Point-of-care-ready nanoscale ISFET arrays for sub-picomolar detection of cytokines in cell cultures.

Rapid and frequent screening of cytokines as immunomodulation agents is necessary for precise interventions in severe pathophysiological conditions. In addition to high-sensitivity detection of such analytes in complex biological fluids such as blood, saliva, and cell culture medium samples, it is also crucial to work out miniaturized bioanalytical platforms with potential for high-density integration enabling screening of multiple analytes. In this work, we show a compact, point-of-care-ready bioanalytical platform for screening of cytokines such as interleukin-4 (IL-4) and interleukin-2 (IL-2) based on one-dimensional ion-sensitive field-effect transistors arrays (nanoISFETs) of silicon fabricated at wafer-scale via nanoimprint lithography. The nanoISFETs biofunctionalized with receptor proteins alpha IL-4 and alpha IL-2 were deployed for screening cytokine secretion in mouse T helper cell differentiation culture media, respectively. Our nanoISFETs showed robust sensor signals for specific molecular binding and can be readily deployed for real-time screening of cytokines. Quantitative analyses of the nanoISFET-based bioanalytical platform was carried out for IL-4 concentrations ranging from 25 fg/mL (1.92 fM) to 2.5 µg/mL (192 nM), showing a limit of detection down to 3-5 fM, which was found to be in agreement with ELISA results in determining IL-4 concentrations directly in complex cell culture media. Graphical abstract.

High Aspect Ratio Fin-Ion Sensitive Field Effect Transistor: Compromises toward Better Electrochemical Biosensing.

The development of next generation medicines demands more sensitive and reliable label-free sensing able to cope with increasing needs of multiplexing and shorter times to results. Field effect transistor-based biosensors emerge as one of the main possible technologies to cover the existing gap. The general trend for the sensors has been miniaturization with the expectation of improving sensitivity and response time but presenting issues with reproducibility and noise level. Here we propose a Fin-Field Effect Transistor (FinFET) with a high height to width aspect ratio for electrochemical biosensing solving the issue of nanosensors in terms of reproducibility and noise, while keeping the fast response time. We fabricated different devices and characterized their performance with their response to the pH changes that fitted to a Nernst-Poisson model. The experimental data were compared with simulations of devices with different aspect ratio, establishing an advantage in linearity and lower device resistance to provide higher current signals for the FinFETs with higher aspect ratio. In addition, these FinFETs promise the optimization of reliability and efficiency in terms of limits of detection for which the interplay of the size and geometry of the sensor with the diffusion of the analytes plays a pivotal role.

Recent Advances in Single Crystal Diamond Device Fabrication for Photonics, Sensing and Nanomechanics.

In the last two decades, the use of diamond as a material for applications in nanophotonics, optomechanics, quantum information, and sensors tremendously increased due to its outstanding mechanical properties, wide optical transparency, and biocompatibility. This has been possible owing to advances in methods for growth of high-quality single crystal diamond (SCD), nanofabrication methods and controlled incorporation of optically active point defects (e.g., nitrogen vacancy centers) in SCD. This paper reviews the recent advances in SCD nano-structuring methods for realization of micro- and nano-structures. Novel fabrication methods are discussed and the different nano-structures realized for a wide range of applications are summarized. Moreover, the methods for color center incorporation in SCD and surface treatment methods to enhance their properties are described. Challenges in the upscaling of SCD nano-structure fabrication, their commercial applications and future prospects are discussed.

The influence of geometry and other fundamental challenges for bio-sensing with field effect transistors.

We present a review of field effect transistors (FET) from the point of view of their applications to label-free sensing in the era of genomics and proteomics. Here, rather than a collection of Bio-FET achievements, we propose an analysis of the different issues hampering the use of these devices into clinical applications. We make a particular emphasis on the influence of the sensor geometry in the phenomena of mass transport of analytes, which is a topic that has been traditionally overlooked in the analysis and design of biosensors, but that plays a central role in the achievement of low limits of detection. Other issues like the screening of charges by the ions in liquids with physiological ionic strength and the non-specific binding are also reviewed. In conclusion, we give an overview of different solutions that have been proposed to address all these challenges, demonstrating the potential of field effect transistors owing to their ease of integration with other semiconductor components for developing cost-effective, highly multiplexed sensors for next-generation medicines.

Single step fabrication of Silicon resistors on SOI substrate used as Thermistors.

Temperature sensing is one of the important features of Micro Electro Mechanical Systems and a monolithic integration provides advantages for both fabrication simplicity and performance. The use of Silicon On Insulator substrates allows simple fabrication of integrated wires that can be used as thermistors. We fabricated rectangular and triangular silicon wires with different dimensions in a single step fabrication process based on the wet etching of a <110> Silicon On Insulator substrate. We determined the experimental resistivity of the two kinds of devices and tested their performance as thermistors in a temperature range between 24 and 100 °C. The accuracy and normalized sensitivities of our devices were 0.4 °C and 0.3-0.5%/°C, respectively. The potential of the proposed method resides in the possibility of having devices with different shapes in a single straightforward process.

Top-Down Fabricated Silicon Nanowire Arrays for Field-Effect Detection of Prostate-Specific Antigen.

Highly sensitive electrical detection of biomarkers for the early stage screening of cancer is desired for future, ultrafast diagnostic platforms. In the case of prostate cancer (PCa), the prostate-specific antigen (PSA) is of prime interest and its detection in combination with other PCa-relevant biomarkers in a multiplex approach is advised. Toward this goal, we demonstrate the label-free, potentiometric detection of PSA with silicon nanowire ion-sensitive field-effect transistor (Si NW-ISFET) arrays. To realize the field-effect detection, we utilized the DNA aptamer-receptors specific for PSA, which were covalently and site-specifically immobilized on Si NW-ISFETs. The platform was used for quantitative detection of PSA and the change in threshold voltage of the Si NW-ISEFTs was correlated with the concentration of PSA. Concentration-dependent measurements were done in a wide range of 1 pg/mL to 1 µg/mL, which covers the clinical range of interest. To confirm the PSA-DNA aptamer binding on the Si NW surfaces, a sandwich-immunoassay based on chemiluminescence was implemented. The electrical approach using the Si NW-ISFET platform shows a lower limit of detection and a wide dynamic range of the assay. In future, our platform should be utilized to detect multiple biomarkers in one assay to obtain more reliable information about cancer-related diseases.

On the Use of Scalable NanoISFET Arrays of Silicon with Highly Reproducible Sensor Performance for Biosensor Applications.

As a prerequisite to the development of real label-free bioassay applications, a high-throughput top-down nanofabrication process is carried out with a combination of nanoimprint lithography, anisotropic wet-etching, and photolithography methods realizing nanoISFET arrays that are then analyzed for identical sensor characteristics. Here, a newly designed array-based sensor chip exhibits 32 high aspect ratio silicon nanowires (SiNWs) laid out in parallel with 8 unit groups that are connected to a very highly doped, Π-shaped common source and individual drain contacts. Intricately designed contact lines exert equal feed-line resistances and capacitances to homogenize the sensor response as well as to minimize parasitic transport effects and to render easy integration of a fluidic layer on top. The scalable nanofabrication process as outlined in this article casts out a total of 2496 nanowires (NWs) on a 4 inch p-type silicon-on-insulator (SOI) wafer, yielding 78 sensor chips based on nanoISFET arrays. The sensor platform exhibiting high-performance transistor characteristics in buffer solutions is thoroughly characterized using state-of-the-art surface and electrical measurement techniques. Deploying a pH sensor in liquid buffers after high-quality gas-phase silanization, nanoISEFT arrays demonstrate typical pH sensor behavior with sensitivity as high as 43 ± 3 mV·pH-1 and a device-to-device variation of 7% at the wafer scale. Demonstration of a high-density sensor platform with uniform characteristics such as nanoISFET arrays of silicon (Si) in a routine and refined nanofabrication process may serve as an ideal solution deployable for real assay-based applications.

Chlorotoxin: a helpful natural scorpion peptide to diagnose glioma and fight tumor invasion.

Chlorotoxin is a small 36 amino-acid peptide identified from the venom of the scorpion Leiurus quinquestriatus. Initially, chlorotoxin was used as a pharmacological tool to characterize chloride channels. While studying glioma-specific chloride currents, it was soon discovered that chlorotoxin possesses targeting properties towards cancer cells including glioma, melanoma, small cell lung carcinoma, neuroblastoma and medulloblastoma. The investigation of the mechanism of action of chlorotoxin has been challenging because its cell surface receptor target remains under questioning since two other receptors have been claimed besides chloride channels. Efforts on chlorotoxin-based applications focused on producing analogues helpful for glioma diagnosis, imaging and treatment. These efforts are welcome since gliomas are very aggressive brain cancers, close to impossible to cure with the current therapeutic arsenal. Among all the chlorotoxin-based strategies, the most promising one to enhance patient mean survival time appears to be the use of chlorotoxin as a targeting agent for the delivery of anti-tumor agents. Finally, the discovery of chlorotoxin has led to the screening of other scorpion venoms to identify chlorotoxin-like peptides. So far several new candidates have been identified. Only detailed research and clinical investigations will tell us if they share the same anti-tumor potential as chlorotoxin.