CMOS‐Compatible Silicon Nanowire Field‐Effect Transistors for Ultrasensitive and Label‐Free MicroRNAs Sensing

MicroRNAs (miRNAs) have been regarded as promising biomarkers for the diagnosis and prognosis of early‐stage cancer as their expression levels are associated with different types of human cancers. However, it is a challenge to produce low‐cost miRNA sensors, as well as retain a high sensitivity, both of which are essential factors that must be considered in fabricating nanoscale biosensors and in future biomedical applications. To address such challenges, we develop a complementary metal oxide semiconductor (CMOS)‐compatible SiNW‐FET biosensor fabricated by an anisotropic wet etching technology with self‐limitation which provides a much lower manufacturing cost and an ultrahigh sensitivity. This nanosensor shows a rapid (< 1 minute) detection of miR‐21 and miR‐205, with a low limit of detection (LOD) of 1 zeptomole (ca. 600 copies), as well as an excellent discrimination for single‐nucleotide mismatched sequences of tumor‐associated miRNAs. To investigate its applicability in real settings, we have detected miRNAs in total RNA extracted from lung cancer cells as well as human serum samples using the nanosensors, which demonstrates their potential use in identifying clinical samples for early diagnosis of cancer.     

Label‐Free Optofluidic Nanobiosensor Enables Real‐Time Analysis of Single‐Cell Cytokine Secretion

Single‐cell analysis of cytokine secretion is essential to understand the heterogeneity of cellular functionalities and develop novel therapies for multiple diseases. Unraveling the dynamic secretion process at single‐cell resolution reveals the real‐time functional status of individual cells. Fluorescent and colorimetric‐based methodologies require tedious molecular labeling that brings inevitable interferences with cell integrity and compromises the temporal resolution. An innovative label‐free optofluidic nanoplasmonic biosensor is introduced for single‐cell analysis in real time. The nanobiosensor incorporates a novel design of a multifunctional microfluidic system with small volume microchamber and regulation channels for reliable monitoring of cytokine secretion from individual cells for hours. Different interleukin‐2 secretion profiles are detected and distinguished from single lymphoma cells. The sensor configuration combined with optical spectroscopic imaging further allows us to determine the spatial single‐cell secretion fingerprints in real time. This new biosensor system is anticipated to be a powerful tool to characterize single‐cell signaling for basic and clinical research.     

Label‐Free Virus Capture and Release by a Microfluidic Device Integrated with Porous Silicon Nanowire Forest

Viral diseases are perpetual threats to human and animal health. Detection and characterization of viral pathogens require accurate, sensitive, and rapid diagnostic assays. For field and clinical samples, the sample preparation procedures limit the ultimate performance and utility of the overall virus diagnostic protocols. This study presents the development of a microfluidic device embedded with porous silicon nanowire (pSiNW) forest for label‐free size‐based point‐of‐care virus capture in a continuous curved flow design. The pSiNW forests with specific interwire spacing are synthesized in situ on both bottom and sidewalls of the microchannels in a batch process. With the enhancement effect of Dean flow, this study demonstrates that about 50% H5N2 avian influenza viruses are physically trapped without device clogging. A unique feature of the device is that captured viruses can be released by inducing self‐degradation of the pSiNWs in physiological aqueous environment. About 60% of captured viruses can be released within 24 h for virus culture, subsequent molecular diagnosis, and other virus characterization and analyses. This device performs viable, unbiased, and label‐free virus isolation and release. It has great potentials for virus discovery, virus isolation and culture, functional studies of virus pathogenicity, transmission, drug screening, and vaccine development.     

Label‐Free Attomolar Detection of Proteins Using Integrated Nanoelectronic and Electrokinetic Devices

High‐sensitivity screening of biomarkers is critical to areas ranging from early disease detection and diagnosis to bioterrorism surveillance. Here the development of integrated nanoelectronic and electrokinetic devices for label‐free attomolar detection of proteins is reported. Electrically addressable silicon nanowire field‐effect transistors and electrodes for electrokinetic transport are integrated onto a common sensor chip platform, and the nanowire devices are subsequently functionalized with receptors for selective biomarker detection. Nanowire devices modified with monoclonal antibody for prostate specific antigen exhibit close to a 10⁴ increase in sensitivity due to streaming dielectrophoresis and corresponding electrostatic contribution to the binding affinity after application of an AC electric field. The devices are also modified with receptors for cholera toxin subunit B and achieve a similar enhancement. These results show general applicability of this method, and could open up opportunities in early stage disease detection and the analysis of proteins from single cells.     

Reduced Graphene Oxide‐Functionalized High Electron Mobility Transistors for Novel Recognition Pattern Label‐Free DNA Sensors

We designed and constructed reduced graphene oxide (rGO) functionalized high electron mobility transistor (HEMT) for rapid and ultra‐sensitive detection of label‐free DNA in real time. The micrometer sized rGO sheets with structural defects helped absorb DNA molecules providing a facile and robust approach to functionalization. DNA was immobilized onto the surface of HEMT gate through rGO functionalization, and changed the conductivity of HEMT. The real time monitor and detection of DNA hybridization by rGO functionalized HEMT presented interesting current responses: a “two steps” signal enhancement in the presence of target DNA; and a “one step” signaling with random DNA. These two different recognition patterns made the HEMT capable of specifically detecting target DNA sequence. The working principle of the rGO functionalized HEMT can be demonstrated as the variation of the ambience charge distribution. Furthermore, the as constructed DNA sensors showed excellent sensitivity of detect limit at 0.07 fM with linear detect range from 0.1 fM to 0.1 pM. The results indicated that the HEMT functionalized with rGO paves a new avenue to design novel electronic devices for high sensitive and specific genetic material assays in biomedical applications.     

Design of Hierarchical Beads for Efficient Label‐Free Cell Capture

Defined hierarchical materials promise cell analysis and call for application‐driven design in practical use. The further issue is to develop advanced materials and devices for efficient label‐free cell capture with minimum instrumentation. Herein, the design of hierarchical beads is reported for efficient label‐free cell capture. Silica nanoparticles (size of ≈15 nm) are coated onto silica spheres (size of ≈200 nm) to achieve nanoscale surface roughness, and then the rough silica spheres are combined with microbeads (≈150–1000 µm in diameter) to assemble hierarchical structures. These hierarchical beads are built via electrostatic interaction, covalent bonding, and nanoparticle adherence. Further, after functionalization by hyaluronic acid (HA), the hierarchical beads display desirable surface hydrophilicity, biocompatibility, and chemical/structural stability. Due to the controlled surface topology and chemistry, HA‐functionalized hierarchical beads afford high cell capture efficiency up to 98.7% in a facile label‐free manner. This work guides the development of label‐free cell capture techniques and contributes to the construction of smart interfaces in bio‐systems.     

Electric‐Field‐Assisted Growth of Functionalized Poly(3,4‐ethylenedioxythiophene) Nanowires for Label‐Free Protein Detection

The construction of functionalized poly(3,4‐ethylenedioxythiophene) (PEDOT) nanowire devices for label‐free protein detection is reported. Direct growth/assembly of PEDOT nanowires with carboxylic acid side‐chain functional groups (poly(EDOT‐COOH)) across the electrode junction is achieved by using an electric‐field‐assisted method. These functionalized PEDOT nanowire devices show typical depletion‐mode p‐type field‐effect transistor (FET) properties. Upon conjugation with a protein‐binding aptamer, the PEDOT nanowire FET devices are used for label‐free electronic detection of a target protein of interest. The binding of a positively charged protein causes a substantial decrease in current flow, attributed to the specific interaction between target protein molecules and aptamer‐conjugated polymer chains.     

Tailored Height Gradients in Vertical Nanowire Arrays via Mechanical and Electronic Modulation of Metal‐Assisted Chemical Etching

In current top‐down nanofabrication methodologies the design freedom is generally constrained to the two lateral dimensions, and is only limited by the resolution of the employed nanolithographic technique. However, nanostructure height, which relies on certain mask‐dependent material deposition or etching techniques, is usually uniform, and on‐chip variation of this parameter is difficult and generally limited to very simple patterns. Herein, a novel nanofabrication methodology is presented, which enables the generation of high aspect‐ratio nanostructure arrays with height gradients in arbitrary directions by a single and fast etching process. Based on metal‐assisted chemical etching using a catalytic gold layer perforated with nanoholes, it is demonstrated how nanostructure arrays with directional height gradients can be accurately tailored by: (i) the control of the mass transport through the nanohole array, (ii) the mechanical properties of the perforated metal layer, and (iii) the conductive coupling to the surrounding gold film to accelerate the local electrochemical etching process. The proposed technique, enabling 20‐fold on‐chip variation of nanostructure height in a spatial range of a few micrometers, offers a new tool for the creation of novel types of nano‐assemblies and metamaterials with interesting technological applications in fields such as nanophotonics, nanophononics, microfluidics or biomechanics.