Magnetic metal-organic frameworks as sensitive aptasensors for coronavirus spike protein.

Electrochemical biosensors, known for their low cost, sensitivity, selectivity, and miniaturization capabilities, are ideal for point-of-care devices. The magnetic metal-organic framework (MMOF), synthesized using the in-situ growth method, consists of ferric salt, magnetic nanoparticles, histidine, and benzene tetracarboxylic acid. MMOF was sequentially modified with aptamer-biotin and streptavidin-horseradish peroxidase, serving as a detector for spike protein and a transducer converting electrochemical signals using H2O2-hydroquinone on a screen-printed electrode. MMOF facilitates easy washing and homogeneous deposition on the working electrode with a magnet, enhancing sensitivity and reducing noise. The physical and electrochemical properties of the modified MMOFs were thoroughly characterized using various analytical techniques. The aptasensors' performance achieved a detection limit of 6 pM for voltammetry and 5.12 pM for impedance spectroscopy in human serum samples. This cost-effective, portable MMOF platform is suitable for rapid point-of-care testing for SARS-CoV-2 spike proteins.

Portable Electrochemical Biosensors Based on Microcontrollers for Detection of Viruses: A Review.

With the rise of zoonotic diseases in recent years, there is an urgent need for improved and more accessible screening and diagnostic methods to mitigate future outbreaks. The recent COVID-19 pandemic revealed an over-reliance on RT-PCR, a slow, costly and lab-based method for diagnostics. To better manage the pandemic, a high-throughput, rapid point-of-care device is needed for early detection and isolation of patients. Electrochemical biosensors offer a promising solution, as they can be used to perform on-site tests without the need for centralized labs, producing high-throughput and accurate measurements compared to rapid test kits. In this work, we detail important considerations for the use of electrochemical biosensors for the detection of respiratory viruses. Methods of enhancing signal outputs via amplification of the analyte, biorecognition of elements and modification of the transducer are also explained. The use of portable potentiostats and microfluidics chambers that create a miniature lab are also discussed in detail as an alternative to centralized laboratory settings. The state-of-the-art usage of portable potentiostats for detection of viruses is also elaborated and categorized according to detection technique: amperometry, voltammetry and electrochemical impedance spectroscopy. In terms of integration with microfluidics, RT-LAMP is identified as the preferred method for DNA amplification virus detection. RT-LAMP methods have shorter turnaround times compared to RT-PCR and do not require thermal cycling. Current applications of RT-LAMP for virus detection are also elaborated upon.

A review on the contamination of SARS-CoV-2 in water bodies: Transmission route, virus recovery and recent biosensor detection techniques.

The discovery of SARS-CoV-2 virus in the water bodies has been reported, and the risk of virus transmission to human via the water route due to poor wastewater management cannot be disregarded. The main source of the virus in water bodies is the sewage network systems which connects to the surface water. Wastewater-based epidemiology has been applied as an early surveillance tool to sense SARS-CoV-2 virus in the sewage network. This review discussed possible transmission routes of the SARS-CoV-2 virus and the challenges of the existing method in detecting the virus in wastewater. One significant challenge for the detection of the virus is that the high virus loading is diluted by the sheer volume of the wastewater. Hence, virus preconcentration from water samples prior to the application of virus assay is essential to accurately detect traceable virus loading. The preparation time, materials and conditions, virus type, recovery percentage, and various virus recovery techniques are comprehensively discussed in this review. The practicability of molecular methods such as Polymer-Chain-Reaction (PCR) for the detection of SARS-CoV-2 in wastewater will be revealed. The conventional virus detection techniques have several shortcomings and the potential of biosensors as an alternative is also considered. Biosensing techniques have also been proposed as an alternative to PCR and have reported detection limits of 10 pg/µl. This review serves to guide the reader on the future designs and development of highly sensitive, robust and, cost effective SARS-CoV-2 lab-on-a-chip biosensors for use in complex wastewater.

Low-cost and rapid prototyping of integrated electrochemical microfluidic platforms using consumer-grade off-the-shelf tools and materials.

We present a low-cost, accessible, and rapid fabrication process for electrochemical microfluidic sensors. This work leverages the accessibility of consumer-grade electronic craft cutters as the primary tool for patterning of sensor electrodes and microfluidic circuits, while commodity materials such as gold leaf, silver ink pen, double-sided tape, plastic transparency films, and fabric adhesives are used as its base structural materials. The device consists of three layers, the silver reference electrode layer at the top, the PET fluidic circuits in the middle and the gold sensing electrodes at the bottom. Separation of the silver reference electrode from the gold sensing electrodes reduces the possibility of cross-contamination during surface modification. A novel approach in mesoscale patterning of gold leaf electrodes can produce generic designs with dimensions as small as 250 µm. Silver electrodes with dimensions as small as 385 µm were drawn using a plotter and a silver ink pen, and fluid microchannels as small as 300 µm were fabricated using a sandwich of iron-on adhesives and PET. Device layers are then fused together using an office laminator. The integrated microfluidic electrochemical platform has electrode kinetics/performance of ΔEp = 91.3 mV, Ipa/Ipc = 0.905, characterized by cyclic voltammetry using a standard ferrocyanide redox probe, and this was compared against a commercial screen-printed gold electrode (ΔEp = 68.9 mV, Ipa/Ipc = 0.984). To validate the performance of the integrated microfluidic electrochemical platform, a catalytic hydrogen peroxide sensor and enzyme-coupled glucose biosensors were developed as demonstrators. Hydrogen peroxide quantitation achieves a limit of detection of 0.713 mM and sensitivity of 78.37 µA mM-1 cm-2, while glucose has a limit of detection of 0.111 mM and sensitivity of 12.68 µA mM-1 cm-2. This rapid process allows an iterative design-build-test cycle in under 2 hours. The upfront cost to set up the system is less than USD 520, with each device costing less than USD 0.12, making this manufacturing process suitable for low-resource laboratories or classroom settings.

RF Remote Blood Glucose Sensor and a Microfluidic Vascular Phantom for Sensor Validation.

Diabetes has become a major health problem in society. Invasive glucometers, although precise, only provide discrete measurements at specific times and are unsuitable for long-term monitoring due to the injuries caused on skin and the prohibitive cost of disposables. Remote, continuous, self-monitoring of blood sugar levels allows for active and better management of diabetics. In this work, we present a radio frequency (RF) sensor based on a stepped impedance resonator for remote blood glucose monitoring. When placed on top of a human hand, this RF interdigital sensor allows detection of variation in blood sugar levels by monitoring the changes in the dielectric constant of the material underneath. The designed stepped impedance resonator operates at 3.528 GHz with a Q factor of 1455. A microfluidic device structure that imitates the blood veins in the human hand was fabricated in PDMS to validate that the sensor can measure changes in glucose concentrations. To test the RF sensor, glucose solutions with concentrations ranging from 0 to 240 mg/dL were injected into the fluidic channels and placed underneath the RF sensor. The shifts in the resonance frequencies of the RF sensor were measured using a network analyzer via its S11 parameters. Based on the change in resonance frequencies, the sensitivity of the biosensor was found to be 264.2 kHz/mg·dL-1 and its LOD was calculated to be 29.89 mg/dL.

Low-cost and cleanroom-free prototyping of microfluidic and electrochemical biosensors: Techniques in fabrication and bioconjugation.

Integrated microfluidic biosensors enable powerful microscale analyses in biology, physics, and chemistry. However, conventional methods for fabrication of biosensors are dependent on cleanroom-based approaches requiring facilities that are expensive and are limited in access. This is especially prohibitive toward researchers in low- and middle-income countries. In this topical review, we introduce a selection of state-of-the-art, low-cost prototyping approaches of microfluidics devices and miniature sensor electronics for the fabrication of sensor devices, with focus on electrochemical biosensors. Approaches explored include xurography, cleanroom-free soft lithography, paper analytical devices, screen-printing, inkjet printing, and direct ink writing. Also reviewed are selected surface modification strategies for bio-conjugates, as well as examples of applications of low-cost microfabrication in biosensors. We also highlight several factors for consideration when selecting microfabrication methods appropriate for a project. Finally, we share our outlook on the impact of these low-cost prototyping strategies on research and development. Our goal for this review is to provide a starting point for researchers seeking to explore microfluidics and biosensors with lower entry barriers and smaller starting investment, especially ones from low resource settings.

Screen-Printed Nickel-Zinc Batteries: A Review of Additive Manufacturing and Evaluation Methods.

The advent of personalized wearable devices has boosted the demand for portable, compact power sources. Compared with lithographic techniques, printed devices have lower fabrication costs, while still maintaining high throughput and precision. These factors make thick film printing or additive manufacturing ideal for the fabrication of low-cost batteries suitable for personalized devices. This article provides comprehensive guidelines for thick-film battery fabrication and characterization, with the focus on printed nickel-zinc (Ni-Zn) batteries. Ni-Zn batteries are a more environmental-friendly option compared with lithium-ion batteries (LIBs) as they are fully recyclable. In this work, important battery fundamentals have been described, especially terms of electrochemistry, basic design approaches, and the printing technology. Different design approaches, such as lateral, concentric, and stacked, are also discussed. Printed batteries can be configured as series or parallel constructions, depending on the power requirements of the application. The fabrication flow of printed battery electrodes for the laboratory-scale prototyping process starts from chemical preparation, mixing, printing, drying, pressing, stacking to finally sealing and testing. Of particular importance is the process of electrolyte injection and pouch sealing for the printed batteries to reduce leakage. This entire process flow is also compared with industrial fabrication flow for LIBs. Criteria for material and equipment selection are also addressed in this article to ensure appropriate electrode consistency and good performance. Two main testing methods cyclic voltammetry for the electrodes and charge-discharge for the battery are also explained in detail to serve as systematic guide for users to validate the functionality of their electrodes. This review article concludes with commercial applications of printed electrodes in the field of health and personalized wearable devices. This work indicates that printed Ni-Zn and other zinc alkaline batteries have a promising future. The success of these devices also opens up different areas of research, such as ink rheology, composition, and formulation of ink using sustainable sources.

MEMS biosensor for monitoring water toxicity based on quartz crystal microbalance.

This paper presents the use of a commercial quartz crystal microbalance (QCM) to investigate live-cell activity in water-based toxic solutions. The QCM used in this research has a resonant frequency of 10 MHz and consists of an AT-cut quartz crystal with gold electrodes on both sides. This QCM was transformed into a functional biosensor by integrating with polydimethylsiloxane culturing chambers. Rainbow trout gill epithelial cells were cultured on the resonators as a sensorial layer. The fluctuation of the resonant frequency, due to the change of cell morphology and adhesion, is an indicator of water toxicity. The shift in the resonant frequency provides information about the viability of the cells after exposure to toxicants. The toxicity result shows distinct responses after exposing cells to 0.526 µM of pentachlorophenol (PCP) solution, which is the Military Exposure Guidelines concentration. This research demonstrated that the QCM is sensitive to a low concentration of PCP and no further modification of the QCM surface was required.

Modeling and development of screen-printed impedance biosensor for cytotoxicity studies of lung carcinoma cells.

Electrical cell-substrate impedance sensing (ECIS) is a powerful technique to monitor real-time cell behavior. In this study, an ECIS biosensor formed using two interdigitated electrode structures (IDEs) was used to monitor cell behavior and its response to toxicants. Three different sensors with varied electrode spacing were first modeled using COMSOL Multiphysics and then fabricated and tested. The silver/silver chloride IDEs were fabricated using a screen-printing technique and incorporated with polydimethylsiloxane (PDMS) cell culture wells. To study the effectiveness of the biosensor, A549 lung carcinoma cells were seeded in the culture wells together with collagen as an extracellular matrix (ECM) to promote cell attachment on electrodes. A549 cells were cultured in the chambers and impedance measurements were taken at 12-h intervals for 120 h. Cell index (CI) for both designs were calculated from the impedance measurement and plotted in comparison with the growth profile of the cells in T-flasks. To verify that the ECIS biosensor can also be used to study cell response to toxicants, the A549 cells were also treated with anti-cancer drug, paclitaxel, and its responses were monitored over 5 days. Both simulation and experimental results show better sensitivity for smaller spacing between electrodes. Graphical abstract The fabricated impedance biosensor used screen-printed silver/silver chloride IDEs. Simulation and experimental results show better sensitivity for smaller between electrodes.

A novel cell-based hybrid acoustic wave biosensor with impedimetric sensing capabilities.

A novel multiparametric biosensor system based on living cells will be presented. The biosensor system includes two biosensing techniques on a single device: resonant frequency measurements and electric cell-substrate impedance sensing (ECIS). The multiparametric sensor system is based on the innovative use of the upper electrode of a quartz crystal microbalance (QCM) resonator as working electrode for the ECIS technique. The QCM acoustic wave sensor consists of a thin AT-cut quartz substrate with two gold electrodes on opposite sides. For integration of the QCM with the ECIS technique a semicircular counter electrode was fabricated near the upper electrode on the same side of the quartz crystal. Bovine aortic endothelial live cells (BAECs) were successfully cultured on this hybrid biosensor. Finite element modeling of the bulk acoustic wave resonator using COMSOL simulations was performed. Simultaneous gravimetric and impedimetric measurements performed over a period of time on the same cell culture were conducted to validate the device's sensitivity. The time necessary for the BAEC cells to attach and form a compact monolayer on the biosensor was 35~45 minutes for 1.5 × 10(4) cells/cm2 BAECs; 60 minutes for 2.0 × 10(4) cells/cm2 BAECs; 70 minutes for 3.0 × 10(4) cells/cm2 BAECs; and 100 minutes for 5.0 × 104 cells/cm2 BAECs. It was demonstrated that this time is the same for both gravimetric and impedimetric measurements. This hybrid biosensor will be employed in the future for water toxicity detection.

Acoustic wave based MEMS devices for biosensing applications.

This paper presents a review of acoustic-wave based MEMS devices that offer a promising technology platform for the development of sensitive, portable, real-time biosensors. MEMS fabrication of acoustic wave based biosensors enables device miniaturization, power consumption reduction and integration with electronic circuits. For biological applications, the biosensors are integrated in a microfluidic system and the sensing area is coated with a biospecific layer. When a bioanalyte interacts with the sensing layer, mass and viscosity variations of the biospecific layer can be detected by monitoring changes in the acoustic wave properties such as velocity, attenuation, resonant frequency and delay time. Few types of acoustic wave devices could be integrated in microfluidic systems without significant degradation of the quality factor. The acoustic wave based MEMS devices reported in the literature as biosensors and presented in this review are film bulk acoustic wave resonators (FBAR), surface acoustic waves (SAW) resonators and SAW delay lines. Different approaches to the realization of FBARs, SAW resonators and SAW delay lines for various biochemical applications are presented. Methods of integration of the acoustic wave MEMS devices in the microfluidic systems and functionalization strategies will be also discussed.