In this report, the fabrication of vertically aligned carbon nanotube nanoelectrode array (VACNT-NEA) by photolithography method is presented. Electrochemical impedance spectroscopy as well as cyclic voltammetry was performed to characterize the arrays with respect to different diffusion regimes. The fabricated array illustrated sigmoidal cyclic voltammogram with steady state current dominated by radial diffusion. The fabricated VACNT-NEA and high density VACNTs were employed as electrochemical glutamate biosensors. Glutamate dehydrogenase is covalently attached to the tip of CNTs. The voltammetric biosensor, based on high density VACNTs, exhibits a sensitivity of 0.976 mA mM(-1) cm(-2) in the range of 0.1-20 μM and 0.182 mA mM(-1) cm(-2) in the range of 20-300 μM glutamate with a low detection limit of 57 nM. Using the fabricated VACNT-NEA, the sensitivity increases approximately to a value of 2.2 Am M(-1) cm(-2) in the range of 0.01 to 20 μM and to 0.1 A mM(-1) cm(-2) in the range of 20-300 μM glutamate. Using this electrode, a record of low detection limit of 10 nM was achieved for glutamate. The results prove the efficacy of the fabricated NEA for low cost and highly sensitive enzymatic biosensor with high sensitivity well suited for voltammetric detection of a wide range of clinically important biomarkers.
We present a portable non-invasive approach for measuring indicators of inflammation and oxidative stress in the respiratory tract by quantifying a biomarker in exhaled breath condensate (EBC). We discuss the fabrication and characterization of a miniaturized electrochemical sensor for detecting nitrite content in EBC using reduced graphene oxide. The nitrite content in EBC has been demonstrated to be a promising biomarker of inflammation in the respiratory tract, particularly in asthma. We utilized the unique properties of reduced graphene oxide (rGO); specifically, the material is resilient to corrosion while exhibiting rapid electron transfer with electrolytes, thus allowing for highly sensitive electrochemical detection with minimal fouling. Our rGO sensor was housed in an electrochemical cell fabricated from polydimethyl siloxane (PDMS), which was necessary to analyze small EBC sample volumes. The sensor is capable of detecting nitrite at a low over-potential of 0.7 V with respect to an Ag/AgCl reference electrode. We characterized the performance of the sensors using standard nitrite/buffer solutions, nitrite spiked into EBC, and clinical EBC samples. The sensor demonstrated a sensitivity of 0.21 μA μM − 1 cm − 2 in the range of 20-100 μM and of 0.1 μA μM − 1 cm − 2 in the range of 100-1000 μM nitrite concentration and exhibited a low detection limit of 830 nM in the EBC matrix. To benchmark our platform, we tested our sensors using seven pre-characterized clinical EBC samples with concentrations ranging between 0.14 and 6.5 μM. This enzyme-free and label-free method of detecting biomarkers in EBC can pave the way for the development of portable breath analyzers for diagnosing and managing changes in respiratory inflammation and disease.
We present a portable system for personalized blood cell counting consisting of a microfluidic impedance cytometer and portable analog readout electronics, feeding into an analog-to-digital converter (ADC), and being transmitted via Bluetooth to a user-accessible mobile application. We fabricated a microfluidic impedance cytometer with a novel portable analog readout. The novel design of the analog readout, which consists of a lock-in-amplifier followed by a high-pass filter stage for subtraction of drift and DC offset, and a post-subtraction high gain stage, enables detection of particles and cells as small as 1 μm in diameter, despite using a low-end 8-bit ADC. The lock-in-amplifier and the ADC were set up to receive and transmit data from a Bluetooth module. In order to initiate the system, as well as to transmit all of the data, a user friendly mobile application was developed, and a proof-of-concept trial was run on a blood sample. Applications such as personalized health monitoring require robust device operation and resilience to clogging. It is desirable to avoid using channels comparable in size to the particles being detected thus requiring high levels of sensitivity. Despite using low-end off-the-shelf hardware, our sensing platform was capable of detecting changes in impedance as small as 0.032%, allowing detection of 3 μm diameter particles in a 300 μm wide channel. The sensitivity of our system is comparable to that of a high-end bench-top impedance spectrometer when tested using the same sensors. The novel analog design allowed for an instrument with a footprint of less than 80 cm. The aim of this work is to demonstrate the potential of using microfluidic impedance spectroscopy for low cost health monitoring. We demonstrated the utility of the platform technology towards cell counting, however, our platform is broadly applicable to assaying wide panels of biomarkers including proteins, nucleic acids, and various cell types.
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