Pyrenetetrasulfonate-grafted 2D ultrathin metal-organic layer as new electrochemiluminescence emitters for ultrasensitive microRNA-21 assay.

The exploration of novel electrochemiluminescence (ECL) luminophores with excellent ECL properties is a current research hotspot in the ECL field. Herein, a novel high-efficiency Ru-complex-free ECL emitter PyTS-Zr-BTB-MOL has been prepared by using porous ultrathin Zr-BTB metal-organic layer (MOL) as carrier to coordinatively graft the cheap and easily available polycyclic aromatic hydrocarbon (PAH) derivative luminophore PyTS whose ECL performance has never been investigated. Gratifyingly, the ECL intensity and efficiency of PyTS-Zr-BTB-MOL were markedly enhanced compared to both PyTS monomers and PyTS aggregates. The main reason was that the distance between pyrene rings was greatly expanded after the PyTS grafting on the Zr6 clusters of Zr-BTB-MOL, which overcame the aggregation-caused quenching (ACQ) effect of PyTS and thus enhanced the ECL emission. Meanwhile, the porous nanosheet structure of PyTS-Zr-BTB-MOL could distinctly increase the exposure of PyTS luminophores and shorten the diffusion paths of coreactants and electrons/ions, which effectively promoted the electrochemical excitation of more PyTS luminophores and thus achieved a further ECL enhancement. In light of the remarkable ECL property of PyTS-Zr-BTB-MOL, it was employed as an ECL indicator to build a novel high-sensitivity ECL biosensor for microRNA-21 determination, possessing a satisfactory response range (100 aM to 100 pM) and an ultralow detection limit (10.4 aM). Overall, this work demonstrated that using MOLs to coordinatively graft the PAH derivative luminophores to eliminate the ACQ effect and increase the utilization rate of the luminophores is a promising and efficient strategy to develop high-performance Ru-complex-free ECL materials for assembling ultrasensitive ECL biosensing platforms.

Acoustofluidics-enhanced biosensing with simultaneously high sensitivity and speed.

Simultaneously achieving high sensitivity and detection speed with traditional solid-state biosensors is usually limited since the target molecules must passively diffuse to the sensor surface before they can be detected. Microfluidic techniques have been applied to shorten the diffusion time by continuously moving molecules through the biosensing regions. However, the binding efficiencies of the biomolecules are still limited by the inherent laminar flow inside microscale channels. In this study, focused traveling surface acoustic waves were directed into an acoustic microfluidic chip, which could continuously enrich the target molecules into a constriction zone for immediate detection of the immune reactions, thus significantly improving the detection sensitivity and speed. To demonstrate the enhancement of biosensing, we first developed an acoustic microfluidic chip integrated with a focused interdigital transducer; this transducer had the ability to capture more than 91% of passed microbeads. Subsequently, polystyrene microbeads were pre-captured with human IgG molecules at different concentrations and loaded for detection on the chip. As representative results, ~0.63, 2.62, 11.78, and 19.75 seconds were needed to accumulate significant numbers of microbeads pre-captured with human IgG molecules at concentrations of 100, 10, 1, and 0.1 ng/mL (~0.7 pM), respectively; this process was faster than the other methods at the hour level and more sensitive than the other methods at the nanomolar level. Our results indicated that the proposed method could significantly improve both the sensitivity and speed, revealing the importance of selective enrichment strategies for rapid biosensing of rare molecules.

Single Molecule Thermodynamic Penalties Applied to Enzymes by Whispering Gallery Mode Biosensors.

Optical microcavities, particularly whispering gallery mode (WGM) microcavities enhanced by plasmonic nanorods, are emerging as powerful platforms for single-molecule sensing. However, the impact of optical forces from the plasmonic near field on analyte molecules is inadequately understood. Using a standard optoplasmonic WGM single-molecule sensor to monitor two enzymes, both of which undergo an open-to-closed-to-open conformational transition, the work done on an enzyme by the WGM sensor as atoms of the enzyme move through the electric field gradient of the plasmonic hotspot during conformational change has been quantified. As the work done by the sensor on analyte enzymes can be modulated by varying WGM intensity, the WGM microcavity system can be used to apply free energy penalties to regulate enzyme activity at the single-molecule level. The findings advance the understanding of optical forces in WGM single-molecule sensing, potentially leading to the capability to precisely manipulate enzyme activity at the single-molecule level through tailored optical modulation.

Electrochemical biosensors in early leukemia detection.

Leukemia, a type of blood cancer marked by an abnormal increase in white blood cells, poses a significant challenge to healthcare. The key to successful treatment lies in early detection. However, traditional methods often fall short. This review investigates the potential of electrochemical biosensors for a more accurate and earlier diagnosis of leukemia. Electrochemical biosensors are compact devices that transform biological interactions into electrical signals. Their small size, ease of use, and minimal sample requirements make them perfectly suited for point-of-care applications. Their remarkable sensitivity and specificity enable the detection of subtle biomolecular changes associated with leukemia, which is crucial for early disease detection. This review delves into studies that have utilized these biosensors to identify various types of leukemia. It examines the roles of electrodes, biorecognition elements, and signal transduction mechanisms. The discussion includes the integration of nanomaterials such as gold nanoparticles and nitrogen-doped graphene into biosensor design. These materials boost sensitivity, enhance signal amplification, and facilitate multi-analyte detection, thereby providing a more holistic view of the disease. Beyond technical advancements, the review underscores the practical benefits of these biosensors. Their portability makes them a promising tool for resource-constrained settings, enabling swift diagnosis in remote areas or at a patient's bedside. The potential for monitoring treatment effectiveness and detecting minimal residual disease to prevent relapse is also explored. This review emphasizes the transformative potential of electrochemical biosensors in combating leukemia. By facilitating earlier and more accurate diagnosis, these biosensors stand to revolutionize patient care and enhance treatment outcomes.

Shifting paradigm in electrochemical biosensing matrices comprising metal organic frameworks and their composites in disease diagnosis.

Metal Organic Frameworks (MOFs) are an evolving category of crystalline microporous materials that have grabbed the research interest for quite some time due to their admirable physio-chemical properties and easy fabrication methods. Their enormous surface area can be a working ground for innumerable molecular adhesions and site for potential sensor matrices. They have been explored in the last decade for incorporation in electrochemical sensor matrices as diagnostic solutions for a plethora of diseases. This review emphasizes on some of the recent advancements in the area of MOF-based electrochemical biosensors with focus on various important diseases and their significance in upgrading the sensor performance. It summarizes MOF-based biosensors for monitoring biomarkers relevant to diabetes, viral and bacterial sepsis infections, neurological disorders, cardiovascular diseases, and cancer in a wide range of real matrices. The discussion has been supplemented with extensive tables elaborating recent trends in the field of MOF-composite probe fabrication strategies with their respective sensing parameters. The article sums up the future scope of these materials in the field of biosensors and enlightens the reader with recent trends for future research scope. This article is categorized under: Diagnostic Tools > Biosensing Diagnostic Tools > Diagnostic Nanodevices.

Two-photon Dye-Based Fluorogenic Organic Nanoparticles as Intracellular Thiols Sensors.

Optical bioimaging is an ever-growing field that benefits both from the fast progress of optical instrumentation and modalities, and from the development of light-emitting probes. The efficacy of molecular fluorescent dyes is crucial, yet hindered by limited brightness and hydrophilicity. Addressing these challenges, self-stabilized fluorogenic organic nanoparticles only made of pure dyes (dFONs) are introduced in this work. Comprising thiol-sensitive fluorogenic chromophores, these dFONs exhibit enhanced brightness exclusively in the presence of biological thiols, notably glutathione, overcoming the need for water-solubilizing moieties. Importantly, these nanoparticles demonstrate large fluorescence and one- and two-photon brightness, enabling sensitive bioimaging of intracellular thiols at micromolar concentrations. Notably, only the pristine fluorogenic nanoparticles can penetrate the cells and does not require to wash the cells before imaging, emphasizing their unique role as dye carriers, fluorogenic probes and ease of use. This work highlights the transformative potential of dFONs in advancing optical bioimaging, paving the way for the use of dFONs not just as tracers, but also now as biosensors and ultimately in the future as biomarkers.

Multiplexed and Quantitative Imaging of Live-Cell Membrane Proteins by a Precise and Controllable DNA-Encoded Amplification Reaction.

Amplifying DNA conjugated affinity ligands can improve the sensitivity and multiplicity of cell imaging and play a crucial role in comprehensively deciphering cellular heterogeneity and dynamic changes during development and disease. However, the development of one-step, controllable, and quantitative DNA amplification methods for multiplexed imaging of live-cell membrane proteins is challenging. Here, we introduce the template adhesion reaction (TAR) method for assembling amplifiable DNA sequences with different affinity ligands, such as aptamers or antibodies, for amplified and multiplexed imaging of live-cell membrane proteins with high quantitative fidelity. The precisely controllable TAR enables proportional amplification of membrane protein targets with variable abundances by modulating the concentration ratios of hairpin templates and primers, thus allowing sensitive visualization of multiple membrane proteins with enhanced signal-to-noise ratios (SNRs) without disturbing their original ratios. Using TAR, we achieved signal-enhanced imaging of six proteins on the same live-cell within 1-2 h. TAR represents an innovative and programmable molecular toolkit for multiplexed profiling of membrane proteins in live-cells.

Architecture design and advanced manufacturing of heart-on-a-chip: scaffolds, stimulation and sensors.

Heart-on-a-chip (HoC) has emerged as a highly efficient, cost-effective device for the development of engineered cardiac tissue, facilitating high-throughput testing in drug development and clinical treatment. HoC is primarily used to create a biomimetic microphysiological environment conducive to fostering the maturation of cardiac tissue and to gather information regarding the real-time condition of cardiac tissue. The development of architectural design and advanced manufacturing for these "3S" components, scaffolds, stimulation, and sensors is essential for improving the maturity of cardiac tissue cultivated on-chip, as well as the precision and accuracy of tissue states. In this review, the typical structures and manufacturing technologies of the "3S" components are summarized. The design and manufacturing suggestions for each component are proposed. Furthermore, key challenges and future perspectives of HoC platforms with integrated "3S" components are discussed. Architecture design concepts of scaffolds, stimulation and sensors in chips.

Pre-Adsorbed H-Mediated Electrochemiluminescence.

In conventional electrochemiluminescence (ECL) systems, the presence of the competitive cathodic hydrogen evolution reaction (HER) in aqueous electrolytes is typically considered to be a side reaction, leading to a reduced ECL efficiency and stability due to H2 generation and aggregation at the electrode surface. However, the significant role of adsorbed hydrogen (H*) as a key intermediate, formed during the Volmer reaction in the HER process, has been largely overlooked. In this study, employing the luminol-H2O2 system as a model, we for the first time demonstrate a novel H*-mediated coreactant activation mechanism, which remarkably enhances the ECL intensity. H* facilitates cleavage of the O-O bond in H2O2, selectively generating highly reactive hydroxyl radicals for efficient ECL reactions. Experimental investigations and theoretical calculations demonstrate that this H*-mediated mechanism achieves superior coreactant activation compared to the conventional direct electron transfer pathway, which unveils a new pathway for coreactant activation in the ECL systems.

Gold nanoparticles-based biosensors: pioneering solutions for bacterial and viral pathogen detection-a comprehensive review.

Gold Nanoparticles (AuNPs) have gained significant attention in biosensor development due to their unique physical, chemical, and optical properties. When incorporated into biosensors, AuNPs offer several advantages, including a high surface area-to-volume ratio, excellent biocompatibility, ease of functionalization, and tunable optical properties. These properties make them ideal for the detection of various biomolecules, including proteins, nucleic acids, and bacterial and viral biomarkers. Traditional methods for detecting bacteria and viruses, such as RT-PCR and ELISA, often suffer from complexities, time consumption, and labor intensiveness. Consequently, researchers are continuously exploring novel devices to address these limitations and effectively detect a diverse array of infectious pathogenic microorganisms. In light of these challenges, nanotechnology has been instrumental in refining the architecture and performance of biosensors. By leveraging advancements in nanomaterials and strategies of biosensor fabrication the sensitivity and specificity of biosensors can be enhanced, enabling more precise detection of pathogenic bacteria and viruses. This review explores the versatility of AuNPs in detecting a variety of biomolecules, including proteins, nucleic acids, and bacterial and viral biomarkers. Furthermore, it evaluates recent advancements in AuNPs-based biosensors for the detection of pathogens, utilizing techniques such as optical biosensors, lateral flow immunoassays, colorimetric immunosensors, electrochemical biosensors, and fluorescence nanobiosensors. Additionally, the study discusses the existing challenges in the field and proposes future directions to improve AuNPs-based biosensors, with a focus on enhancing sensitivity, selectivity, and their utility in clinical and diagnostic applications.

Quantum Dot-Based FRET Nanosensors for Talin-Membrane Assembly and Mechanosensing.

Understanding the mechanisms of assembly and disassembly of macromolecular structures in cells relies on solving biomolecular interactions. However, those interactions often remain unclear because tools to track molecular dynamics are not sufficiently resolved in time or space. In this study, we present a straightforward method for resolving inter- and intra-molecular interactions in cell adhesive machinery, using quantum dot (QD) based Förster resonance energy transfer (FRET) nanosensors. Using a mechanosensitive protein, talin, one of the major components of focal adhesions, we are investigating the mechanosensing ability of proteins to sense and respond to mechanical stimuli. First, we quantified the distances separating talin and a giant unilamellar vesicle membrane for three talin variants. These variants differ in molecular length. Second, we investigated the mechanosensing capabilities of talin, i.e., its conformational changes due to mechanical stretching initiated by cytoskeleton contraction. Our results suggest that in early focal adhesion, talin undergoes stretching, corresponding to a decrease in the talin-membrane distance of 2.5 nm. We demonstrate that QD-FRET nanosensors can be applied for the sensitive quantification of mechanosensing with a sub-nanometer accuracy.

Detection of foodborne pathogens in contaminated food using nanomaterial-based electrochemical biosensors.

Foodborne pathogens are a grave concern for the for food, medical, environmental, and economic sectors. Their ease of transmission and resistance to treatments, such as antimicrobial agents, make them an important challenge. Food tainted with these pathogens is swiftly rejected, and if ingested, can result in severe illnesses and even fatalities. This review provides and overview of the current status of various pathogens and their metabolites transmitted through food. Despite a plethora of studies on treatments to eradicate and inhibit these pathogens, their indiscriminate use can compromise the sensory properties of food and lead to contamination. Therefore, the study of detection methods such as electrochemical biosensors has been proposed, which are devices with advantages such as simplicity, fast response, and sensitivity. However, these biosensors may also present some limitations. In this regard, it has been reported that nanomaterials with high conductivity, surface-to-volume ratio, and robustness have been observed to improve the detection of foodborne pathogens or their metabolites. Therefore, in this work, we analyze the detection of pathogens transmitted through food and their metabolites using electrochemical biosensors based on nanomaterials.

CRISPR-based electrochemical biosensors: an alternative for point-of-care diagnostics?

The combination of CRISPR technology and electrochemical sensors has sparked a paradigm shift in the landscape of point-of-care (POC) diagnostics. This review explores the dynamic convergence between CRISPR and electrochemical sensing, elucidating their roles in rapid and precise biosensing platforms. CRISPR, renowned for its remarkable precision in genome editing and programmability capability, has found a novel application in conjunction with electrochemical sensors, promising highly sensitive and specific detection of nucleic acids and biomarkers associated with diverse diseases. This article navigates through fundamental principles, research developments, and applications of CRISPR-based electrochemical sensors, highlighting their potential to revolutionize healthcare accessibility and patient outcomes. In addition, some key points and challenges regarding applying CRISPR-powered electrochemical sensors in real POC settings are presented. By discussing recent advancements and challenges in this interdisciplinary field, this review evaluates the potential of these innovative sensors as an alternative for decentralized, rapid, and accurate POC testing, offering some insights into their applications across clinical scenarios and their impact on the future of diagnostics.

A review of biocompatible polymer-functionalized two-dimensional materials: Emerging contenders for biosensors and bioelectronics applications.

Bioelectronics, a field pivotal in monitoring and stimulating biological processes, demands innovative nanomaterials as detection platforms. Two-dimensional (2D) materials, with their thin structures and exceptional physicochemical properties, have emerged as critical substances in this research. However, these materials face challenges in biomedical applications due to issues related to their biological compatibility, adaptability, functionality, and nano-bio surface characteristics. This review examines surface modifications using covalent and non-covalent-based polymer-functionalization strategies to overcome these limitations by enhancing the biological compatibility, adaptability, and functionality of 2D nanomaterials. These surface modifications aim to create stable and long-lasting therapeutic effects, significantly paving the way for the practical application of polymer-functionalized 2D materials in biosensors and bioelectronics. The review paper critically summarizes the surface functionalization of 2D nanomaterials with biocompatible polymers, including g-C3N4, graphene family, MXene, BP, MOF, and TMDCs, highlighting their current state, physicochemical structures, synthesis methods, material characteristics, and applications in biosensors and bioelectronics. The paper concludes with a discussion of prospects, challenges, and numerous opportunities in the evolving field of bioelectronics.

FRETting about CRISPR-Cas Assays: Dual-Channel Reporting Lowers Detection Limits and Times-to-Result.

Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR-Associated Protein (CRISPR-Cas) systems have evolved several mechanisms to specifically target foreign DNA. These properties have made them attractive as biosensors. The primary drawback associated with contemporary CRISPR-Cas biosensors is their weak signaling capacity, which is typically compensated for by coupling the CRISPR-Cas systems to nucleic acid amplification. An alternative strategy to improve signaling capacity is to engineer the reporter, i.e., design new signal-generating substrates for Cas proteins. Unfortunately, due to their reliance on custom synthesis, most of these engineered reporter substrates are inaccessible to many researchers. Herein, we investigate a substrate based on a fluorescein (FAM)-tetramethylrhodamine (TAMRA) Förster resonant energy-transfer (FRET) pair that functions as a seamless "drop-in" replacement for existing reporters, without the need to change any other aspect of a CRISPR-Cas12a-based assay. The reporter is readily available and employs FRET to produce two signals upon cleavage by Cas12a. The use of both signals in a ratiometric manner provides for improved assay performance and a decreased time-to-result for several CRISPR-Cas12a assays when compared to a traditional FAM-Black Hole Quencher (BHQ) quench-based reporter. We comprehensively characterize this reporter to better understand the reasons for the improved signaling capacity and benchmark it against the current standard CRISPR-Cas reporter. Finally, to showcase the real-world utility of the reporter, we employ it in a Recombinase Polymerase Amplification (RPA)-CRISPR-Cas12a DNA Endonuclease-Targeted CRISPR Trans Reporter (DETECTR) assay to detect Human papillomavirus in patient-derived samples.

Development of cross-linked glucose oxidase integrated Cu-nanoflower electrode for reusable and stable glucose sensing.

The demand for glucose-sensing devices has increased along with the increasing diabetic population. Here, we aimed to construct a system with a glucose oxidase (GOx)-integrated Cu-nanoflower (Cu-NF) as the underlying electrode. This novel system was successfully developed by creating a cross-linked GOx within a Cu-NF matrix, forming a c-GOx@Cu-NF-coated film on a carbon screen-printed electrode (CSPE). A comparison of the stabilities of the cross-linking methods demonstrated enhanced durability, with an activity level of >88 % maintained after approximately 35 days of storage in room temperature buffer. Regarding the ability of the c-GOx@Cu-NF modified CSPE to detect glucose via electrochemical methods, the redox potential gap (ΔE) and peak current increased in the presence of GOx. In comparison to that of glucose, the sensitivity of c-GOx@Cu-NF was approximately 8 times greater than that of GOx@Cu-NF, with a detection limit of 0.649 µM and a linear range of 5-500 µM. It sustained an average relative activity of 80 % over 20 days. After 10 cycles of repeated use, the activity remained above 75 %. In terms of evaluating the electrode's specificity for glucose, the detection rate for individual similar substances was approximately 1 %. The introduction of a crosslinking strategy to Cu-NF, leading to enhanced mechanical stability and conductivity, improved the detection capability. Furthermore, this approach led to increased long-term storage stability and reusability, allowing for specific glucose detection. To our knowledge, this report represents the first demonstration of a c-GOx@Cu-NF system for integrating electrochemical biosensing devices into digital healthcare pathways, offering enhanced sensing accuracy and mechanical stability.

Revisiting the standards of cancer detection and therapy alongside their comparison to modern methods.

In accordance with the World Health Organization data, cancer remains at the forefront of fatal diseases. An upward trend in cancer incidence and mortality has been observed globally, emphasizing that efforts in developing detection and treatment methods should continue. The diagnostic path typically begins with learning the medical history of a patient; this is followed by basic blood tests and imaging tests to indicate where cancer may be located to schedule a needle biopsy. Prompt initiation of diagnosis is crucial since delayed cancer detection entails higher costs of treatment and hospitalization. Thus, there is a need for novel cancer detection methods such as liquid biopsy, elastography, synthetic biosensors, fluorescence imaging, and reflectance confocal microscopy. Conventional therapeutic methods, although still common in clinical practice, pose many limitations and are unsatisfactory. Nowadays, there is a dynamic advancement of clinical research and the development of more precise and effective methods such as oncolytic virotherapy, exosome-based therapy, nanotechnology, dendritic cells, chimeric antigen receptors, immune checkpoint inhibitors, natural product-based therapy, tumor-treating fields, and photodynamic therapy. The present paper compares available data on conventional and modern methods of cancer detection and therapy to facilitate an understanding of this rapidly advancing field and its future directions. As evidenced, modern methods are not without drawbacks; there is still a need to develop new detection strategies and therapeutic approaches to improve sensitivity, specificity, safety, and efficacy. Nevertheless, an appropriate route has been taken, as confirmed by the approval of some modern methods by the Food and Drug Administration.

In the flow of molecular miniaturized fungal diagnosis.

The diagnosis of fungal infections presents several challenges and limitations, stemming from the similarities in symptomatology, diversity of underlying pathogenic species, complexity of fungal biology, and scarcity of rapid, affordable, and point-of-care approaches. In this review, we assess technological advances enabling the conversion of cutting-edge laboratory molecular diagnostic methods to cost-effective microfluidic devices. The most promising strategies toward the design of DNA sequence-based fungal diagnostic systems, capable of capturing and deciphering the highly informative DNA of the pathogen and adapted for resource-limited settings, are discussed, bridging fungal biology, molecular genetics, microfluidics, and biosensors.

One-Pot Synthesis of Silicon Quantum Dots-Based Fluorescent Nanomaterial and Its Application.

Conventionally obtained silicon quantum dots (Si QDs) generally suffer from the disadvantages of a cumbersome preparation process, large fluctuation in the quality of Si QDs, poor water solubility, and aggregation-caused quenching (ACQ) phenomenon. Here we report a facile one-pot strategy to synthesize a novel Si QDs-based fluorescent nanomaterial in which Si QDs are confined into dendritic mesoporous silica, named as SiQDs@DMSNs. The prepared SiQDs@DMSNs, with adjustable particle sizes ranging from 140 to 300 nm, emit blue fluorescence around 410 nm upon excitation by ultraviolet light at a wavelength of 300 nm. It is found that the addition of sodium salicylate (NaSAL) plays a crucial role in the in situ generation of Si QDs. The obtained SiQDs@DMSNs exhibit excellent fluorescence intensity, water solubility, and stability, facilitating easy surface modification, without being limited by the ACQ phenomenon. It is expected to be widely used in many fields such as biosensors, nanomedicines, in vivo imaging, fingerprint identification, and anticounterfeiting labels.