A dual-emission ratiometric fluorescence sensor (CDs@CdTe@MIP) with a self-calibration function was successfully constructed for AMO detection. In the CDs@CdTe@MIP system, non-imprinted polymer-coated CDs and molecule-imprinted polymer-coated CdTe quantum dots were used as the reference signal and response elements, respectively. The added AMO quenched the fluorescence of the CdTe quantum dots, whereas the fluorescence intensity of the CDs remained almost unchanged. The AMO concentration was monitored using the fluorescence intensity ratio (log(I647/I465)0/(I647/I465)) to reduce interference from the testing environment. The sensor with a low detection limit of 0.15 μg/L enabled detection of the AMO concentration within 6 min. The ratiometric fluorescence sensor was used to detect AMO in spiked pork samples; it exhibited a high recovery efficiency and relative standard deviation (RSD) of 97.94-103.70% and 3.77-4.37%, respectively. The proposed highly sensitive and selective platform opens avenues for sensitive, reliable, and rapid determination of pharmaceuticals in the environment and food safety monitoring using ratiometric sensors.

This work aims at the detection of the important herbicide glyphosate based on the previous modification of glyphosate in two stages and final detection by surface-enhanced Raman spectroscopy (SERS). In a first step, the affinity of glyphosate for metal plasmonic surfaces was increased by inclusion of a sulphur containing group (dithiocarbamate). In a second step, the cyclization of the latter intermediate rendered a thiazole derivative of the herbicide. The latter compound exhibits higher Raman cross section which leads to stronger SERS enhancement factors. The second step was possible thanks to the plasmon catalysis driven by metal nanoparticles, specifically silver adatoms created at the surface, and irradiated at a proper wavelength. This methodology was optimized by selecting the most appropriate experimental conditions for the chemical reactions. Density Functional Theory treatment of all the involved molecules was done in order to obtain the theoretical spectra and to identify the structural marker bands. A key goal of this work was to develop an effective system of glyphosate detection based on portable PickMolTM technology developed and patented by the SAFTRA Photonics Ltd. company to ensure an easy, quick, low cost, in-situ, and univocal detection of glyphosate in the environment.

The incidence of thyroid tumors has been increasing yearly over the past decade, making it the fourth highest tumor in women. This places various biological burdens on those affected. Currently, thyroid tumors are primarily diagnosed using percutaneous fine needle aspiration and ultrasound. However, these methods are complex, expensive, and less accurate, and they may fail to detect some thyroid nodules. As an alternative, researchers are focusing on blood-based biomarkers in addition to the traditional diagnostic methods, assisted predominantly by nanomaterials. Early identification of thyroid cancer is crucial as it is highly treatable. Various sensing systems have been developed using nanomaterial-mediated approaches to enhance the detection system. Nanomaterials are effectively applied in biosensors for surface functionalization and are conjugated with biomolecules to improve the interaction with the target analyte. This review discusses nanomaterial-assisted thyroid tumor detection, with a special focus on nanomaterial-based biosensors.

The intracellular developmental processes in plants, particularly concerning lignin polymer formation and biomass production are regulated by microRNAs (miRNAs). MiRNAs including miR397b are important for developing efficient and cost-effective biofuels. However, traditional methods of monitoring miRNA expression, like PCR, are time-consuming, require sample extraction, and lack spatial and temporal resolution, especially in real-world conditions. We present a novel approach using plasmonics nanosensing to monitor miRNA activity within living plant cells without sample extraction. Plasmonic biosensors using surface-enhanced Raman scattering (SERS) detection offer high sensitivity and precise molecular information. We used the Inverse Molecular Sentinel (iMS) biosensor on unique silver-coated gold nanorods (AuNR@Ag) with a high-aspect ratio to penetrate plant cell walls for detecting miR397b within intact living plant cells. MiR397b overexpression has shown promise in reducing lignin content. Thus, monitoring miR397b is essential for cost-effective biofuel generation. This study demonstrates the infiltration of nanorod iMS biosensors and detection of non-native miRNA 397b within plant cells for the first time. The investigation successfully demonstrates the localization of nanorod iMS biosensors through TEM and XRF-based elemental mapping for miRNA detection within plant cells of Nicotiana benthamiana. The study integrates shifted-excitation Raman difference spectroscopy (SERDS) to decrease background interference and enhance target signal extraction. In vivo SERDS testing confirms the dynamic detection of miR397b in Arabidopsis thaliana leaves after infiltration with iMS nanorods and miR397b target. This proof-of-concept study is an important stepping stone towards spatially resolved, intracellular miRNA mapping to monitor biomarkers and biological pathways for developing efficient renewable biofuel sources.

Nanosensors have become an indispensable tool in the food sector due to their specificity and sensitivity. The biosensor consists of a transducer coupled with a biorecognition component to transform biological signal into digital signal. Nanobiosensors have been widely used for sensing toxic chemicals such as pesticide residues and pathogenic microbes owing to their accurate sensitivity in an affordable manner, which gives more hope to the food industry on their applications. It employs nanocarriers to bind to impurities and pollutants, as well as food-borne microorganisms and their resulting toxins, such as mycotoxins. This modern technology ensures food safety in food processing industries. Nowadays, nanoparticle-immobilized sensors act as spot indicators to improve smart food packing technology. Certain types of nanobiosensors are deployed to monitor food product manufacture till packaging and to check the freshness of the product till spoilage identification. They are mainly using enzyme catalysts, which are highly sensitive to extreme environmental conditions. As a result, there is a greater evaluation requirement in nanosensor technology to adopt any temperature, pH, or other difficult parameters. Its stability, while in contact with food substrates, is another criterion that needs to be regularized. Within this framework, this review delves into the latest developments in nanobiosensors and the obstacles encountered during their use across different food industries.

Fluorescent nanosensors have revolutionized diagnostics and our ability to monitor cellular dynamics. Yet, distinguishing sensor signals from autofluorescence remains a challenge. Here, we merged optode-based sensing with near-infrared-emitting ZnGa2O4:Cr3+ persistent luminescence nanoparticles (PLNPs) to create nanocomposites for autofluorescence-free \"glow-in-the-dark\" sensing. Hydrophobic modification and incorporation of the persistent luminescence nanoparticles into an optode-based nanoparticle core yielded persistent luminescence nanosensors (PLNs) for five analytes (K+, Na+, Ca2+, pH, and O2) via two distinct mechanisms. We demonstrated the viability of the PLNs by quantifying K+ in fetal bovine serum, calibrating the pH PLNs in the same, and ratiometrically monitoring O2 metabolism in cultures of Saccharomyces cerevisiae, all the while overcoming their respective autofluorescence signatures. This highly modular platform allows for facile tuning of the sensing functionality, optical properties, and surface chemistry and promises high signal-to-noise ratios in complex optical environments.

Electrochemical detection of miRNA biomarkers in complex physiological samples holds great promise for accurate evaluation of tumor burden in the perioperative period, yet limited by reproducibility and bias issues. Here, nanosensors installed with hybrid probes that responsively release catalytic DNAzymes (G-quadruplexes/hemin) were developed to solve the fidelity challenge in an immobilization-free detection. miRNA targets triggered toehold-mediated strand displacement reactions on the sensor surface and resulted in amplified shedding of DNAzymes. Subsequently, the interference background was removed by Fe3O4 core-facilitated magnetic separation. Binding aptamers of the electrochemical reporter (dopamine) were tethered closely to the catalytic units for boosting H2O2-mediated oxidation through proximity catalysis. The one-to-many conversion by dual amplification from biological-chemical catalysis facilitated sufficient homogeneous sensing signals on electrodes. Thereby, the nanosensor exhibited a low detection limit (2.08 fM), and high reproducibility (relative standard deviation of 1.99%). Most importantly, smaller variations (RSD of 0.51-1.04%) of quantified miRNAs were observed for detection from cell lysates, multiplexed detection from unprocessed serum, and successful discrimination of small upregulations in lysates of tumor tissue samples. The nanosensor showed superior diagnostic performance with an area under curve (AUC) of 0.97 and 94% accuracy in classifying breast cancer patients and healthy donors. These findings demonstrated the synergy of signal amplification and interference removal in achieving high-fidelity miRNA detection for practical clinical applications.

The correlation between altered extracellular pH and various pathological conditions, including cancer, inflammation and metabolic disorders, is well known. Bulk pH measurements cannot report the extracellular pH value at the cell surface. However, there is a limited number of suitable tools for measuring the extracellular pH of cells with high spatial resolution, and none of them are commonly used in laboratories around the world. In this study, a versatile ratiometric nanosensor for the measurement of extracellular pH was developed. The nanosensor consists of biocompatible polystyrene nanoparticles loaded with the pH-inert reference dye Nile red and is surface functionalized with a pH-responsive fluorescein dye. Equipped with a targeting moiety, the nanosensor can adhere to cell membranes, allowing direct measurement of extracellular pH at the cell surface. The nanosensor exhibits a sensitive ratiometric pH response within the range of 5.5-9.0, with a calculated pKa of 7.47. This range optimally covers the extracellular pH (pHe) of most healthy cells and cells in which the pHe is abnormal, such as cancer cells. In combination with the nanosensors ability to target cell membranes, its high robustness, reversibility and its biocompatibility, the pHe nanosensor proves to be well suited for in-situ measurement of extracellular pH, even over extended time periods. This pH nanosensor has the potential to advance biomedical research by improving our understanding of cellular microenvironments, where extracellular pH plays an important role.

Total bacterial count in food is one of important food safety criteria. The current plate count method (Heterotrophic Plate Count) for food analysis requires microbiology lab facilities and at least 2 days turnover time. We developed a rapid fluorescence-based total bacterial count method that utilises semiconductor nanorods (SNRs) conjugated with a lectin Griffonia simplicifolia II (GSII-SNRs) to stain bacterial cells captured on syringe filters, via the common N-acetylglucosamine molecules on bacterial cell wall. This \"Filter-and-Stain\" detection method has a rapid turnover time of 20 min. The fluorescence emission can be seen under UV light with minimum interference from food sample background. The fluorescence intensity quantified through image analysis is proportional to the bacterial concentration with a limit of detection of 1000 CFU/mL, for total bacterial count assessment in food safety. Moreover, the GSII-SNRs do not bind to heat inactivated bacterial cells, and thus can differentiate live and dead bacteria. Our method has been validated with representative food (coffee powder, raw spinach leaves, and ready-to-eat tomato salsa) to demonstrate its high potential for on-site food safety assessment, especially in places with no immediate access to microbiology labs.

APOBEC3A (A3A) is a cytidine deaminase with critical roles in molecular diagnostics. Herein, we demonstrate the enzymatic DNA repairing amplification-powered construction of an Au nanoparticle-based nanosensor for single-molecule monitoring of A3A activity in cancer cells. Target A3A can convert cytosine (C) in substrate probe to uracil (U), and then the template binds with substrate probe to form a dsDNA containing U/A base pairs. Uracil DNA glycosylase (UDG) excises the U base to produce an apurinic/apyrimidinic (AP) site that can be cleaved by apurinic/apyrimidic endonuclease 1 (APE1) to obtain the substrate fragment with 3\'-OH end. Subsequently, the substrate fragment initiates cyclic enzymatic repairing amplification (ERA), releasing trigger-1 and trigger-2. The resultant trigger-1 can act as the primer to induce multiple cycles of cyclic ERA, producing numerous trigger-1 and trigger-2. The hybridization of trigger-2 with signal probe forms the dsDNA duplexes with an AP site, inducing the cyclic cleavage of signal probes by APE1 to release abundant Cy5 molecules from the AuNPs. Released Cy5 molecules can be easily quantified by single-molecule imaging. This nanosensor allows for specific and sensitive detection of A3A activity with a detection limit of 0.855 aM, and it can further measure kinetic parameters, screen inhibitors, and quantify endogenous A3A activity at the single-cell level, with prospect application in disease diagnostics and therapy.