Phosphate Contaminant Detection in Water Through a Paper-based Microfluidic Device

This report describes a project aimed at developing a low-cost, portable, on-site, user-friendly system for detecting different concentrations of phosphate in drinking water. Phosphate is a natural chemical, but toxic in large concentrations; detection is therefore important to avoid drinking contaminated water. Despite this fact, no cheap, and/or nontoxic system for phosphate detection is yet on the market. The detection system utilizes a paper-based microfluidic device to automate the electrochemical detection process, which normally requires expert use of lab equipment. When combined with a portable potentiostat that works with a mobile app, the device will allow untrained users to determine if any source of drinking water contains unsafe levels of phosphate without equipment or training, and to communicate that information to a central database for further analysis. Those of any background, particularly in developing countries, will be able to maintain health and raise awareness about clean water. Microfluidic devices are useful tools for the detection of water contaminants, but there is a gap in technology for the detection of phosphate. Our phosphate detection system is a paper-based microfluidic device with an already-developed voltammetry device that automates the detection process so that any user can safely find phosphate in water. The system will provide a binary analysis about whether the water is safe to consume or not. Completion of the project provides a valuable tool to both average customers in developing countries and scientific researchers in determining the safety of drinking water

Antimicrobial peptides: Powerful biorecognition elements to detect bacteria in biosensing technologies

Bacterial infections represent a serious threat in modern medicine. In particular, biofilm treatment in clinical settings is challenging, as biofilms are very resistant to conventional antibiotic therapy and may spread infecting other tissues. To address this problem, biosensing technologies are emerging as a powerful solution to detect and identify bacterial pathogens at the very early stages of the infection, thus allowing rapid and effective treatments before biofilms are formed. Biosensors typically consist of two main parts, a biorecognition moiety that interacts with the target (i.e., bacteria) and a platform that transduces such interaction into a measurable signal. This review will focus on the development of impedimetric biosensors using antimicrobial peptides (AMPs) as biorecognition elements. AMPs belong to the innate immune system of living organisms and are very effective in interacting with bacterial membranes. They offer unique advantages compared to other classical bioreceptor molecules such as enzymes or antibodies. Moreover, impedance-based sensors allow the development of label-free, rapid, sensitive, specific and cost-effective sensing platforms. In summary, AMPs and impedimetric transducers combine excellent properties to produce robust biosensors for the early detection of bacterial infectionsPeer ReviewedPostprint (published version

Electrochemical biosensor based on microfabricated electrode arrays for life sciences applications

In developing a biosensor, the utmost important aspects that need to be emphasized are the specificity and selectivity of the transducer. These two vital prerequisites are of paramount in ensuring a robust and reliable biosensor. Improvements in electrochemical sensors can be achieved by using microelectrodes and to modify the electrode surface (using chemical or biological recognition layers to improve the sensitivity and selectivity). The fabrication and characterisations of silicon-based and glass-based gold microelectrode arrays with various geometries (band and disc) and dimension (ranging from 10 μm-100 nm) were reported. It was found that silicon-based transducers of 10 μm gold microelectrode array exhibited the most stable and reproducible electrochemical measurements hence this dimension was selected for further study. Chemical electrodeposition on both 10 μm microband and microdisc were found viable by electro-assisted self-assembled sol-gel silica film and nanoporous-gold electrodeposition respectively. The fabrication and characterisations of on-chip electrochemical cell was also reported with a fixed diameter/width dimension and interspacing variation. With this regard, the 10 μm microelectrode array with interspacing distance of 100 μm exhibited the best electrochemical response. Surface functionalisations on single chip of planar gold macroelectrodes were also studied for the immobilisation of histidine-tagged protein and antibody. Imaging techniques such as atomic force microscopy, fluorescent microscopy or scanning electron microscope were employed to complement the electrochemical characterisations. The long-chain thiol of self-assembled monolayer with NTA-metal ligand coordination was selected for the histidine-tagged protein while silanisation technique was selected for the antibody immobilisation. The final part of the thesis described the development of a T-2 labelless immunosensor using impedimetric approach. Good antibody calibration curve was obtained for both 10 μm microband and 10 μm microdisc array. For the establishment of the T-2/HT-2 toxin calibration curve, it was found that larger microdisc array dimension was required to produce better calibration curve. The calibration curves established in buffer solution show that the microelectrode arrays were sensitive and able to detect levels of T-2/HT-2 toxin as low as 25 ppb (25 μg kg-1) with a limit of quantitation of 4.89 ppb for a 10 μm microband array and 1.53 ppb for the 40 μm microdisc array

Development of on-farm diagnostic devices

The global population, currently 7.7 billion, is expected to grow to 9.7 billion by 2050. This is expected to lead to a 70% increase in demand for animal-based protein. Irish beef and dairy products account for over 50% of our agricultural output and DAFM’s Food Wise 2025 strategy aims to position Ireland as a world leader in sustainable agri-food production. However, the high percentage of livestock that are lost due to infectious diseases (20%), poses a challenge to achieving this sustainability, in addition to more sustainable use of antimicrobials, smarter livestock diagnostics and treatments are therefore required. The goal of this thesis was to develop a low-cost disposable biosensor that would permit point-of-care (POC) detection of diseases in bovines, through cost-effective, scalable microfabrication techniques. Such devices could enable real-time determination of the health status of animals on farm and contribute to more informed therapeutic interventions. Electrochemistry presents a viable option for POC devices in this regard and allows easy integration with portable electronics. Electrochemical Impedance Spectroscopy (EIS) is a surface sensitive technique that measures the resistive and capacitive behaviour of an electrochemical system. It lends itself to serological immunosensor development as it allows label-free detection. For the purposes of this research, silicon devices were fabricated with six microband working electrodes, gold counter, and platinum pseudo-reference electrodes. The microband working electrodes were modified with a biocompatible co-polymer. This co-polymer supported the cross-linking of a bioreceptor (e.g., anti-bovine IgG) to electrode surface, which selectively bound to the target biomolecule (bovine IgG) in serum. This EIS device could distinguish between seronegative and seropositive samples in 15 minutes making it suitable for POC applications. Additionally, the presence of six working electrodes allowed for testing of multiple samples at a time. Often, however, only a single test is required. As such, silicon presents an expensive option for disposable sensors. Hence, polymer replication methods were also investigated in this thesis. This process allowed a single silicon wafer to be repeatedly used to produce polymer structures. A microneedle format was chosen to eliminate the need for taking samples on-farm and provide a pain-free method of in vivo measurements in interstitial fluid in interstitial fluid. The fabrication method used a double-sided micro-moulding process to move towards mass manufacturing. COMSOL simulations were performed to explore the active layer on the microneedle tip surface, ensuring no diffusional overlap between electrodes and providing the most effective tip design. The microneedle structures also presented the opportunity for novel fabrication of nanoring arrays, by removing part of the protruding structure and exposing underlying nanorings. These have the potential to be highly sensitive electrochemical devices due to enhanced mass transport and high current densities, while maintaining the scalable cost-effective fabrication process of the microneedles. Devices produced steady-state CVs in a known redox molecule, with currents in the nA range

Analysis of relevant technical issues and deficiencies of the existing sensors and related initiatives currently set and working in marine environment. New generation technologies for cost-effective sensors

The last decade has seen significant growth in the field of sensor networks, which are currently collecting large amounts of environmental data. This data needs to be collected, processed, stored and made available for analysis and interpretation in a manner which is meaningful and accessible to end users and stakeholders with a range of requirements, including government agencies, environmental agencies, the research community, industry users and the public. The COMMONSENSE project aims to develop and provide cost-effective, multi-functional innovative sensors to perform reliable in-situ measurements in the marine environment. The sensors will be easily usable across several platforms, and will focus on key parameters including eutrophication, heavy metal contaminants, marine litter (microplastics) and underwater noise descriptors of the MSFD. The aims of Tasks 2.1 and 2.2 which comprise the work of this deliverable are: • To obtain a comprehensive understanding and an up-to-date state of the art of existing sensors. • To provide a working basis on “new generation” technologies in order to develop cost-effective sensors suitable for large-scale production. This deliverable will consist of an analysis of state-of-the-art solutions for the different sensors and data platforms related with COMMONSENSE project. An analysis of relevant technical issues and deficiencies of existing sensors and related initiatives currently set and working in marine environment will be performed. Existing solutions will be studied to determine the main limitations to be considered during novel sensor developments in further WP’s. Objectives & Rationale The objectives of deliverable 2.1 are: • To create a solid and robust basis for finding cheaper and innovative ways of gathering data. This is preparatory for the activities in other WPs: for WP4 (Transversal Sensor development and Sensor Integration), for WP(5-8) (Novel Sensors) to develop cost-effective sensors suitable for large-scale production, reducing costs of data collection (compared to commercially available sensors), increasing data access availability for WP9 (Field testing) when the deployment of new sensors will be drawn and then realized

STRATEGIES TO IMPROVE ELECTROCHEMICAL DETECTION OF NITRIC OXIDE IN BIOLOGICAL ENVIRONMENTS

Nitric oxide (NO) is a gaseous molecule of vast biological significance whose activity is likely to be concentration-dependent. As our understanding of this molecule becomes more nuanced and precise, so too must detection strategies evolve to detect NO with greater precision and accuracy. Spectroscopic techniques are able to measure NO with high specificity, but the only technique unhindered by complex instrumentation and the requirement for additional reagents, and able to measure NO directly in situ, is electrochemistry. However, bare electrodes are unable measure NO with sufficient selectivity and sensitivity, particularly in biological environments, necessitating the use of transducer surface modifiers to improve performance. Herein, systematic evaluations and comparisons of electrochemical NO sensor modifications were carried out. Electropolymerized films (EPFs) represent a class of selectivity-enhancing membranes favorable for their reproducible and self-terminating depositions. Six different monomers were evaluated for their permselectivity characteristics for NO against a panel of electroactive biological interferents. After tailored optimizations of their deposition parameters, polymers were also evaluated for their anti-fouling properties in simulated wound fluid. In addition to EPFs, another common strategy to improve NO sensor performance is the incorporation of metallophthalocyanine (MPc) electrocatalysts. Four MPc macrocycles (M = iron, cobalt, nickel, and zinc) previously determined to have the highest electrocatalytic activity towards NO oxidation were evaluated for their selectivity characteristics. The ability to specifically coordinate with NO at the metal center (as opposed to weak physisorption on the aromatic periphery) proved an adequate predictor of selectivity findings. Based on these evaluations of different modifiers, a solid-state electrochemical NO sensor was designed for long-term use in proteinaceous media. With extensive characterizations of sensocompatibility, the final NO sensor was capable of high sensitivity and selectivity retention with continuous operation in culture media. The sensor was then used to successfully interrogate the temporal (> 24 h) and spatial concentration profiles of macrophage NO release under neutral and pro-inflammatory stimulated conditions. Lastly, hydrogen sulfide (H2S) is another gasotransmitter responsible for mediating many of the same biological processes as NO, and may either upregulate or inhibit NO production in a manner likely to be concentration-dependent. An EPF-modified, solid-state electrode was therefore developed for selective detection of H2S for subsequent incorporation into a NO/H2S dual-sensor.Doctor of Philosoph

Sensing at nanostructures for agri-food and enviromental applications

With a predicted population increase of 2.3 billion people, by 2050, agricultural productivity must be vastly improved and made sustainable. Globally, agriculture must deliver a 60% increase in food production to cope with the population demand. Moreover, this needs to be achieved against a changing climate, an exploitation of natural resources, and growing water and land scarcities. New digital technologies can optimise production efficiency and ensure food security and safety while also minimising waste within the production systems and the supply chain. To this end, new sensor technologies are being developed for applications in animal health diagnostics and environmental issues related to the global population, such as food & crop protection, pathogen and toxin detection, and environmental remediation. In this thesis, two new nanosensing diagnostic devices are developed and presented; surface enhanced Raman sensing and electrochemical sensing. Surface-enhanced Raman spectroscopy (SERS) substrates were fabricated by templating a flexible thermoplastic polymer against an aluminium drinks can followed by coating with a silver film, to produce a rough nanostructured metallic surface. SERS is used for both qualitative (molecular fingerprint) and quantitative detection of dye molecules and food toxins. In addition, the SERS technique is also applied in combination with nanoelectrochemical square wave voltammetry to detect nano-concentrations of neonicotinoid pesticides. The enhanced sensitivity and minimum sample preparation requirements provide tremendous opportunities for food safety and security sectors. An impedimetric immunosensor device (with a micro SD style pin-out) was also developed for the serological diagnosis of viruses and antibodies associated with bovine respiratory disease and bovine liver fluke. The silicon chip devices consist of six on-chip nanoband electrodes which can be independently modified with a polymer layer for covalent immobilisation of capture and target biomolecules. This electrochemical biosensor technology provides label-free and cost-efficient sensing capability in a compact size, and demonstrates the potential development of immunoassay-based point-of-use devices for on-farm diagnosis or therapeutic monitoring in animal health applications

An Electrochemical Microsensor Based on a AuNPs-Modified Microband Array Electrode for Phosphate Determination in Fresh Water Samples

This work describes the fabrication, characterization, and application of a gold microband array electrode (MAE) for the determination of phosphate in fresh water samples. The working principle of this MAE is based on the reduction of a molybdophosphate complex using the linear sweep voltammetric (LSV) method. The calibration of this microsensor was performed with standard phosphate solutions prepared with KH2PO4 and pH adjusted to 1.0. The microsensor consists of a platinum counter electrode, a gold MAE as working electrode, and an Ag/AgCl electrode as reference electrode. The microelectrode chips were fabricated by the Micro Electro-Mechanical System (MEMS) technique. To improve the sensitivity, gold nanoparticles (AuNPs) were electrodeposited on the working electrode. With a linear range from 0.02 to 0.50 mg P/L, the sensitivity of the unmodified microsensor is 2.40 µA per (mg P/L) (R2 = 0.99) and that of the AuNPs-modified microsensor is 7.66 µA per (mg P/L) (R2 = 0.99). The experimental results showed that AuNPs-modified microelectrode had better sensitivity and a larger current response than the unmodified microelectrode

An Electrochemical Microsensor Based on a AuNPs-Modified Microband Array Electrode for Phosphate Determination in Fresh Water Samples

This work describes the fabrication, characterization, and application of a gold microband array electrode (MAE) for the determination of phosphate in fresh water samples. The working principle of this MAE is based on the reduction of a molybdophosphate complex using the linear sweep voltammetric (LSV) method. The calibration of this microsensor was performed with standard phosphate solutions prepared with KH2PO4 and pH adjusted to 1.0. The microsensor consists of a platinum counter electrode, a gold MAE as working electrode, and an Ag/AgCl electrode as reference electrode. The microelectrode chips were fabricated by the Micro Electro-Mechanical System (MEMS) technique. To improve the sensitivity, gold nanoparticles (AuNPs) were electrodeposited on the working electrode. With a linear range from 0.02 to 0.50 mg P/L, the sensitivity of the unmodified microsensor is 2.40 µA per (mg P/L) (R2 = 0.99) and that of the AuNPs-modified microsensor is 7.66 µA per (mg P/L) (R2 = 0.99). The experimental results showed that AuNPs-modified microelectrode had better sensitivity and a larger current response than the unmodified microelectrode