Single-Walled Carbon Nanotube-Modified Gold Leaf Immunosensor for Escherichia coli Detection.

The requirement to prevent foodborne illnesses underscores the need for reliable detection tools, stimulating biosensor technology with practical solutions for in-field applications. This study introduces a low-cost immunosensor based on a single-walled carbon nanotube (SWCNT)-modified gold leaf electrode (GLE) for the sensitive detection of Escherichia coli. The immunosensor is realized with a layer-by-layer (LbL) assembly technique, creating an electrostatic bond between positively charged polyethylenimine (PEI) and negatively charged carboxyl-functionalized SWCNTs on the GLE. The structural and functional characterization of the PEI-SWCNT film was performed with Raman spectroscopy, high-resolution scanning electron microscopy (HRSEM), and electrical measurements. The PEI-SWCNT film was used as a substrate for antibody immobilization, and the electrochemical sensing potential was validated using electrochemical impedance spectroscopy (EIS). The results showed a wide dynamic range of E. coli detection, 101-108 cfu/mL, with a limit of detection (LOD) of 1.6 cfu/mL in buffer and 15 cfu/mL in the aqueous solution used for cleansing fresh lettuce leaves, affirming its efficiency as a practical and affordable tool in enhancing food safety.

Mammalian Cell-Growth Monitoring Based on an Impedimetric Sensor and Image Processing within a Microfluidic Platform.

In recent years, advancements in microfluidic and sensor technologies have led to the development of new methods for monitoring cell growth both in macro- and micro-systems. In this paper, a microfluidic (MF) platform with a microbioreactor and integrated impedimetric sensor is proposed for cell growth monitoring during the cell cultivation process in a scaled-down simulator. The impedimetric sensor with an interdigitated electrode (IDE) design was realized with inkjet printing and integrated into the custom-made MF platform, i.e., the scaled-down simulator. The proposed method, which was integrated into a simple and rapid fabrication MF system, presents an excellent candidate for the scaled-down analyses of cell growths that can be of use in, e.g., optimization of the cultivated meat bioprocess. When applied to MRC-5 cells as a model of adherent mammalian cells, the proposed sensor was able to precisely detect all phases of cell growth (the lag, exponential, stationary, and dying phases) during a 96-h cultivation period with limited available nutrients. By combining the impedimetric approach with image processing, the platform enables the real-time monitoring of biomasses and advanced control of cell growth progress in microbioreactors and scaled-down simulator systems.

Low-cost goldleaf electrode as a platform for Escherichia coli immunodetection.

Gold electrodes are one of most prevalent substrates in electrochemical biosensors because they can be easily and highly efficiently functionalized with thiolated biomolecules. However, conventional methods to fabricate gold electrodes are costly, time-consuming and require onerous equipment. Here, an affordable method for rapid fabrication of an electrochemical immunosensor for Escherichia coli detection is presented. The gold electrode was generated using 24-karat gold leaves and lowcost polyvinyl chloride adhesive sheets covered with an insulating PTFE layer. The goldleaf electrode (GLE) was patterned using laser ablation and characterized by cyclic voltammetry, electrochemical impedance spectroscopy, scanning electronic microscopy, contact angle and 3D profiling. The GLEs were modified by a self-assembled mercaptopropionic monolayer, followed by surface activation to allow binding of the specific anti-E. coli antibody via carbodiimide linking. The biosensor showed a detection limit of 2 CFU/mL and a linear dynamic range of 10-107 CFU/mL for E. coli cells. No false positive signals were obtained from control bacteria. The obtained results demonstrated suitability of GLE for use in biosensors with high reliability and reproducibility. It is foreseeable that our work will inspire design of point-of-need biosensors broadly applicable in low-resource settings.

3D-Printed Microfluidic Chip for Real-Time Glucose Monitoring in Liquid Analytes.

The connection of macrosystems with microsystems for in-line measurements is important in different biotechnological processes as it enables precise and accurate monitoring of process parameters at a small scale, which can provide valuable insights into the process, and ultimately lead to improved process control and optimization. Additionally, it allows continuous monitoring without the need for manual sampling and analysis, leading to more efficient and cost-effective production. In this paper, a 3D printed microfluidic (MF) chip for glucose (Glc) sensing in a liquid analyte is proposed. The chip made in Poly(methyl methacrylate) (PMMA) contains integrated serpentine-based micromixers realized via stereolithography with a slot for USB-like integration of commercial DropSens electrodes. After adjusting the sample's pH in the first micromixer, small volumes of the sample and enzyme are mixed in the second micromixer and lead to a sensing chamber where the Glc concentration is measured via chronoamperometry. The sensing potential was examined for Glc concentrations in acetate buffer in the range of 0.1-100 mg/mL and afterward tested for Glc sensing in a cell culturing medium. The proposed chip showed great potential for connection with macrosystems, such as bioreactors, for direct in-line monitoring of a quality parameter in a liquid sample.

Advances in Nanomaterials-Based Electrochemical Biosensors for Foodborne Pathogen Detection.

Electrochemical biosensors utilizing nanomaterials have received widespread attention in pathogen detection and monitoring. Here, the potential of different nanomaterials and electrochemical technologies is reviewed for the development of novel diagnostic devices for the detection of foodborne pathogens and their biomarkers. The overview covers basic electrochemical methods and means for electrode functionalization, utilization of nanomaterials that include quantum dots, gold, silver and magnetic nanoparticles, carbon nanomaterials (carbon and graphene quantum dots, carbon nanotubes, graphene and reduced graphene oxide, graphene nanoplatelets, laser-induced graphene), metal oxides (nanoparticles, 2D and 3D nanostructures) and other 2D nanomaterials. Moreover, the current and future landscape of synergic effects of nanocomposites combining different nanomaterials is provided to illustrate how the limitations of traditional technologies can be overcome to design rapid, ultrasensitive, specific and affordable biosensors.

Microwave Spoof Surface Plasmon Polariton-Based Sensor for Ultrasensitive Detection of Liquid Analyte Dielectric Constant.

In this paper, a microwave microfluidic sensor based on spoof surface plasmon polaritons (SSPPs) was proposed for ultrasensitive detection of dielectric constant. A novel unit cell for the SSPP structure is proposed and its behaviour and sensing potential analysed in detail. Based on the proposed cell, the SSPP microwave structure with a microfluidic reservoir is designed as a multilayer configuration to serve as a sensing platform for liquid analytes. The sensor is realized using a combination of rapid, cost-effective technologies of xurography, laser micromachining, and cold lamination bonding, and its potential is validated in the experiments with edible oil samples. The results demonstrate high sensitivity (850 MHz/epsilon unit) and excellent linearity (R2 = 0.9802) of the sensor, which, together with its low-cost and simple fabrication, make the proposed sensor an excellent candidate for the detection of small changes in the dielectric constant of edible oils and other liquid analytes.

Facile Monitoring of Water Hardness Levels Using Responsive Complex Emulsions.

The cationic content of water represents a major quality control parameter that needs to be followed by a rapid, on-site, and low-cost method. Herein, we report a novel method for a facile monitoring of the mineral content of drinking water by making use of responsive complex emulsions. The morphology of biphasic oil-in-water droplets solely depends on the balance of interfacial tensions, and we demonstrate that changes in the surfactant effectiveness, caused by variations in the mineral content inside the continuous phase, can be visualized by monitoring internal droplet shapes. An addition of metal cations can significantly influence the surfactant critical micelle concentrations and the surface excess values and therefore induce changes in the effectiveness of ionic surfactants, such as sodium dodecyl sulfate. The morphological response of Janus emulsions droplets was tracked via a simple microscopic setup. We observed that the extent of the droplet response was dependent on the salt concentration and valency, with divalent cations (responsive for water hardness), resulting in a more pronounced response. In this way, Ca2+ and Mg2+ levels could be quantitatively measured, which we showcased by determination of the mineral content of commercial water samples. The herein demonstrated device concept may provide a new alternative rapid monitoring of water hardness levels in a simple and cost-effective setup.

Cultivating Multidisciplinarity: Manufacturing and Sensing Challenges in Cultured Meat Production.

Meat cultivation via cellular agriculture holds great promise as a method for future food production. In theory, it is an ideal way of meat production, humane to the animals and sustainable for the environment, while keeping the same taste and nutritional values as traditional meat and having additional benefits such as controlled fat content and absence of antibiotics and hormones used in the traditional meat industry. However, in practice, there is still a number of challenges, such as those associated with the upscale of cultured meat (CM). CM food safety monitoring is a necessary factor when envisioning both the regulatory compliance and consumer acceptance. To achieve this, a multidisciplinary approach is necessary. This includes extensive development of the sensitive and specific analytical devices i.e., sensors to enable reliable food safety monitoring throughout the whole future food supply chain. In addition, advanced monitoring options can help in the further optimization of the meat cultivation which may reduce the currently still high costs of production. This review presents an overview of the sensor monitoring options for the most relevant parameters of importance for meat cultivation. Examples of the various types of sensors that can potentially be used in CM production are provided and the options for their integration into bioreactors, as well as suggestions on further improvements and more advanced integration approaches. In favor of the multidisciplinary approach, we also include an overview of the bioreactor types, scaffolding options as well as imaging techniques relevant for CM research. Furthermore, we briefly present the current status of the CM research and related regulation, societal aspects and challenges to its upscaling and commercialization.

Potential of urban trees for mitigating heavy metal pollution in the city of Novi Sad, Serbia.

The current study analyzed heavy metal (HM) concentrations in the urban soil and tree leaves in the city of Novi Sad, Serbia, with the aim to identify specific tree species with good HM bioaccumulative abilities that can mitigate HM pollution in urban areas. Zinc (Zn), copper (Cu) and lead (Pb) were selected for the analysis of tree leaves and soil at 16 locations close to urban roads, main boulevards and high-traffic streets. Leaf material of four most common tree species: Platanus × acerifolia (Aiton) Willd., Celtis occidentalis L., Tilia argentea L. and Quercus robur L. and soil samples were analyzed with the aim to identify the potential hotspots and sources of pollution. Analysis of variance (ANOVA) and t test was applied to the available data sets to find differences between the HM concentrations in the analyzed trees and soil samples. Geographic information system (GIS) mapping was used for visualization of the concentration distributions and illustration of the HM contaminated hotspots. Bioconcentration factors (BCF) were calculated and they showed that the plants of the species Celtis occidentalis L. have the ability to retain Cu, the species Quercus robur L. retains mostly Zn, Tilia argentea L. accumulates Pb, while Platanus x acerifolia (Aiton) Willd. retains the smallest amount of the tested HMs.

Novel Cost-Effective Microfluidic Chip Based on Hybrid Fabrication and Its Comprehensive Characterization.

Microfluidics, one of the most attractive and fastest developed areas of modern science and technology, has found a number of applications in medicine, biology and chemistry. To address advanced designing challenges of the microfluidic devices, the research is mainly focused on development of efficient, low-cost and rapid fabrication technology with the wide range of applications. For the first time, this paper presents fabrication of microfluidic chips using hybrid fabrication technology-a grouping of the PVC (polyvinyl chloride) foils and the LTCC (Low Temperature Co-fired Ceramics) Ceram Tape using a combination of a cost-effective xurography technique and a laser micromachining process. Optical and dielectric properties were determined for the fabricated microfluidic chips. A mechanical characterization of the Ceram Tape, as a middle layer in its non-baked condition, has been performed and Young's modulus and hardness were determined. The obtained results confirm a good potential of the proposed technology for rapid fabrication of low-cost microfluidic chips with high reliability and reproducibility. The conducted microfluidic tests demonstrated that presented microfluidic chips can resist 3000 times higher flow rates than the chips manufactured using standard xurography technique.

Point-of-Need DNA Testing for Detection of Foodborne Pathogenic Bacteria.

Foodborne pathogenic bacteria present a crucial food safety issue. Conventional diagnostic methods are time-consuming and can be only performed on previously produced food. The advancing field of point-of-need diagnostic devices integrating molecular methods, biosensors, microfluidics, and nanomaterials offers new avenues for swift, low-cost detection of pathogens with high sensitivity and specificity. These analyses and screening of food items can be performed during all phases of production. This review presents major developments achieved in recent years in point-of-need diagnostics in land-based sector and sheds light on current challenges in achieving wider acceptance of portable devices in the food industry. Particular emphasis is placed on methods for testing nucleic acids, protocols for portable nucleic acid extraction and amplification, as well as on the means for low-cost detection and read-out signal amplification.

A Microwave Microfluidic Sensor Based on a Dual-Mode Resonator for Dual-Sensing Applications.

In this paper, we propose a novel microwave microfluidic sensor with dual-sensing capability. The sensor is based on a dual-mode resonator that consists of a folded microstrip line loaded with interdigital lines and a stub at the plane of symmetry. Due to the specific configuration, the resonator exhibits two entirely independent resonant modes, which allows simultaneous sensing of two fluids using a resonance shift method. The sensor is designed in a multilayer configuration with the proposed resonator and two separated microfluidic channels-one intertwined with the interdigital lines and the other positioned below the stub. The circuit has been fabricated using low-temperature co-fired ceramics technology and its performance was verified through the measurement of its responses for different fluids in the microfluidic channels. The results confirm the dual-sensing capability with zero mutual influence as well as good overall performance. Besides an excellent potential for dual-sensing applications, the proposed sensor is a good candidate for application in mixing fluids and cell counting.

Microfluidic EBG Sensor Based on Phase-Shift Method Realized Using 3D Printing Technology.

In this article, we propose a novel microfluidic microstrip electromagnetic band gap (EBG) sensor realized using cost-effective 3D printing technology. Microstrip sensor allows monitoring of the fluid properties flowing in the microchannel embedded between the microstrip line and ground plane. The sensor's operating principle is based on the phase-shift method, which allows the characterization at a single operating frequency of 6 GHz. The defected electromagnetic band gap (EBG) structure is realized as a pattern in the microstrip ground plane to improve sensor sensitivity. The designed microfluidic channel is fabricated using a fused deposition modelling (FDM) 3D printing process without additional supporting layers, while the conductive layers are realized using sticky aluminium tape. The measurement results show that the change of permittivity of the fluid in the microfluidic channel from 1 to 80 results in the phase-shift difference of almost 90°. The potential application is demonstrated through the implementation of a proposed sensor for the detection of toluene concentration in toluene-methanol mixture where various concentrations of toluene were analysed.