Fluorescence based metabisulfite sensing: New aspects of ion sensing by a styryl benzothiazolium dye and understanding nitrite interference.

Detection of specific ions using fluorescent probes has relevance in several areas of therapeutics development and environmental science. Here, we provide new perspectives to the sensing of a styryl benzothiazolium-based fluorescent compound 1 and report that sensing properties are for sulfite ions in general with highest preference for metabisulfite ions (S2O52-) adding to its previously determined role as a bisulfite ion sensor. This probe exhibits its sensing action via an addition reaction in which the styryl double bond gets reduced. The interference studies highlighted that the sequence of addition of nitrite and metabisulfite has a bearing on the overall interference outcome. Spectroscopic studies revealed that the order of preferential sensing of sulfites and sulfide ion is S2O52- > HSO3- > SO32- > S2-. Although this probe displays robust sensing on its own through fluorescence quenching, its fluorescence emission can be enhanced at much lower concentrations in the presence of a G-quadruplex DNA without compromising the outcome of the sensing.

Enhanced enzymatic electrochemical detection of an organophosphate Pesticide: Achieving Wide linearity and femtomolar detection via gold nanoparticles growth within polypyrrole films.

The indiscriminate use of pesticides in agriculture demands the development of devices capable of monitoring contaminations in food supplies, in the environment and biological fluids. Simplicity, easy handling, high sensitivities, and low limits-of-detection (LOD) and quantification are some of the required properties for these devices. In this work, we evaluated the effect of incorporating gold nanoparticles into indigo carmine-doped polypyrrole during the electropolymerization of films for use as an acetylcholinesterase (AChE) enzyme-based biosensor. As proof of concept, the pesticide methyl parathion was tested towards the inhibition of AChE. The enzyme was immobilized simply by drop-casting a solution, eliminating the need for any prior surface modification. The biosensors were characterized with cyclic voltammetry, scanning electron microscopy, transmission electron microscopy, and Raman spectroscopy. The assays for the detection of methyl parathion with films containing polypyrrole, indigo carmine and AChE (PPy-IC-AChE) presented a sensitivity of 5.7 µA cm-2 g-1 mL and a LOD of 12 nmol L-1 (3.0 ng L-1) with a linear range from 1.3 x 10-7 mol L-1 to 1.0 x 10-5 mol L-1. The introduction of gold nanoparticles (AuNP) into the film (PPy-IC-AuNP-AChE) led to remarkable improvements on the overall performance, such as a lower redox potential for the enzymatic reaction, a 145 % increase in sensitivity (14 µA cm-2 g-1 mL), a wider detection dynamic range (from 1.3x10-7 to 1.0x10-3 mol L-1), and a very low LOD of 24 fmol L-1 (64 ag mL-1). These findings underscore the potential of using AuNPs to improve the enzymatic performance of biosensor devices.

Helicoid Grating-Coupled Surface Plasmon Resonance Sensor.

Ultrasensitive, rapid, and reliable biomolecular sensing is essential for biomedical diagnostics, requiring real-time monitoring and detection of trace samples. Optical sensing, particularly plasmonic biosensing, meets these demands through noninvasive, high-sensitivity detection based on the interaction between light and molecules. Here, we present novel plasmonic metamaterial-based sensing strategy, utilizing the circular dichroism (CD) response of grating-coupled surface plasmon resonance (SPR) from chiral nanoparticle grating structure (i.e., 2D helicoid crystal) on gold substrate. Strong chiroptic response of helicoids has been effectively expanded to produce a remarkable CD/greflection response in the SPR mode, achieved by spectral coupling of SPR with localized surface plasmon resonance (LSPR) in helicoids. This CD response, derived from the differential of left and right circularly polarized light, corrects optical fluctuations, enhancing sensitivity and reliability. Our SPR-CD-based approach achieves a sensitivity of 379.2 nm/RIU and detection limit of a few mM for d-glucose, offering a new paradigm for high-performance optical biosensors.

Advances in machine learning-enhanced nanozymes.

Nanozymes, synthetic nanomaterials that mimic the catalytic functions of natural enzymes, have emerged as transformative technologies for biosensing, diagnostics, and environmental monitoring. Since their introduction, nanozymes have rapidly evolved with significant advancements in their design and applications, particularly through the integration of machine learning (ML). Machine learning (ML) has optimized nanozyme efficiency by predicting ideal size, shape, and surface chemistry, reducing experimental time and resources. This review explores the rapid advancements in nanozyme technology, highlighting the role of ML in improving performance across various bioapplications, including real-time monitoring and the development of chemiluminescent, electrochemical and colorimetric sensors. We discuss the evolution of different types of nanozymes, their catalytic mechanisms, and the impact of ML on their property optimization. Furthermore, this review addresses challenges related to data quality, scalability, and standardization, while highlighting future directions for ML-driven nanozyme development. By examining recent innovations, this review highlights the potential of combining nanozymes with ML to drive the development of next-generation diagnostic and detection technologies.

Neurodevelopmental Process Monitoring of Cytosine Arabinoside-Exposed Neurons Using Raman Spectroscopy.

Raman spectroscopy is used to monitor the development of live neurons exposed to cytosine arabinoside (ara-C). Ara-C is widely used to culture neurons and exclude non-neuronal cells. In this study, Raman spectra obtained from neurons exposed to ara-C were plotted using an analytical model of neuronal development to evaluate the impact of ara-C on neuronal development. After two days of culturing, neurons were exposed to ara-C for 24 h at final concentrations of 0 (control), 5, and 25 µM. Principal component analysis (PCA) was performed to build an analytical model for evaluating neurodevelopmental disorders caused by ara-C treatment. We projected the Raman spectra obtained from ara-C-treated cells onto the control group dataset. The distribution of PC1 scores for neurons exposed to ara-C at a final concentration of 5 µM was not significantly different from that of the control group. In contrast, under a final concentration of 25 µM, the data population at 10 and 15 days of culturing overlapped significantly with that of neurons at 4 days of normal culturing. These results suggest that Raman spectroscopy can detect very small physiological alterations in the neurons even after a short-term exposure (24 h) of ara-C. Our analytical method has high potential to evaluate the developmental stages for living neurons under exposure to chemicals.

Analyte-induced hindrance in the RCA-assisted CRISPR/Cas12a system for homogeneous protein assays.

Heterogeneous assays, such as enzyme-linked immunosorbent assays, have become indispensable for in vitro diagnostics. However, the simple, sensitive, and accurate detection is limited by their multiple washing and incubation steps, and limited amplification methods. In this study, we design a novel approach utilizing analyte-induced hindrance within the rolling circle amplification (RCA)-assisted CRISPR/Cas12a system for simple and highly sensitive homogenous protein detection. Streptavidin (SA) and digoxin antibody (anti-Dig) are employed as representative detection models. The specific recognition of target proteins using primers modified with small molecules hinders the RCA process, preventing the activation of Cas12a's trans-cleavage activity, thereby leading to a reduction in fluorescence intensity. Our developed platform exhibites exceptional detection performance characterized by high sensitivity, robust specificity, and significant potential for application in complex samples. By expanding the recognition elements, this platform can evolve into a versatile clinical diagnostic tool with universal applicability. In addition, this platform provides a novel direction for quantifying ultralow-concentration disease biomarkers in clinical practice.

Fast tailoring the ZIF-8 surface microenvironment at ambient temperature to boost glucose oxidase-like activity of AuNPs for biosensing.

Rational design and tailoring of the surface microenvironment surrounding the catalytic sites, such as noble metal nanoparticles, is an effective way to enhance the catalytic activity of mimicking enzymes. However, it remains on-going challenges to regulate the microenvironment of the catalytic sites due to the lack of tunable variability in structural precision of conventional solid catalysts. Herein, three types of zeolitic imidazolate framework-8 (ZIF-8) with different major crystal facet orientations, i.e., cubic with (100) facets (denoted ZIF-8c), truncated dodecahedral with (100), (110) facets (denoted ZIF-8tr), and dodecahedral with (110) facets (denoted ZIF-8r), were developed facilely using an electrochemical method by switching the potential at ambient temperature. Because the Zn2+ nodes were predominantly exposed on the (100) facets of ZIF-8, while the ligands were mainly exposed on the (110) facets. Hence, gold nanoparticles (AuNPs) showed differential glucose oxidase (GOx)-like activities when anchored in situ on different crystal facets of ZIF-8 and obeyed the following order ZIF-8c/Au>ZIF-8tr/Au>ZIF-8r/Au. Notably, both the metal nodes and aromatic linkers of ZIF-8 interacted with AuNPs through coordination and π-π interactions. The Zn2+ nodes facilitated the formation of the electron-deficient Au species. The electron transfer from AuNPs to Zn2+ sites effectively boosted the catalytic activity. It was known that directly tailoring the microenvironment at the supporting sites of noble metal catalysts to boost catalysis through a facile electrochemical method was not reported. Based on the favorable GOx-like activity and long-term stability of ZIF-8tr/Au, a highly sensitive electrochemical biosensing platform for assaying squamous cell carcinoma antigen (SCCA) was developed. It enabled fg-level detection of cancer marker.

Ultrasensitive on-site colorimetric detection of Campylobacter in oyster with a portable biosensing platform based on hydroxamate/Fe3+-violurate chromogenic reaction and smartphone.

The contamination of Campylobacter in shellfish poses a health risk for its pathogenicity associated with campylobacteriosis. However, an efficient method to detect this risk is unavailable. Herein, we introduce a portable colorimetric biosensing platform that comprises three modules: an enrichment module 1, a binding and transduction module 2, and a smartphone-based module 3. Module 1 is an aptamer-modified 96-well plate for the specific capture of Campylobacter in a simple and high-throughput manner. Module 2 features a bifunctional biopolymer of L-glutamic acid γ-hydroxamate-alginate-Fe3+ coordinating fusarinine C, which can bind the captured Campylobacter cells and transduce them into amplified color signals upon reaction with Fe3+-violurate complexes. Module 1 achieves a capture efficiency of 97.24 %, and the subsequent addition of module 2 renders colorimetric indication of Campylobacter ranging from 101 to 106 CFU/mL, achieving an actual limit of detection of 8 CFU/mL validated by Campylobacter single-cells. Moreover, the generated colors can be recognized and converted into cell densities by module 3 with ultrasensitivity. Notably, this biosensor-smartphone platform accomplishe reliable high-throughput colorimetric detection of Campylobacter in real samples with an accuracy of 80 %. This work showcases a proof of principle for efficient on-site detection of Campylobacter contamination regarding shellfish farming.

Noninvasive Injectable Optical Nanosensor-Hydrogel Hybrids Detect Doxorubicin in Living Mice.

While the tissue-transparent fluorescence of single-walled carbon nanotubes (SWCNTs) imparts substantial potential for use in non-invasive biosensors, development of non-invasive systems is yet to be realized. Here, we investigated the functionality of a SWCNT-based nanosensor in several injectable SWCNT-hydrogel systems, ultimately finding SWCNT encapsulation in a sulfonated methylcellulose hydrogel optimal for detection of ions, small molecules, and proteins. We found that the hydrogel system and nanosensor signal were stable for several weeks in live mice. We then found that this system successfully detects local injections of the chemotherapeutic agent doxorubicin in mice. We anticipate future studies to adapt this device for detection of other analytes in animals and, ultimately, patients.

Cutting-Edge Hydrogel Technologies in Tissue Engineering and Biosensing: An Updated Review.

Hydrogels, known for their unique ability to retain large amounts of water, have emerged as pivotal materials in both tissue engineering and biosensing applications. This review provides an updated and comprehensive examination of cutting-edge hydrogel technologies and their multifaceted roles in these fields. Initially, the chemical composition and intrinsic properties of both natural and synthetic hydrogels are discussed, highlighting their biocompatibility and biodegradability. The manuscript then probes into innovative scaffold designs and fabrication techniques such as 3D printing, electrospinning, and self-assembly methods, emphasizing their applications in regenerating bone, cartilage, skin, and neural tissues. In the realm of biosensing, hydrogels' responsive nature is explored through their integration into optical, electrochemical, and piezoelectric sensors. These sensors are instrumental in medical diagnostics for glucose monitoring, pathogen detection, and biomarker identification, as well as in environmental and industrial applications like pollution and food quality monitoring. Furthermore, the review explores cross-disciplinary innovations, including the use of hydrogels in wearable devices, and hybrid systems, and their potential in personalized medicine. By addressing current challenges and future directions, this review aims to underscore the transformative impact of hydrogel technologies in advancing healthcare and industrial practices, thereby providing a vital resource for researchers and practitioners in the field.

LAMP-CRISPR/Cas12a-based impedimetric biosensor powered by Fe3O4@Au-(S-polyA-S)-Au for detection of SARS-CoV-2.

A low-cost, lab-made polytetrafluoroethylene micro-cell, equipped with three electrodes, wasd eveloped for the impedimetric detection of SARS-CoV-2. The gold working electrode was modified with a double-ended thiolated poly-adenine probe, which was conjugated with magnetic Fe3O4@Au nanoparticles (Fe3O4@Au-(S-polyA-S)-Au). After the loop-mediated isothermal amplification (LAMP) of viral RNA, the single-guide RNA (sgRNA), specifically bound to the SARS-CoV-2 target sequence, activates Cas12a. Cas12a then cleaved the immobilized probe. As a result, the magnetic Fe3O4@Au nanoparticles were released and adsorbed onto the gold electrode surface, using an external magnet. This process increased the physical surface area of the gold electrode, facilitating redox ion ([FeIII/II(CN)6]3-/4-) electron transfer. The decrease in the charge transfer resistance was utilized for SARS-CoV-2 detection. Our LAMP-CRISPR/Cas12a-based impedimetric biosensor, powered by Fe3O4@Au-(S-polyA-S)-Au, demonstrated impressive capabilities, including a remarkable detection limit of 0.8 aM (0.48 copies/µL) and a linear range of 0.01 to 36.06 fM.

Double-layer optical fiber interferometer with bio-layer-modified reflector for label-free biosensing of inflammatory proteins.

This work discusses label-free biosensing application of a double-layer optical fiber interferometer where the second layer tailors the reflection conditions at the external plain and supports changes in reflected optical spectrum when a bio-layer binds to it. The double-layer nanostructure consists of precisely tailored thin films, i.e., titanium (TiO2) and hafnium oxides (HfO2) deposited on single-mode fiber end-face by magnetron sputtering. It has been shown numerically and experimentally that the approach besides well spectrally defined interference pattern distinguishes refractive index (RI) changes taking place in a volume and on the sensor surface. These are of interest when label-free biosensing applications are considered. The case of myeloperoxidase (MPO) detection-a protein, which concentration rises during inflammation-is reported as an example of application. The response of the sensor to MPO in a concentration range of 1 × 10-11-5 × 10-6 g/mL was tested. An increase in the MPO concentration was followed by a redshift of the interference pattern and a decrease in reflected power. The negative control performed using ferritin proved specificity of the sensor. The results reported in this work indicate capability of the approach for diagnostic label-free biosensing, possibly also at in vivo conditions.

Deep-Learning Empowered Customized Chiral Metasurface for Calibration-Free Biosensing.

As a 2D metamaterial, metasurfaces offer an unprecedented avenue to facilitate light-matter interactions. The current "one-by-one design" method is hindered by time-consuming, repeated testing within a confined space. However, intelligent design strategies for metasurfaces, limited by data-driven properties, have rarely been explored. To address this gap, a data iterative strategy based on deep learning, coupled with a global optimization network is proposed, to achieve the customized design of chiral metasurfaces. This methodology is applied to precisely identify different chiral molecules in a label-free manner. Fundamentally different from the traditional approach of collecting data purely through simulation, the proposed data generation strategy encompasses the entire design space, which is inaccessible by conventional methods. The dataset quality is significantly improved, with a 21-fold increase in the number of chiral structures exhibiting the desired circular dichroism (CD) response (>0.6). The method's efficacy is validated by a monolayer structure that is easily prepared, demonstrating advanced sensing abilities for enantiomer-specific analysis of bio-samples. These results demonstrate the superior capability of data-driven schemes in photonic design and the potential of chiral metasurface-based platforms for calibration-free biosensing applications. The proposed approach will accelerate the development of complex systems for rapid molecular detection, spectroscopic imaging, and other applications.

Recent Advances in Ocular Drug Delivery: Insights into Lyotropic Liquid Crystals.

Background/Objectives: This review examines the evolution of lyotropic liquid crystals (LLCs) in ocular drug delivery, focusing on their ability to address the challenges associated with traditional ophthalmic formulations. This study aims to underscore the enhanced bioavailability, prolonged retention, and controlled release properties of LLCs that significantly improve therapeutic outcomes. Methods: This review synthesizes data from various studies on both bulk-forming LLCs and liquid crystal nanoparticles (LCNPs). It also considers advanced analytical techniques, including the use of machine learning and AI-driven predictive modeling, to forecast the phase behavior and molecular structuring of LLC systems. Emerging technologies in biosensing and real-time diagnostics are discussed to illustrate the broader applicability of LLCs in ocular health. Results: LLCs are identified as pivotal in promoting targeted drug delivery across different regions of the eye, with specific emphasis on the tailored optimization of LCNPs. This review highlights principal categories of LLCs used in ocular applications, each facilitating unique interactions with physiological systems to enhance drug efficacy and safety. Additionally, novel applications in biosensing demonstrate LLCs' capacity to improve diagnostic processes. Conclusions: Lyotropic liquid crystals offer transformative potential in ocular drug delivery by overcoming significant limitations of conventional delivery methods. The integration of predictive technologies and biosensing applications further enriches the utility of LLCs, indicating a promising future for their use in clinical settings. This review points to continued advancements and encourages further research in LLC technology to maximize its therapeutic benefits.

Recent Advances in Luminophores for Enhanced Electrochemiluminescence Analysis.

Electrochemiluminescence (ECL) detection is widely applied in many fields, including chemical measurement, biological analysis, and clinic tests, due to its high sensitivity. Currently, the fast development of many new electrochemical luminophores is continuously improving the ECL-based detection ability. Besides the enhancement of luminescence emission for a high detection sensitivity, minimizing the effect of co-reactants on ECL detection and achieving multiple analysis in one sample are also the main directions in this field. This review focuses on a summary of recently prepared new luminophores to achieve the three aims mentioned above. Especially, the review is composed by three parts, focusing on the luminophores or materials with high ECL efficiency, self-enhancing properties, and multi-color ECL luminophores. The fabrication of biosensors using these molecules is also reviewed to exhibit the advances in biological applications.

Exploring the landscape of FRET-based molecular sensors: Design strategies and recent advances in emerging applications.

Probing biological processes in living organisms that could provide one-of-a-kind insights into real-time alterations of significant physiological parameters is a formidable task that calls for specialized analytic devices. Classical biochemical methods have significantly aided our understanding of the mechanisms that regulate essential biological processes. These methods, however, are typically insufficient for investigating transient molecular events since they focus primarily on the end outcome. Fluorescence resonance energy transfer (FRET) microscopy is a potent tool used for exploring non-invasively real-time dynamic interactions between proteins and a variety of biochemical signaling events using sensors that have been meticulously constructed. Due to their versatility, FRET-based sensors have enabled the rapid and standardized assessment of a large array of biological variables, facilitating both high-throughput research and precise subcellular measurements with exceptional temporal and spatial resolution. This review commences with a brief introduction to FRET theory and a discussion of the fluorescent molecules that can serve as tags in different sensing modalities for studies in chemical biology, followed by an outlining of the imaging techniques currently utilized to quantify FRET highlighting their strengths and shortcomings. The article also discusses the various donor-acceptor combinations that can be utilized to construct FRET scaffolds. Specifically, the review provides insights into the latest real-time bioimaging applications of FRET-based sensors and discusses the common architectures of such devices. There has also been discussion of FRET systems with multiplexing capabilities and multi-step FRET protocols for use in dual/multi-analyte detections. Future research directions in this exciting field are also mentioned, along with the obstacles and opportunities that lie ahead.

Thermodynamics and Kinetics-Directed Regulation of Nucleic Acid-Based Molecular Recognition.

Nucleic acid-based molecular recognition plays crucial roles in various fields like biosensing and disease diagnostics. To achieve optimal detection and analysis, it is essential to regulate the response performance of nucleic acid probes or switches to match specific application requirements by regulating thermodynamics and kinetics properties. However, the impacts of thermodynamics and kinetics theories on recognition performance are sometimes obscure and the relative conclusions are not intuitive. To promote the thorough understanding and rational utilization of thermodynamics and kinetics theories, this review focuses on the landmarks and recent advances of nucleic acid thermodynamics and kinetics and summarizes the nucleic acid thermodynamics and kinetics-based strategies for regulation of nucleic acid-based molecular recognition. This work hopes such a review can provide reference and guidance for the development and optimization of nucleic acid probes and switches in the future, as well as for advancements in other nucleic acid-related fields.

Highly Stable Biotemplated InP/ZnSe/ZnS Quantum Dots for In Situ Bacterial Monitoring.

Despite their unique optical and electrical characteristics, traditional semiconductor quantum dots (QDs) made of heavy metals or carbon are not ideally suited for biomedical applications. Cytotoxicity and environmental concerns are key limiting factors affecting the adoption of QDs from laboratory research to real-world medical applications. Recently, advanced InP/ZnSe/ZnS QDs have emerged as alternatives to traditional QDs due to their low toxicity and optical properties; however, bioconjugation has remained a challenge due to surface chemistry limitations that can lead to instability in aqueous environments. Here, we report water-soluble, biotemplated InP/ZnSe/ZnS-aptamer quantum dots (QDAPTs) with long-term stability and high selectivity for targeting bacterial membrane proteins. QDAPTs show fast binding reaction kinetics (less than 5 min), high brightness, and high levels of stability (3 months) after biotemplating in aqueous solvents. We use these materials to demonstrate the detection of bacterial membrane proteins on common surfaces using a hand-held imaging device, which attests to the potential of this system for biomedical applications.

Characterization of a Charged Biomimetic Lipid Membrane for Unique Antifouling Effects against Clinically Relevant Matrices in Biosensing.

Clinically relevant matrices such as human blood and serum can cause substantial interference in biosensing measurements, severely compromising the effectiveness of the sensors. We report the characterization of a positively charged lipid membrane that has demonstrated unique features to suppress the nonspecific signal for antifouling effects by using SPR, fluorescence recovery after photobleaching (FRAP), and MALDI-TOF-MS. The ethylphosphocholine (EPC) lipid membrane proved to be exceptionally effective at reducing irreversible interactions from human serum on a Protein A surface. The membrane formation conditions and their effects on membrane fluidity and mobility were characterized for understanding the antifouling functions when various capture molecules were immobilized. Specifically, EPC lipid membranes on a Protein A substrate appear to exhibit a strong interaction, likely through the electrostatic effect with the negatively charged proteins that resulted in a stable hydration layer. The strong interaction also limited lipid mobility, contributing to a robust, protective interface that remained undamaged in undiluted serum. Tailoring a surface with antifouling lipid membranes allows for a range of biosensing applications in highly complex biological media.

Chemical tongues as multipurpose bioanalytical tools for the characterization of complex biological samples.

Chemical tongues are emerging powerful bioanalytical tools that mimic the mechanism of the human taste system to recognize the comprehensive characteristics of complex biological samples. By using an array of chromogenic or fluorogenic probes that interact non-specifically with various components in the samples, this tool generates unique colorimetric or fluorescence patterns that reflect the biological composition of a sample. These patterns are then analyzed using multivariate analysis or machine learning to distinguish and classify the samples. This review focuses on our efforts to provide an overview of the fundamental principles of chemical tongues, probe design, and their applications as versatile tools for analyzing proteins, cells, and bacteria in biological samples. Compared to conventional methods that rely on specific targeting (e.g., antibodies or enzymes) or comprehensive omics analyses, chemical tongues offer advantages in terms of cost and the ability to analyze samples without the need for specific biomarkers. The complementary use of chemical tongues and conventional methods is expected to enable a more detailed understanding of biological samples and lead to the elucidation of new biological phenomena.