorical agreement for VITEK2 and disk diffusion. The eQUANT, combined with a rapid, direct from positive blood culture identification technique, can allow the clinical laboratory to begin antimicrobial susceptibility testing using a standardized inoculum approximately 2-3 h after a blood culture flags positive. This has the potential to improve clinical practice by accelerating conventional antimicrobial susceptibility testing and the resulting targeted antibiotic therapy.

A simple microfluidic aggregation analyzer for the specific, sensitive and multiplexed quantification of proteins in a serum environment.

Portable and low-cost platforms for protein biomarker detection are highly sought after for point of care applications. We demonstrate a simple microfluidic device for the rapid, electrically-based detection of proteins in serum. Our aggregation analyzer relies on detecting the protein-induced aggregation of sub-micron particles, using a one-step procedure followed by a fast, particle-by-particle measurement with a very high count rate. This enables the rapid and precise quantification of C-Reactive protein levels, within the clinically relevant range, using unprocessed human serum and a disposable microfluidic device; no optics are involved in the implementation. Due to the single particle detection format and the use of microfluidics, only a small volume of serum (~50 nL) is needed to complete the analysis. The method can be easily extended to multiplexed biomarker detection by combining an assay using differently sized particles, each targeting a separate protein. We illustrate this by using two sizes of latex beads and demonstrating the simultaneous detection of two different proteins in a serum environment with minimal cross-interference. This confirms that our aggregation analyzer platform provides a simple and straightforward method for multiplexed biomarker detection in a low cost, portable design.

Quantitative probing of surface charges at dielectric-electrolyte interfaces.

The intrinsic charging status at the dielectric-electrolyte interface (DEI) plays a critical role for electrofluidic gating in microfluidics and nanofluidics, which offers opportunities for integration of wet ionics with dry electronics. A convenient approach to quantitatively probe the surface charges at the DEI for material pre-selection purpose has been lacking so far. We report here a low-cost, off-chip extended gate field effect transistor configuration for direct electrostatic probing the charging status at the DEI. Capacitive coupling between the surface charges and the floating extended gate is utilized for signal transducing. The relationship between the surface charge density and the experimentally accessible quantities is given by device modeling. The multiplexing ability makes measuring a local instead of a globally averaged surface charge possible.

Performance limitations for nanowire/nanoribbon biosensors.

Field-effect transistor-based biosensors (bioFETs) have shown great promise in the field of fast, ultra-sensitive, label-free detection of biomolecules. Reliability and accuracy, when trying to measure small concentrations, is of paramount importance for the translation of these research devices into the clinical setting. Our knowledge and experience with these sensors has reached a stage where we are able to identify three main aspects of bioFET sensing that currently limit their applications. By considering the intrinsic device noise as a limitation to the smallest measurable signal, we show how various parameters, processing steps and surface modifications, affect the limit of detection. We also introduce the signal-to-noise ratio of bioFETs as a universal performance metric, which allows us to gain better insight into the design of more sensitive devices. Another aspect that places a limit on the performance of bioFETs is screening by the electrolyte environment, which reduces the signal that could be potentially measured. Alternative functionalization and detection schemes that could enable the use of these charge-based sensors in physiological conditions are highlighted. Finally, the binding kinetics of the receptor-analyte system are considered, both in the context of extracting information about molecular interactions using the bioFET sensor platform and as a fundamental limitation to the number of molecules that bind to the sensor surface at steady-state conditions and to the signal that is generated. Some strategies to overcome these limitations are also proposed. Taken together, these performance-limiting issues, if solved, would bring bioFET sensors closer to clinical applications.

Regenerative electronic biosensors using supramolecular approaches.

A supramolecular interface for Si nanowire FETs has been developed with the aim of creating regenerative electronic biosensors. The key to the approach is Si-NWs functionalized with ß-cyclodextrin (ß-CD), to which receptor moieties can be attached with an orthogonal supramolecular linker. Here we demonstrate full recycling using the strongest biomolecular system known, streptavidin (SAv)-biotin. The bound SAv and the linkers can be selectively removed from the surface through competitive desorption with concentrated ß-CD, regenerating the sensor for repeated use. An added advantage of ß-CD is the possibility of stereoselective sensors, and we demonstrate here the ability to quantify the enantiomeric composition of chiral targets.

Complementary metal oxide semiconductor-compatible silicon nanowire biofield-effect transistors as affinity biosensors.

Affinity biosensors use biorecognition elements and transducers to convert a biochemical event into a recordable signal. They provides the molecule binding information, which includes the dynamics of biomolecular association and dissociation, and the equilibrium association constant. Complementary metal oxide semiconductor-compatible silicon (Si) nanowires configured as a field-effect transistor (NW FET) have shown significant advantages for real-time, label-free and highly sensitive detection of a wide range of biomolecules. Most research has focused on reducing the detection limit of Si-NW FETs but has provided less information about the real binding parameters of the biomolecular interactions. Recently, Si-NW FETs have been demonstrated as affinity biosensors to quantify biomolecular binding affinities and kinetics. They open new applications for NW FETs in the nanomedicine field and will bring such sensor technology a step closer to commercial point-of-care applications. This article summarizes the recent advances in bioaffinity measurement using Si-NW FETs, with an emphasis on the different approaches used to address the issues of sensor calibration, regeneration, binding kinetic measurements, limit of detection, sensor surface modification, biomolecule charge screening, reference electrode integration and nonspecific molecular binding.

Quantification of the affinities and kinetics of protein interactions using silicon nanowire biosensors.

Monitoring the binding affinities and kinetics of protein interactions is important in clinical diagnostics and drug development because such information is used to identify new therapeutic candidates. Surface plasmon resonance is at present the standard method used for such analysis, but this is limited by low sensitivity and low-throughput analysis. Here, we show that silicon nanowire field-effect transistors can be used as biosensors to measure protein-ligand binding affinities and kinetics with sensitivities down to femtomolar concentrations. Based on this sensing mechanism, we develop an analytical model to calibrate the sensor response and quantify the molecular binding affinities of two representative protein-ligand binding pairs. The rate constant of the association and dissociation of the protein-ligand pair is determined by monitoring the reaction kinetics, demonstrating that silicon nanowire field-effect transistors can be readily used as high-throughput biosensors to quantify protein interactions.

Optimal signal-to-noise ratio for silicon nanowire biochemical sensors.

The signal-to-noise ratio (SNR) for silicon nanowire field-effect transistors operated in an electrolyte environment is an essential figure-of-merit to characterize and compare the detection limit of such devices when used in an exposed channel configuration as biochemical sensors. We employ low frequency noise measurements to determine the regime for optimal SNR. We find that SNR is not significantly affected by the electrolyte concentration, composition, or pH, leading us to conclude that the major contributions to the SNR come from the intrinsic device quality. The results presented here show that SNR is maximized at the peak transconductance.

Erratum: "Temperature dependence of 1∕f noise mechanisms in silicon nanowire biochemical field effect transistors" [Appl. Phys. Lett. 97, 243501 (2010)].

[This corrects the article on p. 243501 in vol. 97, PMID: 21221250.][This corrects the article on p. 243501 in vol. 97, PMID: 21221250.].

Multiplexed SOI BioFETs.

Nanoscale Field Effect Transistors have emerged as a promising technology for ultrasensitive, unlabeled diagnostic applications. However, their use as quantitative sensors has been problematic because of the need for individual sensor calibration. In this work we demonstrate an internal calibration scheme for multiplexed nanoribbon field effect sensors by utilizing the initial current rates rather than end point detection. A linear response is observed consistent with initial binding kinetics. Moreover, we are able to show that top-down fabrication techniques yield reproducible device results with minimal fluctuations, enabling internal calibration.

Temperature dependence of 1∕f noise mechanisms in silicon nanowire biochemical field effect transistors.

The 1∕f noise of silicon nanowire biochemical field effect transistors is fully characterized from weak to strong inversion in the temperature range 100-300 K. At 300 K, our devices follow the correlated Δn-Δµ model. As the temperature is lowered, the correlated mobility fluctuations become insignificant and the low frequency noise is best modeled by the Δn-model. For some devices, evidence of random telegraph signals is observed at low temperatures, indicating that fewer traps are active and that the 1∕f noise due to number fluctuations is further resolved to fewer fluctuators, resulting in a Lorentzian spectrum.

Label-free biomarker detection from whole blood.

Label-free nanosensors can detect disease markers to provide point-of-care diagnosis that is low-cost, rapid, specific and sensitive. However, detecting these biomarkers in physiological fluid samples is difficult because of problems such as biofouling and non-specific binding, and the resulting need to use purified buffers greatly reduces the clinical relevance of these sensors. Here, we overcome this limitation by using distinct components within the sensor to perform purification and detection. A microfluidic purification chip simultaneously captures multiple biomarkers from blood samples and releases them, after washing, into purified buffer for sensing by a silicon nanoribbon detector. This two-stage approach isolates the detector from the complex environment of whole blood, and reduces its minimum required sensitivity by effectively pre-concentrating the biomarkers. We show specific and quantitative detection of two model cancer antigens from a 10 microl sample of whole blood in less than 20 min. This study marks the first use of label-free nanosensors with physiological solutions, positioning this technology for rapid translation to clinical settings.